María Camacho

Comparative study of hematologic and plasma biochemical variables in Eastern Atlantic juvenile and adult nesting loggerhead sea turtles (Caretta caretta)

BACKGROUND Plasma biochemical and hematologic variables are important in the management of endangered sea turtles, such as loggerheads. However, studies on blood biochemistry and hematology of loggerheads are limited, and different concentrations according to variable criteria have been reported. OBJECTIVE The purpose of this study was to establish and compare baseline plasma chemistry and hematology values in Eastern Atlantic juvenile and adult nesting loggerhead sea turtles (Caretta caretta). METHODS Blood samples were collected from 69 healthy juvenile loggerhead sea turtles after their rehabilitation in captivity, and from 34 adult nesting loggerheads after oviposition. Fresh blood was used for leukocyte differential count and PCV determination. Heparinized blood was used for RBC and WBC counts. Plasma biochemical concentrations were measured using an automated biochemical analyzer. For the comparative study, nonparametric statistical analysis was done using the Mann-Whitney U-test. RESULTS Minimum, maximum, and median concentrations were obtained for 14 hematologic and 15 plasma chemistry variables. Statistically significant differences between juvenile and adult turtles were found for PCV; RBC, WBC, and leukocyte differential counts; total protein, albumin, globulins, calcium, triglycerides, glucose, total cholesterol and urea concentrations; and lactate dehydrogenase activity. CONCLUSIONS Age, size, and reproductive status cause important variations in the hematologic and plasma biochemical results of loggerheads. The reference values obtained in this study may be used as a standard profile, useful for veterinary surgeons involved in sea turtle conservation.

Monitoring organic and inorganic pollutants in juvenile live sea turtles: results from a study of Chelonia mydas and Eretmochelys imbricata in Cape Verde.

Despite the current environmental concern regarding the risk posed by contamination in marine ecosystems, the concentrations of pollutants in sea turtles have not been thoroughly elucidated. In the current study, we determined the concentrations of 18 organochlorine pesticides (OCPs), 18 polychlorinated biphenyls (PCBs), 16 polycyclic aromatic hydrocarbons (PAHs) and 11 inorganic elements (Cu, Mn, Pb, Zn, Cd, Ni, Cr, As, Al, Hg and Se) for the first time in two sea turtle species (Chelonia mydas and Eretmochelys imbricata). Only five of the 18 analyzed OCPs were detected in both species. The average total OCP concentration was higher in green turtles than in hawksbills (0.33 ng/ml versus 0.20 ng/ml). Higher concentrations of individual congeners and total PCBs were also detected in green turtles than in hawksbills (∑PCBs=0.73ng/ml versus 0.19 ng/ml), and different PCB contamination profiles were observed in these two species. Concerning PAHs, we also observed a different contamination profile and higher levels of contamination in green turtles (∑PAHs=12.06 ng/ml versus 2.95 ng/ml). Di- and tri-cyclic PAHs were predominant in both populations, suggesting a petrogenic origin, rather than urban sources of PAHs. Additionally, all of the samples exhibited detectable levels of the 11 inorganic elements. In this case, we also observed relevant differences between both species. Thus, Zn was the most abundant inorganic element in hawksbills (an essential inorganic element), whereas Ni, a well-known toxicant, was the most abundant inorganic element in green turtles. The presence of contaminants is greater in green turtles relative to hawksbill turtles, suggesting a greater exposure to hazardous chemical contaminants for green turtles. These results provide baseline data for these species that can serve for future monitoring purposes outlined in the EUs Marine Strategy Framework Directive.

Assessment of the exposure to organochlorine pesticides, PCBs and PAHs in six species of predatory birds of the Canary Islands, Spain.

In the present study, we investigated the concentrations and distributions of 57 anthropogenic pollutants, including 23 organochlorine pesticides (OCs), 18 polychlorinated biphenyls (PCBs) and 16 polycyclic aromatic hydrocarbons (PAHs) in liver samples from 102 birds of prey of six species that were found dead or that had died during their stay in the Wildlife Recovery Centers of the Canary Islands (Spain) between 2009 and 2012. The dual goal of this work was to determine the occurrence of these contaminants in these six species of birds of prey, and also whether they can be used as bioindicators for monitoring environmental pollution in the region. We found that Accipiter nisus, Falco pelegrinoides and Falco tinnunculus were the most contaminated species. The profiles of contamination among the species were extremely similar in the case of organochlorine contaminants, with DDT and its metabolites as the most abundant compounds. The contamination by DDT and its metabolites, as well as contamination by dieldrin, could be considered high in these animals, much higher than reports from other regions of the planet, which is in agreement with previous reports from our group regarding humans, food and other animals from this area. In contrast, the contamination by PCBs could be considered extremely low and was probably below the threshold of toxicity for these contaminants. The content of carcinogenic/mutagenic PAHs in these animals was clearly dependent on the feeding pattern of the species; however, the levels were also well below the values that were considered toxic in predictive models. This study represents the first report of contamination by PAHs in all these species and is also the first report of PCB levels in Barbary Falcons.

