Northern map turtles (Graptemys geographica) derive energy from the pelagic pathway through predation on zebra mussels (Dreissena polymorpha)

1. Zebra mussels ( Dreissena polymorpha ) derive their energy from the pelagic energy pathway by filtering plankton. Because zebra mussels occur in high densities in littoral habitats, they potentially constitute an important trophic link between littoral consumers and pelagic energy sources. Northern map turtles ( Graptemys geographica ) are widespread in North America and consume zebra mussels.
2. We used stable isotopes analyses to quantify the flow of energy from the pelagic pathway to northern map turtles and to infer the contribution of zebra mussels to map turtle biomass. We then built a bioenergetic model to estimate the annual intake of zebra mussels by northern map turtles in Lake Opinicon, Ontario, Canada.
3. Stable isotopes analyses indicated that zebra mussels constitute between 0% and 14% of the diet of males and between 4% and 36% of the diet of females. Assuming that zebra mussels account for all of the pelagic contribution, we estimated that map turtles consume 3200 kg of zebra mussels annually. Because female map turtles are much larger than males and consume more zebra mussels, they are responsible for 95% of the zebra mussel biomass ingested annually.
4. The pelagic pathway supports an important part of the standing crop biomass of map turtles in Lake Opinicon. We highlight the importance of freshwater turtles in lake ecosystems. Unravelling the trophic interactions mediated by freshwater turtles will lead to a more integrated picture of lake ecosystems.     

Patterns of chondrification and ossification in the skull of Graptemys pseudogeographica,the false map turtle (Emydidae)

Patterns of ossification and chondrification are well‐described for several species of turtles, but details of the chondrocranial anatomy are known for only a handful of species. Cleared and double‐stained embryos of Graptemys pseudogeographica were used to examine the fully formed chondrocranium and the formation, chondrification, and ossification of the cranium. The chondrocranium of G. pseudogeographica possesses an unusually large, irregularly shaped foramen epiphaniale that is joined with the fenestra olfactoria. As in other emydids, and many turtles generally, the taenia marginalis is present only as a small projection and the taenia medialis is lacking in mature stages of embryonic development. Ossification data for G. pseudogeographica are consistent with those of other Testudines in that the dentary and maxilla (dermal elements of the upper and lower jaws) ossify early, whereas the articular (an endochondral bone of the lower jaw) ossifies relatively late. Additionally, comparative ossification shows that the vomer is quite variable in its relative timing of ossification across Testudines.     

Evolution of the relative width of the head and alveolar surfaces in map turtles (Testudines: Emydidae: Graptemys)

Replicate adaptive radiations are dichotomous morphological patterns related to resource use and repeated in a series of isolated habitats. Biologists have long noted a dichotomy in relative head width and alveolar width among species of map turtles ( Graptemys ), particularly with regard to adult females, which are much larger dian adult males in all 12 species. I measured plastron length (PL), head width (HW), and alveolar width (AW) of nearly 2300 specimens representing the 12 recognized species, and used allometric regressions to investigate this genus as an example of both sexual dimorphism in trophic morphology and replicate adaptive radiation. Sexual dimorphism was noted in HW after correction for PL, with females having wider heads than males of similar body sizes, but AW as corrected for HW was not dimorphic, thus absolute differences in AW are the simple result of differences in HW. Sexual differences in HW probably relate to the dimorphic niches of Graptemys , as females of three species have been reported to occupy deeper water further from shore than conspecific males. Based on predicted HW at maximum PL in adult females, species were categorized as mega-, meso-, or microcephalic. Differences in relative head width are related to differences in degree of molluscivory, with little molluscivory in microcephalic females, moderate to high molluscivory in mesocephalic females, and high molluscivory in megacephalic females. Distributions of Graptemys species in sympatry and allopatry reveal patterns consistent with structuring of distributions via competitive interactions. Superimposing data on relative HW on a phylogeny of the genus suggests there may have been one or two episodes of character displacement, widi subsequent character assortment, in Graptemys evolution. This conclusion is similar to the current model for the evolution of body size during the radation of Anolis lizards in the northern Lesser Antilles.     

