Animal distributions are shaped by the environment and antecedents. Here I show how the temperature dependence of aerobic scope (the difference between maximum and minimum rates of oxygen uptake) is a useful tool to examine the fundamental temperature niches of salmonids and perhaps other fishes. Although the concept of aerobic scope has been recognized for over half a century, only recently has sufficient evidence accumulated to provide a mechanistic explanation for the optimal temperature of salmonids. Evidence suggests that heart rate is the primary driver in supplying more oxygen to tissues as demand increases exponentially with temperature. By contrast, capacity functions (i.e. cardiac stroke volume, tissue oxygen extraction and haemoglobin concentration) are exploited only secondarily if at all, with increasing temperature, and then perhaps only at a temperature nearing that which is lethal to resting fish. Ultimately, however, heart rate apparently becomes a weak partner for the cardiorespiratory oxygen cascade when temperature increases above the optimum for aerobic scope. Thus, the upper limit for heart rate may emerge as a valuable, but simple predictor of optimal temperature in active animals, opening the possibility of using biotelemetry of heart rate in field situations to explore properly the full interplay of environmental factors on aerobic scope. An example of an ecological application of these physiological discoveries is provided using the upriver migration of adult sockeye salmon, which have a remarkable fidelity to their spawning areas and appear to have an optimum temperature for aerobic scope that corresponds to the river temperatures experienced by their antecedents. Unfortunately, there is evidence that this potential adaptation is incompatible with the rapid increase in river temperature presently experienced by salmon as a result of climate change. By limiting aerobic scope, river temperatures in excess of the optimum for aerobic scope directly impact upriver spawning migration and hence lifetime fecundity. Thus, use of aerobic scope holds promise for scientists who wish to make predictions on how climate change may influence animal distributions.

Larvae of the sunburst diving beetle, Thermonectus marmoratus, have a cluster of six stemmata (E1-6) and one eye patch on each side of the head. Each eye has two retinas: a distal retina that is closer to the lens, and a proximal retina that lies directly underneath. The distal retinas of E1 and E2 are made of a dorsal and a ventral stack of at least twelve photoreceptor layers. Could this arrangement be used to compensate for lens chromatic aberration, with shorter wavelengths detected by the distal layers and longer wavelengths by the proximal layers? To answer this question we molecularly identified opsins and their expression patterns in these eyes. We found three opsin-encoding genes. The distal retinas of all six eyes express long-wavelength opsin (TmLW) mRNA, whereas the proximal retinas express ultraviolet opsin (TmUV I) mRNA. In the proximal retinas of E1 and E2, the TmUV I mRNA is expressed only in the dorsal stack. A second ultraviolet opsin mRNA (TmUV II), is expressed in the proximal retinas of E1 and E2 (both stacks). The finding that longer-wavelength opsins are expressed distally to shorter-wavelength opsins makes it unlikely that this retinal arrangement is used to compensate for lens chromatic aberration. In addition, we also described opsin expression patterns in the medial retina of E1 and in the non-tiered retina of the lensless eye patch. To our knowledge, this is also the first report of multiple UV opsins being expressed in the same stemma.

Honeybee (Apis mellifera) society is characterized by a helper caste of essentially sterile female bees called workers. Workers show striking changes in lifespan that correlate with changes in colony demography. When rearing sibling sisters (brood), workers survive for 3-6 weeks. When brood rearing declines, worker lifespan is 20 weeks or longer. Insects can survive unfavorable periods on endogenous stores of protein and lipid. The glyco-lipoprotein vitellogenin extends worker bee lifespan by functioning in free radical defense, immunity and behavioral control. Workers use vitellogenin in brood food synthesis, and the metabolic cost of brood rearing (nurse load) may consume vitellogenin stores and reduce worker longevity. Yet, in addition to consuming resources, brood secretes a primer pheromone that affects worker physiology and behavior. Odors and odor perception can influence invertebrate longevity but it is unknown whether brood pheromone modulates vitellogenin stores and survival. We address this question with a 2-factorial experiment where 12 colonies are exposed to combinations of absence vs presence of brood and brood pheromone. Over an age-course of 24 days, we monitor the amount of vitellogenin stored in workers' fat body (adipose tissue). Thereafter, we track colony survival for 200 days. We demonstrate that brood rearing reduces worker vitellogenin stores and colony long-term survival. Yet also, we establish that the effects can result solely from exposure to brood pheromone. These findings indicate that molecular systems of extreme lifespan regulation are integrated with the sensory system of honeybees to respond to variation in a primer pheromone secreted from larvae.

The biogenic amine tyramine (TA) is a potent diuretic factor when applied to the Malpighian tubule (MT) of Drosophila melanogaster, stimulating both urine production and transepithelial chloride conductance. Isolated MTs can respond not only to TA but also to its precursor, tyrosine; this observation led to the proposal that MTs are able to synthesize TA from applied tyrosine through the action of the enzyme tyrosine decarboxylase (TDC). In the current study it is shown that the non-neuronal isoform of TDC, Tdc1, is expressed in the principal cells of the MT. A mutant allele of Tdc1, Tdc1^f03311, was identified that reduced expression of the mature Tdc1 transcript by greater than 100-fold. MTs isolated from Tdc1^f03311 homozygous flies showed no significant depolarization of their transepithelial potential (TEP) or diuresis in response to tyrosine while retaining normal sensitivity to TA. By contrast, a previously identified null mutant allele of the neuronal TDC isoform Tdc2 had no effect on either tyrosine or TA sensitivity. To determine in which cell type of the MT Tdc1 expression is required, flies were generated carrying a UAS-Tdc1 transgene and cell-type-specific Gal4 drivers on a Tdc1^f03311 homozygous background. Rescue of Tdc1 expression in principal cells fully restored sensitivity to tyrosine whereas expression of Tdc1 in stellate cells had no rescuing effect. It is concluded that synthesis of TA by Tdc1 in the principal cells of the MT is required for physiological responses to tyrosine. TA synthesis in the MT is the first reported physiological role for Drosophila Tdc1.

Learning plays an important role in food acquisition for a wide range of insects and has been demonstrated to be essential during flower foraging in taxa such as bees, parasitoid wasps, butterflies and moths. However, little attention has been focused on differences in floral cue learning abilities among species and sexes. We examined the associative learning of flower colour with nectar in four butterfly species: Idea leuconoe, Argyreus hyperbius, Pieris rapae and Lycaena phlaeas. All butterflies that were trained learned the flower colours associated with food. The flower colour learning rates were significantly higher in I. leuconoe and A. hyperbius than in P. rapae and L. phlaeas. Among the four species examined, the larger and longer-lived species exhibited higher learning rates. Furthermore, female butterflies showed a significantly higher learning rate than males. This study provides the first evidence that learning abilities related to floral cues differ among butterfly species. The adaptive significance of superior learning abilities in the larger and longer-lived butterfly species and in females is discussed.

Songbirds exhibit seasonal plasticity in a broad variety of behavioral and morphological traits associated with reproduction. Changes in song production are well described while changes in song reception are not. In the present study, we test for seasonal variation in auditory processing of the house sparrow (Passer domesticus L.) using auditory brainstem responses (ABRs) to tone bursts. We measured amplitude and latency of the first ABR peak in spring, summer and autumn at stimulus frequencies from 0.8 to 6.4 kHz and intensity levels from 24 to 80 dB SPL. ABR thresholds were determined at each frequency using cross-correlation. Amplitude was greater in spring than in autumn at frequencies from 3.2 to 6.4 kHz whereas latency and thresholds exhibited no seasonal variation. The results indicate an increase in the number or temporal synchrony of responses from peripheral auditory neurons during the early breeding season. Changes in peripheral auditory processing may enhance temporal coding of the fine structure and envelope of song; thereby, improving assessment of encoded information in both sexes (e.g. individual identity and dominance status) and auditory feedback during song production in males. Peripheral auditory changes may be mediated by reproductive hormones, and could involve changes in hair cell density on the basilar papilla. Our results suggest that peripheral auditory processing of songbirds changes seasonally in parallel with other behavioral and morphological traits, such as song production.

Sea turtles are known to detect chemical cues, but in contrast to most marine animals, turtles surface to breathe and thus potentially have access to olfactory cues both in air and in water. To determine whether sea turtles can detect airborne chemical cues, captive loggerhead turtles (Caretta caretta) were placed into a circular, water-filled arena in which odorants could be introduced to the air above the water surface. Air that had passed across the surface of a cup containing food elicited increased activity, diving and other behavior normally associated with feeding. By contrast, air that had passed across the surface of an identical cup containing distilled water elicited no response. Increases in activity during food odor trials occurred only after turtles surfaced to breathe and peaked in the first post-breath minute, implying that the chemical cues eliciting the responses were unlikely to have been detected while the turtles were under water. These results provide the first direct evidence that sea turtles can detect airborne odors. Under natural conditions, this sensory ability might function in foraging, navigation or both.

This study aimed to examine whether the stenohaline freshwater stingray, Potamotrygon motoro, which lacks a functional ornithine—urea cycle, would up-regulate glutamine synthetase (GS) activity and protein abundance, and accumulate glutamine during a progressive transfer from freshwater to brackish (15) water with daily feeding. Our results revealed that, similar to other freshwater teleosts, P. motoro performed hyperosmotic regulation, with very low urea concentrations in plasma and tissues, in freshwater. In 15 water, it was non-ureotelic and non-ureoosmotic, acting mainly as an osmoconformer with its plasma osmolality, [Na⁺] and [Cl^–] comparable to those of the external medium. There were significant increases in the content of several free amino acids (FAAs), including glutamate, glutamine and glycine, in muscle and liver, but not in plasma, indicating that FAAs could contribute in part to cell volume regulation. Furthermore, exposure of P. motoro to 15 water led to up-regulation of GS activity and protein abundance in both liver and muscle. Thus, our results indicate for the first time that, despite the inability to synthesize urea and the lack of functional carbamoyl phosphate synthetase III (CPS III) which uses glutamine as a substrate, P. motoro retained the capacity to up-regulate the activity and protein expression of GS in response to salinity stress. Potamotrygon motoro was not nitrogen (N) limited when exposed to 15 water with feeding, and there were no significant changes in the amination and deamination activities of hepatic glutamate dehydrogenase. In contrast, P. motoro became N limited when exposed to 10 water with fasting and could not survive well in 15 water without food.

