Improving the success of reinforcement programs: effects of a two-week confinement in a field enclosure on the anti-predator behaviour of captive-bred European hamsters.

Captive breeding programs are an important pillar in biodiversity conservation, aiming to prevent the extinction of threatened species. However, the establishment of self-sustaining populations in the wild through the release of captive-bred animals is often hampered by a high mortality upon release. In this study, we investigated how a 2-week confinement period within a large field enclosure affected the anti-predator behaviour of 'naive' captive-bred hamsters and how potential modifications persisted over time. During three consecutive tests, hamsters were confronted with a moving predator model (a red fox mount, Vulpes vulpes) and their behaviour was filmed. After the initial round of confrontation with the predator model, one group of hamsters (field group) was released into a field enclosure protected from predators, while the other group (control) remained in their individual laboratory cages. After 2 weeks, hamsters from the field group were recaptured and individuals of both groups underwent a second confrontation test. A total of 1 month after their return from the field enclosure, field hamsters were subjected to a last confrontation test. Video analysis, investigating four behavioural variables, revealed that field hamsters significantly modified their behavioural response following the 2 weeks confinement in the enclosure, while this was not the case for control hamsters. In addition, most behavioural modifications in field hamsters persisted over 1 month, while others started to revert. We suggest that an appropriate pre-release period inside a field enclosure will enable naive (captive-bred) hamsters to develop an adequate anti-predator behaviour that will increase their immediate survival probability upon release into the wild. We believe that such measure will be of great importance for hamster conservation programs.

The early life of king penguins: ontogeny of dive capacity and foraging behaviour in an expert diver.

The period of emancipation in seabirds, when juveniles change from a terrestrial existence to a life at sea, is associated with many challenges. Apart from finding favourable foraging sites, they have to develop effective prey search patterns and physiological capacities that enable them to capture sufficient prey to meet their energetic needs. Animals that dive to forage, such as king penguins (Aptenodytes patagonicus), need to acquire an adequate breath-hold capacity, allowing them to locate and capture prey at depth. To investigate the ontogeny of their dive capacity and foraging performance, we implanted juvenile king penguins before their first departure to sea and also adult breeders with a data-logger recording pressure and temperature. We found that juvenile king penguins possess a remarkable dive capacity when leaving their natal colony, enabling them to conduct dives in excess of 100 m within their first week at sea. Despite this, juvenile dive/foraging performance, investigated in relation to dive depth, remained below the adult level throughout their first year at sea, probably reflecting physiological limitations as a result of incomplete maturation. A significantly shallower foraging depth of juveniles, particularly during their first 5 months at sea, could also indicate differences in foraging strategy and targeted prey. The initially greater wiggle rate suggests that juveniles fed opportunistically and also targeted different prey from adults and/or that many of the wiggles of juveniles reflect unsuccessful prey-capture attempts, indicating a lower foraging proficiency. After 5 months, this difference disappeared, suggesting sufficient physical maturation and improvement of juvenile foraging skills.

The dive performance of immature king penguins following their annual molt suggests physiological constraints.

Like all birds, penguins undergo periodic molt, during which they replace old feathers. However, unlike other birds, penguins replace their entire plumage within a short period while fasting ashore. During molt, king penguins (Aptenodytes patagonicus) lose half of their initial body mass, most importantly their insulating subcutaneous fat and half of their pectoral muscle mass. The latter might challenge their capacity to generate and sustain a sufficient mechanical power output to swim to distant food sources and propel themselves to great depth for successful prey capture. To investigate the effects of the annual molt fast on their dive/foraging performance, we studied various dive/foraging parameters and peripheral temperature patterns in immature king penguins across two molt cycles, after birds had spent their first and second year at sea, using implanted data-loggers. We found that the dive/foraging performance of immature king penguins was significantly reduced during post-molt foraging trips. Dive and bottom duration for a given depth were shorter during post-molt and post-dive surface interval duration was longer, reducing overall dive efficiency and underwater foraging time. We attribute this decline to the severe physiological changes that birds undergo during their annual molt. Peripheral temperature patterns differed greatly between pre- and post-molt trips, indicating the loss of the insulating subcutaneous fat layer during molt. Peripheral perfusion, as inferred from peripheral temperature, was restricted to short periods at night during pre-molt but occurred throughout extended periods during post-molt, reflecting the need to rapidly deposit an insulating fat layer during the latter period.

