Carotenoid deprivation and beta-carotene's effects on male and female turtle color.

Carotenoid-colored integuments commonly function as sexually selected honest signals because carotenoid pigments can be costly to obtain, ingest, absorb, metabolize or transport before being deposited into the integument. As such, carotenoid pigmentation is often sexually dichromatic, with males being more colorful than females. Sexual dichromatism may also occur in ultraviolet (UV) wavelengths, which is visible to organisms who possess UV-sensitive photoreceptors. The stripes and spots of painted turtles (Chrysemys picta) are carotenoid-based and reflect UV wavelengths. This research describes UV sexual dichromatism in painted turtles and shows how carotenoid deprivation changes spot and stripe color in male and female painted turtles. Adult turtles were fed a diet that was supplemented with carotenoids (i.e., C diet) or deprived of carotenoids (C-). Stripe and spot color were measured with UV-vis spectrometry, and blood was drawn from all turtles before and after the dietary treatment. HPLC analysis revealed five carotenoids (4 xanthophylls and beta-carotene) circulating in turtle blood. C-diet reduced yellow chroma and increased brightness of yellow and red stripes or spots, relative to the C diet, but there was no sexually dimorphic effect of carotenoid deprivation on color, nor did carotenoid deprivation affect UV reflectance. Carotenoid deprivation reduced all circulating carotenoids, but beta-carotene was the only pigment with a significant effect on post-experimental carotenoids, implying that changes in color were due in part to reduction in circulating levels of beta-carotene. Color generation appears to be complex in turtles and have dietary as well as non-dietary components.

PLASMA BIOCHEMISTRY PROFILES OF JUVENILE GREEN TURTLES (CHELONIA MYDAS) FROM THE BAHAMAS WITH A POTENTIAL INFLUENCE OF DIET.

Plasma biochemistry profiles aid health assessment of marine turtles, but knowledge of the influence of regional biological factors (e.g., habitat, diet) on marine turtle blood plasma values is limited. To investigate the influence of diet on plasma biochemistry values in juvenile green turtles (Chelonia mydas), we used carbon and nitrogen stable isotopes to provide a quantitative estimate of forage items in green turtles feeding at two distinct areas (Bonefish Hole and South Flats) in Bimini, Bahamas. Plasma samples were obtained from 13 turtles in Bonefish Hole (a mangrove tidal estuary) and 15 turtles in South Flats (an open water seagrass bed) in 2018. All turtles appeared outwardly healthy. Sessile filter feeders contributed the largest proportion of diet in Bonefish Hole, and seagrass contributed the highest proportion of diet in South Flats. Turtles at Bonefish Hole presented significantly lower cholesterol, total protein, phosphorus, triglycerides, and aspartate transaminase compared to South Flats. Across all turtles, those feeding primarily on red algae presented the highest uric acid and alkaline phosphatase, and turtles with a seagrass-dominated diet had the highest cholesterol. Understanding dietary influence on plasma biochemistry may help explain variances seen in local health and nutritional evaluations, and the trends reported can aid the interpretation of plasma analyte values in marine turtles.

Trace metals in green sea turtles (Chelonia mydas) inhabiting two southern California coastal estuaries.

Foraging aggregations of east Pacific green sea turtles (Chelonia mydas) inhabit the Seal Beach National Wildlife Refuge (SBNWR) and San Diego Bay (SDB), two habitats in southern California, USA, located near urbanized areas. Both juvenile and adult green turtles forage in these areas and exhibit high site fidelity, which potentially exposes green turtles to anthropogenic contaminants. We assessed 21 trace metals (TM) bioaccumulated in green turtle scute and red blood cell (RBC) samples collected from SBNWR (n = 16 turtles) and SDB (n = 20 turtles) using acid digestion and inductively coupled plasma mass spectrometry. Principal component analyses of TM composition indicate that SBNWR and SDB turtles have location-specific contaminant signatures, characterized by differences in cadmium and selenium concentrations: SBNWR turtles had significantly more cadmium and selenium in RBC and more selenium in scute samples, than SDB turtles. Cadmium and selenium concentrations in RBC had a strong positive relationship, regardless of location. SBNWR turtles had higher selenium in RBCs than previously measured in other green turtle populations globally. Due to different retention times in blood vs. scute, these results suggest that SBNWR turtles have high long- and short-term selenium exposure. Turtles from SBNWR and SDB had higher trace metal concentrations than documented in green turtle populations that inhabit non-urbanized areas, supporting the hypothesis that coastal cities can increase trace metal exposure to local green turtles. Our study finds evidence that green turtle TM concentrations can differ between urbanized habitats and that long-term monitoring of these green turtles may be necessary.