Physiology of common map turtles (Graptemys geographica) hibernating in the Lamoille river, Vermont

Common map turtles (Graptemys geographica) were collected from a natural underwater hibernaculum in Vermont at monthly intervals during the winter of 1997-1998. Blood was sampled by cardiac puncture and analyzed for pH, PCO(2), PO(2), and hematocrit; separated plasma was tested for Na(+), K(+), Cl(-), total [Ca], total [Mg], [lactate], and osmolality (mOsm kg(-1) H(2)O). Control (eupneic; 1 degrees C) values for pH, PO(2), PCO(2), [HCO(3)(-)], and [lactate] were 7.98 +/- 0.03, 47.4 +/- 18.7, 10.1 +/- 0.7 (mm Hg), 36.1 +/- 0.2 (mmol liter(-1)), and 2.1 +/- 0.1 (mmol liter(-1)), respectively. Between November 1997 and March 1998, ice covered the river and the turtles rested on the substratum, fully exposed to the water, and were apneic. Blood PO(2) was maintained at less than 3 mm Hg (range 0.9 +/- 0.2 to 2.1 +/- 0.7 mm Hg), PCO(2) decreased slightly, plasma [lactate] was <5 mmol liter(-1), and plasma [HCO(3)(-)] decreased significantly. In March [lactate] rose to 7.5 +/- 1.5 mmol liter(-l), but there was no acidemia. Map turtles meet most of their metabolic demand for O(2) via aquatic respiration and tolerate prolonged submergence at 1 degrees C with little change in acid-base or ionic status. The adaptive significance of remaining essentially aerobic during winter is to avoid the life-threatening progressive acidosis that results from anaerobic metabolism. J. Exp. Zool. 286:143-148, 2000.     

Eimeria juniataensis sp. n. (Protozoa: Eimeriidae) from the map turtle, Graptemys Geographica, in Pennsylvania.

Eimeria juniataensis sp. n. is described from the map turtle, Graptemys geographica, in Pennsylvania. The spherical to subspherical oocysts of E. juniataensis are 11.5 to 18.5 mu by 11.5 to 16.5 mu (mean, 13.5 by 12.9mu). The broadly fusiform sporocysts are 7.5 to 10 mu by 4 to 6.5 mu (mean, 8.3 by 5.0mu) and a Stieda body is present. Oocyst and sporocyst residua are present. This is the first report of an eimerian species from Graptemys.     

Conservation genetics of the yellow-blotched sawback Graptemys flavimaculata (Testudines: Emydidae)

Turtles within the genus Graptemys are increasingly becoming a group of conservation priority due to (1) the number of species endemic to single Gulf of Mexico rivers and (2) human alterations of riverine habitat. The yellow-blotched sawback (Graptemys flavimaculata) is a federally threatened species that is endemic to the Pascagoula River system of southeastern Mississippi, USA. Currently, there is nothing known about genetic variation and population structure in G. flavimaculata. We used microsatellite data to analyze population genetic structure, assess historical demography, and determine effective population size at six sites throughout the Pascagoula River system. Considerable genetic diversity was found within each site (mean allelic richness: 6.65–8.08) and two analyses found no evidence of genetic bottlenecks. All of the pairwise F _ST values, while low (average = 0.026), were significant, with most sites possessing one or more private alleles. Pairwise F _ST values with the Escatawpa River site were larger (0.030–0.047), which likely reflect its historical isolation. Genetic distance was correlated to geographic distance between sites, with the exception of the Escatawpa River site; a similar pattern was also found with estimates of recent rates of migration among sites. While an analysis of molecular variance indicated that most variation was partitioned within rather than among sites, STRUCTURE analysis strongly supported the recognition of two distinct groups (mainstem Pascagoula vs. Escatawpa), with the possibility of additional substructure within the mainstem Pascagoula.     

The physiology of hibernation in common map turtles (Graptemys geographica).

