Trace metal transfer to passerines inhabiting wastewater treatment wetlands.

Wastewater treatment wetlands are cost-effective strategies for remediating trace metals in industrial effluent. However, biogeochemical exchange between wastewater treatment wetlands and adjacent environments provides opportunities for trace metals to cycle in surrounding ecosystems. The transfer of trace metals to wildlife inhabiting treatment wetlands must be considered when evaluating wetland success. Using passerine birds as bioindicators, we conducted a multi-tissue analysis to investigate the mobilization of zinc, copper, and lead derived from wastewater to terrestrial wildlife in treatment wetlands and surrounding habitat. In addition, we evaluate the strength of relationships between metal concentrations in non-lethal (blood and feathers) and lethal (muscle and liver) sample types for estimation of toxicity risk. From July 2020 to August 2021, 177 passerines of seven species were captured at two wetlands constructed to treat industrial wastewater and two reference wetlands in the coastal plain of South Carolina. Feather, blood, liver, and muscle samples from each bird were analyzed for fourteen metals using inductively coupled plasma mass spectrometry and direct mercury analysis. Passerines inhabiting wastewater treatment wetlands accumulated higher concentrations of zinc in liver, copper in blood, and lead in feathers than passerines in reference wetlands, but neither blood nor feather concentrations were correlated with internal tissue concentrations. Of all the detected metals, only mercury in the blood showed a strong predictive relationship with mercury in internal tissues. This study indicates that trace metals derived from wastewater are bioavailable and exported to terrestrial wildlife and that passerine biomonitoring is a valuable tool for assessing metal transfer from treatment wetlands. Regular blood sampling can reveal proximate trace metal exposure but cannot predict internal body burdens for most metals.

Concentrations and biomagnification of persistent organic pollutants in three granivorous food chains from an abandoned e-waste recycling site.

The distinct accumulation patterns of persistent organic pollutants (POPs) among granivorous groups and the biomagnification of POPs from crops to granivorous species are still unclear. In this study, occurrence and biomagnification of POPs in three granivorous species including spotted dove (Spilopelia chinensis), scaly-breasted munia (Lonchura punctulata), and reed vole (Microtus fortis Buechner) from a former e-waste recycling site were investigated. Concentrations of polychlorinated biphenyls (PCBs) and polybrominated diphenyl ethers (PBDEs) in granivorous species ranged from 41.5 to 1370 and 21.1 to 3890 ng/g lipid weight, respectively. PCBs and PBDEs were the main POPs in birds and vole, while decabromodiphenyl ethane (DBDPE) and PBDEs were predominant POPs in crops. The dominance of BDE 209 was observed in samples, with few exceptions. Biomagnification factors (BMFs) of POPs in birds and vole were measured. BMFs of most POPs in vole were higher than those in birds, indicating that POPs had greater biomagnification potential in vole. Species-specific biomagnification of POPs might be affected by many factors, such as physiochemical properties and metabolic capability of POPs. There was significant correlation between concentration ratios of POPs in muscle/air and log KOA, which demonstrated that respiratory elimination to air affects biomagnification of POPs in granivorous birds and vole.

Does capacity to produce androgens underlie variation in female ornamentation and territoriality in White-shouldered Fairywren (Malurus alboscapulatus)?