Evidence of accumulation and elimination of inorganic contaminants from the lachrymal salt glands of leatherback sea turtles (Dermochelys coriacea).

Plasma osmolalities of marine vertebrates are generally lower than the surrounding medium; therefore, marine organisms must cope with the osmoregulatory challenges of life in a salty environment. The salt glands serve to maintain osmotic and ionic homeostasis in a number of lower marine vertebrates. One marine reptile, the leatherback sea turtle (Dermochelys coriacea), ingests excessive amounts of salts due to their diet of gelatinous zooplankton. Outside of the normal osmoregulatory function of the salt gland, little research has been conducted on contaminant accumulation and excretion in this organ. Here, we established arsenic, cadmium, lead, mercury, and selenium concentrations in red blood cells (RBCs) and salt gland secretions (SGSs) of nesting leatherbacks. We also collected salt glands from different life stage classes of dead stranded leatherbacks from the western Atlantic Ocean to determine if inorganic contaminants accumulate in this organ. Using non-metric multidimensional scaling and regression analyses, we determined that RBC and SGS inorganic contaminant concentrations were not correlated. Additionally, RBCs showed significantly higher concentrations of these contaminants in comparison to SGSs, likely due to the affinity of inorganic contaminants for the heme group of RBCs. Lastly, we found that salt gland cadmium and mercury concentrations tended to increase with increasing curved carapace length (CCL) in stranded leatherbacks. Our results indicate that different physiological mechanisms determine the distribution of inorganic contaminants in blood and SGSs. Increases in salt gland contaminant concentrations with increasing CCL suggest this organ as a potential target for accumulation.

Large-scale geographic patterns of mercury contamination in Morocco revealed by freshwater turtles.

Mercury (Hg) is a toxic contaminant present in most aquatic ecosystems. High concentrations pose serious threats to organisms and to human health. Because previous studies focused on few countries, environmental hazard due to Hg contamination remains obscure in many geographic areas, and for example limited information is available in North Africa. We examined total Hg contamination in 13 sites in Morocco (12 rivers and one lake) spread over a large area, 400 km north-south and 350 km west-east, that encompasses different biogeographic zones separated by the Atlas Mountains. Due to their longevity and sedentary habits, we used freshwater turtles as biological probes to monitor Hg exposure. Keratinized tissues reflect long-term Hg exposure; thus, we assayed Hg concentration in the claws of > 200 individuals and supplemented these data with blood Hg concentrations of > 60 individuals (a tissue that provides shorter term Hg exposure integration). The results provide the first large-scale picture of Hg contamination in the aquatic freshwater systems of Morocco. Comparisons with previous studies revealed that some of the sites were highly contaminated (e.g. mean Hg concentrations were above 5 µg g-1, a very high level in keratinized tissues) whereas other sites presented moderate or baseline levels. Unexpectedly, all highly contaminated sites were found in less densely populated areas, while more densely urbanized northern sites, even the sewers of large cities, were not highly contaminated. We hypothesize that silver mining activities in the southern High Atlas and in the Anti-Atlas contaminate rivers of the catchment basins over long distances. These findings indicate that fish, water consumption and contamination levels in local people should be further scrutinized.

Toxic elements and associations with hematology, plasma biochemistry, and protein electrophoresis in nesting loggerhead sea turtles (Caretta caretta) from Casey Key, Florida.

Toxic elements (arsenic, cadmium, lead, mercury, selenium, thallium) are a group of contaminants that are known to elicit developmental, reproductive, general health, and immune system effects in reptiles, even at low concentrations. Reptiles, including marine turtles, are susceptible to accumulation of toxic elements due to their long life span, low metabolic rate, and highly efficient conversion of prey into biomass. The objectives of this study were to (1) document concentrations of arsenic, cadmium, lead, mercury, selenium, and thallium in whole blood and keratin from nesting loggerhead sea turtles (Caretta caretta) from Casey Key, Florida and document correlations thereof and (2) correlate whole blood toxic element concentrations to various hematological and plasma biochemistry analytes. Baselines for various hematological and plasma analytes and toxic elements in whole blood and keratin (i.e., scute) in nesting loggerheads are documented. Various correlations between the toxic elements and hematological and plasma biochemistry analytes were identified; however, the most intriguing were negative correlations between arsenic, cadmium, lead, and selenium with and α- and γ-globulins. Although various extrinsic and intrinsic variables such as dietary and feeding changes in nesting loggerheads need to be considered, this finding may suggest a link to altered humoral immunity. This study documents a suite of health variables of nesting loggerheads in correlation to contaminants and identifies the potential of toxic elements to impact the overall health of nesting turtles, thus presenting important implications for the conservation and management of this species.

