Reproductive parameters in female yellow-blotched map turtles (Graptemys flavimaculata) from a historically contaminated site vs. a reference site.

Graptemys flavimaculata, the yellow-blotched map turtle, is a long-lived, threatened, species, endemic to the Pascagoula River drainage in Mississippi. During the 1980s, one branch of the drainage (i.e. the Leaf River) was impacted by effluent from a wood pulp processing plant known to contain endocrine disrupters. A decade later, we examined seasonal reproductive parameters (i.e. monthly plasma estradiol-17beta (E(2)), testosterone (T), vitellogenin (VTG) and follicular development) in adult female turtles from historically polluted and reference sites in the drainage to determine if legacy exposure to pollution impacts reproduction . We found no seasonal patterns in E(2) or T and these patterns did not differ between sites. However, E(2) differed significantly among ovarian stages for the reference, but not pollutant exposed females. A significantly greater percentage of reference site females were able to produce a second clutch than females from the historically polluted site (50% and 17%). Additionally, there was a significant positive correlation between E(2) with VTG levels for reference, but not pollutant exposed females. Body and yolk tissue contaminant analysis indicated that exposure to pollutants is presently minimal and unlikely the cause of the reproductive differences observed between sites; instead, differences are potentially due to exposure history.

Effect of a Punica granatum enriched diet on immunocompetence in Rhinella marina.

Direct ingestion of plant materials has been shown to have immunomodulatory effects on a variety of herbivores. Studies have also shown that compounds ingested indirectly by predators through prey items can affect the general physiology of the ingesting organism. Relatively little data exists, however, concerning the modulation of a predator's immune system via compounds obtained indirectly through prey. In this study, we sought to determine if the immune-stimulating properties of Punica granatum (pomegranate) could be conveyed from a prey organism, Acheta domestica, to a predator, Rhinella marina, through diet specialization. Experimental crickets were fed a diet of agar supplemented with 10 mg/mL of lyophilized, powdered, whole pomegranate while control crickets were fed unadulterated agar. Experimental toads consumed a diet consisting of crickets fed the pomegranate-enriched diet, while control toads consumed a diet consisting of crickets fed the standard agar diet. Blood samples were taken weekly and leukocyte profiles and neutrophil phagocytic activity were determined for all toads over an 8-week period. Complement activity was measured at 6 weeks. Toads fed the pomegranate-enriched diet showed altered leukocyte profiles as evidenced by an increase in circulating eosinophil number and a decrease in the number of circulating lymphocytes, monocytes, and basophils as compared to controls, indicating an immunomodulatory effect of the pomegranate-enhanced diet. These results suggest that pomegranate-derived immunomodulatory compounds can be transferred from prey to predator, and suggests that the flora in the environment where insectivores forage could have a significant effect on the physiology of the animal.

Characterization and quantification of corticosteroid-binding globulin in a southern toad, Bufo terrestris, exposed to coal-combustion-waste.

Corticosteroid-binding globulin (CBG) is a plasma protein that binds corticosterone and may regulate access of hormone to tissues. The role of CBG during a stress response is not clear. At least two hypotheses have been proposed: 1) CBG levels may increase in response to a stressor, thereby decreasing the amount of circulating free corticosterone, or 2) CBG levels may decline, making corticosterone available for its role in increased metabolic needs during stress. In this study, southern toads, Bufo terrestris, were exposed to a chronic pollutant (coal-combustion-waste), to determine changes in CBG and free corticosterone levels. Since toads exposed to chronic pollutants in previous studies did not exhibit the predicted changes in metabolic rate and mass, but did experience a significant elevation in total corticosterone, we hypothesized that CBG would likewise increase and thus, mitigate the effects of a chronic (i.e. 2 months) pollutant stressor. To conduct this study, we first characterized the properties of CBG in southern toads. Toad CBG has a K(d)=20.6+/-1.0 nM and a B(max)=332.2+/-5.1 nmol/L plasma. The rank order potencies for steroid inhibition of tritiated corticosterone are: dihydrotestosterone > corticosterone >> progesterone=testosterone >>> estrogen=dexamethasone. After characterization, we monitored the changes in CBG, total corticosterone, and free corticosterone in male toads that were exposed to either coal-combustion-waste or control conditions. CBG increased in all groups throughout the experiment. Total corticosterone, on the other hand, was only significantly elevated at four weeks of exposure to coal-combustion-waste. The increase in CBG did not parallel the increase in total corticosterone; as a result, free corticosterone levels were not buffered by CBG, but showed a peak at four weeks similar to total corticosterone. This finding indicates that, in this species, CBG may not provide a protective mechanism during long-term pollution exposure.

Metabolic measures of male southern toads (Bufo terrestris) exposed to coal combustion waste.

Southern toads (Bufo terrestris) are found in coal fly ash collection basins associated with coal-burning electrical power plants. These basins contain large amounts of trace metals and organisms found in these basins are known to accumulate large quantities of metals. Studies on a variety of organisms exposed to trace metals found that they experience a significant increase in standard metabolic rate. We experimentally exposed southern toads to metal-contaminated sediment and food and measured changes in standard and exercise metabolic rates as well as changes in body, liver and muscle mass, blood glucose, and corticosterone. We found that toads exposed to trace metal contamination gained significantly less mass (18.3 %) than control toads (31.3%) when food was limited and experienced significantly decreased RQ after exercise. However, contaminated toads did not experience changes in standard (x control = 0.114 +/- 0.016 mL O2 g(-1) h(-1); x (ash)=0.109 +/- 0.013 mL O2 g(-1) h(-1), p = 0.08) or exercise metabolic rates (x control = 0.53 +/- 0.06 mL O2 g(-1) h(-1); x ash =0.44 +/- 0.04 mL O2 g(-1) h(-1), p = 0.47) plasma glucose levels (p = 0.6), and hepatic or muscle percentage indices (p > 0.2 in all cases) whether food was limited or not.