Map turtles from Wisconsin were submerged at 3 degrees C in normoxic and anoxic water to simulate extremes of potential respiratory microenvironments while hibernating under ice. In predive turtles, and in turtles submerged for up to 150 days, plasma PO2, PCO2) pH, [Cl-], [Na+], [K+], total Mg, total Ca, lactate, glucose, and osmolality were measured; hematocrit and body mass were determined, and plasma [HCO3-] was calculated. Turtles in anoxic water developed a severe metabolic acidosis, accumulating lactate from a predive value of 1.7 to 116 mmol/l at 50 days, associated with a fall in pH from 8.010 to 7.128. To buffer lactate increase, total calcium and magnesium rose from 3.5 and 2.0 to 25.7 and 7.6 mmol/l, respectively. Plasma [HCO3-] was titrated from 44.7 to 4.3 mmol/l in turtles in anoxic water. Turtles in normoxic water had only minor disturbances of their acid-base status and ionic statuses; there was a marked increase in hematocrit from 31.1 to 51.9%. This study and field studies suggest that map turtles have an obligatory requirement for a hibernaculum that provides well-oxygenated water (e.g. rivers and large lakes rather than small ponds and swamps) and that this requirement is a major factor in determining their microdistribution.     

Nesting fidelity and molecular evidence for natal homing in the freshwater turtle, Graptemys kohnii

Numerous studies of sea turtle nesting ecology have revealed that females exhibit natal homing, whereby they imprint on the nesting area from which they hatch and subsequently return there to nest as adults. Because freshwater turtles comprise the majority of reptiles known to display environmental sex determination (ESD), the study of natal homing in this group may shed light on recent evolutionary models of sex allocation that are predicated on natal homing in reptiles with ESD. We examined natal homing in Graptemys kohnii, a freshwater turtle with ESD, using mitochondrial sequencing, microsatellite genotyping and mark and recapture of 290 nesting females. Females showed high fidelity to nesting areas, even after being transplanted several kilometres away. A Mantel test revealed significant genetic isolation by distance with respect to nesting locations (r=0.147; p<0.05), suggesting that related females nest in close proximity to one another. The patterns of fidelity and genotype distributions are consistent with homing at a scale that may affect population sex ratios.     

Intersexual allometry differences and ontogenetic shifts of coloration patterns in two aquatic turtles,Graptemys oculifera and Graptemys flavimaculata

Coloration can play critical roles in a species' biology. The allometry of color patterns may be useful for elucidating the evolutionary mechanisms responsible for shaping the traits. We measured characteristics relating to eight aspects of color patterns from Graptemys oculifera and G. flavimaculata to investigate the allometric differences among male, female, and unsexed juvenile specimens. Additionally, we investigated ontogenetic shifts by incorporating the unsexed juveniles into the male and female datasets. In general, male color traits were isometric (i.e., color scaled with body size), while females and juvenile color traits were hypoallometric, growing in size more slowly than the increase in body size. When we included unsexed juveniles in our male and female datasets, our linear regression analyses found all relationships to be hypoallometric and our model selection analysis found support for nonlinear models describing the relationship between body size and color patterns, suggestive of an ontogenetic shift in coloration traits for both sexes at maturity. Although color is critical for many species' biology and therefore under strong selective pressure in many other species, our results are likely explained by an epiphenomenon related to the different selection pressures on body size and growth rates between juveniles and adults and less attributable to the evolution of color patterns themselves.     