CHARACTERIZING THE 25-HYDROXYVITAMIN D STATUS OF TWO POPULATIONS OF FREE-RANGING EASTERN BOX TURTLES (TERRAPENE CAROLINA CAROLINA).

Ultraviolet B radiation is recommended for captive reptiles to stimulate production of adequate levels of vitamin D; however, little is known regarding the vitamin D status in many free-ranging populations. Current reference ranges for vitamin D in eastern box turtles have not yet been established. Sixty free-ranging eastern box turtles (Terrapene carolina carolina) from two well-studied populations in Illinois (n = 24) and Tennessee (n = 36) were assayed for plasma vitamin D concentration in 2014. There were no significant differences in concentrations between individuals in Illinois (mean: 117.5 nM/L) and Tennessee (mean: 98.7 nM/L) (P = 0.129) populations. Similarly, there were no differences in concentrations based on age class (P = 0.533) or sex (P = 0.532). There was a significant correlation between UV at the time of capture and vitamin D concentrations (R = 0.301, P = 0.030). Vitamin D was not correlated with total calcium (R = 0.018, P = 0.89) or Ca : P ratio (R = 0.025, P = 0.85). Diseases in captive individuals, including secondary nutritional hyperparathyroidism, may commonly be associated with vitamin D deficiencies, and clinical intervention relies on reference data. Vitamin D supplementation may be recommended if animals are deemed to be deficient. Data obtained can be used to improve the care of captive and free-ranging turtles by providing reference ranges, as well as better characterize the health of wild populations.

HEMATOLOGY AND PLASMA CHEMISTRY OF THE PLOUGHSHARE TORTOISE (ASTROCHELYS YNIPHORA) IN A CAPTIVE BREEDING PROGRAM.

Blood samples from 172 captive and 40 wild, healthy, juvenile and adult, ploughshare tortoises ( Astrochelys yniphora ) were analyzed to determine hematological and biochemical reference intervals. Hematological analytes included packed cell volume (PCV), white blood cell count (WBC), and WBC differential estimates. Biochemical analysis included total protein measured by photometry (TP) and by refractometry (TPr), albumin (ALB), creatine kinase (CK), aspartate aminotransferase (AST), glutamate dehydrogenase (GLDH), bile acids (BA), calcium (Ca), phosphorus (P), urea (UREA), and uric acid (UA). The jugular vein was identified as the preferred venipuncture site as subcarapacial vein venipuncture resulted in regular hemodilution. In due consideration of small sample sizes in some of the groups studied, adult tortoises had significantly higher plasma GLDH activity and TPr, TP, ALB, BA, and UREA concentrations and significantly lower AST activity and P concentration than juveniles. Captivity had a significant influence in some reference intervals, with captive adults presenting significantly higher WBC, and estimated counts of all white cell types as well as UREA and TPr than wild counterparts. Captive juveniles also showed significantly higher estimated monocyte and lower estimated eosinophil and basophil counts. Although these differences most likely reflect local environmental or dietary differences, without representing pathology or a deviation from the normal, they question the applicability of reference values from captive animals to wild animals and vice versa. Significant sex differences were only observed for PCV and UA. The reported reference intervals may serve as benchmarks for clinical assessment and conservation of this critically endangered species.

EFFECTS OF "SWIM WITH THE TURTLES" TOURIST ATTRACTIONS ON GREEN SEA TURTLE (CHELONIA MYDAS) HEALTH IN BARBADOS, WEST INDIES.