Carbonic anhydrase (CA) is critical for CO₂ excretion in adult fish, but little is known of the expression or function of CA during early development. The present study examined the hypothesis that, as rates of CO₂ production increased during early development in zebrafish (Danio rerio), CA would become necessary for effective CO₂ excretion, and that the pattern of CA expression during early development would reflect this transition. Real-time RT-PCR was used to examine the mRNA expression of the two main intracellular CA isoforms over a time course of early development ranging from 0 to 120 h post fertilization (h.p.f.). The mRNA expression of zCAb was generally higher than that of zCAc, particularly during the earliest stages of development. Rates of CO₂ excretion increased approximately 15-fold from 24 to 48 h.p.f. whereas rates of O₂ uptake increased only 6.7-fold over the same period, indicating a relative stimulation of CO₂ excretion over O₂ uptake. Treatment of 48 h.p.f. larvae with the CA inhibitor acetazolamide resulted in CO₂ excretion rates that were 52% of the value in control larvae, a significant difference that occurred in the absence of any effect on O₂ uptake. Antisense morpholino oligonucleotides were used to selectively knock down one or both of the main intracellular CA isoforms. Subsequent measurement of gas transfer rates at 48 h.p.f. indicated that CA knockdown caused a significant relative inhibition of CO₂ excretion over O₂ uptake, regardless of which cytosolic CA isoform was targeted for knockdown. These results suggest that between 24 h.p.f. and 48 h.p.f., developing zebrafish begin to rely on CA to meet requirements for increased CO₂ excretion.

While adult zebrafish, Danio rerio, possess ammonia and urea transporters (Rh and UT proteins, respectively) in a number of tissues, they are most heavily concentrated within the gills. UT has a diffuse expression pattern within Na⁺-K⁺-ATPase (NKA)-type mitochondrion-rich cells and Rh proteins form a network similar to the arrangement seen in pufferfish gills (Nakada et al., 2007b). Rhag expression appeared to be limited to the pillar cells lining the blood spaces of the lamellae while Rhbg was localized to the outer layer of both the lamellae and the filament, upon the pavement cells. Exposure to high external ammonia (HEA) or phloretin increased tissue levels of ammonia and urea, respectively, in adult and juvenile zebrafish; however, the responses to these stressors were age dependent. HEA increased mRNA levels for a number of Rh proteins in embryos and larvae but did not elicit similar effects in adult gills, which appear to compensate for the unfavourable ammonia excretory gradient by increasing expression of V-type H⁺-ATPase. Phloretin exposure increased UT mRNA levels in embryos and larvae but was without effect in adult gill tissue. Surprisingly, in both adults and juveniles, HEA increased the mRNA expression of UT and phloretin increased the mRNA expression of Rh proteins. These results imply that, in zebrafish, there may be a tighter link between ammonia and urea excretion than is thought to occur in most teleosts.

Photoperiod is an important cue regulating biological rhythms in mammals, including ‘blind’ subterranean and sighted fossorial rodent species. These species may respond differentially to changes in light quality according to their retinal complexity. The effects of increasing light intensity on daily rhythms of urine excretion and urinary output of 6-sulfatoxymelatonin levels were compared in ‘blind’ mole rats Spalax ehrenbergi and sighted social voles, Microtus socialis. Our results show that the threshold irradiance required to entrain rhythms of voles is three magnitudes greater than that for mole rats. The results suggest that mole rats have an operational photoreceptive pathway with a lower threshold irradiance than voles. Such a low threshold reflects the remarkable capability of this ‘blind’ species to utilize light signals even under challenging light conditions.

The fatty acid composition of muscle membrane phospholipids and fat stores may affect migration performance in birds. The purpose of this study was to investigate seasonal changes in the fatty acid composition of (1) pectoralis muscle phospholipids, (2) intramuscular triglyceride stores and (3) adipose tissue triglycerides in free-living white-throated sparrows (Zonotrichia albicollis). During migratory seasons there was an increase in the n-6:n-3 ratio of muscle membrane phospholipid fatty acids without a change in the proportion of unsaturated fatty acids. This change was driven mainly by an increase in the proportion of 18:2n-6 and a decrease in the proportion of 22:6n-3. An increase in the proportion of 18:2n-6 was also observed in the intramuscular and adipose tissue triglyceride stores during the migratory seasons. These increases in 18:2n-6 were offset by a decrease in 16:0; resulting in an elevated proportion of unsaturated fatty acids and elevated double bond index in both fat stores of migrants. The elevated levels of 18:2n-6 in migrant fat stores indicates a high dietary component of this fatty acid, as white-throated sparrows feed mainly on tree seeds and some insects during migration and may not have access to a diet high in n-3 fatty acids. We suspect that elevated dietary levels of 18:2n-6 also caused the observed increases in the proportion of this fatty acid in muscle phospholipids. Overall, we conclude that seasonal changes in adipose and muscle fatty acid composition are likely attributable to diet more than other factors such as migratory exercise or mitochondrial density.

Intestinal HCO₃^– secretion is essential to marine teleost fish osmoregulation and comprises a considerable source of base efflux attributable to both serosal HCO₃^– and endogenous CO₂ hydration. The role of intestinal HCO₃^– secretion in dynamic acid—base balance regulation appears negligible in studies of unfed fish, but evidence of high intestinal fluid [HCO₃^–] in fed marine teleosts led us to investigate the source of this HCO₃^– and its potential role in offsetting the postprandial ‘alkaline tide’ commonly associated with digestion. Specifically, we hypothesized that elevated metabolic rate and thus endogenous CO₂ production by intestinal tissue as well as increased transepithelial intestinal HCO₃^– secretion occur post-feeding and offset a postprandial alkaline tide. To test these hypotheses changes in HCO₃^– secretion and O₂ consumption by gulf toadfish (Opsanus beta) isolated intestine were quantified 0, 3, 6, 12, 24 and 48 h post-feeding. Intestinal tissue of unfed fish in general showed high rates of HCO₃^– secretion (15.5 µmol g^–1 h^–1) and O₂ consumption (8.9 µmol g^–1 h^–1). Furthermore, postprandial increases in both intestinal HCO₃^– secretion and O₂ consumption (1.6- and 1.9-fold peak increases, respectively) were observed. Elevated intestinal HCO₃^– secretion rates preceded and outlasted those of O₂ consumption, and occurred at a magnitude and duration sufficient to account for the lack of alkaline tide. The dependence of these high rates of postprandial intestinal base secretion on serosal HCO₃^– indicates transepithelial HCO₃^– transport increases disproportionately more than endogenous CO₂ production. The magnitude of postprandial intestinal HCO₃^– secretion indicates the intestine certainly is capable of postprandial acid#x02014;base balance regulation.

We have studied the passive maintenance of high angle of attack and its lift generation during the crane fly's flapping translation using a dynamically scaled model. Since the wing and the surrounding fluid interact with each other, the dynamic similarity between the model flight and actual insect flight was measured using not only the non-dimensional numbers for the fluid (the Reynolds and Strouhal numbers) but also those for the fluid—structure interaction (the mass and Cauchy numbers). A difference was observed between the mass number of the model and that of the actual insect because of the limitation of available solid materials. However, the dynamic similarity during the flapping translation was not much affected by the mass number since the inertial force during the flapping translation is not dominant because of the small acceleration. In our model flight, a high angle of attack of the wing was maintained passively during the flapping translation and the wing generated sufficient lift force to support the insect weight. The mechanism of the maintenance is the equilibrium between the elastic reaction force resulting from the wing torsion and the fluid dynamic pressure. Our model wing rotated quickly at the stroke reversal in spite of the reduced inertial effect of the wing mass compared with that of the actual insect. This result could be explained by the added mass from the surrounding fluid. Our results suggest that the pitching motion can be passive in the crane fly's flapping flight.

This study reveals the peculiar in vivo cell kinetics and cell turnover of the marine sponge Halisarca caerulea under steady-state conditions. The tropical coral reef sponge shows an extremely high proliferation activity, a short cell cycle duration and massive cell shedding. Cell turnover is predominantly confined to a single cell population, i.e. the choanocytes, and in this process apoptosis only plays a minor role. To our knowledge, such fast cell kinetics under steady-state conditions, with high turnover by shedding in the absence of apoptosis, has not been observed previously in any other multicellular organism. The duration of the cell cycle in vivo resembles that of unicellular organisms in culture. Morphological and histochemical studies demonstrate compartmentalization of choanocytes in the sponge tissue, which corresponds well with its remarkable cellular kinetics. Coral reef cavity sponges, like H. caerulea, inhabit low nutrient tropical waters, forcing these organisms to filter large volumes of water and to capture the few nutrients efficiently. Under these oligotrophic conditions, a high cell turnover may be considered as a very useful strategy, preventing permanent damage to the sponge by environmental stress. Halisarca caerulea maintains its body mass and keeps its food uptake system up to date by constantly renewing its filter system. We conclude that studies on cell kinetics and functional morphology provide new and essential information on the growth characteristics and the regulation of sponge growth in vivo as well as in vitro and the role of choanocytes in tissue homeostasis.

The melanocortin 5 receptor (MC5R) plays a key role in the regulation of exocrine secretion in mammalian species. This receptor has also been characterized in some fish species but its function is unknown. We report the molecular and pharmacological characterization, as well as the tissue expression pattern, of sea bass MC5R. Cloning of five active alleles showing different levels of sensitivity to endogenous melanocortin and one non-functional allele demonstrate the allelic complexity of the MC5R locus. The sea bass receptor was activated by all the melanocortins tested, with ACTH and desacetyl-MSH and β-MSH showing the lowest efficiency. The acetylation of the MSH isoforms seems to be critical for the effectiveness of the agonist. Agouti-related protein had no effect on basal or agonist-stimulated activation of the receptor. SbMC5R was mainly expressed in the brain but lower expression levels were found in several peripheral tissues, including liver. Progressive fasting did not induce up- or downregulation of hypothalamic MC5R expression, suggesting that central MC5R is not involved in the regulation of food intake in the sea bass. MTII, a sbMC5R agonist, stimulated hepatic lipolysis in vitro, measured as free fatty acid release into the culture medium after melanocortin agonist exposure of liver fragments, suggesting that MC5R is involved in the regulation of hepatic lipid metabolism. Taken together, the data suggest that different allelic combinations may confer differential sensitivity to endogenous melanocortin in tissues where MC5R is expressed and, by extension, in hepatic lipid metabolism.

