Experimentally elevated corticosterone increases song output and complexity in common mynas.

Vocalization is an important communication tool that can reflect many aspects of an individual's internal and external condition. This is especially true for birds. Previous research has shown that bird calls and songs change in response to a variety of potential stressors, although the extent and direction of the changes depend on the nature of the stressor and the environment. Circulating glucocorticoids, such as corticosterone, often increase in response to stressors and mediate some of the observed changes via alterations of the individual's physiological state. Acute elevations of corticosterone often occur as a physiological response to short-term stressors; however, the effects of this elevation on adult vocalizations have not been well documented. Here, we experimentally elevated corticosterone at two different levels using a noninvasive method and examined the effects on the vocal communication of male and female adult common mynas (Acridotheres tristis). Corticosterone elevation temporarily increased song output and some measures of song complexity, while call output decreased. These effects were dosage dependent (higher corticosterone levels had a stronger effect), most evident 40 min after ingestion, and some vocal changes were sex-specific. Future studies should investigate whether the changes in vocal performance due to elevated glucocorticoids have consequences for the birds' behavior, reproductive success, and survival.

Culex quinquefasciatus (Diptera: Culicidae) survivorship following the ingestion of bird blood infected with Haemoproteus sp. parasites.

Arthropod vectors are frequently exposed to a diverse assemblage of parasites, but the consequence of these infections on their biology and behavior are poorly understood. We experimentally evaluated whether the ingestion of a common protozoan parasite of avian hosts (Haemoproteus spp.; Haemosporida: Haemoproteidae) impacted the survivorship of Culex quinquefasciatus (Say) (Diptera: Culicidae). Blood was collected from wild northern cardinals (Cardinalis cardinalis) in College Station, Texas, and screened for the presence of Haemoproteus spp. parasites using microscopic and molecular methods. Experimental groups of Cx. quinquefasciatus mosquitoes were offered Haemoproteus-positive cardinal blood through an artificial feeding apparatus, while control groups received Haemoproteus-negative cardinal blood or domestic canary (Serinus canaria domestica) blood. Culex quinquefasciatus mosquitoes exposed to Haemoproteus infected cardinal blood survived significantly fewer days than mosquitoes that ingested Haemoproteus-negative cardinal blood. The survival of mosquitoes fed on positive cardinal blood had a median survival time of 18 days post-exposure and the survival of mosquitoes fed on negative cardinal blood exceeded 50% across the 30 day observation period. Additionally, mosquitoes that fed on canary controls survived significantly fewer days than cardinal negative controls, with canary control mosquitoes having a median survival time of 17 days. This study further supports prior observations that Haemoproteus parasites can be pathogenic to bird-biting mosquitoes, and suggests that Haemoproteus parasites may indirectly suppress the transmission of co-circulating vector-borne pathogens by modulating vector survivorship. Our results also suggest that even in the absence of parasite infection, bloodmeals from different bird species can influence mosquito survivorship.

High Seasonal Variation of Plasma Testosterone Levels for a Tropical Grassland Bird Resembles Patterns of Temperate Birds.

AbstractTestosterone (T) is a sexual hormone capable of modulating several traits in birds, including aggressiveness and reproductive behavior. Although variation in T-related traits is well-known for temperate zone birds, this variation has not been extensively studied in tropical species. The campo miner (Geositta poeciloptera) is a threatened bird endemic to the grasslands of the South American Cerrado. We investigated the seasonal variation in plasma T levels and associated behavior in the campo miner, addressing the following questions: (1) Does the species exhibit seasonal variation in T profile? (2) Do males have higher plasma T levels than females, irrespective of season? (3) Are males with higher plasma T levels more aggressive than males with lower T levels? (4) Do males' plasma T levels decrease after females lay eggs? We found that T levels are higher during the breeding season than during the nonbreeding season and that males present higher T levels than females throughout the year. Such high T levels are associated with a higher probability to engage in aggressive behavior; however, T levels decline toward the egg-laying date and keep decreasing afterward. Higher T levels before egg laying are apparently related to territorial defense against invaders and extrapair copulations. With the beginning of parental care, T levels decrease, which is in line with previous observations that the species becomes less aggressive after egg laying. This study contributes to the understanding of environmental endocrinology of tropical birds, filling some knowledge gaps about the diverse Neotropical avifauna.

