Effects of ghrelin on food caching behaviour and body composition in black-capped chickadees (Poecile atricapillus).

Several metabolic hormones signal an organism's energy balance to the brain and modulate feeding behaviours accordingly. These metabolic signals may also regulate other behaviour related to energy balance, such as food caching or hoarding. Ghrelin is one such hormone, but it appears to exert different effects on appetite and fat levels in birds and mammals. Ghrelin treatment inhibits food intake and decreases fat stores in some bird species, but these effects may differ between acylated and unacylated (des-acyl) forms of ghrelin. The effect of ghrelin on food caching in birds has been examined in only one study, that found both leptin and unacylated ghrelin reduced food caching and mass gain in coal tits (Periparus ater). We expanded on this to test how both forms of ghrelin affect food caching and body composition in black-capped chickadees (Poecile atricapillus). We injected each bird with acylated ghrelin, unacylated ghrelin, and a saline control and then measured food caching every 20 min for two hours post-injection. We also measured body mass fat levels the day before, and after treatment using quantitative magnetic resonance (QMR). Contrary to prior work, we found no effects of either form of ghrelin on food caching, or body or fat mass. Future work is required to determine if the difference between our results and those of the prior study stems from species differences in response to ghrelin and/or in the motivation to cache food, or ghrelin effects being modulated by energy reserves.

Neuronal activation in the geomagnetic responsive region Cluster N covaries with nocturnal migratory restlessness in white-throated sparrows (Zonotrichia albicollis).

Cluster N is a region of the visual forebrain of nocturnally migrating songbirds that supports the geomagnetic compass of nocturnal migrants. Cluster N expresses immediate-early genes (ZENK), indicating neuronal activation. This neuronal activity has only been recorded at night during the migratory season. Night-to-night variation in Cluster N activity in relation to migratory behaviour has not been previously examined. We tested whether Cluster N is activated only when birds are motivated to migrate and presumably engage their magnetic compass. We measured immediate-early gene activation in Cluster N of white-throated sparrows (Zonotrichia albicollis) in three conditions: daytime, nighttime migratory restless and nighttime resting. Birds in the nighttime migratory restlessness group had significantly greater numbers of ZENK-labelled cells in Cluster N compared to both the daytime and the nighttime resting groups. Additionally, the degree of migratory restlessness was positively correlated with the number of ZENK-labelled cells in the nighttime migratory restless group. Our study adds to the number of species observed to have neural activation in Cluster N and demonstrates for the first time that immediate early gene activation in Cluster N is correlated with the amount of active migratory behaviour displayed across sampled individuals. We conclude that Cluster N is facultatively regulated by the motivation to migrate, together with nocturnal activity, rather than obligatorily active during the migration season.

Methylmercury effects on avian brains.

Methylmercury (MeHg) is a concerning contaminant due to its ubiquity and harmful effects on organisms. Although birds are important models in the neurobiology of vocal learning and adult neuroplasticity, the neurotoxic effects of MeHg are less understood in birds than mammals. We surveyed the literature on MeHg effects on biochemical changes in the avian brain. Publication rates of papers related to neurology and/or birds and/or MeHg increased with time and can be linked with historical events, regulations, and increased understanding of MeHg cycling in the environment. However, publications on MeHg effects on the avian brain remain relatively low across time. The neural effects measured to evaluate MeHg neurotoxicity in birds changed with time and researcher interest. The measures most consistently affected by MeHg exposure in birds were markers of oxidative stress. NMDA, acetylcholinesterase, and Purkinje cells also seem sensitive to some extent. MeHg exposure has the potential to affect most neurotransmitter systems but more studies are needed for validation in birds. We also review the main mechanisms of MeHg-induced neurotoxicity in mammals and compare it to what is known in birds. The literature on MeHg effects on the avian brain is limited, preventing full construction of an adverse outcome pathway. We identify research gaps for taxonomic groups such as songbirds, and age- and life-stage groups such as immature fledgling stage and adult non-reproductive life stage. In addition, results are often inconsistent between experimental and field studies. We conclude that future neurotoxicological studies of MeHg impacts on birds need to better connect the numerous aspects of exposure from molecular physiological effects to behavioural outcomes that would be ecologically or biologically relevant for birds, especially under challenging conditions.