We found strain differences in the ability of wild Alberta Lymnaea stagnalis to form long-term memory (LTM) following operant conditioning when L. stagnalis were collected from the wild and trained in the laboratory. Lymnaea stagnalis obtained from the Belly River watershed had an enhanced ability to form LTM compared with those from an isolated pond (referred to as Jackson snails). We therefore asked whether the differences in cognitive ability were an epiphenomenon as a result of training in the laboratory. To answer this question we trained each specific strain (Belly and Jackson) in both the laboratory and the field (i.e. in their home pond and in the pond where the other strain resided - referred to as the visitor pond). We found that within each strain there was no difference in the LTM phenotype whether they were trained in the lab or in either their home or visitor pond. That is, the strain differences in the ability to form LTM were still present. Interestingly, we found no strain differences in the ability to learn or the ability to form intermediate-term memory (ITM).

In mammals, pituitary adenylate cyclase-activating polypeptide (PACAP) and vasoactive intestinal peptide (VIP) are involved in cardiovascular and respiratory regulation. Several studies have demonstrated the presence of PACAP, VIP and their receptors in various tissues of teleost fish, including the brain, but little is known about their respiratory and cardiovascular effects. The present study was undertaken to compare the central and peripheral actions of graded doses (25-100 pmol) of trout PACAP and trout VIP on ventilatory and cardiovascular variables in the unanaesthetized rainbow trout. Compared with vehicle, only intracerebroventricular injection of PACAP significantly (P<0.05) elevated the ventilation frequency and the ventilation amplitude, but both peptides significantly increased the total ventilation ( ). However, the maximum hyperventilatory effect of PACAP was approximately 2.5-fold higher than the effect of VIP at the 100 pmol dose (PACAP, =+5407±921 arbitrary units, a.u.; VIP, =+2056±874 a.u.; means ± s.e.m.). When injected centrally, only PACAP produced a significant increase in mean dorsal aortic blood pressure (P_DA) (100 pmol: +21%) but neither peptide affected heart rate (f_H). Intra-arterial injections of either PACAP or VIP were without effect on the ventilatory variables. PACAP was without significant action on P_DA and f_H while VIP significantly elevated P_DA (100 pmol: +36%) without changing f_H. In conclusion, the selective central hyperventilatory actions of exogenously administered trout PACAP, and to a lesser extent VIP, suggest that the endogenous peptides may be implicated in important neuroregulatory functions related to the central control of ventilation in trout.

Honeybee foragers are exposed to thermal stimuli when collecting food outside and receiving food rewards inside the nest. In both contexts, there is an opportunity for foragers to associate warmth with food rewards. However, honeybee thermal learning is poorly understood. Using an associative learning paradigm (the proboscis extension reflex), we show that honeybees can learn to associate a nectar reward with a heated stimulus applied to the antenna to mimic natural contact with a warm flower or nectar-offering forager. Conditioning with longer inter-trial intervals (ITI) significantly improved learning acquisition. We also trained bees to discriminate between temperatures above (warm) and below (cold) ambient air temperature. Learning acquisition improved by 38% per 10°C increase in absolute stimulus intensity (difference between the rewarded temperature and unrewarded ambient air temperature). However, bees learned positive temperature (warm) significantly better than negative temperature (cold) differences, approximately twice as well for 10°C as compared with a –10°C difference. Thus, thermosensation, a sensory modality that is relatively unexplored in honeybees, could play a role in the acquisition of information from nestmates (social learning) and in foraging decisions influenced by associations between floral temperature and nectar rewards.

Nowadays few people consider finding their way in unfamiliar areas a problem as a GPS (Global Positioning System) combined with some simple map software can easily tell you how to get from A to B. Although this opportunity has only become available during the last decade, recent experiments show that long-distance migrating animals had already solved this problem. Even after displacement over thousands of kilometres to previously unknown areas, experienced but not first time migrant birds quickly adjust their course toward their destination, proving the existence of an experience-based GPS in these birds. Determining latitude is a relatively simple task, even for humans, whereas longitude poses much larger problems. Birds and other animals however have found a way to achieve this, although we do not yet know how. Possible ways of determining longitude includes using celestial cues in combination with an internal clock, geomagnetic cues such as magnetic intensity or perhaps even olfactory cues. Presently, there is not enough evidence to rule out any of these, and years of studying birds in a laboratory setting have yielded partly contradictory results. We suggest that a concerted effort, where the study of animals in a natural setting goes hand-in-hand with lab-based study, may be necessary to fully understand the mechanism underlying the long-distance navigation system of birds. As such, researchers must remain receptive to alternative interpretations and bear in mind that animal navigation may not necessarily be similar to the human system, and that we know from many years of investigation of long-distance navigation in birds that at least some birds do have a GPS – but we are uncertain how it works.

The crayfish swimmeret system undergoes transitions between a silent state and an active state. In the silent state, no patterned firing occurs in swimmeret motor neurons. In the active state, bursts of spikes in power stroke motor neurons alternate periodically with bursts of spikes in return stroke motor neurons. In preparations of the isolated crayfish central nervous system (CNS), the temporal structures of motor patterns expressed in the active state are similar to those expressed by the intact animal. These transitions can occur spontaneously, in response to stimulation of command neurons, or in response to application of neuromodulators and transmitter analogues. We used single-electrode voltage clamp of power-stroke exciter and return-stroke exciter motor neurons to study changes in membrane currents during spontaneous transitions and during transitions caused by bath-application of carbachol or octopamine (OA). Spontaneous transitions from silence to activity were marked by the appearance of a standing inward current and periodic outward currents in both types of motor neurons. Bath-application of carbachol also led to the development of these currents and activation of the system. Using low Ca²⁺–high Mg²⁺ saline to block synaptic transmission, we found that the carbachol-induced inward current included a direct response by the motor neuron and an indirect component. Spontaneous transitions from activity to silence were marked by disappearance of the standing inward current and the periodic outward currents. Bath-application of OA led promptly to the disappearance of both currents, and silenced the system. OA also acted directly on both types of motor neurons to cause a hyperpolarizing outward current that would contribute to silencing the system.

This study investigated the role of Na⁺/H⁺ exchanger (NHE) and signalling molecules, such as cAMP, PKC, PI 3-kinase, and immune defence enzymes, NADPH oxidase and nitric oxide synthase, in the induction of protein glutathionylation and carbonylation in cadmium-treated haemocytes of mussel Mytilus galloprovincialis. Glutathionylation was detected by western blot analysis and showed actin as its main target. A significant increase of both actin glutathionylation and protein carbonylation, were observed in haemocytes exposed to micromolar concentration of cadmium chloride (5 µmol l^–1). Cadmium seems to cause actin polymerization that may lead to its increased glutathionylation, probably to protect it from cadmium-induced oxidative stress. It is therefore possible that polymerization of actin plays a signalling role in the induction of both glutathionylation and carbonylation processes. NHE seems to play a regulatory role in the induction of oxidative damage and actin glutathionylation, since its inhibition by 2 µmol l^–1 cariporide, significantly diminished cadmium effects in each case. Similarly, attenuation of cadmium effects were observed in cells pre-treated with either 11 µmol l^–1 GF-109203X, a potent inhibitor of PKC, 50 nmol l^–1 wortmannin, an inhibitor of PI 3-kinase, 0.01 mmol l^–1 forskolin, an adenylyl cyclase activator, 10 µmol l^–1 DPI, a NADPH oxidase inhibitor, or 10 µmol l^–1 L-NAME, a nitric oxide synthase inhibitor, suggesting a possible role of PKC, PI 3-kinase and cAMP, as well as NADPH oxidase and nitric oxide synthase in the enhancement of cadmium effects on both actin glutathionylation and protein carbonylation.

Among potential agents that might damage bird feathers are certain microorganisms which secrete enzymes that digest keratin, as is the case of the ubiquitous bacterium Bacillus licheniformis, present in both the feathers and skin of wild birds. It is therefore a good candidate for testing the effects of bird defences against feather-degrading microorganisms. One of these defences is the oil secreted by the uropygial gland, which birds use to protect their feathers against parasites. In previous studies we have shown how Enterococcus faecalis strains isolated from nestling hoopoes exert antagonistic effects against B. licheniformis, mediated by the production of bacteriocins. Consequently we hypothesized that this enterococcus and the bacteriocins it engenders might act as a defence against feather-degrading microorganisms in hoopoes. We investigated this hypothesis in a series of laboratory experiments and evaluated the extent to which the keratinolytic effects caused by B. licheniformis were reduced by the E. faecalis MRR10-3 strain, isolated from hoopoes, and its bacteriocins. In different treatments, feathers or pure keratin was incubated with B. licheniformis, B. licheniformis together with E. faecalis MRR10-3, and B. licheniformis together with the bacteriocins produced by E. faecalis MRR10-3. Our results were in accordance with the predicted effects on hoopoe feathers. There was a significant decrease both in pure keratin loss and in feather degradation in the presence of the symbiotic bacterium or its bacteriocin. These results suggest that by preening their feathers hoopoes benefit from their symbiotic relationship with bacteriocin-producing enterococci, which constitute a chemical defence against feather degradation.