Comparative study of polycyclic aromatic hydrocarbons (PAHs) in plasma of Eastern Atlantic juvenile and adult nesting loggerhead sea turtles (Caretta caretta).

Polycyclic aromatic hydrocarbons (PAHs) were measured in plasma samples of 162 juvenile loggerhead sea turtles (Caretta caretta) from Canary Islands, and 205 adult nesting loggerheads from Cape Verde. All the 367 samples showed detectable values of some type of PAH. Phenanthrene was the PAH most frequently detected and at the highest concentration in both populations. Median concentrations of ∑PAHs in the plasma of loggerheads from the Canary Islands and Cape Verde were similar (5.5 and 4.6 ng/ml, respectively). Di- and tri-cyclic PAHs were predominant in both populations suggesting petrogenic origin rather than urban sources of PAHs. In the group of turtles from Canary Islands, there was evident an increasing level of contamination over the last few years. The present study represents the first data of contamination by PAHs in sea turtles from the studied areas.

Potential adverse effects of inorganic pollutants on clinical parameters of loggerhead sea turtles (Caretta caretta): results from a nesting colony from Cape Verde, West Africa.

A large number of nesting loggerhead sea turtles (n = 201) were sampled to establish the blood levels of 11 elements (Cu, Mn, Pb, Zn, Cd, Ni, Cr, As, Al, Hg, and Se). Almost all of the samples showed detectable levels of these 11 elements, and Zn and Se exhibited the highest concentrations (median values as high as 6.05 and 2.28 μg/g, respectively). The median concentrations of the most toxic compounds, As, Cd, Pb, and Hg, were relatively low (0.38, 0.24, 0.06, and 0.03 μg/g, respectively). We also determined the haematological and biochemical parameters in a subsample of 50 turtles to evaluate the potential effects of these contaminants on clinical parameters and found several associations. Our study reinforces the usefulness of blood for the monitoring of the levels of contaminating elements and their adverse effects on blood parameters in sea turtles.

Consumption of foods of animal origin as determinant of contamination by organochlorine pesticides and polychlorobiphenyls: Results from a population-based study in Spain

The level of contamination with persistent organic pollutants (POPs) and dietary habits and food consumption was extensively studied in the population from the Canary Islands (Spain). Because foodstuffs of animal origin are well known to be prominent contributors to these contaminants, the current study aimed to assess the role of the dietary intake of animal products as a probability factor for increased serum POPs. The intake of animal products (dietary variables) as a determining factor for serum POP levels was investigated using multivariate statistical models. Our results showed that while poultry, rabbit, and cheese consumption increases the probability of having high levels of non-DDT-derivative pesticides, sausage, yogurt, lard, and bacon consumption decreases the probability of having high levels of these pesticides. In addition, poultry, rabbit, eggs, cream, and butter consumption increased the probability of having detectable levels of marker PCB, while dairy desserts decreased the probability of having detectable levels of these PCBs. On the contrary, sausage and meat consumption increased the probability of having detectable levels of dioxin-like PCBs (DL-PCBs). The current results confirm that dietary intake of foodstuffs of animal origin is a relevant risk factor for the accumulation of POPs (and therefore their serum levels). Our study indicates that the analysis of dietary patterns may be useful for identifying those individuals that will probably present a high body burden of POPs. Because POPs can exert deleterious effects on human health, the identification of populations at risk of being highly contaminated is mandatory in order to implement policies that minimize the exposure to these compounds.

Continued implication of the banned pesticides carbofuran and aldicarb in the poisoning of domestic and wild animals of the Canary Islands (Spain)

Although nowadays the intentional poisoning of domestic and wild animals is a crime in EU, in the past the poison was used in rural areas of a number of European countries to kill animals that were considered harmful for human activities. In Spain evidences indicate that intentional poisonings continue to occur throughout the entire country nowadays. This situation seems to be of particular concern in the Canary Islands (Spain), where this study was performed. Our results confirmed that 225 animals were poisoned by pesticides over the study period (32 months; 2010-2013). The intentionality of the poisoning was confirmed in 117 cases. It has to be highlighted that the other 108 animals also died by pesticide poisoning, although the intentionality was only suspected. This incidence is currently the highest reported in any region from European Union. The pesticides carbofuran, bromadiolone, brodifacoum and aldicarb were the most frequently detected involved. Among the affected species, it has to be highlighted that endangered species are frequently affected in poisoning incidents. Notably, chemicals banned in the EU (carbofuran and aldicarb) were identified in approximately 75% of cases, and in almost 100% of baits, which suggests that these pesticides are still available to the population. Several circumstances may explain these results. Firstly, little control over the sale and possession of pesticide products, and the potential existence of an illegal market of pesticides banned in the European Union in the neighbouring African continent. In addition, the limited awareness of the population about the dangerousness of these compounds, for the environment, animals, or even people, make the situation very worrying in these islands. Stronger regulations, control of legal and illegal pesticide use, development of educational programs and legal action in poisoning incidents are needed to decrease the impact of pesticide misuse on wildlife and domestic animals.