Why implantation of bio-loggers may improve our understanding of how animals cope within their natural environment.

Bio-loggers are miniaturized autonomous devices that record quantitative data on the state of free-ranging animals (e.g. behavior, position and physiology) and their natural environment. This is especially relevant for species where direct visual observation is difficult or impossible. Today, ongoing technical development allows the monitoring of numerous parameters in an increasing range of species over extended periods. However, the external attachment of devices might affect various aspects of animal performance (energetics, thermoregulation, foraging as well as social and reproductive behavior), which ultimately affect fitness. External attachment might also increase entanglement risk and the conspicuousness of animals, leaving them more vulnerable to predation. By contrast, implantation of devices can mitigate many of these undesirable effects and might be preferable, especially for long-term studies, provided that the many challenges associated with surgical procedures can be mastered. Implantation may then allow us to gather data that would be impossible to obtain otherwise and thereby may provide new and ecologically relevant insights into the life of wild animals. Here, we: (i) discuss the pros and cons of attachment methods; (ii) highlight recent field studies that used implanted bio-loggers to address eco-physiological questions in a wide range of species; and (iii) discuss logger implantation in light of ethical considerations.

Almost like a whale - first evidence of suction feeding in a seabird.

Little auks (Alle alle) are one of the most numerous seabird species in the world and feed primarily on copepods in arctic waters. Their high daily energy requirements leave them vulnerable to current changes in the arctic plankton community, where a smaller, less-profitable copepod species (Calanus finmarchicus) becomes increasingly abundant. Little auks have been estimated to require ∼60,000 copepods per day, necessitating prey capture rates of ∼6 copepods per second underwater. To achieve such performance, it has been suggested that little auks capture their prey by (non-visual) filter feeding. We tested this hypothesis by exposing little auks to varying copepod densities within a shallow experimental pool and filming their prey capture behaviour. At none of the copepod densities tested did birds filter feed. Instead, all birds captured copepods by what we identified as visually guided suction feeding, achieved through an extension of their sub-lingual pouch. Suction feeding is very common in fish and marine mammals, but to the best of our knowledge, this is the first time that it has been specifically identified in a seabird species. While presumably less efficient than filter feeding, this behaviour may allow little auks to foster higher resilience when facing the consequences of arctic climate change.

Thermal strategies of king penguins during prolonged fasting in water.

Most animals experience periods of unfavourable conditions, challenging their daily energy balance. During breeding, king penguins fast voluntarily for up to 1.5 months in the colony, after which they replenish their energy stores at sea. However, at sea, birds might encounter periods of low foraging profitability, forcing them to draw from previously stored energy (e.g. subcutaneous fat). Accessing peripheral fat stores requires perfusion, increasing heat loss and thermoregulatory costs. Hence, how these birds balance the conflicting demands of nutritional needs and thermoregulation is unclear. We investigated the physiological responses of king penguins to fasting in cold water by: (1) monitoring tissue temperatures, as a proxy of tissue perfusion, at four distinct sites (deep and peripheral); and (2) recording their oxygen consumption rate while birds floated inside a water tank. Despite frequent oscillations, temperatures of all tissues often reached near-normothermic levels, indicating that birds maintained perfusion to peripheral tissues throughout their fasting period in water. The oxygen consumption rate of birds increased with fasting duration in water, while it was also higher when the flank tissue was warmer, indicating greater perfusion. Hence, fasting king penguins in water maintained peripheral perfusion, despite the associated greater heat loss and, therefore, thermoregulatory costs, probably to access subcutaneous fat stores. Hence, the observed normothermia in peripheral tissues of king penguins at sea, upon completion of a foraging bout, is likely explained by their nutritional needs: depositing free fatty acids (FFA) in subcutaneous tissues after profitable foraging or mobilizing FFA to fuel metabolism when foraging success was insufficient.

