Ulf Bauchinger

Differential catabolism of muscle protein in garden warblers (Sylvia borin): flight and leg muscle act as a protein source during long-distance migration.

Abstract. Samples of flight and leg muscle tissue were taken from migratory garden warblers at three different stages of migration: (1) pre-flight: when birds face an extended flight phase within the next few days, (2) post-flight: when they have just completed an extended flight phase, and (3) recovery: when they are at the end of a stop-over period following an extended flight phase. The changes in body mass are closely related to the changes in flight (P<0.001) and leg muscle mass (P<0.001), suggesting that the skeletal muscles are involved in the protein metabolism associated with migratory flight. From pre- to post-flight, the flight and the leg muscle masses decrease by about 22%, but are restored to about 12% above the pre-flight masses during the recovery period. Biochemical analyses show that following flight a selective reduction occurred in the myofibrillar (contractile) component of the flight muscle (P<0.01). As this selective reduction accounts only for a minor part of the muscle mass changes, sarcoplasmic (non-contractile) and myofibrillar proteins of both the flight and leg muscle act as a protein source during long-distance migration. As a loss of leg muscle mass is additionally observed besides the loss in flight muscle mass, mass change seems not to be strictly associated with the mechanical power output requirements during flight. Whereas the specific content of sarcoplasmic proteins in the flight muscle is nearly twice as high as that in the leg muscle (P<0.001), the specific content of myofibrillar proteins differs only slightly (P<0.05), being comparably low in both muscles. The ratio of non-contractile to contractile proteins in the flight muscle is one of the highest observed in muscles of a vertebrate.

Carbon Turnover in Tissues of a Passerine Bird: Allometry, Isotopic Clocks, and Phenotypic Flexibility in Organ Size

Stable isotopes are an important tool for physiological and behavioral ecologists, although their usefulness depends on a thorough understanding of the dynamics of isotope incorporation into tissue(s) over time. In contrast to hair, claws, and feathers, most animal tissues continuously incorporate carbon (and other elements), and so carbon isotope values may change over time, depending on resource use and tissue‐specific metabolic rates. Here we report the carbon turnover rate for 12 tissues from a passerine bird, the zebra finch (Taeniopygia guttata). We measured average carbon retention time (τ) for 8 d for small intestine; 10–13 d for gizzard, kidney, liver, pancreas, and proventriculus; 17–21 d for heart, brain, blood, and flight muscle; and 26–28 d for leg muscle and skin. We used these data, along with the few other published estimates, to confirm that the fractional rate of isotopic turnover for red blood cells, whole blood, liver, and leg muscle scales with body mass to approximately the −1/4 power. Our data also support several key assumptions of the “isotopic‐clock” model, which uses differences in isotope value between tissues, along with estimates of turnover rate of these tissues, to predict time elapsed since a diet shift. Finally, we show that between‐tissues differences in turnover rate largely, but not entirely, explain the extent of phenotypic flexibility in organs of garden warblers during their long‐distance flight across the Sahara Desert during spring. More studies that measure tissue‐specific protein synthesis, metabolic rate, and elemental turnover in many tissues from a variety of animals are needed.

Constitutive immune function in European starlings, Sturnus vulgaris, is decreased immediately after an endurance flight in a wind tunnel