Androgen and prolactin manipulation do not induce changes in immunocompetence measures in a fish with male parental care.

Prolactin and 11-ketotestosterone (11-KT) are important reproductive hormones in fishes, which may also influence immunocompetence. The immunocompetence handicap hypothesis states that higher androgen concentrations that support secondary sex traits are traded off against a decrease in immune system function. To test the relationships between these hormones and immunocompetence, we experimentally manipulated 11-ketotestosterone and prolactin in the freshwater fish, bluegill (Lepomis macrochirus) during parental care using implants that contained either 11-KT, prolactin, or an inert control. We vaccinated individuals to stimulate the acquired immune response, then measured immunocompetence as the number of granulocytes, lymphocytes and monocytes, and the expression of interleukin 8 in each sample. We did not observe any significant differences in the immune measures among the hormone treatments. Our results indicate that in bluegill, there is no trade-off between androgens or prolactin and immunocompetence.

Opening the black box of bird-window collisions: passive video recordings in a residential backyard.

Collisions with windows on buildings are a major source of bird mortality. The current understanding of daytime collisions is limited by a lack of empirical data on how collisions occur in the real world because most data are collected by recording evidence of mortality rather than pre-collision behaviour. Based on published literature suggesting a causal relationship between bird collision risk and the appearance of reflections on glass, the fact that reflections vary in appearance depending on viewing angle, and general principles of object collision kinematics, we hypothesized that the risk and lethality of window collisions may be related to the angle and velocity of birds' flight. We deployed a home security camera system to passively record interactions between common North American bird species and residential windows in a backyard setting over spring, summer and fall seasons over 2 years. We captured 38 events including 29 collisions and nine near-misses in which birds approached the glass but avoided impact. Only two of the collisions resulted in immediate fatality, while 23 birds flew away immediately following impact. Birds approached the glass at variable flight speeds and from a wide range of angles, suggesting that the dynamic appearance of reflections on glass at different times of day may play a causal role in collision risk. Birds that approached the window at higher velocity were more likely to be immediately killed or stunned. Most collisions were not detected by the building occupants and, given that most birds flew away immediately, carcass surveys would only document a small fraction of window collisions. We discuss the implications of characterizing pre-collision behaviour for designing effective collision prevention methods.

Sex differences in myelination of the zebra finch vocal control system emerge relatively late in development.

The role of myelination in the development of motor control is widely known, but its role in the development of cognitive abilities is less understood. Here, we examined sex differences in the development of myelination of structures and tracts that support song learning and production in songbirds. We collected brains from 63 young male and female zebra finches (Taeniopygia guttata) over four stages of development that correspond to different stages of song learning. Using a myelination marker (myelin basic protein), we measured the development of myelination in three different nuclei of the vocal control system (HVC, RA, and lateral magnocellular nucleus of the anterior nidopallium [LMAN]) and two tracts (HVC-RA and lamina mesopallium ventralis [LMV]). We found that the myelination of the vocal control nuclei and tracts is sex related and male biased. In males, the patterns of myelination were age-dependent, asynchronous in rate and progression and associated with the development of song learning and production. In females, myelination of vocal control nuclei was low or absent and did not significantly change with age. Sex differences in myelination of the HVC-RA tract were large and emerged late in development well after sex differences in the size of vocal control brain regions are established. Myelination of this tract in males coincides with the age of song crystallization. Overall, the changes in myelination in the vocal control areas and tracts measured are region-, age-, and sex-specific and are consistent with sex differences in song development.

Effects of methylmercury and food stress on migratory activity in song sparrows, Melospiza melodia.