Apparent changes in body insulation of juvenile king penguins suggest an energetic challenge during their early life at sea.

Little is known about the early life at sea of marine top predators, like deep-diving king penguins (Aptenodytes patagonicus), although this dispersal phase is probably a critical phase in their life. Apart from finding favourable foraging sites, they have to develop effective prey search patterns as well as physiological capacities that enable them to capture sufficient prey to meet their energetic needs. To investigate the ontogeny of their thermoregulatory responses at sea, we implanted 30 juvenile king penguins and 8 adult breeders with a small data logger that recorded pressure and subcutaneous temperature continuously for up to 2.5 years. We found important changes in the development of peripheral temperature patterns of foraging juvenile king penguins throughout their first year at sea. Peripheral temperature during foraging bouts fell to increasingly lower levels during the first 6 months at sea, after which it stabilized. Most importantly, these changes re-occurred during their second year at sea, after birds had fasted for ∼4 weeks on land during their second moult. Furthermore, similar peripheral temperature patterns were also present in adult birds during foraging trips throughout their breeding cycle. We suggest that rather than being a simple consequence of concurrent changes in dive effort or an indication of a physiological maturation process, these seasonal temperature changes mainly reflect differences in thermal insulation. Heat loss estimates for juveniles at sea were initially high but declined to approximately half after ∼6 months at sea, suggesting that juvenile king penguins face a strong energetic challenge during their early oceanic existence.

High peripheral temperatures in king penguins while resting at sea: thermoregulation versus fat deposition.

Marine endotherms living in cold water face an energetically challenging situation. Unless properly insulated, these animals will lose heat rapidly. The field metabolic rate of king penguins at sea is about twice that on land. However, when at sea, their metabolic rate is higher during extended resting periods at the surface than during foraging, when birds descend to great depth in pursuit of their prey. This is most likely explained by differences in thermal status. During foraging, peripheral vasoconstriction leads to a hypothermic shell, which is rewarmed during extended resting bouts at the surface. Maintaining peripheral perfusion during rest in cold water, however, will greatly increase heat loss and, therefore, thermoregulatory costs. Two hypotheses have been proposed to explain the maintenance of a normothermic shell during surface rest: (1) to help the unloading of N2 accumulated during diving; and (2) to allow the storage of fat in subcutaneous tissue, following the digestion of food. We tested the latter hypothesis by maintaining king penguins within a shallow seawater tank, while we recorded tissue temperature at four distinct sites. When king penguins were released into the tank during the day, their body temperature immediately declined. However, during the night, periodic rewarming of abdominal and peripheral tissues occurred, mimicking temperature patterns observed in the wild. Body temperatures, particularly in the flank, also depended on body condition and were higher in 'lean' birds (after 10 days of fasting) than in 'fat' birds. While not explicitly tested, our observation that nocturnal rewarming persists in the absence of diving activity during the day does not support the N2 unloading hypothesis. Rather, differences in temperature changes throughout the day and night, and the effect of body condition/mass supports the hypothesis that tissue perfusion during rest is required for nutritional needs.

Diving physiology of seabirds and marine mammals: Relevance, challenges and some solutions for field studies.

To fully understand how diving seabirds and marine mammals balance the potentially conflicting demands of holding their breath while living their lives underwater (and maintaining physiological homeostasis during exercise, feeding, growth, and reproduction), physiological studies must be conducted with animals in their natural environments. The purpose of this article is to review the importance of making physiological measurements on diving animals in field settings, while acknowledging the challenges and highlighting some solutions. The most extreme divers are great candidates for study, especially in a comparative and mechanistic context. However, physiological data are also required of a wide range of species for problems relating to other disciplines, in particular ecology and conservation biology. Physiological data help with understanding and predicting the outcomes of environmental change, and the direct impacts of anthropogenic activities. Methodological approaches that have facilitated the development of field-based diving physiology include the isolated diving hole protocol and the translocation paradigm, and while there are many techniques for remote observation, animal-borne biotelemetry, or "biologging", has been critical. We discuss issues related to the attachment of instruments, the retrieval of data and sensing of physiological variables, while also considering negative impacts of tagging. This is illustrated with examples from a variety of species, and an in-depth look at one of the best studied and most extreme divers, the emperor penguin (Aptenodytes forsteri). With a variety of approaches and high demand for data on the physiology of diving seabirds and marine mammals, the future of field studies is bright.