The material properties of food can exert a significant influence on tooth morphology. Although the stiffness or toughness of a material is usually of prime concern, other aspects of material properties (such as extensibility) can be of equal importance. Previous experimental work on the effect blade shape has on fracturing biological materials indicated a notched blade greatly reduced the work required to cut tough tissue. As a notched blade both traps materials and cuts at an angle, it is not clear which of these features leads to increased cutting efficiency. This paper tests whether the ability to cut at an angle or trap the material has the greater effect on the work to fracture required to cut tough tissues with different levels of extensibility (asparagus and fish muscle). Results show that the work to fracture required to cut more extensible materials is reduced by up to 50% when a trapping mechanism alone is used in comparison with an angled blade alone. For less extensible materials, the trapping ability of a notch seems to have no effect, whereas the angled blade reduces work to fracture by up to 25% relative to a straight blade. The aspects of blade shape most important to the breaking down of foods depend upon the relative stiffness or toughness, as well as other material properties.

Birds adjust their flight behaviour to the physical properties of the air. Lift and drag, the two major properties in aerodynamics, are highly dependent on air density. With decreasing air density drag is reduced and lift per wingbeat decreases. According to flight mechanical theory, wingbeat frequency and air speed should increase with decreasing air density, i.e. increasing flight altitude. Although wind tunnel experiments have shed light on many aspects of avian flight, the effect of air density remained ambiguous, because air density could not be adjusted in wind tunnels, until now. By means of radar we recorded tracks of several thousand free-flying individual birds during nocturnal migration. From these tracks we derived wingbeat frequencies and air speeds covering air densities from 0.84 kg m^–3 to 1.13 kg m^–3, corresponding to an altitudinal range of about 3000 m. We demonstrate here with this sample of nocturnal migrants that: (1) wingbeat frequency decreases with air density (which corresponds to an increase in flap-gliding flyers by 0.4 Hz km^–1 and in bounding flyers by 1.1 Hz km^–1), (2) reducing wingbeat frequency to equivalent sea level values did not abolish the dependency on air density, as expected by flight mechanical theory, and (3) bounding flyers show a higher response in their flight behavioural adjustments to changes in air density than flap-gliding flyers. With respect to air speed flap-gliding flyers increase their air speed by 1.0 m s^–1 km^–1 and bounding flyers by 1.4 m s^–1 km^–1.

The ability of sponges to feed in diverse (including oligotrophic) ecosystems significantly contributes to their ubiquitous aquatic distribution. It was hypothesized that sponges that harbour small amounts of symbiotic bacteria in their mass feed mainly on particulate organic matter (POM). We examined the nearly symbiont-free (by microscopic observation) filter-feeding Red Sea sponge Negombata magnifica in order to: (a) study removal efficiency of naturally occurring organic particles, (b) measure the total amount of absorbed particulate organic carbon (POC) and nitrogen (PON), and (c) estimate organic carbon and nitrogen flux in this sponge. Total amount of organic carbon and nitrogen in the Gulf of Aqaba was found to be 48.46±5.69 µg l^–1 and 6.45±0.7 µg l^–1, respectively. While detritus contributed 54% of POC, most PON (84%) came from planktonic microorganisms, mainly prokaryotes. Particle removal efficiency ranged from 99% (the cyanobacterium Synechococcus sp.) to 37% (for eukaryotic cells >8 µm). On average, N. magnifica ingested 480 µg C day^–1 g^–1 (wet mass, WM) sponge and 76.6 µg N day^–1 g^–1 sponge. Ingested POC balanced 85% of the sponge's energetic demand but more is needed for biomass production because it cannot digest all of the carbon. 54.4±16.1 µg N day^–1 g^–1 (WM) nitrogen was excreted as total ammonia nitrogen (TAN); however, nitrogen allowance should be higher because more nitrogen is deposited for sponge biomass during growth. It is hypothesized that the discrepancy in the nutritional requirements should be covered by the sponge absorbing carbon and nitrogen from sources that are not dealt with in the present research, such as dissolved organic carbon and nitrogen. This study highlights the significance of detritus as a carbon source, and prokaryotes as a PON source in sponge feeding.

The role of the two distinct retinal photoreceptor organs in photoreception for photoperiodism was examined in the carabid beetle, Leptocarabus kumagaii, by surgical removal. This beetle shows long-day and short-day photoperiodic responses in the larval and adult stages, respectively. Larval diapause in the final instar is induced under short-day conditions whereas pupation occurs without diapause under long-day conditions. Adult reproductive diapause is terminated under short-day conditions but maintained under long-day conditions. The stemmata of the larvae and compound eyes of the adults were removed and the responses of the animals to photoperiod were compared to those of intact beetles. When all the stemmata were removed, larvae pupated without entering diapause under both long-day and short-day conditions, indicating that the larvae lacking stemmata were incapable of photoreception for photoperiodism. As in other holometabolous insects, the stemmata migrated into the brain during metamorphosis and remained rudimentarily in the optic lobe of the adult brain. However, these stemmata-derived organs were found to be no longer necessary for photoperiodism, because adults lacking the stemmata-derived organs responded to photoperiod normally. By contrast, removal of the compound eyes in adults resulted in the termination of reproductive diapause under both long-day and short-day conditions, indicating that photoreception for photoperiodism in the adult stage is performed by the compound eyes. Therefore, the site of photoperiodic photoreception in L. kumagaii appear to change from the stemmata to the compound eyes during metamorphosis.

Although green striped burrowing frogs (Cyclorana alboguttata) experience large reductions in the mass and absorptive surface area of the small intestine (SI) during aestivation, little is known about how this may affect the functional capacity of the SI. We examined changes in the function (l-proline uptake rate and capacity) and metabolism of the SI (in vitro oxygen consumption, Na⁺/K⁺-ATPase activity and abundance) of C. alboguttata following 6 months of aestivation. l-Proline uptake rate was significantly higher in aestivating frogs, but overall uptake capacity was lower than in active frogs. Total SI oxygen consumption rate (V_O2) was also lower in aestivating frogs, despite no difference in mass-specific V_O2. The proportion of intestinal V_O2 associated with Na⁺/K⁺-ATPase activity and protein synthesis was equivalent between active and aestivating frogs, suggesting these processes were unaffected by aestivation. Indeed, the activity of Na⁺/K⁺-ATPase transporters in the SI of aestivating frogs was not different from that of active animals. Aestivating frogs maintained Na⁺/K⁺-ATPase activity, despite experiencing a reduction in the density of Na⁺/K⁺-ATPase transporters, by increasing the molecular activity of the remaining pumps to 2–3 times that of active frogs. These results show that functionality of the SI is maintained at the cellular level, potentially facilitating the reclamation of nutrients from the intestinal lumen while in aestivation. Despite this, the functional capacity of the SI in aestivating C. alboguttata is significantly reduced due to a reduction in tissue mass, helping frogs to conserve energy while in aestivation.

We investigated the effect of prolonged immobilisation of six and nine months duration on the morphology and antioxidant biochemistry of skeletal muscles in the amphibian aestivator Cyclorana alboguttata. We hypothesised that, in the event of atrophy occurring during aestivation, larger jumping muscles were more likely to be preserved over smaller non-jumping muscles. Whole muscle mass (g), muscle cross-sectional area (CSA) (µm²), water content (%) and myofibre number (per mm²) remained unchanged in the cruralis muscle after six to nine months of aestivation; however, myofibre area (µm²) was significantly reduced. Whole muscle mass, water content, myofibre number and myofibre CSA remained unchanged in the gastrocnemius muscle after six to nine months of aestivation. However, iliofibularis dry muscle mass, whole muscle CSA and myofibre CSA was significantly reduced during aestivation. Similarly, sartorius dry muscle mass, water content and whole muscle CSA was significantly reduced during aestivation. Endogenous antioxidants were maintained at control levels throughout aestivation in all four muscles. The results suggest changes to muscle morphology during aestivation may occur when lipid reserves have been depleted and protein becomes the primary fuel substrate for preserving basal metabolic processes. Muscle atrophy as a result of this protein catabolism may be correlated with locomotor function, with smaller non-jumping muscles preferentially used as a protein source during fasting over larger jumping muscles. Higher levels of endogenous antioxidants in the jumping muscles may confer a protective advantage against oxidative damage during aestivation; however, it is not clear whether they play a role during aestivation or upon resumption of normal metabolic activity.

Proteins belonging to the family of neprilysins are typically membrane bound M13 endopeptidases responsible for the inactivation and/or activation of peptide signaling events on cell surfaces. Mammalian neprilysins are known to be involved in the metabolism of various regulatory peptides especially in the nervous, immune, cardiovascular and inflammatory systems. Although there is still much to learn about their participation in various diseases, they are potential therapeutic targets. Here we report on the identification and first characterization of neprilysin 4 (NEP4) from Drosophila melanogaster. Reporter lines as well as in situ hybridization combined with immunolocalization demonstrated NEP4 expression during embryogenesis in pericardial cells, muscle founder cells, glia cells and male gonads. Western blot analysis confirmed the prediction of one membrane bound and one soluble isoform, a finding quite unusual among neprilysins with presumably strong physiological relevance. At least one NEP4 isoform was found in every developmental stage indicating protein activities required throughout the whole life cycle of Drosophila. Heterologously expressed NEP4 exhibited substrate preferences comparable to human neprilysin 2 with distinct cleavage of substance P and angiotensin I.

The velvet belly lantern shark (Etmopterus spinax) emits a blue luminescence from thousands of tiny photophores. In this work, we performed a pharmacological study to determine the physiological control of luminescence from these luminous organs. Isolated photophore-filled skin patches produced light under melatonin (MT) and prolactin (PRL) stimulation in a dose-dependent manner but did not react to classical neurotransmitters. The -melanocyte-stimulating hormone (-MSH) had an inhibitory effect on hormonal-induced luminescence. Because luzindole and 4P-PDOT inhibited MT-induced luminescence, the action of this hormone is likely to be mediated through binding to the MT2 receptor subtype, which probably decreases the intracellular concentration of cyclic AMP (cAMP) because forskolin (a cAMP donor) strongly inhibits the light response to MT. However, PRL seems to achieve its effects via janus kinase 2 (JAK2) after binding to its receptor because a specific JAK2 inhibitor inhibits PRL-induced luminescence. The two stimulating hormones showed different kinetics as well as a seasonal variation of light intensity, which was higher in summer (April) than in winter (December and February). All of these results strongly suggest that, contrary to self-luminescent bony fishes, which harbour a nervous control mechanism of their photophore luminescence, the light emission is under hormonal control in the cartilaginous E. spinax. This clearly highlights the diversity of fish luminescence and confirms its multiple independent apparitions during the course of evolution.