Avian migration is a challenging life stage susceptible to the adverse effects of stressors, including contaminants like methylmercury (MeHg). Although birds often experience stressors and contaminants concurrently in the wild, no study to date has investigated how simultaneous exposure to MeHg and food stress affects migratory behavior. Our objectives were to determine if MeHg or food stress exposure during summer, alone or combined, has carry-over effects on autumn migratory activity, and if hormone levels (corticosterone, thyroxine) and body condition were related to these effects. We tested how exposure to dietary MeHg and/or food stress (unpredictable temporary food removal) affected migratory behavior in captive song sparrows, Melospiza melodia. Nocturnal activity was influenced by a 3-way interaction between MeHg × stress × nights of the study, indicating that activity changed over time in different ways depending on prior treatments. Thyroxine was not affected by treatment or sampling date. During the migratory season, fecal corticosterone metabolite concentrations increased in birds co-exposed to MeHg and food stress compared to controls, suggesting an additive carry-over effect. As well, during the period of behavioral recording, body condition increased with time in unstressed birds, but not in stressed birds. Fecal corticosterone metabolite concentrations were positively correlated to duration of nocturnal activity, but thyroxine levels and body condition were not. The differences in nocturnal activity between groups suggest that food stress and MeHg exposure on breeding grounds could have direct and indirect carry-over effects that have the potential to affect the fall migration journey.

Leptin administration does not influence migratory behaviour in white-throated sparrows (Zonotrichia albicollis).

Migratory flights by birds are among the most energetically demanding forms of animal movement, and are primarily fueled by fat as an energy source. Leptin is a critical fat-regulation hormone associated with energy balance in non-avian species but its function in birds is highly controversial. Prior research indicated the effects of leptin differed between birds in migratory condition or not, but no research has assessed the effect of leptin on migratory behaviour itself. In this study, our objective was to determine if leptin affects migratory restlessness and fat deposition in migratory songbirds. We used photoperiod manipulation to induce spring migratory condition, and measured migratory restlessness in leptin-injected and saline-injected white-throated sparrows (Zonotrichia albicollis). Leptin treatment had no effect on migratory restlessness nor fat deposition, providing evidence that leptin does not influence avian migratory motivation or behaviour. Our results also further support the idea that birds in a hyperphagic migratory condition may be insensitive to leptin.

Experimental ghrelin administration affects migratory behaviour in a songbird.

Twice a year, billions of birds take on drastic physiological and behavioural changes to migrate between breeding and wintering areas. On migration, most passerine birds regularly stop over along the way to rest and refuel. Endogenous energy stores are not only the indispensable fuel to complete long distance flights, but are also important peripheral signals that once integrated in the brain modulate crucial behavioural decisions, such as the decision to resume migration after a stopover. A network of hormones signals metabolic fuel availability to the brain in vertebrates, including the recently discovered gut-hormone ghrelin. Here, we show that ghrelin takes part in the control of migratory behaviour during spring migration in a wild migratory passerine. We manipulated blood concentrations of ghrelin of 53 yellow-rumped warblers (Setophaga coronata coronata) caught during stopover and automatically radio-tracked their migratory behaviour following release. We found that injections of acylated and unacylated ghrelin rapidly induced movements away from the release site, indicating that the ghrelin system acts centrally to mediate stopover departure decisions. The effects of the hormone manipulation declined within 8 h following release, and did not affect the overall rate of migration. These results provide experimental evidence for a pivotal role of ghrelin in the modulation of behavioural decisions during migration. In addition, this study offers insights into the regulatory functions of metabolic hormones in the dialogue between gut and brain in birds.

European starlings (sturnus vulgaris) discriminate rhythms by rate, not temporal patterns.

Humans can perceive a regular psychological pulse in music known as the beat. The evolutionary origins and neural mechanisms underlying this ability are hypothetically linked to imitative vocal learning, a rare trait found only in some species of mammals and birds. Beat perception has been demonstrated in vocal learning parrots but not in songbirds. We trained European starlings (Sturnus vulgaris) on two sound discriminations to investigate their perception of the beat and temporal structure in rhythmic patterns. First, we trained birds on a two-choice discrimination between rhythmic patterns of tones that contain or lack a regular beat. Despite receiving extensive feedback, the starlings were unable to distinguish the first two patterns. Next, we probed the temporal cues that starlings use for discriminating rhythms in general. We trained birds to discriminate a baseline set of isochronous and triplet tone sequences. On occasional probe trials, we presented transformations of the baseline patterns. The starlings' responses to the probes suggest they relied on absolute temporal features to sort the sounds into "fast" and "slow" and otherwise ignored patterns that were present. Our results support that starlings attend to local features in rhythms and are less sensitive to the global temporal organization.

