Differences in THg in wood frog vs. boreal chorus frog tadpoles
Body burden and THg concentration in all tadpoles ranged from 0.5 to 2300.5 ng/frog dry weight (dw) and 25.9 to 7889.3 ng/g dw, respectively (
Tables S1 and
S2). Lower values were not outside the range of those measured in other studies; however, upper values exceeded those of any previous study, including in amphibians from some contaminated studies (see
Bradford et al. 2012 for a summary). The variables species, stage, and mass explained 21% of total variation for body burden in our models but had negligible predictive ability for THg concentration (
Table S3). Nearly all the remaining variation in both body burden and THg concentration was attributable to differences among ponds (
R 2 = 96% when including the random pond effect in both cases;
Table S3).
The THg body burden of boreal chorus frog tadpoles (
n = 32, mean ± standard error: 33 ± 17 ng/frog) was lower than that of wood frog tadpoles (
n = 61, mean = 329 ± 76 ng;
Fig. 2a). THg concentration showed a similar trend (mean = 597.2 ± 132.1 ng/g and 1668.9 ± 351.6 ng/g for boreal chorus frogs and wood frogs, respectively;
Fig. 2b). Bioaccumulation of MeHg through trophic levels has been well documented, as prey items are the primary source of MeHg for aquatic organisms (
Hall et al. 1997;
Rimmer et al. 2005;
Ward et al. 2010). Boreal chorus frog tadpoles are herbivorous and feed on algae, whereas wood frog tadpoles are omnivorous, and likely supplement a primary diet of algae with invertebrates (e.g., Odonata and Dytiscidae) and eggs of amphibians including wood frog tadpoles (
McDiarmid and Altig 1999;
Wells 2007). Thus, the lower values observed in boreal chorus frogs were not surprising considering the trophic position of both species.
There was also high variation within species within ponds, especially for wood frog tadpoles. This may be due to life history parameters such as stage of development, because greater development leads to more opportunity for Hg biomagnification and a change in diet as the organism approaches metamorphosis (
Bradford et al. 2012). To address differences in development timelines, we tested the effect of metamorphic stage on body burden and THg concentration. Models that included both species and stage were unambiguously supported over other models for body burden (Σω
i > 0.999 for both variables;
Table 2), providing further evidence of Hg bioaccumulation with metamorphic stage. There was also some evidence that tadpoles with greater mass had a higher body burden, after accounting for effects of both species and metamorphic stage (Σω
i > 0.77;
Table 2). We could not definitively conclude that any of the predictors had an effect on THg concentration (Σω
i = 0.661, 0.727, and 0.407 for species, stages, and mass, respectively), but there was some evidence that more developed wood frogs had a higher THg concentration than less developed boreal chorus frogs (
Fig. S1).
Given the lack of study of Hg in amphibians in prairie wetlands, we decided to compare our results with lower trophic organisms from other studies in similar systems. Gastropods and some Corixidae feed on algae and are therefore at a similar trophic level as boreal chorus frog tadpoles (
Clarke 1981;
Merritt and Cummins 1996). The THg concentration in pond snails (Gastropods and Lymnaeidae) and aquatic insects (Corixidae, Odonata, Dytiscidae, and Notonectidae) from wetlands in the PPR ranged from 11.7 to 161.2 ng/g (
Bates 2010) and 9.7 and 507.1 ng/g (
Bates and Hall 2012), respectively. These values were lower than our boreal chorus frog tadpoles (25.9–2736.4 ng/g). The potentially higher accumulation in tadpoles compared with other organisms at similar trophic levels suggests that tadpoles may be an important vector for Hg within aquatic and terrestrial food webs as the Hg accumulated by tadpoles is retained throughout metamorphosis (
Bergeron et al. 2010a). Elevated THg levels may be problematic in areas where tadpoles are abundant prey sources for higher trophic levels, as tadpoles may increase the amount of MeHg accumulated through food webs.
Why were concentrations in these ponds so high?
Factors that influence HgII methylation and MeHg bioavailability are important controls in MeHg bioaccumulation in aquatic organisms (
Wiener 2010;
Li and Cai 2013;
Hsu-Kim et al. 2013;
Paranjape and Hall 2017). We explored water chemistry and physical parameters that are often useful in explaining the variation of MeHg and THg in whole water (such as DOC, pH, total anion concentrations; see Methods section for complete list), and thus Hg in aquatic animals, using PCA (
Fig. 4). We did not identify any water chemistry parameters, including THg and MeHg concentrations in water, as being strongly associated with the concentration of Hg in tadpoles in our systems.
Differences in Hg concentration in the ponds were apparent for both species of tadpoles, suggesting that Hg concentration in algae and lower trophic organisms must also have been elevated because diet is the main vector of Hg in most vertebrates (
Burger et al. 2014). Another explanation of high values in certain ponds is that the gastrointestinal tracts (GT) of tadpoles sampled from the high Hg ponds contained sediment with an elevated Hg concentration.
Burger and Snodgrass (1998) showed that the concentration of Hg in tadpoles GTs was elevated compared with Hg concentration in the tail or rest of the body, suggesting that gut contents can contribute to overall body concentration. In addition, tadpoles have been shown to accumulate metals from sediment only (
Snodgrass et al. 2004). We did not measure THg concentration in sediments in our ponds. However, our previous work has shown that Hg concentration in sediments can vary in these types of systems (
Hoggarth et al. 2015), and it is therefore conceivable that the high Hg ponds had elevated Hg concentration in the sediments. If sediments were a significant source of Hg for tadpoles, we would expect to find a high inorganic Hg to MeHg ratio in the organisms. A further exploration of Hg speciation in tissues is required, and clearly a more thorough study on the biogeochemical cycling of Hg, including concentration in the sediments, in these high Hg ponds is warranted.
Wetland geomorphology is likely important in the determination of Hg levels in aquatic organisms. For example, natural areas tend to retain more water in soils and thus input less water into wetlands compared with agricultural areas (
van der Kamp et al. 2003), leading to a greater diversity in hydrological patterns, especially in prairie wetland ponds (
Hayashi et al. 2016). Wet–dry cycling is important for the methylation process as it reoxidizes sulfur for metabolism by sulfate reducing bacteria, and therefore, methylation of HgII, and subsequent bioaccumulation of MeHg in biota, may be influenced by the frequency of drying and flooding in wetlands (
Sando et al. 2007;
Eckley et al. 2015;
Singer et al. 2016). Consequently, intermittent wetlands will contain higher amounts of MeHg in the water and in residing fish (
Ackerman and Eagles-Smith 2010;
Coleman Wasik et al. 2015). Second, the vegetation composition of riparian zones may also be an important factor in determining MeHg levels in aquatic habitats because a higher amount of shading by riparian zones is associated with higher MeHg levels in aquatic organisms (
Balogh et al. 2002;
Ward et al. 2010). Riparian vegetation of our ponds was typical of PPR wetlands and could provide shading of sediments where methylation could be occurring. These two examples demonstrate that the processes controlling Hg accumulation in these tadpoles are much more complex than inorganic Hg deposition alone.