SUMMARY Life-history theory predicts that animals face a trade-off in energy allocation between performing strenuous exercise, such as migratory flight, and mounting an immune response. We experimentally tested this prediction by studying immune function in European starlings, Sturnus vulgaris, flown in a wind tunnel. Specifically, we predicted that constitutive immune function decreases in response to training and, additionally, in response to immediate exercise. We compared constitutive immune function among three groups: (1) ‘untrained’ birds that were kept in cages and were not flown; (2) ‘trained’ birds that received flight training over a 15 day period and performed a 1-4 h continuous flight, after which they rested for 48 h before being sampled; and (3) ‘post-flight’ birds that differed from the ‘trained’ group only in being sampled immediately after the final flight. A bird in our trained group represents an individual during migration that has been resting between migratory flights for at least 2 days. A bird in our post-flight group represents an individual that has just completed a migratory flight and has not yet had time to recover. Three of our four indicators (haptoglobin, agglutination and lysis) showed the predicted decrease in immune function in the post-flight group, and two indicators (haptoglobin, agglutination) showed the predicted decreasing trend from the untrained to trained to post-flight group. Haptoglobin levels were negatively correlated with flight duration. No effect of training or flight was detected on leukocyte profiles. Our results suggest that in European starlings, constitutive immune function is decreased more as a result of immediate exercise than of exercise training. Because of the recent emergence of avian-borne diseases, understanding the trade-offs and challenges faced by long-distance migrants has gained a new level of relevance and urgency.

Testicular development during long-distance spring migration.

In birds, gonadal size varies between fully functional and maximally sized during reproduction and a regressed state with limited function during the non-reproductive periods. Recent findings show that testicular mass of the long-distance migratory garden warbler begins to increase during spring migration. Therefore, we sampled garden warblers during spring migration from Tanzania to Ethiopia, and finally to Egypt to determine if this mass increase is functional in terms of sperm and hormone production. In addition, we compared these birds, with garden warblers sampled after a 9-day recovery period following the crossing of the Sahara Desert (simulation of stopover) in Egypt and a group sampled in breeding condition in the laboratory. During migration there was a significant increase in testicular mass that was correlated with seminiferous tubule area and the stage of spermatogenesis. Plasma testosterone levels were low during migration, but were significantly correlated with testicular mass. LH concentration was not related to testicular mass, and, surprisingly, remained constant during migration. We suggest that previous experiments that found a tight relationship between testicular growth and LH level in the laboratory may not reflect the situation during long-distance migration where the great physical and energetic demands of migration likely affect the reproductive axis and, consequently, the relationship between testicular growth and LH titer. However, testes do develop to a considerable extent during spring migration and the present data suggest that initiation of testicular maturation during spring migration is necessary to ensure full spermatogenetic development soon after arrival at the breeding area.

Migratory stopover conditions affect the developmental state of male gonads in garden warblers (Sylvia borin)

Long-distance migrants face the challenge of a short window for reproduction that requires optimal timing and full functional gonads. Male garden warblers (Sylvia borin) meet these demands by initiating testicular recrudescence during spring migration, enabling them to reproduce immediately after arrival at the breeding grounds. In a combined field and laboratory study, we investigated testicular size, plasma levels of luteinizing hormone (LH), androstenedione (AE), 5alpha-dihydrotestosterone (DHT), testosterone and nocturnal migratory restlessness (Zugunruhe) under different stopover conditions. We manipulated food availability, the duration of stopover and simulated migration by food deprivation. Garden warblers showed significantly retarded testicular development after nine days of stopover under limited food conditions compared to birds that had ad libitum access to food. However, there was no significant difference in Zugunruhe between the two groups. Thus, the degree of Zugunruhe was unaffected by the quality of the stopover site and migration continued independent of the developmental state of the testis. We suggest that male garden warblers face the necessity to either compensate for slowed testicular recrudescence during the subsequent leg of migration and delay arrival at the breeding grounds, or arrive with less developed testes. Either of these may reduce annual reproductive success.