Assessment of anticoagulant rodenticide exposure in six raptor species from the Canary Islands (Spain).

Anticoagulant rodenticides are highly toxic compounds that are widely used for pest control of rodents, but that also may threaten the wildlifes health. This work aimed to assess the exposure to first- and second-generation anticoagulant rodenticides (ARs) in six birds of prey species from the Canary Islands (Spain). The concentrations of seven widely used ARs were determined by LC-MS/MS in 104 liver samples of six species of birds of prey (Buteo buteo, Accipiter nisus, Falco pelegrinoides, Falco tinnunculus, Asio otus, and Tyto alba). We determined that 61% of the livers had detectable residues of at least one AR. The most frequently detected AR was bromadiolone, which was detected in 60.3% of the positive cases. The detection frequencies of these compounds varied widely, depending on the species. More than 75% of the A. nisus, T. alba, and A. otus individuals had detectable rodenticide residues in the liver. However, F. tinnunculus exhibited the highest concentrations of AR, with median values above 100 ng/g w.w. We did not detect first-generation ARs in any of the samples. When grouped, nocturnal species exhibited higher AR concentrations than diurnal species (P<0.001). The residue levels were higher among small mammal-eaters than bird-eaters (P<0.01). While most animals exhibited no macroscopic signs of coagulation disorders, approximately 35% exceeded the threshold levels of toxicity, which suggests that these compounds could weaken these animals in their natural environment. In conclusion, the control of rodent populations by ARs suggests that these compounds will enter the food chain and thus threaten the vulnerable populations of raptors on the Canary Islands. Our findings require authorities to ban or strictly control the use of these rodenticides in the natural environment for the conservation of raptors and other predatory species.

Assessment of human health hazards associated with the dietary exposure to organic and inorganic contaminants through the consumption of fishery products in Spain.

In this work we have evaluated the potential carcinogenic and acutely toxic risks associated to the exposure to highly prevalent organic and inorganic contaminants through the consumption of fishery products by the Spanish population. The concentrations of 8 organochlorine pesticides (OCPs), 18 polychlorinated biphenils (PCBs), 7 polycyclic aromatic hydrocarbons (expressed as benzo[a]pyrene toxic equivalents (B[a]Peq)), and three inorganic toxic elements [arsenic (As), cadmium (Cd), and mercury (Hg)] were determined in 93 samples of the most consumed species of white fish, blue fish, cephalopods and seafood species, which were acquired directly in markets and supermarkets in the Canary Islands, Spain. The chemical concentration data were combined with the pattern of consumption of these foodstuffs in order to calculate the daily intake of these contaminants, and on this basis the risk quotients for carcinogenicity and acute toxicity were determined for Spanish adults and children. Our results showed that the daily intake of OCPs, PCBs and B[a]Peq, which is associated to blue fish consumption was the highest within the fish group. The estimated intake of pollutants can be considered low or very low for the individual contaminants, when compared to reference values, except in the case of HCB and As. All the estimated intakes were below the reported Tolerable Daily Intakes. Considering the additive effects of multiple contaminants, the risk of acute toxic effects can also be considered as low or very low. However, our results reflect that the current consumption of white fish in adults and children, and also the blue fish in the case of adults, poses a moderate carcinogenic risk to Spanish consumers, mainly related to their concentrations of As. The conclusions of this research may be useful for the design of appropriate risk communication campaigns.

Plasma levels of pollutants are much higher in loggerhead turtle populations from the Adriatic Sea than in those from open waters (Eastern Atlantic Ocean)

In this paper we determined the levels of 63 environmental contaminants, including organic (PCBs, organochlorine pesticides, and PAHs) and inorganic (As, Cd, Cu, Pb, Hg and Zn) compounds in the blood of loggerhead turtles (Caretta caretta) from two comparable populations that inhabit distinct geographic areas: the Adriatic Sea (Mediterranean basin) and the Canary Islands (Eastern Atlantic Ocean). All animals were sampled at the end of a period of rehabilitation in centers of wildlife recovery, before being released back into the wild, so they can be considered to be in good health condition. The dual purpose of this paper is to provide reliable data on the current levels of contamination of this species in these geographic areas, and secondly to compare the results of both populations, as it has been reported that marine biota inhabiting the Mediterranean basin is exposed to much higher pollution levels than that which inhabit in other areas of the planet. According to our results it is found that current levels of contamination by organic compounds are considerably higher in Adriatic turtles than in the Atlantic ones (∑PCBs, 28.45 vs. 1.12ng/ml; ∑OCPs, 1.63 vs. 0.19ng/ml; ∑PAHs, 13.39 vs. 4.91ng/ml; p<0.001 in all cases). This is the first time that levels of PAHs are reported in the Adriatic loggerheads. With respect to inorganic contaminants, although the differences were not as great, the Adriatic turtles appear to have higher levels of some of the most toxic elements such as mercury (5.74 vs. 7.59μg/ml, p<0.01). The results of this study confirm that the concentrations are larger in turtles from the Mediterranean, probably related to the high degree of anthropogenic pressure in this basin, and thus they are more likely to suffer adverse effects related to contaminants.