Diversity of North American map and sawback turtles (Testudines: Emydidae: Graptemys)

Map turtles of the genus Graptemys are native to North America, where a high degree of drainage endemism is believed to have shaped current diversity. With 14 species and one additional subspecies, Graptemys represents the most diverse genus in the family Emydidae. While some Graptemys species are characterized by pronounced morphological differences, previous phylogenetic analyses have failed yet to confirm significant levels of genetic divergence for many taxa. As a consequence, it has been debated whether Graptemys is taxonomically inflated or whether the low genetic divergence observed reflects recent radiations or ancient hybridization. In this study, we analysed three mtDNA blocks (3228 bp) as well as 12 nuclear loci (7844 bp) of 89 specimens covering all species and subspecies of Graptemys. Our analyses of the concatenated mtDNA sequences reveal that the widespread G. geographica constitutes the sister taxon of all other Graptemys species. These correspond to two clades, one comprised of all broad‐headed Graptemys species and another clade containing the narrow‐headed species. Most species of the broad‐headed clade are reciprocally monophyletic, except for G. gibbonsi and G. pearlensis, which are not differentiated. By contrast, in the narrow‐headed clade, many currently recognized species are not monophyletic and divergence is significantly less pronounced. Haplotype networks of phased nuclear loci show low genetic divergence among taxa and many shared haplotypes. Principal component analyses using coded phased nuclear DNA sequences revealed eight distinct clusters within Graptemys that partially conflict with the terminal mtDNA clades. This might be explained by male‐mediated gene flow across drainage basins and female philopatry within drainage basins. Our results support that Graptemys is taxonomically oversplit and needs to be revised.     

Myxidium mackiei (Myxosporea) in Indo-Gangetic flap-shelled turtles Lissemys punctata andersonii: parasite-host interaction and ultrastructure

Myxosporeans are common parasites of fish, and uncommon parasites of amphibians, reptiles and invertebrates, that can cause significant morbidity and mortality. The common genus Myxidium infects the excretory system of turtles, yet knowledge of its pathogenicity in these hosts is limited. We offer new knowledge of morphological and ultrastructural aspects of host-parasite interactions in Myxidium infections from our recent diagnostic investigations on captive freshwater turtles listed in CITES (Appendix II). We investigated the cause of death of 2 adult Indo-Gangetic flap-shelled turtles Lissemys punctata andersonii from a zoo collection. After post-mortem examination, tissues were processed for histopathology, and special stains were used to demonstrate morphology of myxosporean spores. Additional kidney tissue, immersion-fixed in formalin, was processed for transmission electron microscopy. Both turtles were infected with a myxosporidian, Myxidium mackiei, in the kidney, which occluded 5 to 10% of the renal proximal convoluted tubules. The polysporic plasmodia contained pairs of developing and mature spores. Each mature, spindle-shaped spore had 2 asymmetric valves (1 overlapping, 1 overlapped), with 10 to 13 and 10 to 14 longitudinal ridges per valve, and 2 polar capsules each containing a polar filament with 4 to 5 turns. A pair of spores, each surrounded by a membrane-bound electron-lucent matrix, lay in an enclosing cell within the plasmodium. Regions of the parasite-host interface consisted of undulations of the parasite surface, with intense pinocytotic activity beneath, intermingled with the hosts' microvilli, and endocytotic channels at the apex of renal epithelial cells. The microvilli of the renal epithelial cells of infected tubules were frequently sheared or compressed, or occasionally missing; we did not detect other pathology induced by the parasite. Our report of M. mackiei in L. punctata is a new host record. Both individuals also had disseminated pale yellow nodules (bacterial granulomas) present in lung, heart, kidney, and skeletal muscle, and both were infected with coccidia (tentatively identified as Eimeria sp.) in multiple organs. The cause of death for one turtle was septicemia, but remained unknown for the other individual.     

Detection of low plasma estradiol concentrations in nesting green turtles (Chelonia mydas) by HPLC/Ms-Ms

In previous studies on nesting green turtles under natural conditions from different geographical regions, 17-β-estradiol (E(2) ) was either undetectable or detected at very low levels. RIA and other related techniques were not sensitive enough to measure low E(2) values in the green turtles. In this study, a sensitive method was used in detecting low hormone concentrations: high performance liquid chromatography with tandem quadruple mass spectrometry (HPLC-MS/MS). Using this technique, estradiol for the first time was detected in nesting green turtles during the peak season (June-October) at Ras Al-Hadd Reserve, Oman. The E(2) values recorded from this study were the highest ever recorded from nesting green turtles in any geographical region, but the levels did not vary significantly throughout different phases of nesting. The presence of E(2) during nesting presumably plays a role in the physiology and behavior of this species. Ras Al-Hadd hosts one of the largest nesting populations of green turtles in the world, and an understanding of their nesting patterns may be of value in conservation and management programs for this endangered species.     