On the relationship between baseline corticosterone levels and annual survival of the thorn-tailed rayadito.

Baseline concentrations of glucocorticoids (i.e., cortisol and/or corticosterone) can moderately increase with the degree of energy demands that an individual faces. This could be a mechanism based on which glucocorticods (GCs) can mediate life history trade-offs, and therefore fitness. The 'cort-fitness hypothesis' predicts a negative relationship between GCs and fitness, meanwhile the 'cort-adaptation hypothesis' predicts the opposite pattern. Field studies on the relation between baseline GCs and survival rate have shown mixed results, supporting both positive and negative effect. These ambiguous results could be partially consequence of the short time frame in that most of the studies are carried on. In this study, we tested the predictions of the 'cort-fitness hypothesis' and 'cort-adaptation hypothesis' by using long-term data (eight-year of capture-mark-recapture) of Thorn-tailed Rayadito (Aphrastura spinicauda) in two populations at different latitudes. We assessed whether survival varied as a function of Cort levels and whether it varied in a linear (positive: 'cort-adaptation hypothesis' or negative: 'cort-fitness hypothesis') or curvilinear way. The two populations in our study had different baseline Cort levels, then we evaluated whether the association between baseline Cort and survival probability varied between them. In the high latitude population (i.e., lower baseline Cort levels), we observed a marginally quadratic relationship that is consistent with the cort-fitness hypothesis. In contrast, in the low altitude population we did not find this relation. Our findings suggests that the association between baseline Cort and survival probability is context-dependent, and highlights the importance of comparing different populations and the use of long-term data.

Testing the developmental hypothesis of the HPA axis in a tropical passerine: Dampened corticosterone response and faster negative feedback in nestling lance-tailed manakins (Chiroxiphia lanceolata).

When vertebrates are exposed to stressors, the subsequent acute increase in glucocorticoids by the hypothalamic-pituitaryadrenal (HPA) axis triggers a suite of adaptive responses, including mobilization of stored energy and repression of non-essential processes. However, chronic exposure to high concentrations of glucocorticoids can lead to metabolic dysregulation, impaired immune function, and cognitive decline. In developing young, this hormonal stress response shows considerable variation. Generally, the physiological stress response of young of precocial species is comparable to that of adults, whereas offspring of altricial species exhibit an attenuated response compared to adults. The developmental hypothesis of the HPA axis proposes that the dampened stress response in dependent offspring is an adaptive response to avoid the negative effects of elevated glucocorticoids, particularly in altricial species where young lack the ability to mitigate stressful stimuli.We aimed to test the developmental hypothesis in a tropical avian species, the lance-tailed manakin (Chiroxiphia lanceolata). We predicted that nestlings of this altricial species should have a dampened corticosterone response, in both magnitude and duration, compared to that of adults. We also predicted that recently fledged hatch-year birds would display a response intermediate to that of adults and nestlings. We quantified circulating corticosterone levels in adults, recently fledged hatch-year birds, and 11-day-old nestlings using a standardized capture and restraint protocol. Nestlings showed a lower maximal corticosterone response and faster negative feedback compared to adults. Further, five post-fledging hatch-year birds showed a feedback response intermediate to those of nestlings and adults. However, we caution against generalizing about fledgling responses beyond this study due to the small sample (n = 5). Interestingly, lance-tailed manakin nestlings appear to return to baseline concentrations faster than nestlings of temperate species. These results support the developmental hypothesis of the HPA axis explaining variation in stress response. This study is the first to assess the development of the hormonal stress response in nestlings of a tropical bird, which is of interest because of our still-developing understanding of how tropical and temperate species differ physiologically. Finally, findings here underscore the importance of validating and adjusting sampling protocols that quantify nestling stress responses, as sampling timelines identified for adults may underestimate the magnitude of the nestling stress response.

Chronic stress reverses enhanced neophobia following an acute stressor in European starlings.

