Soil, water, and amphibian tissues collected between 1995 and 1999 from 15 study sites in Bermuda were analysed for pesticides and heavy metals. The most abundant pesticide residue in soil was p,p'-dichlorodiphenyldichloroethylene (DDE) which was found at all sites in concentrations ranging from 0.003 to 4.023 p.p.m. No pesticide residues were found in water. DDE was also recovered from the livers and fat bodies of marine toads (Bufo marinus) and whistling frogs (Eleutherodactylus johnstonei). Analyses of food sources consumed by these anuran species revealed residue levels of p, p'-DDE ranging from 0.05 to 0.217 p.p.m. Other soil residues included dichlorodiphenyltrichloroethane (DDT) at eight study sites, Dicofol(kelthane) at eight sites, dieldrin at five sites, and polychlorinated biphenyls (PCBs) as Arochlor 1254 and Arochlor 1260 at seven sites. Analyses of toad livers revealed significant concentrations of cadmium, chromium, copper and zinc. Livers of Bermuda toads exhibited altered hepatocytic morphology and an increased number of melanomacrophages and possible granulomas, while spleens showed a marked decrease in white pulp. Spleen cells from Bufo marinus collected at one site having high levels of cadmium exhibited a decreased B cell response to lipopolysaccharide. The incidence of trematode infection in Bufo marinus increased from 53.8% in 1995 to 90% in 1999. Deformity rates in the limbs of subadult and adult toads ranged between 15 and 25%. Examination of 1,995 newly-metamorphosed toads revealed deformity rates as high as 47%. The current comprehensive study suggests that environmental pollutants may account for immunosuppression, increased susceptibility to infections, limb malformations and possible decline in amphibian populations from Bermuda.

The interactive effects of contaminants and ultraviolet light (UV)-exposure on the incidence and types of abnormalities observed were measured in newly metamorphosed cane toads (Rhinella marina) from four Bermuda ponds contaminated with petrochemicals and metals. Abnormalities were compared in toadlets that were field-collected, reared in predator exclusion cages, reared in laboratory microcosms exposed to control media or corresponding pond media, and reared in laboratory microcosms exposed to UV-light and control media or media from two ponds. Percent abnormal for field-collected, cage-reared, and microcosm-reared toadlets were equivalent per site and ranged between 14% and 63%. All treatments produced similar limb abnormalities but the percentage of hind versus forelimb defects was statistically greater only in field-collected toadlets. UV-exposed control media did not induce abnormalities in larvae exhibiting no maternal effect, and did not alter the types of abnormalities observed in larvae exhibiting a maternal or latent effect. Site media treatments without UV exposure induced significant cephalic and limb abnormalities, proved additive to the observed maternal/latent effect, and produced limb defects predominantly in forelimbs. Concurrent exposure to site media and UV-light induced similar types of abnormalities but a significantly higher percentage of hind limb abnormalities (68-89%) than exposure to site media alone (7-13%). Our results suggest that the types of abnormalities expressed were principally determined by direct and/or transgenerational contaminant exposure, but that UV-light exposure caused limb abnormalities to occur primarily in the hind limbs, mirroring field observations. Our field observations also suggest that ectromelia and brachydactyly in some field-collected specimens may be predator-induced.

Fetal and maternal heart rates were studied in unrestrained, pregnant harbor seals during the last third of gestation. Heart rates were recorded while the mothers were resting on land or performing trained simulated dives of up to 2.25 min. Data from resting mothers showed the development of a bimodal or two‐speed fetal heart rate pattern during late gestation. The mean faster and slower fetal heart rates at term were 125 ± 3.7 and 79 ± 3.1 (mean + SEM) respectively. The amount of fetal bradycardia observed increased steadily towards term, and fetal heart rate changes were not correlated with changes in maternal heart rate or maternal respiration. The bimodal fetal heart rate was also seen during the simulated dives, and no decrease in either the faster or slower heart rate was found. Heart rates from resting, unrestrained harbor seal pups were also studied. The pups displayed a bimodal heart rate similar to the fetuses' with the slower rate occurring during breath‐holds. The bradycardia in the pups was equivalent to the slower fetal heart rate. These findings suggest that the regulatory mechanism that determines the apneic bradycardia in young harbor seals during non‐stressful conditions develops in the last quarter of gestation.

Short-term embryo-larval, limb development, and a battery of toxicity identification studies using sediment and water collected from selected ponds found that developmentally toxic samples contained elevated levels of petroleum hydrocarbons and As, Cu, Pb, Cr, Cd, Hg, Fe, and Zn. Sediments spiked with diesel fuel or metals confirmed that the levels of petroleum hydrocarbons and metals found in the sediment were capable of inducing developmental toxicity individually. However, joint mixture interaction studies indicated that the metals and petroleum hydrocarbons acted synergistically. To more rigorously evaluate exposure pathways and the effects induced, chronic laboratory-based microcosm experiments were performed using B. marinus from early embryo to metamorphosis. Results from these microcosm studies indicated that reasonably consistent frequencies of malformations were induced by exposure to contaminated media samples from four contaminated sites compared with frequencies found in field-collected specimens. The frequencies of specific types of deformities in newly metamorphosed organisms from the microcosm studies and in free-roaming field-collected specimens were also similar. Exposure to contaminated media from three of the four sites decreased the frequency of metamorphic completion and at one site reduced the metamorphic rate, compared to exposure to laboratory controls or reference media. Elevated whole body petroleum hydrocarbons and metals tissue residues were detected in organisms from the laboratory microcosms, as well as in the free-roaming specimens collected. To evaluate the impact of maternal contaminant transfer as an exposure route, cross-over exposure studies in which reference organisms were raised, from embryos through metamorphosis, in contaminated media and organisms from a contaminated site were cultured in reference media were conducted. Results from these studies indicated that environmental exposure and maternal toxicant transfer were the potential exposure routes. Overall, the present studies suggested the petroleum hydrocarbons and metals that accumulated in B. marinus during development in microcosms corresponded with increased frequencies of malformation.

Widespread introduced species can be leveraged to investigate the genetic, ecological and adaptive processes underlying rapid evolution and range expansion, particularly the contributions of genetic diversity to adaptation. Rhinella marina, the cane toad, has been a focus of invasion biology for decades in Australia. However, their introduction history in North America is less clear. Here, we investigated the roles of introduction history and genetic diversity in establishment success of cane toads across their introduced range. We used reduced representation sequencing (ddRAD) to obtain 34,000 SNPs from 247 toads in native (