Along the West Coast of Barbados a unique relationship has developed between endangered green sea turtles (Chelonia mydas) and humans. Fishermen began inadvertently provisioning these foraging turtles with fish offal discarded from their boats. Although initially an indirect supplementation, this activity became a popular attraction for visitors. Subsequently, demand for this activity increased, and direct supplementation or provisioning with food began. Food items offered included raw whole fish (typically a mixture of false herring [Harengula clupeola] and pilchard [Harengula humeralis]), filleted fish, and lesser amounts of processed food such as hot dogs, chicken, bread, or various other leftovers. Alterations in behavior and growth rates as a result of the provisioning have been documented in this population. The purpose of this study was to determine how tourism-based human interactions are affecting the overall health of this foraging population and to determine what potential health risks these interactions may create for sea turtles. Juvenile green sea turtles (n=29) were captured from four sites off the coast of Barbados, West Indies, and categorized into a group that received supplemental feeding as part of a tour (n=11) or an unsupplemented group (n=18) that consisted of individuals that were captured at sites that did not provide supplemental feeding. Following capture, a general health assessment of each animal was conducted. This included weight and morphometric measurements, a systematic physical examination, determination of body condition score and body condition index, epibiota assessment and quantification, and clinical pathology including hematologic and biochemical testing and nutritional assessments. The supplemented group was found to have changes to body condition, vitamin, mineral, hematologic, and biochemical values. Based on these results, recommendations were made to decrease negative behaviors and health impacts for turtles as a result of this provisioning.

Metals and metalloids in whole blood and tissues of Olive Ridley turtles (Lepidochelys olivacea) from La Escobilla Beach (Oaxaca, Mexico).

Concentrations of eight metals and metalloids (Pb, Cd, Cu, Zn, Mn, Se, Ni and As) were evaluated from 41 nesting females (blood) and 13 dead (tissues) Olive Ridley turtles (Lepidochelys olivacea), a species classified as vulnerable and also listed in Appendix I of the Convention of International Trade in Endangered Species (CITES). The mean blood, liver and kidney lead concentration were 0.02 ± 0.01, 0.11 ± 0.08 and 0.06 ± 0.03 µ gg(-1) ww respectively, values lower than other turtle species and locations, which it could be due to the gradual disuse of leaded gasoline in Mexico and Central America since the 1990s. Mean concentration of cadmium was 0.17 ± 0.08 (blood), 82.88 ± 36.65 (liver) and 150.88 ± 110.9 9µg g(-1) (kidney). To our knowledge, the mean renal cadmium levels found is the highest ever reported worldwide for any sea turtle species, while other six elements showed a concentration similar to other studies in sea turtles.

Influence of the rehabilitation of injured loggerhead turtles (Caretta caretta) on their blood levels of environmental organic pollutants and elements.

We monitored the levels of 57 organic pollutants and 11 elements in the blood of 61 live stranded loggerhead sea turtles at the beginning and end of the rehabilitation period with the goal of evaluating whether recuperation of normal physiological conditions has an influence on the circulating levels of these pollutants from the blood. According to our results, several pollutants (OCPs, PCBs and toxic elements) are mobilized from storage tissues to the blood in sick turtles, and this is more evident in the subgroup of cachectic animals. We observed a significant decrease in the concentrations of some contaminants at the end of the rehabilitation period, probably due to redistribution processes. In contrast, an increase in the levels of the essential elements selenium and zinc was observed after a period of correct feeding and supplementation during hospitalization.

Selected heavy metals and selenium in the blood of black sea turtle (Chelonia mydas agasiizzi) from Sonora, Mexico.

The concentration of heavy metals (Zn, Cd, Ni, Cu, Mn) and selenium (Se) was analyzed in blood collected from 12 black turtles (Chelonia mydas agasiizzi) captured in Canal del Infiernillo, Punta Chueca, Mexico. The most abundant metals were Zn (63.58 µg g(-1)) and Se (7.66 µg g(-1)), and Cd was the lower (0.99 µg g(-1)). The sequential concentrations of trace metals were Zn > Se > Cu > Mn > Ni > Cd. In conclusion, this information is important as a baseline when using blood as tissue analysis of heavy metals; however, these levels could represent recent exposure in foraging grounds of black turtles in the Sea of Cortez.

Characterization and quantification of the influence of season and gender on plasma chemistries of Hermann's tortoises (Testudo hermanni, Gmelin 1789).

The present study is the first to monitor plasma chemistries of a group of 30 Hermann's tortoises (HTs) over two summer seasons to characterize and quantify seasonal and gender-related influences. The following analytes were analyzed: ALT, ALP, AST, BA, CA, CHO, CK, LDH, GLU, GLDH, P, TP, TRIG, urea and uric acid. Two-way repeated-measures ANOVA, mean values, confidence intervals were calculated. The overall results showed distinct patterns and somewhat unexpected results concerning the plasma chemistry changes induced by season and gender in a majority of routine biochemical analytes. TRIG, CHO, CA, and P were significantly higher in females. AST, ALT, BA, LDH and GLDH showed an increase in males in midsummer. ALP showed a significant and analogous seasonal variation for both sexes. GLU showed a moderate increase during midsummer. Uric acid and urea concentrations showed a distinct seasonal variation. For some analytes, fitted curves could be calculated describing the seasonal variation.