Historic bias toward study of sex hormones and sexual ornamentation in males currently constrains our perspective of hormone-behavior-phenotype relationships. Resolving how ornamented female phenotypes evolve is particularly important for understanding the diversity of social signals across taxa. Studies of both males and females in taxa with variable female phenotypes are needed to establish whether sexes share mechanisms underlying expression of signaling phenotypes and behavior. White-shouldered Fairywren (Malurus alboscapulatus) subspecies vary in female ornamentation, baseline circulating androgens, and response to territorial intrusion. The moretoni ornamented female subspecies is characterized by higher female, but lower male baseline androgens, and a stronger pair territorial response relative to pairs from the lorentzi unornamented female subspecies. Here we address whether subspecific differences in female ornamentation, baseline androgens, and pair territoriality are associated with ability to elevate androgens following gonadotropin releasing hormone (GnRH) challenge and in response to simulated territorial intrusion. We find that subspecies do not differ in their capacity to produce androgens in either sex following GnRH or simulated territorial intrusion (STI) challenges. STI-induced androgens were predictive of degree of response to territorial intrusions in females only, but the direction of the effect was mixed. GnRH-induced androgens did not correlate with response to simulated intruders, nor did females sampled during intrusion elevate androgens relative to flushed controls, suggesting that increased androgens are not necessary for the expression of territorial defense behaviors. Collectively, our results suggest that capacity to produce androgens does not underlie subspecific patterns of female ornamentation, territoriality, and baseline plasma androgens.

Biomarkers of oxidative stress, metabolic processes, and lysosomal activity in the muscle tissue of the great tit (Parus major) living in sodium industry and agricultural areas in Inowroclaw region (central part of northern Poland).

An region of ecological hazard (EHR) is an area where, as a result of intensive human economic activity, degradation of components of the natural environment has taken place, leading to the deterioration of the ecological balance. EHR management is a globalised ever-increasing challenge. To eliminate the hazardous effect of these pollutants, research has been accelerated worldwide. The current study analyzed the specific biomarkers of the lipid and protein oxidation, total antioxidative status, activities of antioxidant enzymes (superoxide dismutase, catalase, glutathione reductase and peroxidase), lysosomal enzymes (alanyl aminopeptidase, leucyl aminopeptidase, acid phosphatase), and biomarkers of aerobic and anaerobic metabolic pathways (activities of alanine and aspartate aminotransferases, succinate dehydrogenase, lactate dehydrogenase, lactate and pyruvate levels) in skeletal muscle tissue of wild great tit nestlings in environments with different levels of anthropogenic pressure such as sodium industry and agriculture in the central part of northern Poland (Inowroclaw EHR). Control samples were collected from Tuchola Forest and Borkowo village, where no direct sources of contamination were found. The relevance of the study was to assess the changes in lysosomal functioning caused by pollution-induced oxidative stress that may indicate multidirectional adaptative mechanisms of metabolic processes occurring in the wild birds to compensate for the negative effects of contamination. It was shown that the initiation of oxidative stress caused by anthropogenic pollution shifted the balance of the normal functioning of lysosomal enzymes towards their increased activity. A general tendency towards an increase in the intensity of lipid peroxidation processes with an increasing level of oxidatively modified proteins (aldehydic and ketonic derivatives) and a simultaneous reduction in the TAS was observed in the muscle tissue of great tits living in the anthropogenically modified areas. The intensity of lipid peroxidation and protein damage caused changes and reorganization of the energy-related metabolic pathways in the muscle tissue of wild great tits living in the sodium industry and agricultural areas of the EHR.

Longer Sperm Swim More Slowly in the Canary Islands Chiffchaff.

Sperm swimming performance affects male fertilization success, particularly in species with high sperm competition. Understanding how sperm morphology impacts swimming performance is therefore important. Sperm swimming speed is hypothesized to increase with total sperm length, relative flagellum length (with the flagellum generating forward thrust), and relative midpiece length (as the midpiece contains the mitochondria). We tested these hypotheses and tested for divergence in sperm traits in five island populations of Canary Islands chiffchaff (Phylloscopus canariensis). We confirmed incipient mitochondrial DNA differentiation between Gran Canaria and the other islands. Sperm swimming speed correlated negatively with total sperm length, did not correlate with relative flagellum length, and correlated negatively with relative midpiece length (for Gran Canaria only). The proportion of motile cells increased with relative flagellum length on Gran Canaria only. Sperm morphology was similar across islands. We thus add to a growing number of studies on passerine birds that do not support sperm morphology-swimming speed hypotheses. We suggest that the swimming mechanics of passerine sperm are sufficiently different from mammalian sperm that predictions from mammalian hydrodynamic models should no longer be applied for this taxon. While both sperm morphology and sperm swimming speed are likely under selection in passerines, the relationship between them requires further elucidation.