We have used behavioural tests to determine the intensity thresholds of colour vision in Bourke's parrots (Neopsephotus bourkii) and budgerigars (Melopsittacus undulatus). We have also examined the relationship between these thresholds and the optical sensitivities of single photoreceptors using morphological methods. Bourke's parrots lose colour vision in brighter light (0.4 cd m^–2) than budgerigars (0.1 cd m^–2) and both birds lose colour vision in brighter light (`end of civil twilight') than humans (0.02 cd m<sup>–2</sup>, `moonlight'). The optical sensitivities of single cones are similar in both birds (budgerigar 0.27 µm² sr, Bourke's parrot 0.25 µm² sr) but Bourke's parrots have more (cone to rod ratio, 1.2:1.0), thinner (2.8 µm) and longer rods (18.5 µm) than budgerigars (2.1:1.0, 3.4 µm, 13.3 µm). Bourke's parrots thus have an eye type that, with a flexible pooling mechanism, allows for high resolution or high absolute sensitivity depending on the light conditions. The results nicely agree with the activity patterns of the birds, Bourke's parrots being active during the day and in twilight while budgerigars are not normally active before sunrise and after sunset. However, Bourke's parrots have fewer cones than budgerigars, which implies that a smaller number of cones are pooled within each retinal integration area. That could explain why Bourke's parrots have a higher intensity threshold of colour vision than budgerigars. Furthermore, the study emphasises the need to expand the sensitivity measure so that photoreceptor integration units are used rather than single receptors.

The musculoskeletal structure of the foot and ankle has the potential to influence human sprinting performance in complex ways. A large Achilles' tendon moment arm improves the mechanical advantage of the triceps surae but also produces larger shortening velocity during rapid plantarflexion, which detracts from the force-generating capacity of the plantarflexors. The lever arm of the ground reaction force that resists the muscular plantarflexor moment during propulsive push-off is constrained in part by the skeletal structure of the foot. In this study, we measured the plantarflexion moment arms of the Achilles' tendon, lateral gastrocnemius fascicle lengths and pennation angles, and anthropometric characteristics of the foot and lower leg in collegiate sprinters and height-matched non-sprinters. The Achilles' tendon moment arms of the sprinters were 25% smaller on average in sprinters than in non-sprinters (P<0.001) whereas the sprinters' fascicles were 11% longer on average (P=0.024). The ratio of fascicle length to moment arm was 50% larger in sprinters (P<0.001). Sprinters were found to have longer toes (P=0.032) and shorter lower legs (P=0.026) than non sprinters. A simple computer simulation of the sprint push-off demonstrated that shorter plantarflexor moment arms and longer toes, like those measured in sprinters, permit greater generation of forward impulse. Simulated propulsion was enhanced in both cases by increasing the `gear ratio' of the foot, thus maintaining plantarflexor fibre length and reducing peak fibre shortening velocity. Longer toes especially prolonged the time of contact, giving greater time for forward acceleration by propulsive ground reaction force.

We developed a 2D heat flux model to elucidate routes and rates of heat transfer within bigeye tuna Thunnus obesus Lowe 1839 in both steady-state and time-dependent settings. In modeling the former situation, we adjusted the efficiencies of heat conservation in the red and the white muscle so as to make the output of the model agree as closely as possible with observed cross-sectional isotherms. In modeling the latter situation, we applied the heat exchanger efficiencies from the steady-state model to predict the distribution of temperature and heat fluxes in bigeye tuna during their extensive daily vertical excursions. The simulations yielded a close match to the data recorded in free-swimming fish and strongly point to the importance of the heat-producing and heat-conserving properties of the white muscle. The best correspondence between model output and observed data was obtained when the countercurrent heat exchangers in the blood flow pathways to the red and white muscle retained 99% and 96% (respectively) of the heat produced in these tissues. Our model confirms that the ability of bigeye tuna to maintain elevated muscle temperatures during their extensive daily vertical movements depends on their ability to rapidly modulate heating and cooling rates. This study shows that the differential cooling and heating rates could be fully accounted for by a mechanism where blood flow to the swimming muscles is either exclusively through the heat exchangers or completely shunted around them, depending on the ambient temperature relative to the body temperature. Our results therefore strongly suggest that such a mechanism is involved in the extensive physiological thermoregulatory abilities of endothermic bigeye tuna.

Greater sage-grouse, Centrocercus urophasianus, have been a model system in studies of sexual selection and lek evolution. Mate choice in this species depends on acoustic displays during courtship, yet we know little about how males produce these sounds. Here we present evidence for previously undescribed two-voiced sound production in the sage-grouse. We detected this `double whistle' (DW) using multi-channel audio recordings combined with video recordings of male behavior. Of 28 males examined, all males produced at least one DW during observation; variation in DW production did not correlate with observed male mating success. We examined recordings from six additional populations throughout the species' range and found evidence of DW in all six populations, suggesting that the DW is widespread. To examine the possible mechanism of DW production, we dissected two male and female sage-grouse; the syrinx in both sexes differed noticeably from that of the domestic fowl, and notably had two sound sources where the bronchi join the syrinx. Additionally, we found males possess a region of pliable rings at the base of the trachea, as well as a prominent syringeal muscle that is much reduced or absent in females. Experiments with a live phonating bird will be necessary to determine how the syrinx functions to produce the whistle, and whether the DW might be the result of biphonation of a single sound source. We conclude that undiscovered morphological and behavioral complexity may exist even within well-studied species, and that integrative research approaches may aid in the understanding of this type of complexity.

The role of serotonin in the regulation of larval Aedes aegypti hemolymph composition was investigated in vivo using two reuptake inhibitors (SSRIs), alaproclate HCl and 6-nitroquipazine maleate, and the receptor antagonist methiothepin mesylate. Larvae were placed in media differing in pH and salinity in the presence and absence of serotonergic agents. The toxicity of each agent was strongly influenced by ambient pH. For each agent, toxicity was negligible in acidic media, intermediate in neutral media and greatest in alkaline media. By contrast, toxicity of all agents was independent of salinity. No effects on mass-specific body water or hemolymph volume were observed whereas hemolymph osmotic pressure, Na⁺ concentrations and pH differed significantly among treatments. 6-nitroquipazine caused a decrease in Na⁺ from 115±1.7 to 103±0.9 mmol l^–1, and alaproclate caused alkalosis of the hemolymph from pH 7.55±0.026 to pH 7.72±0.044. Methiothepin decreased hemolymph osmotic pressure from 329±9.9 to 304±8.8 and showed the greatest overall toxicity. Control larvae excreted net base in pH 4 media (1.4 µmol g^–1 h^–1) and net acid in pH 7 (1.2 µmol g^–1 h^–1) and pH 11 (5.1 µmol g^–1 h^–1) media. In pH 4 media, alaproclate and methiothepin caused a shift to net H⁺ excretion (1.1 and 1.5 µmol g^–1 h¹, respectively) whereas these agents did not influence acid excretion rates in pH 7 or pH 11 media. The hypothesis that serotonin is involved in hemolymph acid–base balance is discussed.

Maximum power production during multi-joint tasks increases as children grow older. Previous research suggests that in adults, maximum power production in jumping is related to lower limb stiffness. In a developmental context, the question arises as to whether the relationship between maximum power production and lower limb stiffness is age-dependent. The purpose of this study was to investigate the relationship between lower limb stiffness and peak power production in adolescents (AD) and pre-adolescents (PA). With institutional approval, two groups of pre-adults (pre-adolescents: 11–13 years of age, N=43; adolescents: 16–18 years of age, N=30) performed 30 two-legged hops at their preferred frequency and three maximum counter-movement jumps. AD produced significantly greater peak power during the counter-movement jump than PA (t₇₁=–5.28, P<0.001) even when body mass was accounted for. Lower limb stiffness was significantly correlated with peak power production during the counter-movement jump in AD (R=0.62, P<0.001) but not in PA (R=0.26, P=0.10). When normalised to body mass, the relationship between lower limb stiffness and peak power also differed between the two age groups (R=0.30, P=0.11 for AD and R=0.02, P=0.88 for PA). In addition, we found that during hopping, both PA and AD behaved like a simple spring-mass system. Our findings highlight the importance of lower limb stiffness in the context of muscular power production during multi-joint tasks. They let us speculate that during adolescence, children acquire the ability to take greater advantage of elastic energy storage in the musculotendinous system when performing maximum counter-movement jumps.

Closely related species often specialize on different types of prey, but little is known about the fitness consequences of making an evolutionary transition to a novel diet. Spadefoot toad larvae provide a unique opportunity to reconstruct these evolutionary events. Although most anuran larvae feed on detritus or plankton, Spea larvae have also evolved the ability to consume large anostracan fairy shrimp. To investigate the changes that may have accompanied the shift to shrimp prey, we compared shrimp-induced physiological responses of Spea larvae with those of its sister genus, Scaphiopus, that has not made this transition. Although Spea larvae performed equally well on either diet, shrimp-fed Scaphiopus larvae experienced reduced growth and developmental rates, as well as elevated levels of the stress hormone corticosterone when compared with those that ate the ancestral detritus diet. These results suggest that ancestral Spea likely experienced reduced fitness when they first adopted a carnivorous feeding strategy.

Studies of the functional morphology of feeding have typically not included an analysis of the potential for the kinematics of the gape cycle to vary based on the material properties of the prey item being consumed. Variation in prey properties is expected not only to reveal variation in feeding function, but allows testing of the functional role of the phases of the gape cycle. The jaw kinematics of two species of lizards are analyzed when feeding trials are conducted using quantitative control of prey mass, hardness and mobility. For both species, there were statistically significant prey effects on feeding kinematics for all the prey properties evaluated (i.e. prey mass, hardness and mobility). Of these three prey properties, prey mass had a more significant effect on feeding kinematics than prey hardness or mobility. Revealing the impact of varying prey properties on feeding kinematics helps to establish the baseline level of functional variability in the feeding system. Additionally, these data confirm the previously hypothesized functional role of the slow open (SO) phase of the gape cycle as allowing for physical conformation of the tongue to the surface of the food bolus in preparation for further intraoral transport.