Relationships between carapace sizes and plasma major and trace element status in captive hawksbill sea turtles (Eretmochelys imbricata).

The aim of this study was to evaluate the relationships between carapace parameters as indicators of age and plasma elements in 25 captive hawksbill sea turtles. Particle-induced X-ray emission allowed detection of 23 trace and major elements. There were significant but weak correlations between the virtual carapace surface area and plasma bromide (r = -0.552, P<0.01), phosphorus (r = 0.547, P<0.01), lead (r =-0.434, P<0.05) and strontium (r = 0.599, P<0.01), while there were no significant correlations with other elements. These results suggest that major and trace plasma elements in captive sea turtles show almost no variation with carapace parameters, suggesting that the increase in plasma elements seen in wild sea turtles might be the result of marine pollution.

Measuring the level of agreement in hematologic and biochemical values between blood sampling sites in leatherback sea turtles (Dermochelys coriacea).

Conservation programs to protect endangered sea turtles are being instituted worldwide. A common practice in these programs is to collect blood to evaluate the health of the turtles. Several different venipuncture sites are used to collect blood from sea turtles for hematologic and biochemistry tests, depending on the species. To date, it is unknown what affect venipuncture site may have on sample results. The purpose of this study was to measure the level of agreement between hematologic and biochemistry values collected from the dorsal cervical sinus and the interdigital vein of leatherback (Dermochelys coriacea) sea turtles. Paired heparinized blood samples were obtained from the dorsal cervical sinus and the interdigital vein of 12 adult female nesting leatherback sea turtles on Keys Beach, St. Kitts, West Indies. Even though the sample population was small, the data for each chemistry were normally distributed, except for creatine kinase (CK). There was no significant difference when comparing biochemistry or hematologic values by venipuncture site, except for CK (P = 0.02). The level of agreement between sampling sites was considered good for albumin, calcium, globulin, glucose, packed cell volume, phosphorus, potassium, sodium, total protein, total solids, uric acid, white blood cell count, and all of the individual white cell types, while the level of agreement for aspartate aminotransferase and CK were considered poor. This information, coupled with the fact that the interdigital vein affords a less-invasive procedure, demonstrates that the interdigital vein is an appropriate location to use when establishing a hematologic and biochemical profile for leatherback sea turtles.

Cadmium toxicokinetics and bioaccumulation in turtles: trophic exposure of Trachemys scripta elegans.

Ecotoxicological data in reptiles are mainly represented by field studies reporting the tissue burden of wild-captured individuals but much less is known regarding the processes of uptake, depuration, accumulation and the effects of inorganic contaminants in these species. In the present study, the accumulation, the path and the effects of exposure to cadmium (Cd) through diet intake were investigated in female red eared slider turtles, Trachemys scripta elegans. In the first phase of the experiment, turtles underwent an acclimatization period during which they were fed a control diet. In the second phase, the turtles were exposed to cadmium through a CdCl(2) supplemented-diet with increased environmentally relevant concentrations for a period of 13 weeks. Following this, the turtles went through a third phase, a recovery phase of 3 weeks, during which they were fed uncontaminated food. Blood and feces were collected during the three phases of the experiment. The turtles were euthanized at the end of the experiment and organ samples collected. The Cd-concentrations in blood remained stable over the course of the experiment while Cd-concentrations in feces increased with time and with the amount of Cd ingested. The proportional accumulation in liver and kidney together was comprised between 0.7 and 6.1% and they represented the main organs of accumulation. Cd accumulated in the organs in the following order of concentration: kidney > liver > pancreas > muscle. In terms of burden in organs, the Cd-burden was the highest in liver followed by kidney and pancreas. The proportional accumulation decreased as Cd ingestion increased, suggesting that at a higher dose of Cd, assimilation decreased. Mineral content of the liver and pancreas became modified according to Cd level; increasing dietary Cd exposure increased concentrations of zinc and iron in liver and copper in pancreas in a dose-dependent manner. Accumulation of Cd had no effect on survival, food consumption, growth, weight or length suggesting no effect of the treatment on female turtle body condition.

Baseline heavy metals and metalloid values in blood of loggerhead turtles (Caretta caretta) from Baja California Sur, Mexico.