Seasonal metabolic adjustments in an avian evolutionary relict restricted to mountain habitat.

For endotherms, maintaining body temperature during cold winters is energetically costly.Greater increase in winter maximum thermogenic capacity (Msum) has typically been correlated with improved cold tolerance. However, seasonal studies have shown equivocal direction change in basal metabolic rate (BMR) in winter, perhaps explained by latitude or phylogeny. We examined seasonal metabolic responses in the Cape rockjumper (Chaetops frenatus; "rockjumper"), a range-restricted mountain bird. We hypothesized that, given their mountain habitat preference, rockjumpers would be physiologically specialized for cooler air temperatures compared to other subtropical passerines. We measured body condition (using the ratio of Mb/tarsus), BMR, and Msum, in wild-living rockjumpers during winter and summer (n = 12 adults in winter -- 4 females, 8 males; n = 12 adults in summer -- 6 females, 6 males). We found birds had lesser BMR and thermal conductance, and greater Msum and body condition, in winter compared to summer. These changes may help rockjumpers conserve energy in winter while still allowing birds to produce more metabolic heat during the coldest air temperatures. When compared with existing data on avian seasonal metabolic adjustments, rockjumper BMR fit general patterns observed in passerines, but their Msum was low compared with other members of the oscine Passeriformes. These patterns may be explained by the narrow temperature range of their habitat not requiring cold-adjustment, or perhaps by their basal placement within passerine phylogeny. Further work on the physiological phenotypic plasticity in habitat specialists across different latitudinal zones and taxa is needed to better understand the relationship between metabolism, habitat, and phylogeny.

Turdoides affinis mitogenome reveals the translational efficiency and importance of NADH dehydrogenase complex-I in the Leiothrichidae family.

Mitochondrial genome provides useful information about species concerning its evolution and phylogenetics. We have taken the advantage of high throughput next-generation sequencing technique to sequence the complete mitogenome of Yellow-billed babbler (Turdoides affinis), a species endemic to Peninsular India and Sri Lanka. Both, reference-based and de-novo assemblies of mitogenome were performed and observed that de-novo assembled mitogenome was most appropriate. The complete mitogenome of yellow-billed babbler (assembled de-novo) was 17,672 bp in length with 53.2% AT composition. Thirteen protein-coding genes along with two rRNAs and 22 tRNAs were detected. The arrangement pattern of these genes was found conserved among Leiothrichidae family mitogenomes. Duplicated control regions were found in the newly sequenced mitogenome. Downstream bioinformatics analysis revealed the effect of translational efficiency and purifying selection pressure over thirteen protein-coding genes in yellow-billed babbler mitogenome. Ka/Ks analysis indicated the highest synonymous substitution rate in the nad6 gene. Evolutionary analysis revealed the conserved nature of all the protein-coding genes across Leiothrichidae family mitogenomes. Our limited phylogeny results placed T. affinis in a separate group, a sister group of Garrulax. Overall, our results provide a useful information for future studies on the evolutionary and adaptive mechanisms of birds belong to the Leiothrichidae family.

Wintering Snow Buntings Elevate Cold Hardiness to Extreme Levels but Show No Changes in Maintenance Costs.