Neurogenesis and the development of neural sex differences in vocal control regions of songbirds.

The brain regions that control the learning and production of song and other learned vocalizations in songbirds exhibit some of the largest sex differences in the brain known in vertebrates and are associated with sex differences in singing behavior. Song learning takes place through multiple stages: an early sensory phase when song models are memorized, followed by a sensorimotor phase in which auditory feedback is used to modify song output through subsong, plastic song, to adult crystalized song. However, how patterns of neurogenesis in these brain regions change through these learning stages, and differ between the sexes, is little explored. We collected brains from 63 young male and female zebra finches (Taeniopygia guttata) over four stages of song learning. Using neurogenesis markers for cell division (proliferating cell nuclear antigen), neuron migration (doublecortin), and mature neurons (neuron-specific nuclear protein), we demonstrate that there are sex-specific changes in neurogenesis over song development that differ between the caudal motor pathway and anterior forebrain pathway of the vocal control circuit. In many of these regions, sex differences emerged very early in development, by 25 days post hatch, at the beginning of song learning. The emergence of sex differences in other components of the system was more gradual and had specific trajectories depending on the brain region and its function. In conclusion, we found that sex differences occurred early and continued during song learning. Moreover, transitions from the different phases of song development do not seem to depend on large changes in neurogenesis in the vocal control areas measured.

Feathers accurately reflect blood mercury at time of feather growth in a songbird.

Methylmercury (MeHg) is a globally distributed pollutant that can negatively affect wildlife. Bird feathers are often used as a monitoring tool of contaminant exposure, but variability in total mercury (THg) content in flight feathers has raised concerns over their utility. The objective of this study was to quantify blood and feather THg depuration through the progression of primary feather molt in order to clarify the relationship between blood and feather mercury concentration, and test the reliability of feather THg measurements as a monitoring tool in wild songbirds. Song sparrows (Melospiza melodia) were experimentally exposed to dietary MeHg and their blood and primary feather THg concentrations were measured during exposure and post-exposure periods of three months each. A rapid decrease in feather and blood THg concentration through molt progression was observed. Primary feather THg content was higher in feathers grown during the MeHg exposure period compared to those grown during the post-exposure period. Primary feather THg concentration was highly correlated with blood THg measured at the time of feather growth (R = 0.98), indicating that, although THg concentration is variable among flight feathers, this reflects temporally sequential molting patterns and declining blood concentration during depuration. Primary flight feathers thus provide an accurate and useful tool for estimating the mercury burden of birds at the time a chosen feather was grown, and have the potential to be an effective and reliable biomonitoring tool for species with well-characterized molt patterns.

Black-capped chickadees (Poecile atricapillus) use temperature as a cue for reproductive timing.

Reliable environmental cues, such as photoperiod, act as initial predictive cues that allow birds to time reproduction to match peak food abundance for their offspring. More variable local cues, like temperature, may, however, provide more precise information about the timing of food abundance. Non-migratory birds, in particular, should be sensitive to temperature cues and use them to modulate their reproductive timing. We conducted two experiments to examine the effect of temperature on reproductive condition (gonad size and circulating androgen levels) in non-migratory black-capped chickadees (Poecile atricapillus). First, we exposed groups of birds in outdoor aviaries to three different over-winter temperature treatments and assessed gonad size in the spring. Second, we manipulated temperature in environmental chambers under photostimulatory and non-photostimulatory photoperiodic conditions and assessed gonad size and circulating testosterone levels. Temperature had no independent effect on gonad size or testosterone levels, but when photostimulated birds exposed to warmer conditions became reproductively ready earlier than birds experiencing cooler conditions. We conclude that temperature acts as a supplementary cue that modulates the photoperiod-driven timing of reproduction.

Predator-induced fear causes PTSD-like changes in the brains and behaviour of wild animals.