Neophobia is an animal's avoidance of novelty. Animals tend to respond to novel objects by increasing their latency to approach the objects, and they eventually habituate after repeated exposure by attenuating this increased approach latency. Interestingly, the physiological stress response does not appear to have a causal link to neophobia, although acute stress can prevent animals from habituating to novel objects, possibly through a permissive effect. Chronic stress can induce an anxiety-like state in animals, while often disrupting the ability to respond to acute stress. We thus hypothesized that chronic stress may increase neophobia and tested this by inducing chronic stress in wild-caught European starlings (Sturnus vulgaris). Four distinct anthropogenic stressors were administered daily for 30 min each in a randomized order for 21 days. We then evaluated whether exposure to chronic stress altered the latency to approach a novel object placed on or near a food dish presented after overnight fasting. Chronically stressed birds and nonstressed controls exhibited similar initial neophobic responses to novel objects and showed similar habituation in response to repeated exposure. However, when birds were exposed to 15 min of restraint before repeated exposure to the same object, habituation was eliminated in control birds (i.e., they continued to respond with neophobia), whereas chronically stressed birds continued to show habituation as measured by attenuated approach latencies. These results demonstrate that an acute stress response (restraint) has a different impact on neophobia depending upon whether the bird is or is not concurrently exposed to chronic stress.

Recovery of hematocrit and fat deposits varies by cage size in food-restricted captive red crossbills (Loxia curvirostra).

Hematocrit-or the percent volume of red blood cells in whole blood-is thought to fluctuate adaptively in response to changing oxygen demands that occur during different life activities and in different environments. Because red blood cells are made from materials that can be limiting, however, it is thought that hematocrit may also reflect general body condition and access to resources. We tested the effect of hydration state, resource restriction (i.e., time available to forage), and activity (i.e., different cage sizes) on hematocrit in captive red crossbills (Loxia curvirostra). We found no evidence that a mild dehydration protocol impacts hematocrit and only weak support that mild food restriction impacts hematocrit. Food restriction did, however, reduce fat deposits and fat loss was more significant in birds that were also sampled for hematocrit. Furthermore, food-restricted birds housed in flight aviaries recovered hematocrit but not fat stores following repeated blood sampling, whereas birds housed in small cages lost additional hematocrit but mitigated fat loss following successive bleeds. Together these results suggest that different flight demands may determine response to blood loss during food restriction, potentially revealing a trade-off between fat storage and red blood cell development. Our results also demonstrate the need for scientists to carefully record hematocrit data and the time course across which multiple tubes of blood are collected to avoid confounding real patterns with variation generated by sampling protocol.

Effect of sleep loss on executive function and plasma corticosterone levels in an arctic-breeding songbird, the Lapland longspur (Calcarius lapponicus).

Sleep is a fundamental component of vertebrate life, although its exact functions remain unclear. Animals deprived of sleep typically show reduced neurobiological performance, health, and in some cases, survival. However, a number of vertebrate taxa exhibit adaptations that permit normal activities even when sleep is reduced. Lapland longspurs (Calcarius lapponicus), arctic-breeding passerine birds, exhibit around-the-clock activity during their short breeding season, with an inactive period of ca. 4 h/day. Whether behavioral or physiological costs occur from sleep loss (SL) in this species is unknown. To assess the effects of SL, wild-caught male longspurs were placed in captivity (12L:12D) and trained for one month to successfully learn color association and spatial memory tasks. Birds were then placed in automated sleep fragmentation cages that utilize a moving wire to force movement every 1 min (60 arousals/h) during 12D (inactive period) or control conditions (during 12L; active period). After SL (or control) treatment, birds were presented with color association and spatial memory tasks a final time to assess executive function. Baseline plasma corticosterone concentration, body mass, and satiety were also measured. SL significantly elevated corticosterone levels and increased accuracy during color association recall but did not affect the overall time required to complete the task. SL had no effect upon spatial memory, body mass, or satiety. Taken together, these results suggest that Lapland longspurs exhibit a degree of behavioral, but not physiological, insensitivity to acute SL. Whether elevated plasma concentrations of corticosterone play a direct role in ameliorating cognitive deficits from SL require additional study.

Geographic variation in baseline innate immune function does not follow variation in aridity along a tropical environmental gradient.

