Tobias Landberg

Lung ventilation during treadmill locomotion in a terrestrial turtle, Terrapene carolina.

SUMMARY The limb girdles and lungs of turtles are both located within the bony shell, and therefore limb movements during locomotion could affect breathing performance. A mechanical conflict between locomotion and lung ventilation has been reported in adult green sea turtles, Chelonia mydas, in which breathing stops during terrestrial locomotion and resumes during pauses between bouts of locomotion. We measured lung ventilation during treadmill locomotion using pneumotach masks in three individual Terrapene carolina (mass 304-416 g) and found no consistent mechanical effects of locomotion on breathing performance. Relatively small tidal volumes (2.2±1.4 ml breath-1; mean ± s.d., N=3 individuals) coupled with high breath frequencies (36.6±26.4 breaths min-1; mean ± s.d., N=3 individuals) during locomotion yield mass-specific minute volumes that are higher than any previously reported for turtles (264±64 ml min kg-1; mean ± s.d., N=3 individuals). Minute volume was higher during locomotion than during recovery from exercise (P<0.01; paired t-test), and tidal volumes measured during locomotion were not significantly different from values measured during brief pauses between locomotor bouts or during recovery from exercise (P>0.05; two-way ANOVA). Since locomotion does not appear to conflict with breathing performance, the mechanism of lung ventilation must be either independent of, or coupled to, the stride cycle. The timing of peak airflow from breaths occurring during locomotion does not show any fixed phase relationship with the stride cycle. Additionally, the peak values of inhalatory and exhalatory airflow rates do not differ consistently with respect to the stride cycle. Together, these data indicate that T. carolina is not using respiratory-locomotor coupling and limb and girdle movements do not contribute to lung ventilation during locomotion. X-ray video recordings indicate that lung ventilation is achieved via bilateral activity of the transverse (exhalatory) and oblique (inhalatory) abdominal muscles. This specialized abdominal ventilation mechanism may have originally circumvented a mechanical conflict between breathing and locomotion in the ancestor of turtles and subsequently allowed the ribs to abandon their role in lung ventilation and to fuse to form the shell.

Prey Responses to Predator Chemical Cues: Disentangling the Importance of the Number and Biomass of Prey Consumed

To effectively balance investment in predator defenses versus other traits, organisms must accurately assess predation risk. Chemical cues caused by predation events are indicators of risk for prey in a wide variety of systems, but the relationship between how prey perceive risk in relation to the amount of prey consumed by predators is poorly understood. While per capita predation rate is often used as the metric of relative risk, studies aimed at quantifying predator-induced defenses commonly control biomass of prey consumed as the metric of risk. However, biomass consumed can change by altering either the number or size of prey consumed. In this study we determine whether phenotypic plasticity to predator chemical cues depends upon prey biomass consumed, prey number consumed, or both. We examine the growth response of red-eyed treefrog tadpoles (Agalychnis callidryas) to cues from a larval dragonfly (Anax amazili). Biomass consumed was manipulated by either increasing the number of prey while holding individual prey size constant, or by holding the number of prey constant and varying individual prey size. We address two questions. (i) Do prey reduce growth rate in response to chemical cues in a dose dependent manner? (ii) Does the magnitude of the response depend on whether prey consumption increases via number or size of prey? We find that the phenotypic response of prey is an asymptotic function of prey biomass consumed. However, the asymptotic response is higher when more prey are consumed. Our findings have important implications for evaluating past studies and how future experiments should be designed. A stronger response to predation cues generated by more individual prey deaths is consistent with models that predict prey sensitivity to per capita risk, providing a more direct link between empirical and theoretical studies which are often focused on changes in population sizes not individual biomass.

Lung ventilation during treadmill locomotion in a semi-aquatic turtle, Trachemys scripta.