Predator-induced fear is both, one of the most common stressors employed in animal model studies of post-traumatic stress disorder (PTSD), and a major focus of research in ecology. There has been a growing discourse between these disciplines but no direct empirical linkage. We endeavoured to provide this empirical linkage by conducting experiments drawing upon the strengths of both disciplines. Exposure to a natural cue of predator danger (predator vocalizations), had enduring effects of at least 7 days duration involving both, a heightened sensitivity to predator danger (indicative of an enduring memory of fear), and elevated neuronal activation in both the amygdala and hippocampus - in wild birds (black-capped chickadees, Poecile atricapillus), exposed to natural environmental and social experiences in the 7 days following predator exposure. Our results demonstrate enduring effects on the brain and behaviour, meeting the criteria to be considered an animal model of PTSD - in a wild animal, which are of a nature and degree which can be anticipated could affect fecundity and survival in free-living wildlife. We suggest our findings support both the proposition that PTSD is not unnatural, and that long-lasting effects of predator-induced fear, with likely effects on fecundity and survival, are the norm in nature.

The neural response of female zebra finches (Taeniopygia guttata) to conspecific, heterospecific, and isolate song depends on early-life song exposure.

The auditory forebrain regions caudo-medial nidopallium (NCM) and caudo-medial mesopallium (CMM) of songbirds exhibit differential expression of the immediate-early gene ZENK in response to playback of different song stimuli, and dependent on early-life auditory experience. Similarly, song preferences depend both on auditory experience and unlearned biases for particular song features. We explored the contributions of early-life auditory experience and the type of song stimuli on the Zenk response in the auditory forebrain of female zebra finches. Females were raised in three different early tutoring conditions: conspecific tutors that sang isolate song, heterospecific tutors, or conspecific tutors that sang wild-type song. At maturity, these females were exposed to one of five different playback conditions: wild-type song, isolate song, tutor song, heterospecific song, or white noise. Subsequently, the number of cells immunoreactive for ZENK in CMM and NCM was measured. We predicted that birds exposed to conspecific song early in life, and during the song playback in adulthood, would have the highest neural response. Instead, we found that the Zenk response varied across playback conditions with the highest response to conspecific wild-type and conspecific isolate song. In addition, we found a main effect of tutoring, with the lowest overall Zenk response in females tutored by males singing isolate song. Most importantly, there was a significant interaction in that females tutored by wild-type conspecific or heterospecific songs showed a similar increased response to zebra finch songs (wild-type or isolate), but females tutored by isolate song showed no differential response to conspecific song and only showed elevated Zenk response to the particular songs they were tutored with. Combined, our results indicate that unlearned response biases to conspecific song elements depend on previous auditory experience. That is, early experience appears to modulate the expression of innate biases.

Embryonic exposure to environmentally relevant concentrations of a brominated flame retardant reduces the size of song-control nuclei in a songbird.

Environmental contaminants have the potential to act as developmental stressors and impair development of song and the brain of songbirds, but they have been largely unstudied in this context. 2,2',4,4',5-Pentabromodiphenyl ether (BDE-99) is a brominated flame retardant congener that has demonstrated endocrine disrupting effects, and has pervaded the global environment. We assessed the effects of in ovo exposure to environmentally relevant levels of BDE-99 on the neuroanatomy of the song-control system in a model songbird species, the zebra finch (Taeniopygia guttata). Embryos were exposed via egg injection to a vehicle control (DMSO), 10, 100, or 1000 ng BDE-99/g egg on the day the egg was laid. Chicks were raised to sexual maturity to investigate long-term effects of BDE-99 on the adult male brain. Three key song-control nuclei (Area X, HVC, RA) all showed a dose-dependent trend toward decreasing volume as BDE-99 concentration increased, and birds exposed to 1000 ng/g in ovo BDE-99 had significantly smaller song-control nuclei volume compared to control birds. High environmental concentrations of BDE-99 in avian tissues can be within that range and thus could affect development of the song-control system in birds, and potentially other processes. We previously found that BDE-99 exposure during the nestling period had no effect of on the song-control system, although it did have significant effects on some behaviural endpoints. Taken together, these results suggest that exposure to polybrominated diphenyl ether (PBDEs) during critical developmental windows can significantly alter neurological development. © 2018 Wiley Periodicals, Inc. Develop Neurobiol, 2018.

Exposing migratory sparrows to Plasmodium suggests costs of resistance, not necessarily of infection itself.