AbstractResident temperate passerines adjust their phenotypes to cope with winter constraints, with peak performance in metabolic traits typically occurring during the coldest months. However, it is sparsely known whether cold-adapted northern species make similar adjustments when faced with variable seasonal environments. Life in near-constant cold could be associated with limited flexibility in traits underlying cold endurance. We investigated this by tracking individual physiological changes over five consecutive winters in snow buntings (Plectrophenax nivalis), an Arctic-breeding migratory passerine typically confronted with nearly constant cold. Buntings were held in an outdoor aviary and exposed to seasonal temperature variation typical of temperate zone climates. We measured phenotypic changes in body composition (body, fat, and lean mass, pectoralis muscle thickness), oxygen transport capacity (hematocrit), metabolic performance (basal metabolic rate [BMR] and summit metabolic rate [Msum]), thermogenic endurance (time to reach Msum), and cold tolerance (temperature at Msum). Snow buntings showed flexibility in functions underlying thermogenic capacity and cold endurance comparable to that observed in temperate resident passerines wintering at similar latitudes. Specifically, they increased body mass (13%), fat mass (246%), hematocrit (23%), pectoralis muscle thickness (8%), and Msum (27%). We also found remarkable cold tolerance in these birds, with individuals reaching Msum in helox at temperatures equivalent to less than -90°C in air. However, in contrast with resident temperate passerines, lean mass decreased by 12%, and there was no clear increase in maintenance costs (BMR). Our results show that the flexibility of traits underlying thermal acclimatization in a cold-adapted northern species is comparable to that of temperate resident species living at lower latitudes and is therefore not limited by life in near-constant cold.

Body condition and poxvirus infection predict circulating glucose levels in a colorful songbird that inhabits urban and rural environments.

There is widespread contemporary interest in causes and consequences of blood glucose status in humans (e.g., links to diabetes and cardiovascular disease), but we know comparatively less about what underlies variation in glucose levels of wild animals. Several environmental factors, including diet, disease status, and habitat quality, may regulate glucose circulation, and we are in need of work that assesses many organismal traits simultaneously to understand the plasticity and predictability of glucose levels in ecological and evolutionary contexts. Here, we measured circulating glucose levels in a species of passerine bird (the house finch, Haemorhous mexicanus) that has served as a valuable model for research on sexual selection, disease, and urban behavioral ecology, as these animals display sexually dichromatic ornamental coloration, harbor many infectious diseases (e.g., poxvirus, coccidiosis, mycoplasmal conjunctivitis), and reside in both natural habitats and cities. We tested the effects of sex, habitat type, body condition, coccidiosis and poxvirus infections, and expression of carotenoid plumage coloration on blood glucose concentrations and found that the body condition and poxvirus infection significantly predicted circulating glucose levels. Specifically, birds with higher blood glucose levels had higher body condition scores and were infected with poxvirus. This result is consistent with biomedical, domesticated-animal, and wildlife-rehabilitation findings, and the premise that glucose elevation is a physiological response to or indicator of infection and relative body weight. The fact that we failed to find links between glucose and our other measurements suggests that blood glucose levels can reveal some but not all aspects of organismal or environmental quality.

Seasonal Metabolic Acclimatization Varies in Direction and Magnitude among Years in Two Arid-Zone Passerines.

Adaptive plasticity in avian thermal physiology is increasingly apparent, with a well-studied example being metabolic upregulation during cold winters in small birds inhabiting temperate and boreal latitudes. Recent studies have revealed greater variation in the direction and magnitude of seasonal metabolic adjustments among subtropical/tropical birds experiencing milder winters compared with higher-latitude counterparts, suggesting that patterns could vary among years within populations. We quantified seasonal metabolic variation (summer vs. winter) in Kalahari Desert populations of two Afrotropical passerines, the white-browed sparrow-weaver (WBSW; Plocepasser mahali; ∼40 g) and the scaly-feathered weaver (SFW; Sporopipes squamifrons; ∼10 g) over subsequent years (2014-2017). We used flow-through respirometry to measure basal metabolic rate (BMR) and summit metabolism (Msum; maximum cold-induced resting metabolic rate) and quantified seasonal fluctuations in air temperature (Ta) and food abundance (arthropod and grass seed abundance) at the study site. Our data reveal that the direction and magnitude of seasonal metabolic acclimatization vary among years in both species, with the winter BMR of WBSWs ranging from ∼20% lower to 68% higher compared with the summer BMR. In contrast to higher-latitude species, Msum was not related to the cold-limit temperature of birds or to winter minimum Ta at the study site, but interannual variation in BMR and Msum was significantly lower in seasons with lower food abundance in both WBSWs and SFWs. Our data support the idea that patterns of seasonal acclimatization are more variable in birds from lower latitudes and that there is considerable phenotypic flexibility in avian thermal physiology.

