New Method for Imputation of Unquantifiable Values Using Bayesian Statistics for a Mixture of Censored or Truncated Distributions: Application to Trace Elements Measured in Blood of Olive Ridley Sea Turtles from Mexico.

One recurring difficulty in ecotoxicological studies is that a substantial portion of concentrations are below the limits of detection established by analytical laboratories. This results in censored distributions in which concentrations of some samples are only known to be below a threshold. The currently available methods have several limitations because they cannot be used with complex situations (e.g., different lower and upper limits in the same dataset, mixture of distributions, truncation and censoring in a single dataset). We propose a versatile method to fit the most diverse situations using conditional likelihood and Bayesian statistics. We test the method with a fictive dataset to ensure its correct description of a known situation. Then we apply the method to a dataset comprising 25 element concentrations analyzed in the blood of nesting marine turtles. We confirm previous findings using this dataset, and we also detect an unexpected new relationship between mortality and strontium concentration.

Hatching Success Rather Than Temperature-Dependent Sex Determination as the Main Driver of Olive Ridley (Lepidochelys olivacea) Nesting Activity in the Pacific Coast of Central America.

In marine turtles, sex is determined during a precise period during incubation: males are produced at lower temperatures and females at higher temperatures, a phenomenon called temperature-dependent sex determination. Nest temperature depends on many factors, including solar radiation. Albedo is the measure of the proportion of reflected solar radiation, and in terms of sand color, black sand absorbs the most energy, while white sand reflects more solar radiation. Based on this observation, darker sand beaches with higher temperatures should produce more females. As marine turtles show a high degree of philopatry, including natal homing, dark beaches should also produce more female hatchlings that return to nest when mature. When sand color is heterogeneous in a region, we hypothesize that darker beaches would have the most nests. Nevertheless, the high incubation temperature on beaches with a low albedo may result in low hatching success. Using Google Earth images and the SWOT database of nesting olive ridleys (Lepidochelys olivacea) in the Pacific coast of Mexico and Central America, we modeled sand color and nesting activity to test the hypothesis that darker beaches host larger concentrations of females because of feminization on darker beaches and female philopatry. We found the opposite result: the lower hatching success at beaches with a lower albedo could be the main driver of nesting activity heterogeneity for olive ridleys in Central America.

Inorganic elements in live vs dead nesting olive ridley marine turtles in the Mexican Pacific: Introducing a new statistical methodology in ecotoxicology.

This study reports the largest inorganic elements database in the blood of live marine turtles (Lepidochelys olivacea), with 241 live as well as 38 dead nesting turtles sampled and analyzed for 26 inorganic elements, including essential (Al, As, B, Ca, Co, Cr, Cu, Fe, K, Mg, Mn, Mo, Na, Ni, P, Se, S, V, and Zn) and non-essential elements (Cd, Li, Pb, Sr, Ti, Tl, and Hg). We compared inorganic element concentrations in live and dead olive ridleys from the arribada beach "La Escobilla" located on the Pacific coast of southeastern Mexico. The most outstanding result of our study is the higher Cd concentration in dead (mean 4.27 µg g-1 ww: min 0.01-max 81.5) compared with live animals (mean 0.14 µg g-1 ww: min 0.02-max 0.52). This population has been previously reported to have the highest Cd concentration worldwide in kidney and liver samples from marine turtles (with 150.88 ±â€¯110.99 and 82.88 ±â€¯36.65 µg g-1 ww, respectively). Other important findings of this study include the low Hg concentration along with the decrease in Pb over the years in this population. The study also uses a new statistical method - the iconography of correlations - in which all available information is used without removing individuals or variables with missing information for the whole analysis, which is a common problem in ecotoxicology. A major advantage of this method compared to other multivariate methods is that the missing information can be easily handled, because the correlations (2 variables) and partial correlations (3 variables) are estimated only with the available data using a one-at-a-time strategy.

Relationship between plasma biochemistry values and metal concentrations in nesting olive ridley sea turtles.

A hundred nesting olive ridley turtles were sampled to determine biochemical parameters (ALP, AST, ALT, creatinine, albumin, cholesterol, glucose, proteins, triglycerides, urea, and P-nitrophenyl acetate esterase activity). Esterase activity (EA) is a new biomarker very sensitive to metals. Most of the samples showed detectable levels. We also analyzed the concentration of 11 inorganic elements (As, Cd, Cr, Cu, Mn, Ni, Pb, Sr, Ti, Se, and Zn), some of them previously reported with very high concentrations in this population (especially cadmium with 82 and 150 µg g-1 ww in liver and kidney, respectively). Cadmium presented two negative relationships with creatinine and glucose. Some other understudied elements, Sr and Ti, for instance, presented five and four significant relationships with some biochemical parameters, respectively (most of them positive). EA was the parameter with most negative relationships (with Pb, Ti, As, Cr, and Se), reinforcing the results of other researchers in humans regarding the possible inhibition of EA by metals.