It is reasonable to presume that locomotion should have a mechanical effect on breathing in turtles. The turtle shell is rigid, and when the limbs protract and retract, air in the lungs should be displaced. This expectation was met in a previous study of the green sea turtle, Chelonia mydas; breathing completely ceased during terrestrial locomotion (Jackson and Prange, 1979. J Comp Physiol 134:315-319). In contrast, another study found no direct effect of locomotion on ventilation in the terrestrial box turtle, Terrapene carolina (Landberg et al., 2003. J Exp Biol 206:3391-3404). In this study we measured lung ventilation during treadmill locomotion in a semi-aquatic turtle, the red-eared slider, Trachemys scripta. Sliders breathed almost continuously during locomotion and during brief pauses between locomotor bouts. Tidal volume was relatively small (approximately 1 mL) during locomotion and approximately doubled during pauses. Minute ventilation was, however, not significantly smaller during locomotion because breath frequency was higher than that during the pauses. We found no consistent evidence for phase coupling between breathing and locomotion indicating that sliders do not use locomotor movements to drive breathing. We also found no evidence for a buccal-pump mechanism. Sliders, like box turtles, appear to use abdominal musculature to breathe during locomotion. Thus, locomotion affects lung ventilation differently in the three turtle species studied to date: the terrestrial Te. carolina shows no measurable effect of locomotion on ventilation; the semi-aquatic Tr. scripta breathes with smaller tidal volumes during locomotion; and the highly aquatic C. mydas stops breathing completely during terrestrial locomotion.

Environmental context shapes immediate and cumulative costs of risk-induced early hatching

In animals with complex life cycles, fitness trade-offs across life stages determine the optimal time for transitions between stages. If these trade-offs vary predictably, adaptive plasticity in the timing of life history transitions may evolve. For instance, embryos of many species are capable of accelerating hatching to escape from egg predation and other hazards, but for plasticity in hatching timing to be selectively maintained, early hatching must also entail costs, probably in subsequent life stages. However the post-hatching environment, which influences this cost, is variable in nature. We assessed how two elements of the post-hatching environment, predator species and age structure created by hatching age plasticity, affect costs of hatching early in red-eyed treefrogs, Agalychnis callidryas. Red-eyed treefrog embryos were induced to hatch at the onset of hatching competence or near the peak of spontaneous hatching and exposed to one of three insect predators in single or mixed hatching-age treatments. Age structure created by hatching-age plasticity did not affect tadpole survivorship or growth; however, the consequences of hatching timing depended on predator species and foraging mode. Tadpoles that were induced to hatch early experienced initially higher mortality rates only with the more actively foraging predator. Nonetheless, mortality costs of accelerated hatching were apparent with all predators once we factored in the longer duration of exposure that early hatchlings experience in nature. This study suggests that extended exposure of young larvae to predators may be a general cost of early hatching, explaining why spontaneous hatching occurs later in life across variable environmental contexts.

The evolution of locomotor rhythmicity in tetrapods.

Differences in rhythmicity (relative variance in cycle period) among mammal, fish, and lizard feeding systems have been hypothesized to be associated with differences in their sensorimotor control systems. We tested this hypothesis by examining whether the locomotion of tachymetabolic tetrapods (birds and mammals) is more rhythmic than that of bradymetabolic tetrapods (lizards, alligators, turtles, salamanders). Species averages of intraindividual coefficients of variation in cycle period were compared while controlling for gait and substrate. Variance in locomotor cycle periods is significantly lower in tachymetabolic than in bradymetabolic animals for datasets that include treadmill locomotion, non‐treadmill locomotion, or both. When phylogenetic relationships are taken into account the pooled analyses remain significant, whereas the non‐treadmill and the treadmill analyses become nonsignificant. The co‐occurrence of relatively high rhythmicity in both feeding and locomotor systems of tachymetabolic tetrapods suggests that the anatomical substrate of rhythmicity is in the motor control system, not in the musculoskeletal components.