Migratory birds move through multiple habitats and encounter a diverse suite of parasites. This raises concern over migrants' role in transporting infectious disease between breeding and wintering grounds, and along migratory flyways. Trade-offs between flight and immune defenses could interfere with infected individuals' migratory timing and success, potentially affecting infection dynamics. However, experimental evidence that parasitic infection affects migratory preparation or timing remains scant. We hypothesized that birds encountering hematozoan parasites shortly before migration incur physical costs (reduced body condition) and behavioral costs (delayed migration), due to the infection itself and/or to the demands of mounting an immune response. We experimentally inoculated song sparrows (Melospiza melodia) with Plasmodium shortly before fall migration. We monitored infection and body composition for 2 weeks after inoculation, and used radiotelemetry to track timing of migratory departure for another 7 weeks after release. Inoculated individuals that resisted infection had lower lean mass 12 days post exposure, relative to controls and infected individuals. This suggests trade-offs between body composition and immune defenses that might reduce migration success of resistant individuals. Despite group differences in body composition prior to release, we did not detect significant differences in timing of migration departure several weeks later. Thus, malarial infection did not appear to incur detectable costs to body composition or to migratory timing, at least when exposure occurs several weeks before migration. This study is novel considering not only the costs of infection, but also the costs of resisting infection, in an experimental context.

Symposium Overview: Integrating Cognitive, Motivational, and Sensory Biases Underlying Acoustic- and Multimodal-Based Mate Choice.

The goal of this symposium "Integrating Cognitive, Motivational and Sensory Biases Underlying Acoustic and Multimodal-Based Mate Choice" was to build a more complete framework in which to understand the mate choice brain. The presentations and papers within this symposium incorporate studies of motivational, cognitive, sensory and salience components of mate choice and highlight future directions that are needed to understand the biological basis of mate choice decision-making.

Male song sparrows have elevated testosterone in response to neighbors versus strangers.

Upon hearing a conspecific signal, animals must assess their relationship with the signaller and respond appropriately. Territorial animals usually respond more aggressively to strangers than neighbors in a phenomenon known as the "dear enemy effect". This phenomenon likely evolved because strangers represent a threat to an animal's territory tenure and parentage, whereas neighbors only represent a threat to an animal's parentage because they already possess a territory (providing territory boundaries are established and stable). Although the dear enemy effect has been widely documented using behavioral response variables, little research has been conducted on the physiological responses of animals to neighbors versus strangers. We sought to investigate whether the dear enemy effect is observed physiologically by exposing territorial male song sparrows (Melospiza melodia) to playback simulating a neighbor or a stranger, and then collecting blood samples to measure plasma testosterone levels. We predicted that song sparrows would exhibit increased testosterone levels after exposure to stranger playback compared to neighbor playback, due to the role testosterone plays in regulating aggression. Contrary to our prediction, we found that song sparrows had higher testosterone levels after exposure to neighbor playback compared to stranger playback. We discuss several explanations for our result, notably that corticosterone may regulate the dear enemy effect in male song sparrows and this may inhibit plasma testosterone. Future studies will benefit from examining corticosterone in addition to testosterone, to better understand the hormonal underpinnings of the dear enemy effect.

The avian hippocampus and the hypothetical maps used by navigating migratory birds (with some reflection on compasses and migratory restlessness).

The homology between the avian hippocampal formation (HF) and mammalian hippocampus nurtures the expectation that HF plays a fundamental role in navigation by migratory birds. Indeed, HF of migratory birds displays anatomical properties that differ from non-migratory species. Using a hypothetical framework of multiple maps of differing spatial resolution and range, homing pigeon data suggest that HF is important for navigating by landscape features near familiar breeding, over-wintering, and stop-over sites. By contrast, HF would be unimportant for an olfactory navigational map, which could be operational over unfamiliar space farther away from a goal location, nor is there any evidence for HF involvement in the sun or geomagnetic compass. The most intriguing question that remains open is what role HF may play in navigation when a migrant is thousands kms away from a familiar area, where homing pigeon data are uninformative and a geomagnetic map may be operational. Beyond navigation, successful migration depends on seasonal timing and often becoming nocturnally active. There is little evidence that HF plays a role in the timing of circannual and circadian cycles. Rather, circadian pacemakers including the pineal gland may control circadian timing of nocturnal restlessness and photoperiodic seasonal pacemakers likely control circannual expression.