Carapace asymmetry: A possible biomarker for metal accumulation in adult olive Ridleys marine turtles?

The Olive Ridley marine turtle (Lepidochelys olivacea) is characterized by individual morphological variability in the number and shape of scutes. The influence of pollutants on developmental instability and one of its consequences, the asymmetry of individuals, has been demonstrated in several species, especially invertebrates and some birds. However, the use of this asymmetry as a biomarker of contamination in adult individuals has never been explored. We developed an index to quantify developmental instability (DIx) based on the number and relative size of costal carapace scutes. The link between DIx and inorganic elements concentrations was explored in various tissues of stranded turtles from the Southern Mexican Pacific. The relationships between adult contamination and DIx could directly or indirectly reflect (i) the disruption of metal elimination in the adult stage dependent on embryonic perturbation and thus determining DIx, (ii) the difference in metal absorption dependent on DIx status, or (iii) DIx linked to other unknown factors.

Molecular oxidative stress markers in olive ridley turtles (Lepidochelys olivacea) and their relation to metal concentrations in wild populations.

Due to their longevity and extensive migration areas, marine turtles are able to accumulate diverse contaminants over many years and as a consequence they represent an interesting bioindicator species for marine ecosystem pollution. Metals provoke toxicological effects in many aquatic animal species, but marine turtles have been under-investigated in this area. Thus, we have determined the presence of certain inorganic elements (As, Cd, Cu, Ni, Pb, Se and Zn) in olive ridley turtles (Lepidochelys olivacea) and related them to metallothionein (MT), superoxide dismutase (SOD), catalase (CAT) and glutathione reductase (GR) transcription and/or enzymatic activities. Gene expression of sod, cat and gr was found to be higher in blood than liver or kidney but most of the significant relationships were found in liver, not only for gene expression but also for enzyme activities. This must be related to the role the liver has as the first filter organ. Several positive relationships of sod, cat and gr gene expression in the different tissues were found in this population, as well as very high Cd concentrations. This could mean that these turtles are adapting to the metals-production of ROS and damage through a high transcription of these antioxidants. Multiple positive relationships with GR seem to be part of its compensatory effect due to the decrease of SOD production against the high and chronic exposure to certain xenobiotics. CAT, on the other hand, seems not to be used much, and glutathione detoxification of H2O2 may be more important in this species. Finally, despite the very high Cd concentrations found in this population, no significant relationship was found in any tissue with metallothionein gene expression. These results, along with very high Cd concentrations and a negative relationship with Cu, lead us to consider some kind of disruption in mt gene expression in these turtles.

p-Nitrophenyl Acetate Esterase Activity and Cortisol as Biomarkers of Metal Pollution in Blood of Olive Ridley Turtles (Lepidochelys olivacea).

This study was designed to determine the concentrations of p-nitrophenyl acetate esterase activity (EA) and cortisol in serum of marine Olive Ridley turtles (Lepidochelys olivacea) from a Mexican Pacific population ("La Escobilla" beach) and to evaluate the possible relationship of inorganic elements with these biomarkers. EA, cortisol, and selected chemical elements (Cd, Pb, Ti, Sr, Se, Al, As, and Zn) were measured in the blood of 44 sea turtles from the Eastern Pacific (Southeast Mexico). Serum EA ranged from 0.4 to 3.9 UI mL-1, and cortisol concentrations ranged from 0.07 to 2.5 µL dL-1. A strong negative correlation between EA and cortisol was observed (r = - 0.59, p < 0.01), and significant correlations also were found between EA and important metals, such as Cd (r = - 0.31, p < 0.05) and Pb (r = - 0.27, p < 0.05), and elements of growing concern like Ti (r = - 0.37, p < 0.01) or Al (r = - 0.34, p < 0.05) and between cortisol and Sr (r = 0.29, p < 0.05), Se (r = - 0.38, p < 0.01), and As (r = - 0.26, p < 0.05). These results suggest that turtles chronically exposed to different inorganic elements (such as Pb and Cd), driving to a highly consume of esterase and to a prolonged cortisol elevation. The obtained results indicate the usefulness of these biomarkers in the assessment of inorganic elements pollution in this species.