Geographic variation in aridity determines environmental productivity patterns, including large-scale variability in pathogens, vectors and associated diseases. If disease risk decreases with increasing aridity and is matched by immune defense, we predict a decrease in innate immune function along a gradient of increasing aridity from the cool-wet forest to the hot-dry Sahel, from south to north in Nigeria. We sampled blood and measured five innate immune indices from 286 Common Bulbuls Pycnonotus barbatus between 6 and 13°N. We sampled in the dry season; we resampled the first location (Jos) also as the last sample location to test temporal change in immune function. Immune indices did not decrease with aridity. One immune index, nitric oxide concentration showed a weak quadratic pattern. In Jos, ovotransferrin concentration, haemagglutination and haemolysis titres increased 12 weeks into the dry season, contrary to expectations that immune indices should decrease with increased dryness. In this tropical system, innate immune function does not decrease with increasing aridity but temporal factors within a location may influence immune function more strongly than spatial variation in aridity, suggesting that immune variation does not follow a simple environmental productivity pattern. Consequently, caution should probably be exercised in predicting effects of climate variability on immune function or disease risk.

Availability of specific prey types impact pied flycatcher (Ficedula hypoleuca) nestling health in a moderately lead contaminated environment in northern Sweden.

Anthropogenic metal contamination can cause increased stress in exposed organisms, but it can be difficult to disentangle the anthropogenic influence from natural variation in environmental conditions. In the proximity of a closed lead (Pb)/zinc (Zn) mine in northern Sweden, the health effects of Pb exposure, essential element (calcium [Ca] and Zn) uptake, and prey availability and composition were estimated on pied flycatcher (Ficedula hypoleuca) nestlings, using hemoglobin (Hb) level as a proxy for health. Pb concentration in nestling blood range between 0.00034 and 2.21 µg/g (ww) and nestlings close to the mine had higher Pb concentrations and lower Hb, but contrary to our hypothesis, Hb was not directly related to Pb accumulation. Proportions of flying terrestrial and aquatic insects in available prey and availability of flying terrestrial insects were positively associated with nestling Hb, whereas the proportion of terrestrial ground living prey, the most common prey type, showed a negative association. This suggests that positive influence of certain prey, which does not have to be the most common in the surroundings, can counteract the negative effects from Pb contamination on bird health. Nestlings inhabiting sites adjacent to lakes had an advantage in terms of prey composition and availability of preferred prey, which resulted in higher Hb. As such, our results show that during moderate exposure to metals, variation in natural conditions, such as prey availability, can have great impact on organism health compared to Pb exposure.

Proximity to roads, but not exposure to metal pollution, is associated with accelerated developmental telomere shortening in nestling great tits.

Comprehensively understanding the factors affecting physiology and fitness in urban wildlife requires concurrently considering multiple stressors. To this end, we simultaneously assessed how metal pollution and proximity to roads affect body condition and telomere shortening between days 8 and 15 of age in nestling great tits (Parus major), a common urban bird. We employed a repeated-measures sampling design to compare telomere shortening and body condition between nestlings from four urban study sites south of Antwerp, Belgium, which are located at different distances from a metal pollution point source. In addition, we explored associations between metal exposure and telomere dynamics on the individual level by measuring blood concentrations of five metals/metalloids, of which lead, copper and zinc were present at concentrations above the limit of detection. To assess whether roadway-associated stressors (e.g. noise and air pollution) might affect nestling condition and telomere shortening, we measured the proximity of nest boxes to roads. Metal exposure was not associated with nestling telomere length or body condition, despite elevated blood lead concentrations close to the metal pollution source (mean ±â€¯SE = 0.270 ±â€¯0.095 µg/g wet weight at the most polluted study site), suggesting that nestlings may have some capacity to detoxify metals. However, nestlings from nest boxes near roads exhibited more telomere shortening between days 8 and 15 of age, and shorter telomeres at day 15. Nestlings in poorer condition also had shorter telomeres, but proximity to the road was unrelated to body condition. Thus, nutritional stress is unlikely to mediate the relationship between proximity to roads and telomere length. Rather, proximity to roads could have affected telomere shortening by exposing nestlings to air or noise pollution. Our study highlights that traffic-related pollution, which is implicated in human health problems, might also affect urban wildlife.

Patterns of blood mercury variation in two long-distance migratory thrushes on Mount Mansfield, Vermont.