Reassessment of the cardio-respiratory stress response, using the king penguin as a model.

Research in to short-term cardio-respiratory changes in animals in reaction to a psychological stressor typically describes increases in rate of oxygen consumption (V̇(O2)) and heart rate. Consequently, the broad consensus is that they represent a fundamental stressor response generalizable across adult species. However, movement levels can also change in the presence of a stressor, yet studies have not accounted for this possible confound on heart rate. Thus the direct effects of psychological stressors on the cardio-respiratory system are not resolved. We used an innovative experimental design employing accelerometers attached to king penguins (Aptenodytes patagonicus) to measure and thus account for movement levels in a sedentary yet free-to-move animal model during a repeated measures stress experiment. As with previous studies on other species, incubating king penguins (N = 6) exhibited significant increases in both V̇(O2) and heart rate when exposed to the stressor. However, movement levels, while still low, also increased in response to the stressor. Once this was accounted for by comparing periods of time during the control and stress conditions when movement levels were similar as recorded by the accelerometers, only V̇(O2) significantly increased; there was no change in heart rate. These findings offer evidence that changing movement levels have an important effect on the measured stress response and that the cardio-respiratory response per se to a psychological stressor (i.e. the response as a result of physiological changes directly attributable to the stressor) is an increase in V̇(O2) without an increase in heart rate.

The effects of experimentally induced hyperthyroidism on the diving physiology of harbor seals (Phoca vitulina).

Many phocid seals are expert divers that remain submerged longer than expected based on estimates of oxygen storage and utilization. This discrepancy is most likely due to an overestimation of diving metabolic rate. During diving, a selective redistribution of blood flow occurs, which may result in reduced metabolism in the hypoperfused tissues and a possible decline in whole-body metabolism to below the resting level (hypometabolism). Thyroid hormones are crucial in regulation of energy metabolism in vertebrates and therefore their control might be an important part of achieving a hypometabolic state during diving. To investigate the effect of thyroid hormones on diving physiology of phocid seals, we measured oxygen consumption, heart rate, and post-dive lactate concentrations in five harbor seals (Phoca vitulina) conducting 5 min dives on command, in both euthyroid and experimentally induced hyperthyroid states. Oxygen consumption during diving was significantly reduced (by 25%) in both euthyroid and hyperthyroid states, confirming that metabolic rate during diving falls below resting levels. Hyperthyroidism increased oxygen consumption (by 7-8%) when resting in water and during diving, compared with the euthyroid state, illustrating the marked effect of thyroid hormones on metabolic rate. Consequently, post-dive lactate concentrations were significantly increased in the hyperthyroid state, suggesting that the greater oxygen consumption rates forced seals to make increased use of anaerobic metabolic pathways. During diving, hyperthyroid seals also exhibited a more profound decline in heart rate than seals in the euthyroid state, indicating that these seals were pushed toward their aerobic limit and required a more pronounced cardiovascular response. Our results demonstrate the powerful role of thyroid hormones in metabolic regulation and support the hypothesis that thyroid hormones play a role in modulating the at-sea metabolism of phocid seals.

Energy expenditure of freely swimming adult green turtles (Chelonia mydas) and its link with body acceleration.