Sperm Gatekeeping: 3D Imaging Reveals a Constricted Entrance to Zebra Finch Sperm Storage Tubules.

Females across many internally fertilizing taxa store sperm, often in specialized storage organs in their reproductive tracts. In birds, several hundred sperm storage tubules exist in the utero-vaginal junction of the oviduct, and there is growing evidence that sperm storage in these tubules is selective. The mechanisms underlying female sperm storage in birds remain unknown because of our limited ability to make three-dimensional, live observations inside the large, muscular avian oviduct. Here, we describe a new application of fluorescence selective plane illumination microscopy to optically section oviduct tissue from zebra finch Taeniopygia guttata females label free by harnessing tissue autofluorescence. Our data provide the first description of the three-dimensional structure of sperm storage organs in any vertebrate to the best of our knowledge and reveal the presence of gate-like constricted openings that may play a role in sperm selection.

Persistent organic pollutants (POPs) in oriental magpie-robins from e-waste, urban, and rural sites: Site-specific biomagnification of POPs.

Plenty of banned and emerging persistent organic pollutants (POPs), including dichlorodiphenyltrichloroethane and its metabolites (DDTs), polychlorinated biphenyls (PCBs), polybrominated diphenyl ethers (PBDEs), dechlorane plus (DP), and decabromodiphenyl ethane (DBDPE), were measured in oriental magpie-robins from an e-waste recycling site, an urban site (Guangzhou City), and a rural site in South China. Median concentrations of DDTs, PCBs, PBDEs, DP, and DBDPE ranged from 1,000-1,313, 800-59,368, 244-5,740, 24.1-127, and 14.7-36.0 ng/g lipid weight, respectively. Birds from the e-waste site had significantly higher concentrations of PCBs and PBDEs than those from urban and rural sites (p < 0.05), implying contamination of PCBs and PBDEs brought by e-waste recycling activities. DDTs were the predominant POPs in birds from urban and rural sites. The values of δ15N were significantly and positively correlated with concentrations of p,p'-DDE and low-halogenated chemicals in samples from the e-waste site (p < 0.05), indicating the trophic magnification of these chemicals in birds. However, concentrations of most POPs were not significantly correlated with the δ15N values in birds from urban and rural sites. PCBs and PBDEs in birds from urban and rural sites were not likely from local sources, and the biomagnification of POPs in different sites needed to be further investigated with caution.

Physiological condition of nestling great tits (Parus major) declines with the date of brood initiation: a long term study of first clutches.

In seasonal environments, a temporal decline in breeding performance (e.g. clutch size, nestling condition, and fledging success) of altricial bird species is a well-known phenomenon. In this study, we present the effect of laying phenology on the physiological condition of nestling great tits (Parus major) in 14 consecutive breeding seasons. We used blood haemoglobin and baseline glucose concentrations as indicators of nestling physiological condition. Nestling blood haemoglobin reflects food base quality and availability during the breeding period. Blood glucose concentration can be used as a supplementary reverse index of condition, since it is negatively related to environmental quality. It might be indicative of the stress caused by unfavourable extrinsic factors, though, due to potential confounding factors such as adverse weather conditions, low food quality, or feeding interruptions, glucose levels should be used in this ecological context with caution. Great tit nestlings from earlier broods were characterised by higher mean haemoglobin concentrations, indicating a seasonal decline in food quality and availability. The blood glucose concentration displayed an opposite pattern, with nestlings from earlier broods being characterised by lower mean concentrations than those from later broods. However, very little of the variation in blood glucose concentration can be explained by the variation in laying date, which suggests that blood glucose concentration is of little importance in the context of breeding phenology. Our results show that the physiological condition of nestlings of this species decreases as the breeding season progresses, most probably due to environmental factors.