Visual displays often play a large role in animal communication, particularly in sexual interactions. The blue crab Callinectes sapidus is both colorful and highly visually responsive, yet almost all studies of their courtship have focused on chemical cues. In the blue crab's underwater environment, however, visual cues may function more rapidly and over a longer distance than chemical cues. Given that blue crabs are aggressive and cannibalistic, visual cues may therefore allow blue crabs to quickly evaluate potential mates from safer distances. In the present study we show that courtship and mate choice behavior in C. sapidus can be stimulated by visual cues alone. Further, we show that males have a preference for females with red claw dactyls. In binary choice experiments, males displayed more often to photographs of females with red claws than to those with white claws or to those with black claws that were isoluminant to the red ones. This strongly suggests that male blue crabs made their choices based on the hue of the red claws, further suggesting that blue crabs are capable of color vision and use color in mate choice.

Vertebrate red blood cells (RBCs) seem to serve tissue oxygen delivery in two distinct ways. Firstly, RBCs enable the adequate transport of O₂ between respiratory surfaces and metabolizing tissues by means of their high intracellular concentration of hemoglobin (Hb), appropriate allosteric interactions between Hb ligand-binding sites, and an adjustable intracellular chemical environment that allows fine-tuning of Hb O₂ affinity. Secondly, RBCs may sense tissue O₂ requirements via their degree of deoxygenation when they travel through the microcirculation and release vasodilatory compounds that enhance blood flow in hypoxic tissues. This latter function could be important in matching tissue O₂ delivery with local O₂ demand. Three main mechanisms by which RBCs can regulate their own distribution in the microcirculation have been proposed. These are: (1) deoxygenation-dependent release of ATP from RBCs, which stimulates production of nitric oxide (NO) and other vasodilators in the endothelium; (2) release of vasoactive NO from S-nitroso-Hb upon deoxygenation; and (3) reduction of naturally occurring nitrite to vasoactive NO by deoxygenated Hb. This Commentary inspects all three hypotheses with regard to their mechanisms, experimental evidence in their support and details that remain unresolved. The prime focus is on human/mammalian models, where most evidence for a role of erythrocyte ATP and NO release in blood flow regulation have accumulated. Information from other vertebrate groups is integrated in the analysis and used to discuss the evolutionary origin and general relevance of each hypothesis.

Although acoustic communication is an integral part of cichlid behaviour, its mechanism has never been identified before. In the present study, a combination of approaches was used to investigate the sound-producing mechanism of Oreochromis niloticus. Synchronisation of high-speed video data (500 frames s^–1) and cineradiographies (250 frames s^–1) with the sound recordings made it possible to locate the different body parts involved in sound production in territorial males. Sounds are made during a backward movement of the pelvic and pectoral girdles and a forward movement of the second pterygiophore of the anal fin. Various electrostimulation experiments, dissections and observation of histological cross-sections revealed a set of bundles (that we call the vesica longitudinalis) situated in the hypaxial musculature, ventro-laterally to the swimbladder. Contraction of these bundles should result in compression of the rib cage and also of the swimbladder, because of its close association with the serosa and ribs. Deflation of the swimbladder resulted in a reduced sound intensity.

Calsequestrin (CASQ) is the main Ca²⁺ binding protein within the sarcoplasmic reticulum (SR) of the vertebrate heart. The contribution of SR Ca²⁺ stores to contractile activation is larger in atrial than ventricular muscle, and in ectothermic fish hearts acclimation to low temperatures increases the use of SR Ca²⁺ in excitation–contraction coupling. The hypotheses that chamber-specific and temperature-induced differences in SR function are due to the increased SR CASQ content were tested in rainbow trout (Oncorhynchus mykiss) acclimated at either 4°C (cold acclimation, CA) or 18°C (warm acclimation, WA). To this end, the trout cardiac CASQ (omCASQ2) was cloned and sequenced. The omCASQ2 consists of 1275 nucleotides encoding a predicted protein of 425 amino acids (54 kDa in molecular mass, MM) with a high (75–87%) sequence similarity to other vertebrate cardiac CASQs. The transcript levels of the omCASQ2 were 1.5–2 times higher in CA than WA fish and about 2.5 times higher in the atrium than ventricle (P<0.001). The omCASQ2 protein was measured from western blots using a polyclonal antibody against the amino acid sequence 174–315 of the omCASQ2. Unlike the omCASQ2 transcripts, no differences were found in the abundance of the omCASQ2 protein between CA and WA fish, nor between the atrium and ventricle (P>0.05). However, a prominent qualitative difference appeared between the acclimation groups: two CASQ isoforms with apparent MMs of 54 and 59 kDa, respectively, were present in atrial and ventricular muscle of the WA trout whereas only the 54 kDa protein was clearly expressed in the CA heart. The 59 kDA isoform was a minor CASQ component representing 22% and 13% of the total CASQ proteins in the atrium and ventricle of the WA fish, respectively. In CA hearts, the 59 kDa protein was present in trace amounts (1.5–2.4%). Collectively, these findings indicate that temperature-related and chamber-specific differences in trout cardiac SR function are not related to the abundance of luminal Ca²⁺ buffering by cardiac CASQ.

Rod absorbance spectra, characterized by the wavelength of peak absorbance (_max) were related to the rod opsin sequences of individual sand gobies (Pomatoschistus minutus) from four allopatric populations [Adriatic Sea (A), English Channel (E), Swedish West Coast (S) and Baltic Sea (B)]. Rod _max differed between populations in a manner correlated with differences in the spectral light transmission of the respective water bodies [_max: (A)503 nm; (E and S)505–506 nm; (B)508 nm]. A distinguishing feature of B was the wide within-population variation of _max (505.6–511.3 nm). The rod opsin gene was sequenced in marked individuals whose rod absorbance spectra had been accurately measured. Substitutions were identified using EMBL/GenBank X62405 English sand goby sequence as reference and interpreted using two related rod pigments, the spectrally similar one of the Adriatic P. marmoratus (_max507 nm) and the relatively red-shifted Baltic P. microps (_max515 nm) as outgroups. The opsin sequence of all E individuals was identical to that of the reference, whereas the S and B fish all had the substitution N151N/T or N151T. The B fish showed systematic within-population polymorphism, the sequence of individuals with _max at 505.6–507.5 nm were identical to S, but those with _max at 509–511.3 nm additionally had F261F/Y. The substitution F261Y is known to red-shift the rod pigment and was found in all P. microps. We propose that ambiguous selection pressures in the Baltic Sea and/or gene flow from the North Sea preserves polymorphism and is phenotypically evident as a wide variation in _max.

Clupeiform fish species, including the Gulf menhaden (Brevoortia patronus) that belong to the subfamily Alosinae, can detect ultrasound. Clupeiform fishes are unique in that they have specialized gas-filled bullae in the head associated with the ear via the bulla membrane and with the lateral line via the lateral recess membrane. It has been hypothesized that the utricle of the inner ear is responsible for ultrasound detection through a specialized connection to the gas-filled bullae complex. Here, we show that the lateral line and its connection to the gas-filled bullae complex via the lateral recess are involved in ultrasound detection in Gulf menhaden. Removal of a small portion of the lateral line overlying the lateral recess membrane eliminates the ability of Gulf menhaden to detect ultrasound. We further show that the gas-filled bullae vibrates in response to ultrasound, that the gas-filled bullae are necessary for detecting ultrasound, and that the bullae connections to the lateral line via the lateral recess membrane play an important role in ultrasound detection. These results add a new dimension to the role of the lateral line and bullae as part of the ultrasonic detection system in Gulf menhaden.

Callinectes sapidus (Rathbun), the Atlantic blue crab, commonly harbors low to moderate amounts of bacteria in hemolymph and other tissues. These bacteria are typically dominated by Vibrio spp., which are known to cause mortality in the blue crab. The dose-dependent lethality of an isolate of Vibrio campbellii was determined in crabs; the mean 48 h LD₅₀ (half-maximal lethal dose) was 6.2x10⁵ colony forming units g^–1 crab. Injection of a sublethal dose of V. campbellii into the hemolymph of the crab resulted in a rapid and large depression (30–42%) of metabolic rate, which persisted for 24 h. Because gills are an organ of immune function as well as respiration, we were interested in how bacteria injected into the crab would affect the energetic costs associated with walking. Overall metabolism (aerobic and anaerobic) more than doubled in crabs walking for 30 min at 8 m min^–1. The metabolic depression resulting from bacterial injection persisted throughout the exercise period and patterns of phosphagen and adenylate consumption within walking leg muscle were not affected by treatment. The ability of crabs to supply required energy for walking is largely unaffected by exposure to Vibrio; however, Vibrio-injected crabs are less aerobic while doing so. This depressed metabolic condition in response to bacteria, present during moderate activity, could be a passive result of mounting an immune response or may indicate an actively regulated metabolic depression. A compromised metabolism can affect the performance of daily activities, such as feeding and predator avoidance or affect the ability to cope with environmental stressors, such as hypoxia.

Adult leatherbacks are large animals (300–500 kg), overlapping in size with marine pinniped and cetacean species. Unlike marine mammals, they start their aquatic life as 40–50 g hatchlings, so undergo a 10,000-fold increase in body mass during independent existence. Hatchlings are limited to the tropics and near-surface water. Adults, obligate predators on gelatinous plankton, encounter cold water at depth (<1280 m) or high latitude and are gigantotherms that maintain elevated core body temperatures in cold water. This study shows that there are great ontogenetic changes in tracheal structure related to diving and exposure to cold. Hatchling leatherbacks have a conventional reptilian tracheal structure with circular cartilaginous rings interspersed with extensive connective tissue. The adult trachea is an almost continuous ellipsoidal cartilaginous tube composed of interlocking plates, and will collapse easily in the upper part of the water column during dives, thus avoiding pressure-related structural and physiological problems. It is lined with an extensive, dense erectile vascular plexus that will warm and humidify cold inspired air and possibly retain heat on expiration. A sub-luminal lymphatic plexus is also present. Mammals and birds have independently evolved nasal turbinates to fulfil such a respiratory thermocontrol function; for them, turbinates are regarded as diagnostic of endothermy. This is the first demonstration of a turbinate equivalent in a living reptile.