Marine turtles are globally threatened. Crucial for the conservation of these large ectotherms is a detailed knowledge of their energy relationships, especially their at-sea metabolic rates, which will ultimately define population structure and size. Measuring metabolic rates in free-ranging aquatic animals, however, remains a challenge. Hence, it is not surprising that for most marine turtle species we know little about the energetic requirements of adults at sea. Recently, accelerometry has emerged as a promising tool for estimating activity-specific metabolic rates of animals in the field. Accelerometry allows quantification of the movement of animals (ODBA/PDBA, overall/partial dynamic body acceleration), which, after calibration, might serve as a proxy for metabolic rate. We measured oxygen consumption rates (V(O(2))) of adult green turtles (Chelonia mydas; 142.1±26.9 kg) at rest and when swimming within a 13 m-long swim channel, using flow-through respirometry. We investigated the effect of water temperature (T(w)) on turtle and tested the hypothesis that turtle body acceleration can be used as a proxy for V(O(2)). Mean mass-specific V(O(2)) (sV(O(2))) of six turtles when resting at a T(w) of 25.8±1.0°C was 0.50±0.09 ml min(-1) kg(-0.83). sV(O(2))increased significantly with T(w) and activity level. Changes in sV(O(2)) were paralleled by changes in respiratory frequency (f(R)). Deploying bi-axial accelerometers in conjunction with respirometry, we found a significant positive relationship between sV(O(2)) and PDBA that was modified by T(w). The resulting predictive equation was highly significant (r(2)=0.83, P<0.0001) and associated error estimates were small (mean algebraic error 3.3%), indicating that body acceleration is a good predictor of V(O(2)) in green turtles. Our results suggest that accelerometry is a suitable method to investigate marine turtle energetics at sea.

Heat increment of feeding in double-crested cormorants (Phalacrocorax auritus) and its potential for thermal substitution.

Diving endotherms inhabiting polar regions face potentially high thermoregulatory costs. Unless properly insulated, these animals will lose vast amounts of heat when diving in cold water, which has to be balanced by heat production. Heat generated as a by-product of digestion (heat increment of feeding, HIF) or from exercising muscles might be important in maintaining thermal balance under such conditions, as it would reduce the need for shivering thermogenesis. Recording the rate of oxygen consumption (V(O(2))), respiratory exchange ratio (RER), and stomach temperature, we studied the magnitude and duration of HIF in seven double-crested cormorants (Phalacrocorax auritus) following the voluntary ingestion of a single herring (Clupea pallasi) while birds rested in air. Conducting trials at thermoneutral (21.1+/-0.2 degrees C) and sub-thermoneutral temperatures (5.5+/-0.7 degrees C), we investigated the potential of HIF for thermal substitution. After the ingestion of a 100 g herring at thermoneutral conditions, V(O(2))was elevated for an average of 328+/-28 min, during which time birds consumed 2697+/-294 ml O(2) in excess of the resting rate. At sub-thermoneutral conditions, duration (228+/-6 min) and magnitude (1391+/-271 ml O(2)) of V(O(2))elevation were significantly reduced. This indicates that cormorants are able to use the heat generated as by-product of digestion to substitute for regulatory thermogenesis, if heat loss is sufficiently high. Altering meal size during sub-thermoneutral trials, we also found that HIF in cormorants was significantly greater after larger food intake. Based on these experimental results, a simple calculation suggests that substitution from HIF might reduce the daily thermoregulatory costs of double-crested cormorants wintering in coastal British Columbia by approximately 38%. Magnitude of HIF and its potential for thermal substitution should be integrated into bioenergetic models to avoid overestimating energy expenditure in these top predators.

When cormorants go fishing: the differing costs of hunting for sedentary and motile prey.

Cormorants hunt both benthic (sedentary) and pelagic (motile) prey but it is not known if the energy costs of foraging on these prey differ. We used respirometry to measure the costs of diving in double-crested cormorants (Phalacrocorax auritus) foraging either for sedentary (fish pieces) or motile (juvenile salmon) prey in a deep dive tank. Short dives for sedentary prey were more expensive than dives of similar duration for motile prey (e.g. 20% higher for a 10s dive) whereas the reverse was true for long dives (i.e. long dives for motile prey were more expensive than for sedentary prey). Across dives of all durations, the foraging phase of the dive was more expensive when the birds hunted motile prey, presumably due to pursuit costs. The period of descent in all the dives undertaken appears to have been more expensive when the birds foraged on sedentary prey, probably due to a higher swimming speed during this period.