Exposure to cold but not exercise increases carbon turnover rates in specific tissues of a passerine

SUMMARY Carbon turnover differs between tissues within an animal, but the extent to which ecologically relevant increases in metabolism affect carbon turnover rates is largely unknown. We tested the energy expenditure and protein turnover hypotheses that predict increased carbon turnover, either in association with increased daily energy expenditure, or in concert with tissue-specific increased protein metabolism. We used stable-isotope-labeled diets to quantify the rate of carbon turnover in 12 different tissues for three groups of zebra finches (Taeniopygia guttata): cold-exposed birds kept at ambient temperatures below their thermoneutral zone, exercised birds that were flown for 2 h per day in a flight arena, and control birds that were kept at ambient temperatures within their thermoneutral zone and that were not exercised. We found that increases in metabolism associated with cold-exposure but not exercise produced measurable increases in carbon turnover rate of, on average, 2.4±0.3 days for pectoral muscle, gizzard, pancreas and heart, even though daily energy intake was similar for exercised and cold-exposed birds. This evidence does not support the energy expenditure hypothesis, and we invoke two physiological processes related to protein metabolism that can explain these treatment effects: organ mass increase and tissue-specific increase in activity. Such changes in carbon turnover rate associated with cold temperatures translate into substantial variation in the estimated time window for which resource use is estimated and this has important ecological relevance.

Migrating songbirds on stopover prepare for, and recover from, oxidative challenges posed by long-distance flight

Managing oxidative stress is an important physiological function for all aerobic organisms, particularly during periods of prolonged high metabolic activity, such as long-distance migration across ecological barriers. However, no previous study has investigated the oxidative status of birds at different stages of migration and whether that oxidative status depends on the condition of the birds. In this study, we compared (1) energy stores and circulating oxidative status measures in (a) two species of Neotropical migrants with differing migration strategies that were sampled at an autumn stopover site before an ecological barrier; and (b) a species of trans-Saharan migrant sampled at a spring stopover site after crossing an ecological barrier; and (2) circulating oxidative measures and indicators of fat metabolism in a trans-Saharan migrant after stopovers of varying duration (0–8 nights), based on recapture records. We found fat stores to be positively correlated with circulating antioxidant capacity in Blackpoll Warblers and Red-eyed Vireos preparing for fall migration on Block Island, USA, but uncorrelated in Garden Warblers on the island of Ponza, Italy, after a spring crossing of the Sahara Desert and Mediterranean Sea. In all circumstances, fat stores were positively correlated with circulating lipid oxidation levels. Among Garden Warblers on the island of Ponza, fat anabolism increased with stopover duration while oxidative damage levels decreased. Our study provides evidence that birds build antioxidant capacity as they build fat stores at stopover sites before long flights, but does not support the idea that antioxidant stores remain elevated in birds with high fuel levels after an ecological barrier. Our results further suggest that lipid oxidation may be an inescapable hazard of using fats as the primary fuel for flight. Yet, we also show that birds on stopover are capable of recovering from the oxidative damage they have accrued during migration, as lipid oxidation levels decrease with time on stopover. Thus, the physiological strategy of migrating songbirds may be to build prophylactic antioxidant capacity in concert with fuel stores at stopover sites before a long-distance flight, and then repair oxidative damage while refueling at stopover sites after long-distance flight.

Blackcap warblers maintain digestive efficiency by increasing digesta retention time on the first day of migratory stopover.

Almost all of the internal organs of migrating birds undergo pronounced mass changes, but the digestive tract changes most and fastest. The masses of the small intestine and the liver may be reduced by as much as 50% during migratory flight, indicating extreme phenotypic flexibility. Birds must rebuild these organs during stopovers to facilitate rapid mass gain and fuel deposition for continuation of migration. Laboratory studies indicate that birds may vary mean gut retention time to maintain high digestive efficiency, even while intestine length is substantially reduced. We examined migratory blackcaps (Sylvia atricapilla) after they performed a northward flight across the Sahara Desert in spring, and we subjected them to a 5‐d artificial stopover. Body mass (mb) changes over the 5‐d period resembled the typical mass change pattern of blackcaps stopping over naturally, with a small increase on the first day and a subsequent peak in the rate of mb gain on day 2 of the stopover. By day 5 of the stopover, the rate of mb gain had decreased to the point that it was not significantly different from that on day 1, presumably because the digestive tract had been rebuilt by this time. The same pattern was observed for food intake rates and amount of excreta produced, while digestive efficiency remained constant throughout the five experimental days. In contrast, mean retention time on stopover day 1 was significantly higher than it was on day 5 (106 min vs. 53 min; P = 0.031). Thus, after a sustained migration period, during a stopover blackcaps apparently compensate for reduced digestive tract size by processing food for twice as long, thereby maintaining high digestive efficiency.