Environmental pollution due to heavy metals is having an increased impact on marine wildlife accentuated by anthropogenic changes in the planet including overfishing, agricultural runoff and marine emerging infectious diseases. Sea turtles are considered sentinels of ecological health in marine ecosystems. The objective of this study was to determine baseline concentrations of zinc, cadmium, copper, nickel, selenium, manganese, mercury and lead in blood of 22 clinically healthy, loggerhead turtles (Caretta caretta), captured for several reasons in Puerto López Mateos, Baja California Sur, Mexico. Zinc was the most prevalent metal in blood (41.89 µg g⁻¹), followed by Selenium (10.92 µg g⁻¹). The mean concentration of toxic metal Cadmium was 6.12 µg g⁻¹ and 1.01µg g⁻¹ respectively. Mean concentrations of metals followed this pattern: Zn>Se>Ni>Cu>Mn>Cd>Pb and Hg. We can conclude that blood is an excellent tissue to measure in relatively non-invasive way baseline values of heavy metals in Caretta caretta.

Why are hatching and emergence success low? Mercury and selenium concentrations in nesting leatherback sea turtles (Dermochelys coriacea) and their young in Florida.

Leatherback sea turtles (Dermochelys coriacea) have low hatching and emergence success compared to other sea turtle species. Postmortem examinations of hatchlings showed degeneration of heart and skeletal muscle that was similar to that found in other neonates with selenium deficient mothers. Selenium deficiency can result from elevated concentrations of bodily mercury. Ingested mercury is detoxified by the liver through mercury-selenium compound formation. In animals persistently exposed to mercury, the liver's ability to detoxify this element may decrease, especially if dietary selenium is insufficient. We measured mercury and selenium concentrations in nesting female leatherbacks and their hatchlings from Florida and compared the levels to hatching and emergence success. Both liver selenium and the liver selenium-to-mercury ratio positively correlated with leatherback hatching and emergence success. This study provides the first evidence for the roles of mercury and selenium in explaining low reproductive success in a globally imperiled species, the leatherback sea turtle.

Calcium and phosphorus values and their derivatives in captive tortoises (Testudo species).

OBJECTIVE: To evaluate the relationships between total calcium and phosphorus and ionised calcium and phosphorus values in clinically healthy tortoises. METHODS: Jugular blood samples were obtained from 25 tortoises, as part of a health screen of the population. These comprised Hermann's tortoises, Testudo hermanni boettgeri, spur-thighed tortoises, Testudo graeca ibera, marginated tortoises, Testudo marginata, and horsfield tortoises, Testudo horsfieldi. Plasma from these samples were analysed for total calcium, ionised calcium and phosphorus levels. These samples were taken in the immediate posthibernation period, before the onset of reproductive activity. RESULTS: Females exhibited statistically significantly higher levels of phosphorus. Ionised calcium and total calcium levels were elevated in females compared with males, but this was not statistically significant. Females did have statistically significantly higher calculated solubility indexes and statistically significantly lower ratios compared with males. CLINICAL SIGNIFICANCE: This study has provided an insight into the ratios and solubility indexes in tortoises, and this information may lead to further understanding of the significance of these parameters in chelonians.

Arsenic, cadmium, chromium, lead, mercury, and selenium levels in blood of four species of turtles from the Amazon in Brazil.

Using blood as a method of assessing metal levels in turtles may be useful for populations that are threatened or endangered or are decreasing. In this study the levels of arsenic (As), cadmium (Cd), chromium (Cr), lead (Pb), mercury (Hg), and selenium (Se) in blood of four species of turtles from the tributaries of the Rio Negro in the Amazon of Brazil were examined. The turtles included the six-tubercled Amazon (river) turtle (Podocnemis sextuberculata), red-headed Amazon (river) turtle (Podocnemis erythrocephala), big-headed Amazon (river) turtle (Peltocephalus dumerilianus), and matamata turtle (Chelus fimbriatus). Blood samples were taken from the vein in the left hind leg of each turtle. There were significant interspecific differences in the sizes of the turtles from the Rio Negro, and in concentrations of Pb, Hg, and Se; the smallest species (red-headed turtles) had the highest levels of Pb in their blood, while Se levels were highest in big-headed turtles and lowest in red-headed turtles. Hg in blood was highest in matamata, intermediate in big-headed, and lowest in the other two turtles. Even though females were significantly larger than males, there were no significant differences in metal levels as a function of gender, and the only relationship of metals to size was for Cd. Variations in metal levels among species suggest that blood may be a useful bioindicator. Metal levels were not high enough to pose a health risk to the turtles or to consumers, such as humans.