A comparative study of inorganic elements in the blood of male and female Caspian pond turtles (Mauremys caspica) from the southern basin of the Caspian Sea.

Due to their bioaccumulation and biomagnification pathways, inorganic elements can accumulate in high-level aquatic organisms in the food web. Then, this species can be used to monitor the quality of the environment. Blood concentration of nine inorganic elements, including possible toxic metals (An, Cu, Mn, Se, As, Ni, Cd, Pb, and Hg), in 20 males and 20 females from eight different locations with high industry and agriculture activities in Iran were evaluated in this work. Additionally, size, sex, condition index, and locations were also included and analyzed. Among the essential elements, Zn and Se presented very high concentrations (56.14 ± 2.66 and 8.44 ± 0.77 µg/g ww, respectively) in all locations. Regarding possible toxic elements, Pb and Cd presented concerning concentrations as well (0.52 and 0.58 µg/g ww); this is especially true for Pb, an element found in very high concentrations in tissues of turtles from the same area in a previous study. The sex and the size of the individuals also had significant differences in concentration of Pb, Cd, As, and Hg.

The current situation of inorganic elements in marine turtles: A general review and meta-analysis.

Inorganic elements (Pb, Cd, Hg, Al, As, Cr, Cu, Fe, Mn, Ni, Se and Zn) are present globally in aquatic systems and their potential transfer to marine turtles can be a serious threat to their health status. The environmental fate of these contaminants may be traced by the analysis of turtle tissues. Loggerhead turtles (Caretta caretta) are the most frequently investigated of all the sea turtle species with regards to inorganic elements, followed by Green turtles (Chelonia mydas); all the other species have considerably fewer studies. Literature shows that blood, liver, kidney and muscle are the tissues most frequently used for the quantification of inorganic elements, with Pb, Cd, Cu and Zn being the most studied elements. Chelonia mydas showed the highest concentrations of Cr in muscle (4.8 ± 0.12), Cu in liver (37 ± 7) and Mg in kidney (17 µg g-1 ww), Cr and Cu from the Gulf of Mexico and Mg from Japanese coasts; Lepidochelys olivacea presented the highest concentrations of Pb in blood (4.46 5) and Cd in kidney (150 ± 110 µg g-1 ww), both from the Mexican Pacific; Caretta caretta from the Mediterranean Egyptian coast had the highest report of Hg in blood (0.66 ± 0.13 µg g-1 ww); and Eretmochelys imbricata from Japan had the highest concentration of As in muscle (30 ± 13 13 µg g-1 ww). The meta-analysis allows us to examine some features that were not visible when data was analyzed alone. For instance, Leatherbacks show a unique pattern of concentration compared to other species. Additionally, contamination of different tissues shows some tendencies independent of the species with liver and kidney on one side and bone on the other being different from other tissues. This review provides a general perspective on the accumulation and distribution of these inorganic elements alongside existing information for the 7 sea turtle species.

Levels of perfluorinated acids (PFCAs) in different tissues of Lepidochelys olivacea sea turtles from the Escobilla beach (Oaxaca, Mexico).

Lepidochelys olivacea is the most abundant and globally distributed sea turtle species in the world and thus, monitoring this species for persistent organic pollutants, such as perfluorinated chemicals, is fundamental for their protection. This study was the first to evaluate the occurrence of five PFCAs (PFOA, PFNA, PFDA, PFUnA, PFDoA) in liver and blood samples of Olive Ridley turtle population from the Escobilla beach (Oaxaca, Mexico). PFDA and PFUnA were the predominant PFCs in blood samples (detected in 93% and 84% of samples, respectively) and were also present in the highest concentrations. Liver samples showed higher PFCA concentrations than whole blood samples, with PFNA and PFDA the most abundant PFCs congeners in liver samples, detected in 65% and 47% of the samples, respectively. The measured levels of contaminants in the blood samples of Lepidochelys olivacea sea turtles were compared to the levels reported in the literature for other turtle species. While linear significant correlations between PFNA, PFDA and PFUnA concentrations in blood samples and curved carapace lengths were determined, no correlation was found for PFOA, supporting the hypothesis that sea turtles could have a higher ability to eliminate this perfluorinated chemical from their blood than other PFCAs. However, we do not know if the concentrations are species or sampling areas dependent.

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.