Changes in the material properties of the shell during simulated aquatic hibernation in the anoxia-tolerant painted turtle.

Western painted turtles (Chrysemys picta bellii) tolerate anoxic submergence longer than any other tetrapod, surviving more than 170 days at 3°C. This ability is due, in part, to the shell and skeleton simultaneously releasing calcium and magnesium carbonates, and sequestering lactate and H+ to prevent lethal decreases in body fluid pH. We evaluated the effects of anoxic submergence at 3°C on various material properties of painted turtle bone after 60, 130 and 167-170 days, and compared them with those of normoxic turtles held at the same temperature for the same time periods. To assess changes in the mechanical properties, beams (4×25 mm) were milled from the plastron and broken in a three-point flexural test. Bone mineral density, CO2 concentration (a measure of total bone HCO3-/CO32-) and elemental composition were measured using microcomputed tomography, HCO3-/CO32- titration and inductively coupled plasma mass spectrometry (ICP-MS), respectively. Tissue mineral density of the sampled bone beams was not significantly altered by 167-170 days of aquatic overwintering in anoxic or normoxic water, but bone CO2 and Mg were depleted in anoxic compared with normoxic turtles. At this time point, the plastron beams from anoxic turtles yielded at stresses that were significantly smaller and strains that were significantly greater than the plastron beams of normoxic turtles. When data from anoxic and normoxic turtles were pooled, plastron beams had a diminished elastic modulus after 167-170 days compared with those of control turtles sampled on day 1, indicating an effect of prolonged housing of the turtles in 3°C water without access to basking sites. There were no changes in the mechanical properties of the plastron beams at any of the earlier time points in either group. We conclude that anoxic hibernation can weaken the painted turtle's plastron, but likely only after durations that exceed what it might naturally experience. The duration of aquatic overwintering, regardless of oxygenation state, is likely to be an important factor determining the mechanical properties of the turtle shell during spring emergence.

Barcoding Techniques Help Tracking the Evolutionary History of the Introduced Species Pennaria disticha (Hydrozoa, Cnidaria).

The Christmas tree hydroid Pennaria disticha is listed as one of the most common introduced species in Hawaii. Firstly reported in Kaneohe Bay (Oahu) in 1928, it is now established throughout the entire archipelago, including the Northwestern Hawaiian Islands, a U.S. National Monument and World Heritage site. The Hawaiian population of P. disticha has also been reported as being the source of further introductions to Palmyra Atoll in the U.S. Line Islands. Using a phylogenetic hypothesis based on a 611 base pair fragment of the mitochondrial 16S barcoding gene, we demonstrate that P. disticha is a complex of cryptic species, rather than one species with cosmopolitan distribution. We also show that in Hawaii there are three species of Pennaria, rather than one introduced species. Two of these species share haplotypes with specimens from distant locations such as Florida and Panama and may have been introduced, possibly from the Atlantic Ocean. A third species could either represent a lineage with nearly cosmopolitan distribution, or another introduced species. Our dataset refutes the widely accepted idea that only one lineage of P. disticha is present in Hawaii. On the contrary, P. disticha in Hawaii may be the outcome of multiple independent introductions of several morphologically undistinguishable cryptic lineages. Our results uncover an unsuspected complexity within the very common hydroid P. disticha, and highlight the need for routine use of molecular tools, such as DNA barcoding, to improve the identification and recognition of non-indigenous species.