Virtually all aspects of insect biology are affected by body temperature, and many taxa have evolved sophisticated temperature-control mechanisms. All insects, however, begin life as eggs and lack the ability to thermoregulate. Eggs laid on leaves experience a thermal environment, and thus a body temperature, that is strongly influenced by the leaves themselves. Because plants can maintain leaf temperatures that differ from ambient, e.g. by evapotranspiration, plant hosts may protect eggs from extreme ambient temperatures. We examined the degree to which leaves buffer ambient thermal variation and whether that buffering benefits leaf-associated insect eggs. In particular, we: (1) measured temperature variation at oviposition sites in the field, (2) manipulated temperatures in the laboratory to determine the effect of different thermal conditions on embryo development time and survival, and (3) tested embryonic metabolic rates over increasing temperatures. Our results show that Datura wrightii leaves buffer Manduca sexta eggs from fatally high ambient temperatures in the southwestern USA. Moreover, small differences in temperature profiles among leaves can cause large variation in egg metabolic rate and development time. Specifically, large leaves were hotter than small leaves during the day, reaching temperatures that are stressfully high for eggs. This study provides the first mechanistic demonstration of how this type of leaf-constructed thermal refuge interacts with egg physiology.

The maximum rate of sustained energy intake (SusEI) may limit reproductive effort, thermoregulatory capability and other aspects of an animal's energy expenditure. Consequently, factors that limit SusEI are of interest. The `heat dissipation limitation hypothesis' suggests that maximum SusEI during lactation is limited by the capacity to dissipate body heat generated as a by-product of processing food and producing milk. In the present study, we tested the heat dissipation limitation hypothesis in lactating Brandt's voles (Lasiopodomys brandtii). Female voles were mated and pregnant at 21(±1)°C. A random sample of animals was transferred into a hot room 30(±1)°C on the day of parturition. The energy intake of lactating voles at 30°C was always lower than that at 21°C. At peak lactation food intake was 3.3 g day^–1 lower at 30°C than at 21°C. There was no significant difference in digestibility. With similar mean litter sizes (7.26±0.46 pups at 21°C and 7.78±0.39 pups at 30°C at the beginning of parturition, 6.83±0.51 pups at 21°C and 7.73±0.50 pups at 30°C at weaning), the milk energy output of mothers, evaluated from the difference between metabolizable energy intake and daily energy expenditure measured by doubly labelled water, at 30°C was 23.3 kJ day^–1 lower than that at 21°C on days 14–16 of lactation. As for reproductive performance, there was a difference in the response to the higher temperature between mothers raising large and those raising small litters. For small litters (<7) there was no significant change in litter mass, but for large litters (<7) there was a significant decrease at the higher temperature. On average, in larger litters the pups were 15.5 g heavier on day 12 of lactation when raised at 21°C. Our data from Brandt's voles support the suggestion that SusEI at peak lactation is limited by heat dissipation capacity, particularly for those voles raising large litters. In smaller litters the peripheral limitation hypothesis may be more relevant. The importance of heat dissipation limits in species raising exclusively small litters needs to be investigated.

The interaction and hierarchy of celestial and magnetic compass cues used by migratory songbirds for orientation has long been the topic of an intense debate. We have previously shown that migratory Savannah sparrows, Passerculus sandwichensis, use polarized light cues near the horizon at sunrise and sunset to recalibrate their magnetic compass. Birds exposed to a ±90 deg. shifted artificial polarization pattern at sunrise or sunset recalibrated their magnetic compass, but only when given full access to celestial cues, including polarized light cues near the horizon. In the current study, we carried out cue conflict experiments with white-throated sparrows, Zonotrichia albicollis, during both spring and autumn migration in a transition zone between the species' breeding and wintering areas on the south shore of Lake Ontario. We show that white-throated sparrows also recalibrate their magnetic compass by polarized light cues at sunrise and sunset. Sunrise exposure to an artificial polarization pattern shifted relative to the natural magnetic field or exposure to a shift of the magnetic field relative to the natural sky both led to recalibration of the magnetic compass, demonstrating that artificial polarizing filters do not create an anomalous, unnatural orientation response. Our results further indicate that there is no evidence for a difference in compass hierarchy between different phases of migration, confirming previous work showing that polarized light cues near the horizon at sunrise and sunset provide the primary calibration reference both in the beginning and at the end of migration.

The sense that allows birds to orient themselves by the Earth's magnetic field can be disabled by an oscillating magnetic field whose intensity is just a fraction of the geomagnetic field intensity and whose oscillations fall into the medium or high frequency radio wave bands. This remarkable phenomenon points very clearly at one of two existing alternative magnetoreception mechanisms in terrestrial animals, i.e. the mechanism based on the radical pair reactions of specific photosensitive molecules. As the first such study in invertebrates, our work offers evidence that geomagnetic field reception in American cockroach is sensitive to a weak radio frequency field. Furthermore, we show that the `deafening' effect at Larmor frequency 1.2 MHz is stronger than at different frequencies. The parameter studied was the rise in locomotor activity of cockroaches induced by periodic changes in the geomagnetic North positions by 60 deg. The onset of the disruptive effect of a 1.2 MHz field was found between 12 nT and 18 nT whereas the threshold of a doubled frequency field 2.4 MHz fell between 18 nT and 44 nT. A 7 MHz field showed no impact even in maximal 44 nT magnetic flux density. The results indicate resonance effects rather than non-specific bias of procedure itself and suggest that insects may be equipped with the same magnetoreception system as the birds.

This article shows that differences in the waveforms of the electric organ discharges (EODs) from two taxa are due to the different responsiveness of their electric organs (EOs) to their previous activity (auto-excitability). We compared Gymnotus omarorum endemic to Uruguay (35° South, near a big estuary), which has four components in the head to tail electric field (V₁ to V₄), with Gymnotus sp. endemic to the south of Brazil, Paraguay and Argentinean Mesopotamia (25° South, inland), which shows a fifth component in addition to the others (V₅). We found that: (a) the innervation pattern of the electrocytes, (b) the three earlier, neurally driven, EOD components (V₁ to V₃), and (c) their remnants after curarisation were almost identical in the two taxa. The equivalent electromotive forces of late components (V₄ and V₅) increased consistently as a function of the external current associated with the preceding component and were abolished by partial curarisation in both taxa. Taken together these data suggest that these components are originated in the responses of the electrocytes to longitudinal currents through the EO. By using a differential load procedure we showed that V₄ in G. omarorum responded to experimental changes in its excitation current with larger amplitude variations than V₄ in Gymnotus sp. We conclude that the differences in the EOD phenotype of the two studied taxa are due to the different EO auto-excitability. This, in turn, is caused either by the different expression of a genetic repertoire of conductance in the electrocyte membrane or in the wall of the tubes forming the EO.

Fish typically use a rostro-caudal wave of head expansion to generate suction, which is assumed to cause a uni-directional, anterior-to-posterior flow of water in the expanding head. However, compared with typical fish, syngnathid fishes have a remarkably different morphology (elongated snout, small hyoid, immobile pectoral girdle) and feeding strategy (pivot feeding: bringing the small mouth rapidly close to the prey by neurocranial dorsorotation). As a result, it is unclear how suction is generated in Syngnathidae. In this study, lateral and ventral expansions of the head were quantified in Hippocampus reidi and linked to the kinematics of the mouth, hyoid and neurocranium. In addition, the flow velocities inside the bucco-pharyngeal cavity and in front of the mouth were calculated. Our data suggest that the volume changes caused by lateral expansion are dominant over ventral expansion. Maximum gape, neurocranium rotation and hyoid depression are all reached before actual volume increase and before visible prey movement. This implies that, unlike previously studied teleosts, hyoid rotation does not contribute to ventral expansion by lowering the floor of the mouth during prey capture in H. reidi. The lateral volume changes show a rostro-caudal expansion, but the maximal flow velocity is not near the mouth aperture (as has been demonstrated for example in catfish) but at the narrow region of the buccal cavity, dorsal to the hyoid.

Enzymes and biochemical mechanisms essential to survival are under extreme selective pressure and are highly conserved through evolutionary time. We applied this evolutionary concept to barnacle cement polymerization, a process critical to barnacle fitness that involves aggregation and cross-linking of proteins. The biochemical mechanisms of cement polymerization remain largely unknown. We hypothesized that this process is biochemically similar to blood clotting, a critical physiological response that is also based on aggregation and cross-linking of proteins. Like key elements of vertebrate and invertebrate blood clotting, barnacle cement polymerization was shown to involve proteolytic activation of enzymes and structural precursors, transglutaminase cross-linking and assembly of fibrous proteins. Proteolytic activation of structural proteins maximizes the potential for bonding interactions with other proteins and with the surface. Transglutaminase cross-linking reinforces cement integrity. Remarkably, epitopes and sequences homologous to bovine trypsin and human transglutaminase were identified in barnacle cement with tandem mass spectrometry and/or western blotting. Akin to blood clotting, the peptides generated during proteolytic activation functioned as signal molecules, linking a molecular level event (protein aggregation) to a behavioral response (barnacle larval settlement). Our results draw attention to a highly conserved protein polymerization mechanism and shed light on a long-standing biochemical puzzle. We suggest that barnacle cement polymerization is a specialized form of wound healing. The polymerization mechanism common between barnacle cement and blood may be a theme for many marine animal glues.