Advancing the integration of multi-marker metabarcoding data in dietary analysis of trophic generalists.

The application of DNA metabarcoding to dietary analysis of trophic generalists requires using multiple markers in order to overcome problems of primer specificity and bias. However, limited attention has been given to the integration of information from multiple markers, particularly when they partly overlap in the taxa amplified, and vary in taxonomic resolution and biases. Here, we test the use of a mix of universal and specific markers, provide criteria to integrate multi-marker metabarcoding data and a python script to implement such criteria and produce a single list of taxa ingested per sample. We then compare the results of dietary analysis based on morphological methods, single markers, and the proposed combination of multiple markers. The study was based on the analysis of 115 faeces from a small passerine, the Black Wheatears (Oenanthe leucura). Morphological analysis detected far fewer plant taxa (12) than either a universal 18S marker (57) or the plant trnL marker (124). This may partly reflect the detection of secondary ingestion by molecular methods. Morphological identification also detected far fewer taxa (23) than when using 18S (91) or the arthropod markers IN16STK (244) and ZBJ (231), though each method missed or underestimated some prey items. Integration of multi-marker data provided far more detailed dietary information than any single marker and estimated higher frequencies of occurrence of all taxa. Overall, our results show the value of integrating data from multiple, taxonomically overlapping markers in an example dietary data set.

Within-winter flexibility in muscle and heart lipid transport and catabolism in passerine birds.

Small birds in cold climates may show within-winter metabolic flexibility to match metabolic capacities to prevailing weather conditions. This flexibility may occur over periods of days to weeks, but the underlying mechanisms for such flexibility are not well understood. Because lipids are the primary fuel for sustained thermogenesis, we examined whether lipid transport and catabolism can mediate within-winter metabolic flexibility in two small temperate-zone wintering passerine birds, dark-eyed juncos (Junco hyemalis) and house sparrows (Passer domesticus). We used simple and multiple regression analyses to test for correlations of several lipid transporters in pectoralis muscle (plasma membrane-bound and cytosolic fatty acid-binding proteins, FABP; fatty acyl translocase, FAT/CD36) and regulatory enzymes (carnitine acyl transferase, CPT; ß-hydroxyacyl CoA dehydrogenase, HOAD) in pectoralis and heart with short-term (ST, 0-7 days), medium-term (MT, 14-30 days) and long-term (LT, 30-year mean daily and extreme minimum temperatures, day of winter season) temperature variables. We hypothesized negative correlations between these regulators and temperature variables. Juncos showed negative correlations for FABPs with ST or MT temperature variables, but other lipid transporters and regulatory enzymes showed positive correlations with ST or MT temperatures for juncos, suggesting no consistent pathway-wide response to within-winter temperatures. LT temperature variables showed several significant associations with lipid transporters and enzymes for juncos, but also not in consistent directions. House sparrows showed the expected negative correlations with LT temperatures for FABPpm, but positive correlations with temperature variables for FABPc, CPT and HOAD. Different species-specific patterns of variation and the absence of consistent pathway-wide responses to temperature suggest that the lipid transport and catabolism pathway is not a uniform mediator of within-winter metabolic flexibility among small birds.

Snow buntings (Plectrophenax nivealis) as bio-indicators for exposure differences to legacy and emerging persistent organic pollutants from the Arctic terrestrial environment on Svalbard.