Adaptations to terrestrial overwintering of hatchling northern map turtles,<Emphasis Type="Italic"> Graptemys geographica</Emphasis>

We conducted a 3-year field and laboratory study of winter biology in hatchlings of the northern map turtle (Graptemys geographica). At our study area in northern Indiana, hatchlings routinely overwintered in their natal nests, emerging after the weather warmed in spring. Winter survival was excellent despite the fact that hatchlings were exposed frequently to subfreezing temperatures (to –5.4 °C). In the laboratory, cold-acclimated hatchlings exhibited low rates of evaporative water loss (mean=2.0 mg g^–1 day^–1), which would enable them to conserve body water during winter. Laboratory-reared hatchlings were intolerant of freezing at –2.5 °C for 24 h, conditions that are readily survived by freeze-tolerant species of turtles. Winter survival of hatchling G. geographica probably depended on their extensive capacity for supercooling (to –14.8 °C) and their well-developed resistance to inoculative freezing, which may occur when hatchlings contact ice and ice-nucleating agents present in nesting soil. Supercooled hatchlings survived a brief exposure to –8 °C. Others, held at –6 °C for 5 days, maintained ATP concentrations at control levels, although they did accumulate lactate and glucose, probably in response to tissue hypoxia. Therefore, anoxia tolerance, as evidenced by the viability of hatchlings exposed to N₂ gas for 8 days, may promote survival during exposure to subfreezing temperatures.Abbreviations EWL evaporative water loss - FP_eq equilibrium freezing point - INA ice-nucleating agents - T_c temperature of crystallizationCommunicated by L.C.-H. Wang     

Relationships among turtles of the genus Clemmys (Reptilia, Testudines, Emydidae) as suggested by plastron scute morphology

The Conservative morphology of hardshelled turtles has fostered the use of size relationships between epidermal scutes (scales) on the shell to differentiate between species and subspecies of many taxa. The size relationship of the six major pairs of plastral scutes were used to compare the four currently recognized species of the genus Clemmys with each other. as well as with the distantly related Graptemys barbouri using Jaccard Coefficients. Shannon-Weiner diversity indices, and multivariate analysis. Results were concordant among the three techniques used and confirm our prediction that plastral morphology varies little among closely related species and widely among distantly related taxa. Clemmys muhlenbergii appears to he more different from Clemmys guttata than previously suggested. Analysis of plastral morphology shows promise as a taxonomic tool for turtle systematists.     

Blood oxygen transport in common map turtles during simulated hibernation

We assessed the effects of cold and submergence on blood oxygen transport in common map turtles (Graptemys geographica). Winter animals were acclimated for 6-7 wk to one of three conditions at 3 degrees C: air breathing (AB-3 degrees C), normoxic submergence (NS-3 degrees C), and hypoxic (PO2=49 Torr) submergence (HS-3 degrees C). NS-3 degrees C turtles exhibited a respiratory alkalosis (pH 8.07; PCO2=7.9 Torr; [lactate]=2.2 mM) relative to AB-3 degrees C animals (pH 7.89; PCO2=13.4 Torr; [lactate]=1.1 mM). HS-3 degrees C animals experienced a profound metabolic acidosis (pH 7.30; PCO2=7.9 Torr; [lactate]=81 mM). NS-3 degrees C turtles exhibited an increased blood O2 capacity; however, isoelectric focusing revealed no seasonal changes in the isohemoglobin (isoHb) profile. Blood O2 affinity was significantly increased by cold acclimation; half-saturation pressures (P50's) for air-breathing turtles at 3 degrees and 22 degrees C were 6.5 and 18.8 Torr, respectively. P50's for winter animals submerged in normoxic and hypoxic water were 5.2 and 6.5 Torr, respectively. CO2 Bohr slopes (Delta logP50/Delta pH) were -0.15, -0.16, and -0.07 for AB-3 degrees C, NS-3 degrees C, and HS-3 degrees C turtles, respectively; the corresponding value for AB-22 degrees C was -0.37. The O2 equilibrium curve (O2EC) shape was similar for AB-3 degrees C and NS-3 degrees C turtles; Hill plot n coefficients ranged from 1.8 to 2.0. The O2EC shape for HS-3 degrees C turtles was anomalous, exhibiting high O2 affinity below P50 and a right-shifted segment above half-saturation. We suggest that increases in Hb-O2 affinity and O2 capacity enhance extrapulmonary O2 uptake by turtles overwintering in normoxic water. The anomalous O2EC shape and reduced CO2 Bohr effect of HS-3 degrees C turtles may also promote some aerobic metabolism in hypoxic water.     