We investigated mercury (Hg) blood concentrations in Bicknell's thrush (Catharus bicknelli) and Swainson's thrush (C. ustulatus), congeneric long-distance migratory songbirds, from 2000-2017 at a montane forest site in north-central Vermont. We analyzed variation in blood Hg of both species using mixed-effects models, incorporating atmospheric wet Hg deposition data from a nearby sampling location. Although Hg deposition varied among years and seasonally, we detected no temporal trend in either atmospheric deposition or blood Hg, nor evidence of a relationship between the two. Sampling date had the strongest effect on blood Hg concentration, which declined seasonally, followed by age and sex of the individual. The data did not support an effect of species. We believe that the absence of a clear relationship between local atmospheric deposition and thrush blood Hg concentrations suggests that Hg cycling dynamics, mechanisms of transfer, and timing of uptake by montane forest biota are complex and poorly understood. The blood Hg concentrations of ~0.07-0.1 µg/g we documented in Bicknell's and Swainson's thrush are below those found to negatively impact physiological or reproductive endpoints in other invertivorous terrestrial passerines. To better evaluate the validity of Bicknell's thrush as a bioindicator of MeHg availability in montane forest ecosystems, we recommend (1) effects-based investigations, (2) a more robust understanding of Hg and MeHg cycling, (3) more clear geospatial and temporal links between Hg deposition and biotic uptake, and (4) more thorough documentation of Hg burdens across the species' annual cycle.

Sex-biased parasitism in vector-borne disease: Vector preference?

Sex-biased infections are a recurrent observation in vertebrates. In many species, males are more parasitized than females. Two potentially complementary mechanisms are often suggested to explain this pattern: sexual differences in susceptibility mainly caused by the effect of sex hormones on immunity and differential exposure to parasites. Exposure is mostly a consequence of host behavioural traits, but vector-borne parasitic infections involve another degree of complexity due to the active role of vectors in transmission. Blood-sucking insects may make choices based on cues produced by hosts. Regarding malaria, several studies highlighted a male-biased infection by Plasmodium sp in great tits (Parus major). We hypothesize that the mosquito vector, Culex pipiens, might at least partially cause this bias by being more attracted to male birds. Intrinsic variation associated to bird sex would explain a preference of mosquitoes for males. To test this hypothesis, we provide uninfected mosquitoes with a choice between uninfected male and female nestlings. Mosquito choice is assessed by sex typing of the ingested blood. We did not observe any preference for a given sex. This result does not support our prediction of a preference of mosquitoes for male great tits during the nestling period. In conclusion, mosquitoes do not seem to have an intrinsic preference for male nestlings. However, sexually divergent traits (e.g. behaviour, odour, metabolic rate) present in adults may play a role in the attraction of mosquitoes and should be investigated.

Feeding Success and Host Selection by Culex quinquefasciatus Say Mosquitoes in Experimental Trials.

Arthropod vector feeding preferences are defined as an overutilization of a particular host species given its abundance in relationship to other species in the community. Numerous methods exist to quantify vector feeding preferences; however, controlled host choice experiments are generally an underutilized approach. In this report, we present results from controlled vector host choice experiments using Culex quinquefasciatus Say (Diptera: Culicidae) mosquitoes and wild avian hosts identified as important contributors to West Nile virus (WNv) transmission in Atlanta, Georgia, United States. In each experiment, we allowed lab-reared F1Cx. quinquefasciatus to feed freely overnight on two avian individuals of a different species (i.e., northern cardinals, American robins, blue jays, brown thrashers, and gray catbirds). We then estimated WNv transmission potential using vectorial capacity and R0. We found that mosquito blood feeding success was extremely variable among experimental replicates and that patterns of host choice only occasionally aggregated to a particular bird species. Vectorial capacity was highest for American robins and blue jays due to these species' higher reservoir competence for WNv and greater probabilities of mosquito selection of these species. Despite species-specific differences in vectorial capacity, total community capacity was similar among species pairs. R0 estimates were qualitatively similar to capacity, and R0 was below and above unity across species pairs. Our results provide empirical evidence that C. quinquefasciatus is an opportunistic blood feeder and highlight how variability in vector-host contact rates as well as host community composition can influence the likelihood of WNv transmission in avian communities.