Eggs of snow buntings (Plectrophenax nivealis) were applied as a bio-indicator to examine differences in exposure to legacy persistent organic pollutants (POPs) and perflouroalkyl subtances (PFAS) from the terrestrial environment surrounding the settlements of Longyearbyen, Barentsburg and Pyramiden on Svalbard, Norway. Significantly higher concentrations of summed polychlorinated biphenyls (sumPCB7) in eggs collected from Barentsburg (2980 ng/g lipid weight (lw)) and Pyramiden (3860 ng/g lw) compared to Longyearbyen (96 ng/g lw) are attributed to local sources of PCBs within these settlements. Similar findings were observed for p,p'-dichlorodiphenyldichloroethylene (p,p'-DDE) where higher median concentrations observed in Pyramiden (173 ng/g lw) and Barentsburg (75 ng/g lw) compared to Longyearbyen (48 ng/g lw) may be influenced by guano inputs from breeding seabird populations, although other point sources cannot be ruled out. Concentrations of perfluorooctane sulphonate (PFOS) and several perfluorinated carboxylic acids (PFCAs) in snow bunting eggs were found to be statistically higher in the populated settlements of Longyearbyen and Barentsburg compared to the abandoned Pyramiden. Narrow foraging ranges of snow buntings during breeding season was useful in assessing point sources of exposure for PCBs and PFAS at particular sites with extreme differences observed between nest locations. SumPCB7 concentrations ranged from 2 µg/g ww to below detection limits between nest sites located less than a kilometer from each other in Pyramiden. Similar findings were observed in Longyearbyen, where several PFCAs ranged from 2 to 55 times higher between nest sites with similar spatial distances. These findings indicate that snow buntings can be a useful bio-indicator offering high spatial resolution for contaminant source apportionment in terrestrial environments on Svalbard.

Socially regulated estrogen in an eavesdropping brood parasite.

Social regulation of reproductive hormones is a means by which conspecific males and females orchestrate successful reproductive efforts. We investigate whether social cues modify activity within the hypothalamic-pituitary-gonadal (HPG) axis and the specificity of this response in a social parasite that is known to eavesdrop on the communication signals of other species: the brown-headed cowbird (Molothrus ater). Brown-headed cowbirds are obligate brood parasites that do not build nests or care for their own young. Instead, obligate brood parasites leave their eggs in the nest of a host species and therefore must coordinate their breeding attempts with conspecifics as well as potential heterospecific hosts. Here, we explore whether the vocal signals of potential host species can also be used as a social cue that modifies the HPG axis of female brown-headed cowbirds. Results reveal that both conspecific and heterospecific song-exposed females exhibit significantly greater circulating estradiol concentrations as compared to silence-exposed females. While conspecific song induces the greatest elevation in circulating estradiol, there is no significant difference in circulating estradiol levels in females exposed to either conspecific or heterospecific songs. This pattern suggests both song types are effective at evoking a reproductive physiological response. On the other hand, circulating progesterone concentrations did not differ among the song- and silence-exposed groups nor did the size of the female's ovarian follicles. These results indicate that heterospecific vocal communication signals can effectively be used as a social cue that simultaneously provides necessary information regarding breeding status of hosts and modifies breeding condition of the eavesdropper.

Dose-response effects of light at night on the reproductive physiology of great tits (Parus major): Integrating morphological analyses with candidate gene expression.

Artificial light at night (ALAN) is increasingly recognized as a potential threat to wildlife and ecosystem health. Among the ecological effects of ALAN, changes in reproductive timing are frequently reported, but the mechanisms underlying this relationship are still poorly understood. Here, we experimentally investigated these mechanisms by assessing dose-dependent photoperiodic responses to ALAN in the great tit (Parus major). We individually exposed photosensitive male birds to one of three nocturnal light levels (0.5, 1.5, and 5 lux), or to a dark control. Subsequent histological and molecular analyses on their testes indicated a dose-dependent reproductive response to ALAN. Specifically, different stages of gonadal growth were activated after exposure to different levels of light at night. mRNA transcript levels of genes linked to the development of germ cells (stra8 and spo11) were increased under 0.5 lux compared to the dark control. The 0.5 and 1.5 lux groups showed slight increases in testis size and transcript levels associated with steroid synthesis (lhr and hsd3b1) and spermatogenesis (fshr, wt1, sox9, and cldn11), although spermatogenesis was not detected in histological analysis. In contrast, all birds under 5 lux had 10 to 30 times larger testes than birds in all other groups, with a parallel strong increase in mRNA transcript levels and clear signs of spermatogenesis. Across treatments, the volume of the testes was generally a good predictor of testicular transcript levels. Overall, our findings indicate that even small changes in nocturnal light intensity can increase, or decrease, effects on the reproductive physiology of wild organisms.