Biomechanics and neurophysiology studies suggest whole limb function to be an important locomotor control parameter. Inverted pendulum and mass-spring models greatly reduce the complexity of the legs and predict the dynamics of locomotion, but do not address how numerous limb elements are coordinated to achieve such simple behavior. As a first step, we hypothesized whole limb kinematics were of primary importance and would be preferentially conserved over individual joint kinematics after neuromuscular injury. We used a well-established peripheral nerve injury model of cat ankle extensor muscles to generate two experimental injury groups with a predictable time course of temporary paralysis followed by complete muscle self-reinnervation. Mean trajectories of individual joint kinematics were altered as a result of deficits after injury. By contrast, mean trajectories of limb orientation and limb length remained largely invariant across all animals, even with paralyzed ankle extensor muscles, suggesting changes in mean joint angles were coordinated as part of a long-term compensation strategy to minimize change in whole limb kinematics. Furthermore, at each measurement stage (pre-injury, paralytic and self-reinnervated) step-by-step variance of individual joint kinematics was always significantly greater than that of limb orientation. Our results suggest joint angle combinations are coordinated and selected to stabilize whole limb kinematics against short-term natural step-by-step deviations as well as long-term, pathological deviations created by injury. This may represent a fundamental compensation principle allowing animals to adapt to changing conditions with minimal effect on overall locomotor function.

Given the great range of visual systems, tasks and habitats, there is surprisingly little experimental evidence of how visual limitations affect behavioural strategies under natural conditions. Analysing this relationship will require an experimental system that allows for the synchronous measurement of visual cues and visually guided behaviour. The first step in quantifying visual cues from an animal's perspective is to understand the filter properties of its visual system. We examined the first stage of visual processing – sampling by the ommatidial array – in the compound eye of the fiddler crab Uca vomeris. Using an in vivo pseudopupil method we determined sizes and viewing directions of ommatidia and created a complete eye map of optical and sampling resolution across the visual field. Our results reveal five distinct eye regions (ventral, dorsal, frontal, lateral and medial) which exhibit clear differences in the organisation of the local sampling array, in particular with respect to the balance of resolution and contrast sensitivity. We argue that, under global eye space constraints, these regional optimisations reflect the information content and behavioural relevance of the corresponding parts of the visual field. In demonstrating the tight link between visual sampling, visual cues and behavioural strategies, our analysis highlights how the study of natural behaviour and natural stimuli is essential to our understanding and interpretation of the evolution and ecology of animal behaviour and the design of sensory systems.

The ears of insects exhibit a broad functional diversity with the ability to detect sounds across a wide range of frequencies and intensities. In tympanal ears, the membrane is a crucial step in the transduction of the acoustic stimulus into a neural signal. The tropical butterfly Morpho peleides has an oval-shaped membrane at the base of the forewing with an unusual dome in the middle of the structure. We are testing the hypothesis that this unconventional anatomical arrangement determines the mechanical tuning properties of this butterfly ear. Using microscanning laser Doppler vibrometry to measure the vibrational characteristics of this novel tympanum, the membrane was found to vibrate in two distinct modes, depending on the frequency range: at lower frequencies (1–5 kHz) the vibration was focused at the proximal half of the posterior side of the outer membrane, while at higher frequencies (5–20 kHz) the entire membrane contributed to the vibration. The maximum deflection points of the two vibrational modes correspond to the locations of the associated chordotonal organs, suggesting that M. peleides has the capacity for frequency partitioning because of the different vibrational properties of the two membrane components. Extracellular nerve recordings confirm that the innervating chordotonal organs respond to the same frequency range of 1–20 kHz, and are most sensitive between 2 and 4 kHz, although distinct frequency discrimination was not observed. We suggest that this remarkable variation in structure is associated with function that provides a selective advantage, particularly in predator detection.

Both the swimbladder and sonic muscles of the oyster toadfish Opsanus tau (Linnaeus) increase in size with fish growth making it difficult to distinguish their relative contributions to sound production. We examined acoustics of the swimbladder independent of the sonic muscles by striking it with a piezoelectric impact hammer. Amplitude and timing characteristics of bladder sound and displacement were compared for strikes of different amplitudes. Most of the first cycle of sound occurred during swimbladder compression, indicating that the bladder rapidly contracted and expanded as force increased during the strike. Harder hits were shorter in duration and generated a 30 dB increase in amplitude for a 5-fold or 14 dB range in displacement. For an equivalent strike dominant frequency, damping, bladder displacement and sound amplitude did not change with fish size, i.e. equal input generated equal output. The frequency spectrum was broad, and dominant frequency was driven by the strike and not the natural frequency of the bladder. Bladder displacement decayed rapidly ( averaged 0.33, equivalent to an automobile shock absorber), and the bladder had a low Q (sharpness of tuning), averaging 1.8. Sound output of an acoustic source is determined by volume velocity (surface area x velocity), and bladder surface area, muscle dimensions and contraction amplitude increase with fish size. Therefore, larger fish will be capable of producing more intense sound. Because the bladder is a low Q resonator, its output will follow muscle contraction rates independent of its size and natural frequency.

Crocodilians have complete anatomical separation between the ventricles, similar to birds and mammals, but retain the dual aortic arch system found in all non-avian reptiles. This cardiac anatomy allows surgical modification that prevents right-to-left (R–L) cardiac shunt. A R–L shunt is a bypass of the pulmonary circulation and recirculation of oxygen-poor blood back to the systemic circulation and has often been observed during the frequent apnoeic periods of non-avian reptiles, particularly during diving in aquatic species. We eliminated R–L shunt in American alligators (Alligator mississippiensis) by surgically occluding the left aorta (LAo; arising from right ventricle) upstream and downstream of the foramen of Panizza (FoP), and we tested the hypotheses that this removal of R–L shunt would cause afterload-induced cardiac remodelling and adversely affect diving performance. Occlusion of the LAo both upstream and downstream of the FoP for ~21 months caused a doubling of RV pressure and significant ventricular enlargement (average ~65%) compared with age-matched, sham-operated animals. In a separate group of recovered, surgically altered alligators allowed to dive freely in a dive chamber at 23°C, occlusion of the LAo did not alter oxygen consumption or voluntary apnoeic periods relative to sham animals. While surgical removal of R–L shunt causes considerable changes in cardiac morphology similar to aortic banding in mammals, its removal does not affect the respiratory pattern or metabolism of alligators. It appears probable that the low metabolic rate of reptiles, rather than pulmonary circulatory bypass, allows for normal aerobic dives.

Controversy exists over the scaling of oxygen consumption with body mass in vertebrates. A combination of biochemical and structural analyses were used to examine whether individual elements influencing oxygen delivery and demand within locomotory muscle respond similarly during ontogenetic growth of striped bass. Mass-specific metabolic enzyme activity confirmed that glycolytic capacity scaled positively in deep white muscle (regression slope, b=0.1 to 0.8) over a body mass range of ~20–1500 g, but only creatine phosphokinase showed positive scaling in lateral red muscle (b=0.5). Although oxidative enzymes showed negative allometry in red muscle (b=–0.01 to –0.02), mass-specific myoglobin content scaled positively (b=0.7). Capillary to fibre ratio of red muscle was higher in larger (1.42±0.15) than smaller (1.20±0.15) fish, suggesting progressive angiogenesis. By contrast, capillary density decreased (1989±161 vs 2962±305 mm^–2) as a result of larger fibre size (658±31 vs 307±24 µm² in 1595 g and 22.9 g fish, respectively). Thus, facilitated and convective delivery of O₂ show opposite allometric trends. Relative mitochondrial content of red muscle (an index of O₂ demand) varied little with body mass overall, but declined from ~40% fibre volume in the smallest to ~30% in the largest fish. However, total content per fibre increased, suggesting that mitochondrial biogenesis supported aerobic capacity during fibre growth. Heterogeneous fibre size indicates both hypertrophic and hyperplastic growth, although positive scaling of fibre myofibrillar content (b=0.085) may enhance specific force generation in larger fish. Modelling intracellular P_O2 distribution suggests such integrated structural modifications are required to maintain adequate oxygen delivery (calculated P_O2 5.15±0.02 kPa and 5.21±0.01 kPa in small and large fish, respectively).

The extent to which the expression of melanin-based plumage colouration in birds is genetically or environmentally determined is controversial. Here, we performed a between-nest design supplementation with either the sulphur amino acid dl-methionine or with water to investigate the importance of the non-genetic component of melanin-based plumage colouration in the Eurasian kestrel, Falco tinnunculus. Methionine affects growth and immunity, thus we aimed to modify nestling growth and immunity before feather development. Then, we measured the effect of the experiment on colouration of two melanin-based plumage patches of nestling kestrels. We found that methionine slowed down nestling growth through treatment administration and that nestlings compensated by speeding up their growth later. We did not find any effects of methionine on nestling immunity (i.e. lymphocyte counts, natural antibody levels or complement-mediated immunity). Effects on growth seemed to be mirrored by changes in nestling colouration in the two sexes: methionine-nestlings showed less intense brown plumage on their backs compared with control nestlings. These results provide support for a non-genetic determination of a melanin-based plumage patch in the two sexes of nestling kestrels.

Elasmobranch fishes utilise their vision as an important source of sensory information, and a range of visual adaptations have been shown to reflect the ecological diversity of this vertebrate group. This study investigates the hypotheses that visual optics can predict differences in habitat and behaviour and that visual optics change with ontogenetic growth of the eye to maintain optical performance. The study examines eye structure, pupillary movement, transmission properties of the ocular media, focal properties of the lens, tapetum structure and variations in optical performance with ontogenetic growth in two elasmobranch species: the carcharhinid sandbar shark, Carcharhinus plumbeus, inhabiting nearshore coastal waters, and the squalid shortspine spurdog, Squalus mitsukurii, inhabiting deeper waters of the continental shelf and slope. The optical properties appear to be well tuned for the visual needs of each species. Eyes continue to grow throughout life, resulting in an ontogenetic shift in the focal ratio of the eye. The eyes of C. plumbeus are optimised for vision under variable light conditions, which change during development as the animal probes new light environments in its search for food and mates. By contrast, the eyes of S. mitsukurii are specifically adapted to enhance retinal illumination within a dim light environment, and the detection of bioluminescent prey may be optimised with the use of lenticular short-wavelength-absorbing filters. Our findings suggest that the light environment strongly influences optical features in this class of vertebrates and that optical properties of the eye may be useful predictors of habitat and behaviour for lesser-known species of this vertebrate group.