Evolution of maternal egg size effects in sister salamander species

Egg size varies genetically and with the maternal environment. It is correlated with and can act as a resource fueling variation in many other key life history traits. This study examined hypotheses about how plastic responses of offspring to yolk variation evolve (and contribute to phenotypic evolution) when maternal investment in egg size evolves. I used a split-clutch, controlled, surgical experiment with a longitudinal (repeated-measures) design to examine the effects of yolk removal on sister salamander species with distinct egg and larval phenotypes. Yolk removal had large effects in the derived larger-egged species, A. barbouri, and greatly reduced effects in A. texanum. Early hatching and smaller larval body size was only found in A. barbouri and survival rates decreased more in A. barbouri. These results provide strong experimental evidence that as female salamanders evolve greater yolk investment in each egg, offspring coevolve an increased magnitude of phenotypic plasticity in response to yolk variation across a suite of life history traits. Yolk therefore acts as an integrator of phenotypes that allows females to modify modules of life history traits together (facilitating adaptation). When organisms invade new environments, complex integrated phenotypes may evolve via correlated responses to increased maternal investment, yet individual traits can be coupled or decoupled to yolk quantity variation in different species.

Embryonic yolk removal affects a suite of larval salamander life history traits.

Egg size is a key life history trait affecting fitness, and it varies abundantly. The value of egg size to a mother and her offspring is often determined by a trade-off between investing more yolk in a few large eggs or less yolk into many more, smaller eggs. Smaller eggs are generally expected to be phenotypically inferior or females could increase their fitness by making more smaller eggs. However, many females produce a mix of egg sizes and natural yolk variation induces normal developmental responses which may persist into subsequent stages of a complex life history. Since sources of phenotypic variation are easily confounded, I surgically removed yolk from embryonic spotted salamanders (Ambystoma maculatum) using a sham surgery as a control and a split-clutch design to isolate the effects of yolk reserve variation from genetic sources of variation. Yolk removal induced early hatching, reduced developmental stage and hatchling body size. Small hatchlings stayed relatively small through the early larval period, but 17 weeks later the correlation with early larval body size was lost. When the experiment ended, larger individuals were further along in metamorphic development but mortality was independent of early larval body size. Variation in spotted salamander yolk reserves affects a suite of hatchling life history traits that persists into the larval period. Outside the laboratory, egg size effects may cascade throughout complex amphibian life histories. Applied experimentally and comparatively, this simple yolk removal technique may help identify how traits increase or decrease their response to maternal yolk investment.

Embryonic yolk removal affects neither morphology nor escape performance of larval axolotls

Maternal effects, the influences of maternal phenotype on the phenotypes of her offspring, mediate early ontogenetic traits through maternal investment. In amphibians, provisioning eggs with yolk is the main source of maternal investment. While larger eggs generally result in larger, higher-quality offspring, the relationship between egg size and offspring phenotype is complicated because offspring can evolve to be more or less responsive to variation in yolk provisions. Previous studies of several ambystomatid salamanders suggest that the effects of embryonic yolk reserve reduction on hatchling life history traits increase with egg size. In this study, a similar controlled experimental yolk removal technique in Ambystoma mexicanum was used to determine the effects of reduced yolk reserves on phenotypes including hatching time and stage, hatchling and larval size and performance in predation trials with fish. Surprisingly, yolk reduction revealed no effects on any traits. These findings suggest that larval morphology in A. mexicanum is highly canalized and larval phenotypes are decoupled from yolk reserve variation. This surprising lack of yolk removal effects in hatchling and larval axolotls illustrates the evolutionary flexibility of early life history traits. Traits can evolve to increase or decrease their response to resources and can even become completely unresponsive. Since we found no effects in early life history, we hypothesize that domestication of the axolotl may have altered yolk properties or allocation dynamics and that maternal investment in yolk reserves may manifest at later life stages by reducing the time to reproductive maturity or increasing fecundity.