Woodpecker drumming behavior is linked to the elevated expression of genes that encode calcium handling proteins in the neck musculature.

Many animals perform elaborate physical displays for social communication. Identifying molecular mechanisms that co-evolve with these complex behavioral signals can therefore help reveal how forces of selection shape animal design. To study this issue, we examined gene expression profiles in select skeletal muscles that actuate woodpecker drum displays. This remarkable whole-body signal is produced when individuals rapidly hammer their bill against trees. We found that, compared with muscles that play no part in producing this behavior, the main muscle used to drum abundantly expresses two genes that encode proteins that support myocytic calcium (Ca2+) handling dynamics - namely parvalbumin (PV) and sarcoplasmic reticulum Ca2+ ATPase 1 (SERCA1). Meanwhile, we found no such difference in the expression of another gene similarly vital to Ca2+ handling, ryanodine receptor 1 (RYR1). These differences are not present in a non-woodpecker species, which readily produce much slower drum-like movements for foraging (but not social signaling). Our data therefore point to an association between the fast drum displays of woodpeckers and muscle-specific expression of genes whose protein products enhance select aspects of myocytic Ca2+ handling.

Scotostimulation of reproductive neural pathways and gonadal maturation are not correlated with hypothalamic expression of deiodinases in subtropical spotted munia.

Circannual rhythm regulates the annual timing of reproduction in spotted munia, with sex differences in its relationship with the external photoperiod environment. Interestingly, munia show an atypical photosensitivity and exhibit gonadal maturation when acutely exposed to an unnatural short photoperiod (eg 3 hours of light per day; ie a long scotoperiod). The proximate mechanisms regulating scotoperiod-induced hypothalamic-pituitary-gonadal (HPG) activation are unclear. Because thyroid hormone signalling plays a central role in photoperiodic induction, we hypothesised the involvement of similar mechanism, comprising alterations in hypothalamic deiodinases, under long scotoperiod-induced HPG activation. To test this, several endpoints of cellular and molecular correlates were assayed in male and female munias after 1 and 4 weeks of exposure to an 3:21 hour light/dark cycle (3L:21D), with controls on a 21:3 hour light/dark cycle (21L:3D). We measured the hypothalamic expression of mRNA and protein of light-sensitive (neuropsin, OPN5) and reproductive (vasoactive intestinal peptide [VIP], neuropeptide Y [NPY], gonadotrophin-releasing hormone [GnRH], gonadotrophin-inhibiting hormone [GnIH]) neuropeptides by quantitative polymerase chain reaction (PCR) and immunohistochemistry, respectively. In addition, we also measured mRNA expression of types 2 (DIO2) and 3 (DIO3) deiodinases that regulate triiodothyronine-mediated GnRH release and gonadal maturation in photoperiodic species. The quantitative PCR and immunohistochemistry results were consistent. Higher OPN5 levels under 21L:3D than under 3L:21D suggested its role in sensing the length of the light period. Similarly, low VIP and high NPY expression under 3L:21D than under 21L:3D were consistent with their roles as cellular correlates of photic and nonphotic environment, respectively. High GnRH-I/low GnIH levels and gonadal recrudescence under 3L:21D, and an inverse pattern under 21L:3D, confirmed the scotostimulation of HPG axis in spotted munia. However, DIO2 and DIO3 mRNA levels did not differ between 2 scotoperiods, in contrast to their reciprocal expression pattern found during long-day photostimulation. We demonstrate for the first time sex-dependent scotostimulation of reproductive neural pathways and suggest the involvement of molecules other than hypothalamic deiodinases in the regulation of gonad development cycle in 'nonphotoperiodic' seasonally breeding vertebrates.