Non-invasive monitoring of physiological stress in an afrotropical arid-zone passerine bird, the southern pied babbler.

Using faecal matter to monitor stress levels in animals non-invasively is a powerful technique for elucidating the effects of biotic and abiotic stressors on free-living animals. To validate the use of droppings for measuring stress in southern pied babblers (Turdoides bicolor) we performed an ACTH challenge on captive individuals and determined the effect of temporary separation from their social group on their faecal glucocorticoid metabolite (fGCM) concentration. Additionally, we compared fGCM concentrations of captive babblers to those of wild conspecifics and examined the effects of dominance rank on fGCM concentration. We found droppings to be a suitable matrix for measuring physiological stress in babblers and that individual separation from the group caused an increase in fGCM levels. In addition, babblers temporarily held in captivity had substantially higher fGCM concentrations than wild individuals, indicating that babblers kept in captivity experience high levels of stress. In wild, free-living individuals, dominant males showed the highest levels of stress, suggesting that being the dominant male of a highly territorial social group is stressful. Non-invasive sampling allows field-based researchers to reduce disturbance related to monitoring adrenocortical function, thereby avoiding artificially increasing circulating corticosterone concentration as it is not necessary to physically restrain study animals.

An exception to the rule: Captivity does not stress wild migrating northern wheatears.

Wild animals typically suffer from stress when brought into captivity. This stress is characterized by elevated circulating glucocorticoid levels and weight loss. We here describe for the first time a case where a wild animal, the long-distance migrating northern wheatear, does not show signs of stress when caged. We captured these birds on a stopover site during their spring migration and caged them individually with ad libitum access to food and water. The birds were divided into four groups and were blood-sampled immediately in the field, a few hours after caging, one day after caging, or three days after caging, respectively. From these blood-samples we determined circulating corticosterone level. Food intake and body mass were also monitored. We found that, with very few exceptions, corticosterone levels were low and did not differ among the groups. Accordingly, almost all birds consumed huge quantities of food and substantially increased their body mass. Together these results clearly show that caging does not result in indications of stress in wild migrating northern wheatears. Confinement-specific conditions such as restricted movement normally stress animals. We suggest migratory birds may not perceive such conditions as stressors due to their hyperphagic state, a notion that requires further testing.

Sampling baseline androgens in free-living passerines: Methodological considerations and solutions.

Obtaining baseline hormone samples can be challenging because circulating hormone levels often change rapidly due to the acute stress of capture. Although field protocols are established for accurately sampling baseline glucocorticoid concentrations, fewer studies have examined how common sampling techniques affect androgens levels. Indeed, many studies focused on understanding the functional significance of baseline androgen levels use sampling methods known to activate the endocrine responses to stress. To understand how different field sampling protocols affect plasma androgen levels, we measured the androgen response to two types of capture stressors in a free-living tropical bird, the wire-tailed manakin (Pipra filicauda). First, we subjected males to a standardized capture and restraint protocol lasting either 15 or 30 min. Second, males were passively captured in nets that were filmed (to establish exact duration of time between capture and blood sampling) and checked every 30 min. The first study showed that circulating plasma androgen levels decreased significantly following both 15 and 30 min of restraint in a cloth bag, with a trend for the 30 min samples to be lower than the 15 min samples. Further, the change in androgen levels was dependent on an individual's initial androgen levels, with the individuals with the highest initial levels registering the largest declines. The results of the second study suggest that hanging in a mist net for extended periods of time also leads to a decrease in circulating androgen levels, but this effect was weaker than that of capture and restraint in a cloth bag. Our findings demonstrate that, overall, circulating androgen levels decrease in response to common sampling techniques; a finding that has important implications for studies measuring baseline androgen levels in free-living birds. Future studies should prioritize sampling individuals immediately upon removal from the mist net, as handling and restraint have a strong negative effect on circulating androgen levels. When constant monitoring of the mist net is not possible, investigators should use video cameras to record the amount of time an individual spends in the net prior to blood sampling and then statistically control for the effect of this variable in analyses.

Corticosterone profiles in northern cardinals (Cardinalis cardinalis): Do levels vary through life history stages?