During Stopover, Migrating Blackcaps Adjust Behavior and Intake of Food Depending on the Content of Protein in Their Diets

During migration, birds undergo alternating periods of fasting and re-feeding that are associated with dynamic changes in body mass (m(b)) and in organ size, including that of the digestive tract. After arrival at a migratory stopover site, following a long flight, a bird must restore the tissues of its digestive tract before it can refuel. In the present study we examined how the availability of dietary protein influences refueling of migrating blackcaps (Sylvia atricapilla) during a migratory stopover. We tested the following predictions in blackcaps deprived of food and water for 1-2 days to induce stopover behavior: (1) birds provided with a low-protein diet will gain m(b), lean mass and fat mass, and increase in pectoral muscle size slower than do birds fed a high-protein diet; (2) since stopover time is shorter in spring, birds will gain m(b) and build up fat tissue and lean tissue faster than in autumn; and (3) if low dietary protein limits a birds ability to gain m(b) and fat reserves, then birds that do not obtain enough protein will initiate migratory restlessness (Zugunruhe) earlier than will birds with adequate dietary protein. These predictions were tested by providing captured migrating blackcaps with semisynthetic isocaloric diets differing only in their protein content. Each day, we measured m(b), and food intake; also lean mass and fat mass were measured using dual energy X-ray absorptiometry. In addition, we monitored nocturnal activity with a video recording system. In both spring and autumn, birds fed diets containing either 3 or 20% protein increased in m(b), lean mass and fat mass at similar rates during the experiment. However, the group receiving 3% protein ate more than did the group receiving 20% protein. In support of our predictions, m(b), lean mass, fat mass, and intake of food all were higher in spring than in autumn. We also found that in spring all birds had higher levels of migratory restlessness, but birds fed 3% protein were less active at night than were birds fed 20% protein, possibly an adaptation conserving energy and protein. We conclude that protein requirements of migrating blackcaps during stopover are lower than expected, and that birds can compensate for low dietary protein by behavioral responses, i.e. hyperphagia and decreased migratory restlessness, that ensure rapid refueling.

Site-specific regulation of adult neurogenesis by dietary fatty acid content, vitamin E and flight exercise in European starlings

Exercise is known to have a strong effect on neuroproliferation in mammals ranging from rodents to humans. Recent studies have also shown that fatty acids and other dietary supplements can cause an upregulation of neurogenesis. It is not known, however, how exercise and diet interact in their effects on adult neurogenesis. We examined neuronal recruitment in multiple telencephalic sites in adult male European starlings (Sturnus vulgaris) exposed to a factorial combination of flight exercise, dietary fatty acids and antioxidants. Experimental birds were flown in a wind tunnel following a training regime that mimicked the birds natural flight behaviour. In addition to flight exercise, we manipulated the composition of dietary fatty acids and the level of enrichment with vitamin E, an antioxidant reported to enhance neuronal recruitment. We found that all three factors – flight exercise, fatty acid composition and vitamin E enrichment – regulate neuronal recruitment in a site‐specific manner. We also found a robust interaction between flight training and vitamin E enrichment at multiple sites of neuronal recruitment. Specifically, flight training was found to enhance neuronal recruitment across the telencephalon, but only in birds fed a diet with a low level of vitamin E. Conversely, dietary enrichment with vitamin E upregulated neuronal recruitment, but only in birds not flown in the wind tunnel. These findings indicate conserved modulation of adult neurogenesis by exercise and diet across vertebrate taxa and indicate possible therapeutic interventions in disorders characterized by reduced adult neurogenesis.