Phylogeny and paraphyly among tetrapod blood flukes (Digenea: Schistosomatidae and Spirorchiidae)

The blood flukes of turtles (Digenea: Spirorchiidae) and the blood flukes of crocodilians, birds and mammals (Digenea: Schistosomatidae) have long been considered as closely related, but distinct evolutionary lineages. Recent morphological and molecular studies have considered these families as sister taxa within the Schistosomatoidea. Representatives of both families have similar furcocercous cercariae and similar two-host life cycles, but have different definitive hosts, distinct reproductive patterns and different morphologies. Sequences including approximately 1800 bases of the small subunit ribosomal DNA and 1200 bases of the large subunit ribosomal DNA were generated from representatives of eight spirorchiid genera. These sequences were aligned with pre-existing sequences of Schistosomatidae and other representatives of the Diplostomida and analysed for phylogenetic signal using maximum parsimony and Bayesian inference. These analyses revealed that the Spirorchiidae is paraphyletic and that the turtle blood flukes are basal to the highly derived schistosomatids. Three genera of spirorchiids from marine turtles form a sister group to the Schistosomatidae and five genera of spirorchiids from freshwater turtles occupy basal positions in the phylogeny of tetrapod blood flukes. Marine turtles are considered to be derived from freshwater turtles and the results of the current study indicate that the spirorchiid parasites of marine turtles are similarly derived from a freshwater ancestor. The close relationship of the marine spirorchiids to schistosomatids and the basal position of the marine transmitted Austrobilharzia and Ornithobilharzia in the schistosomatid clade suggests that schistosomatids arose after a marine turtle blood fluke ancestor successfully colonised birds.     

Cytonuclear equilibrium following interspecific introgression in a turtle lacking sex chromosomes

When reproductive barriers break down, interspecific hybridization can lead to gene flow between evolutionarily distinct species. Studying the fate of these introgressing elements can offer valuable insights into the factors contributing to reproductive isolation. We have identified a population of false map turtles (Graptemys pseudogeographica) that hybridized historically with the common map turtle (Graptemys geographica), but were subsequently isolated from interbreeding for several generations by unique geological events. Although many studies conclude that genic interactions involving sex chromosomes impact the introgression of mitochondrial or nuclear genomes, Graptemys turtles have environmental sex determination, and thus introgression can be explored while controlling for the effects of sex‐specific heterogameity. We identified and sequenced a species‐specific mitochondrial control region marker, as well as two nuclear markers (ODC and HNFAL), in turtles from across the ranges of these species. We found both nuclear and mitochondrial introgression in our study population, and present evidence consistent with the proposed time range of reproductive contact and isolation. We also report an absence of cytonuclear or linkage disequilibrium among markers, indicating that some important pre‐ and postzygotic barriers to gene flow that characterize other systems are absent in Graptemys. Finally, we show that Graptemys turtles have a complex molecular evolutionary history, and that leaks in reproductive barriers probably occur frequently. © 2012 The Linnean Society of London, Biological Journal of the Linnean Society, 2012, 106 , 405–417.