As animals move through life history stages, energy requirements for each stage will vary. Both daily and annual variation in the glucocoriticoid hormones (specifically corticosterone, or CORT, in birds) helps provide the variable energy needed through life history stages. In many bird species, CORT is higher in the breeding season when energy demands can be quite high and is often lower in the non-breeding season. Additionally, CORT has a role to play in the response to stressful stimuli and the level to which CORT is elevated following stressful events can vary through the annual cycle as well. Here we report on baseline and stress-induced CORT levels in both sexes of northern cardinals, Cardinalis cardinalis, a non-migrating, year-round territorial species across life history stages. Corticosterone is overall higher in the non-breeding season than the breeding season in both sexes. Males tend to have higher levels of stress-induced CORT than females, although the observed patterns are complex. Our findings differ from one of the more common profile reported in songbirds where breeding season CORT tends to be higher than non-breeding levels. A strong influence may be the prolonged breeding season seen in cardinals; lower levels of CORT during breeding may guard against adverse maternal effects, interruptions in breeding behaviors, or egg production. Additional investigation of species with similar ecologies to northern cardinals, and more populations of cardinals, may show that annual glucocorticoid profiles are more labile than previously appreciated.

Divergent and parallel routes of biochemical adaptation in high-altitude passerine birds from the Qinghai-Tibet Plateau.

When different species experience similar selection pressures, the probability of evolving similar adaptive solutions may be influenced by legacies of evolutionary history, such as lineage-specific changes in genetic background. Here we test for adaptive convergence in hemoglobin (Hb) function among high-altitude passerine birds that are native to the Qinghai-Tibet Plateau, and we examine whether convergent increases in Hb-O2 affinity have a similar molecular basis in different species. We documented that high-altitude parid and aegithalid species from the Qinghai-Tibet Plateau have evolved derived increases in Hb-O2 affinity in comparison with their closest lowland relatives in East Asia. However, convergent increases in Hb-O2 affinity and convergence in underlying functional mechanisms were seldom attributable to the same amino acid substitutions in different species. Using ancestral protein resurrection and site-directed mutagenesis, we experimentally confirmed two cases in which parallel substitutions contributed to convergent increases in Hb-O2 affinity in codistributed high-altitude species. In one case involving the ground tit (Parus humilis) and gray-crested tit (Lophophanes dichrous), parallel amino acid replacements with affinity-enhancing effects were attributable to nonsynonymous substitutions at a CpG dinucleotide, suggesting a possible role for mutation bias in promoting recurrent changes at the same site. Overall, most altitude-related changes in Hb function were caused by divergent amino acid substitutions, and a select few were caused by parallel substitutions that produced similar phenotypic effects on the divergent genetic backgrounds of different species.

Accurate identification of Anopheles gambiae Giles trophic preferences by MALDI-TOF MS.

The determination of the trophic preferences of the Anopheles gambiae Giles (Diptera: Culicidae) is a decisive parameter for the monitoring and the prevention of malaria risk transmission. Currently, arthropod blood feeding sources are identified using immunological or molecular biology traditional techniques. Despite the effectiveness of these methods, they present several limitations, and notably, they are time-consuming and costly techniques. A recent study demonstrated that MALDI-TOF MS could be a useful tool for the identification of blood meal origins in freshly engorged mosquitoes. However, the limited number of blood vertebrate species tested to date, did not allow an assessment of the efficiency of MALDI-TOF MS in distinguishing blood MS spectra among close host species, such as humans versus primates. Therefore, in the present study, blood from ten distinct vertebrate host species, including four domestic species, four wild species, and two primates, was selected to control the reliability of MALDI-TOF MS based identification. Host blood species-specific MS profiles, up to 24h post-feeding in engorged Anopheles abdomens, were confirmed. Blind tests underlined the high specificity of MS spectra for the recognition of each host species, preventing misidentification. Nevertheless, an accurate analysis of the results from MS spectra queried against the MS database revealed that the reliability of identification is directly linked to the comprehensiveness of the MS reference database. Finally, the rapidity, the low-cost reagents, the simplicity of data analysis, and the accuracy of the tool for blood origin determination, make this proteomic strategy a promising complementary method for the elucidation of host/vector interactions.