Plasma metabolite profiling offers a potential means to assess stopover refueling performance of migratory birds from a single capture. However, this method has not previously been validated where site quality has been determined independently using analysis of capture data. We captured and blood sampled six passerine bird species refueling at known high-quality (BASE) and low-quality (TIP) sites at Long Point, Ontario, Canada. Plasma triglyceride, an indicator of fat deposition, was higher at the BASE in three early-season species: the hermit thrush, the American robin, and the white-throated sparrow. Plasma B-OH-butyrate, an indicator of fasting and lipid utilization, was lower at the BASE in the same three species. Plasma glycerol was lower at the BASE in American robins, and plasma phospholipid did not differ between sites. No metabolite suggested better conditions at the TIP in any species. Regression of size-corrected mass on time of day also indicated better refueling performance at the BASE in some species, but metabolite profiling was generally more sensitive to site differences. The relationship between plasma glycerol and triglyceride was U-shaped, indicating high glycerol production during both lipolysis (as was previously known) and rapid fat deposition. Our results confirm the validity of metabolite profiling to assess stopover habitat quality and individual performance in refueling migrants.

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SUMMARYSeasonal adjustments to muscle size in migratory birds may result from preparatory physiological changes or responses to changed workloads. The mechanisms controlling these changes in size are poorly understood. We investigated some potential mediators of flight muscle size (myostatin and insulin-like growth factor, IGF1) in pectoralis muscles of wild wintering or migrating white-throated sparrows (Zonotrichia albicollis), captive white-throated sparrows that were photoperiod manipulated to be in a 'wintering' or 'migratory' (Zugunruhe) state, and captive European starlings (Sturnus vulgaris) that were either exercised for 2 weeks in a wind tunnel or untrained. Flight muscle size increased in photo-stimulated 'migrants' and in exercised starlings. Acute exercise but not long-term training caused increased expression of IGF1, but neither caused a change in expression of myostatin or its metalloprotease activator TLL1. Photo-stimulated 'migrant' sparrows demonstrated increased expression of both myostatin and IGF1, but wild sparrows exhibited no significant seasonal changes in expression of either myostatin or IGF1. Additionally, in both study species we describe several splice variants of myostatin that are shared with distantly related bird species. We demonstrate that their expression patterns are not different from those of the typical myostatin, suggesting that they have no functional importance and may be mistakes of the splicing machinery. We conclude that IGF1 is likely to be an important mediator of muscle phenotypic flexibility during acute exercise and during endogenous, seasonal preparation for migration. The role of myostatin is less clear, but its paradoxical increase in photo-stimulated 'migrants' may indicate a role in seasonal adjustments of protein turnover.

In order to apply plasma metabolite profiling to assess the quality of stopover habitat for migrant birds it is crucial to know how quickly metabolite concentrations change in response to variation in energy intake rate. We fed live mealworms to Wilson's warblers Wilsonia pusilla at rates designed to mimic a low quality (0.58 wet g/h) and a high quality (0.75 wet g/h) habitat. Plasma concentrations of triglycerides and B‐OH‐butyrate were higher and lower, respectively, on the high feeding rate, but plasma glycerol levels did not differ between feeding rates. Birds were then fed at the low rate for five hours, switched to the high rate and blood sampled at various times after the switch. Plasma triglycerides and B‐OH‐butyrate reflected the increase in feeding rate within 10 and 20 min, respectively. Plasma glycerol did not change significantly following the change in feeding rate. We conclude that plasma metabolite concentrations can change very quickly in response to variation in feeding rate, and thus provide a sensitive index of fueling rates near a capture site.