The effects of depth, temperature and food ingestion on the foraging energetics of a diving endotherm, the double-crested cormorant (Phalacrocorax auritus).

Avian divers are confronted with a number of physiological challenges when foraging in cold water, especially at depth. Besides the obvious constraint imposed by the necessity to return to the surface for gas exchange, cold water temperatures and a reduction in body insulation due to the increase in pressure with dive depth will elevate the energetic costs of foraging in these endotherm divers. The complex effect that depth has on the diving energetics of aquatic birds has largely been ignored. To date, no study has assessed the impact of depth on diving energetics over a significant depth range, naturally encountered by the diver. We used open-circuit respirometry to study the energetic requirements of a foot-propelled pursuit diver, the double-crested cormorant (Phalacrocorax auritus albociliatus), when diving in a shallow (1 m) and deep (10 m) dive tank and when resting in air and water. We also investigated the modifying effects of air or water temperature and feeding status on the costs associated with diving and resting. Of all factors investigated, dive depth exercised the strongest influence on diving metabolic rate. Diving to 10 m depth increased metabolic rate on average by 22% when compared with shallow diving. Declining temperatures in air and water significantly elevated metabolic rate of cormorants resting in air and water as well as during diving. Feeding before resting in water or diving increased metabolic rate by 5-8% for at least 2 h. Cormorants maintained an elevated stomach temperature (>42 degrees C) when resting in water and during diving, even at cold temperatures. The elevated dive costs during deep diving, when compared with shallow diving, are most likely a consequence of the increased thermoregulatory costs associated with a greater heat loss to the water at depth. Nevertheless, our study shows that dive costs in double-crested cormorants are similar to those of other foot-propelled avian divers.

Energetic costs of diving and thermal status in European shags (Phalacrocorax aristotelis).

Diving is believed to be very costly in cormorants (Phalacrocoracidae) when compared with other avian divers because of their poor insulation and less-efficient foot propulsion. It was therefore suggested that cormorants might employ a behavioural strategy to reduce daily energy expenditure by minimizing the amount of time spent in water. However, European shags (Phalacrocorax aristotelis) have been observed to spend up to 7 h day(-1) diving in water of around 5-6 degrees C. To gain a better understanding of the energetic requirements in European shags, we measured their metabolic rates when resting in air/water and during shallow diving using respirometry. To investigate the effects of water temperature and feeding status on metabolic rate, birds dived at water temperatures ranging from 5 to 13 degrees C in both post-absorptive and absorptive states. In parallel with respirometry, stomach temperature loggers were deployed to monitor body temperature. Basal metabolic rate (BMR) was almost identical to allometric predictions at 4.73 W kg(-1). Metabolic rate when resting on water, during diving and after feeding was significantly elevated when compared with the resting-in-air rate. During diving, the metabolic rate of post-absorptive shags increased to 22.66 W kg(-1), which corresponds to 4.8x BMR. Minimum cost of transport (COT) was calculated at 17.8 J kg(-1) m(-1) at a swim speed of 1.3 m s(-1). Feeding before diving elevated diving metabolic rate by 13% for up to 5 h. There was a significant relationship between diving metabolic rate and water temperature, where metabolic rate increased as water temperature declined. Thermal conductance when resting in air at 10-19 degrees C was 2.05 W m(-2) degrees C(-1) and quadrupled during diving (7.88 W m(-2) degrees C(-1)). Stomach temperature when resting in air during the day was 40.6 degrees C and increased during activity. In dive trials lasting up to 50 min, stomach temperature fluctuated around a peak value of 42.0 degrees C. Hence, there is no evidence that European shags might employ a strategy of regional hypothermia. The energetic costs during shallow diving in European shags are considerably lower than has previously been reported for great cormorants (Phalacrocorax carbo) and are comparable to other foot-propelled divers. The lower dive costs in shags might be the consequence of a more streamlined body shape reducing hydrodynamic costs as well as a greater insulative plumage air layer (estimated to be 2.71 mm), which reduces thermoregulatory costs. The latter might be of great importance for shags especially during winter when they spend extended periods foraging in cold water.