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Avian eggs develop outside of the female body, and therefore embryonic development is subject to multiple internal (physiological) and external (ecological) factors. Embryonic developmental rate has important consequences for survival. Within species, embryos that develop too quickly often experience deformities, disorders, or mortality, while embryos that develop slowly risk inviability and increase the time they are exposed to various sources of mortality in the nest. These contrasting forces may lead to interspecific variation in developmental rates. We investigated potential factors affecting embryonic heart rate (EHR), a proxy of development, across 14 passerine species in the field. More specifically, we investigated if nest predation risk, clutch size, seasonality, and egg volume influenced EHR. From previous research, we expected, and found, that EHR was positively associated with embryonic age and egg temperature. Species with greater nest predation risk had higher EHR, shorter incubation periods, and lower nest temperature variance. EHR increased as the season progressed and with egg volume, while EHR declined with clutch size. Bird species exhibit varying strategies to increase nestling and fledgling survival in response to predation risk, and these results suggest that variation in embryonic development may be related to species‐specific differences in nest predation risk. [ABSTRACT FROM AUTHOR]

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Abstract Avian eggs develop outside of the female body, and therefore embryonic development is subject to multiple internal (physiological) and external (ecological) factors. Embryonic developmental rate has important consequences for survival. Within species, embryos that develop too quickly often experience deformities, disorders, or mortality, while embryos that develop slowly risk inviability and increase the time they are exposed to various sources of mortality in the nest. These contrasting forces may lead to interspecific variation in developmental rates. We investigated potential factors affecting embryonic heart rate (EHR), a proxy of development, across 14 passerine species in the field. More specifically, we investigated if nest predation risk, clutch size, seasonality, and egg volume influenced EHR. From previous research, we expected, and found, that EHR was positively associated with embryonic age and egg temperature. Species with greater nest predation risk had higher EHR, shorter incubation periods, and lower nest temperature variance. EHR increased as the season progressed and with egg volume, while EHR declined with clutch size. Bird species exhibit varying strategies to increase nestling and fledgling survival in response to predation risk, and these results suggest that variation in embryonic development may be related to species‐specific differences in nest predation risk.

Weather conditions play a pivotal role in embryo development and parental incubation costs, potentially impacting the clutch size and incubation behavior of birds. Understanding these effects is crucial for bird conservation. Reeves's Pheasant (Syrmaticus reevesii) is a threatened species endemic to China, which is characterized by female-only incubation. However, there is a lack of information regarding the impact of weather conditions on clutch size and incubation behavior in this species. Using satellite tracking, we tracked 27 wild female Reeves's Pheasants from 2020 to 2023 in Hubei Province, China. We explored their clutch size and incubation behavior, as well as their responses to ambient temperature and precipitation. Clutch size averaged 7.75 ± 1.36, had an association with average ambient temperature and average daily precipitation during the egg-laying period, and was potentially linked to female breeding attempts. Throughout the incubation period, females took an average of 0.73 ± 0.46 recesses every 24 h, with an average recess duration of 100.80 ± 73.37 min and an average nest attendance of 92.98 ± 5.27%. They showed a unimodal recess pattern in which nest departures peaked primarily between 13:00 and 16:00. Furthermore, females rarely left nests when daily precipitation was high. Recess duration and nest attendance were influenced by the interaction between daily mean ambient temperature and daily precipitation, as well as day of incubation. Additionally, there was a positive correlation between clutch size and recess duration. These results contribute valuable insights into the life-history features of this endangered species.

Noise pollution can degrade the behavioral, physiological, and psychological health of humans and other creatures. We used breeding pairs of Eastern Bluebirds ( Sialia sialis ) to assess behavioral and reproductive responses to both chronic roadway noise and experimental intermittent playbacks of construction noise. Active nests in boxes placed near and far from large roads were randomly assigned as treatments or controls for experimental playbacks during incubation. Using temperature signatures from iButtons placed within nest cups we quantified certain female incubation behaviors (# and length of bouts, # of small temperature fluctuations, and total warming minutes per day) and hatching success was recorded for 40 nests in spring of 2019. Nests in quiet areas that received no additional playback treatments of construction noise had markedly higher nest success than any exposed to noise. Nests exposed to chronic traffic noise only, and quiet nests that received 3–4 days of construction noise had the lowest hatching success. Females in traffic-quiet nests increased restlessness (small temperature fluctuations) and experienced decreasing hatching success as the number of days of construction noise playback increased. Thus, birds choosing either quiet or noisy boxes had contrasting responses to bouts of construction noise. Other female incubation behaviors we could detect were unaffected by noise but changed in expected ways with seasonal progression. In sum, both types of noise can decrease hatch rate, but with intermittent noise this is likely due to female restlessness, or too many small drops in temperature to maintain optimal embryo development.

The incubation behavior of the Japanese Nagoya chicken breed is a commercial issue because it often causes a sudden and sharp drop in egg production. In this study, whether the incidence of incubation behavior in Nagoya laying hens was associated with calls and the presence of roosters in the same laying house was investigated. Four experiments were conducted using commercial layer-type Nagoya hens where the hatching time of the experimental birds and the treatment order in the presence of males were changed . In Experiment 1, the proportion of incubation behavior in the presence of roosters kept in another pen located between pen-rearing hens (51.3%) was higher than that in their absence (15.9%) or with only rooster calls (23.8%). In Experiments 2, 3, and 4, the proportion of incubation behavior in the presence of roosters (47.3%, 33.3%, and 37.9%, respectively) was higher than that in their absence (33.3%, 17.4%, and 25.6%, respectively). In all experiments, approximately 70% of the incubating hens observed in the absence of roosters exhibited incubation behavior, even in the presence of roosters. Therefore, the presence of roosters may enhance egg incubation behavior in Nagoya laying hens.

The incubation behavior of geese seriously affects their egg production performance. Studies on incubation behavior have identified functional genes, but the regulatory architecture relationship between functional genes and chromatin accessibility remains poorly understood. Here, we present an integrated analysis of open chromatin profiles and transcriptome to identify the cis-regulatory element and their potential transcription factors involved in regulating incubation behavior in goose pituitary. Assay for transposase-accessible chromatin sequencing (ATAC-seq) revealed that open chromatin regions increased in the pituitary during the transition from incubation behavior to laying. We identified 920 significant differential accessible regions (DARs) in the pituitary. Compared to the laying stage, most DARs had higher chromatin accessibility in the brooding stage. Motif analysis of open DARs showed that the most significant transcription factor (TF) occupied sites predominantly enriched in motifs binding to the RFX family (RFX5, RFX2, and RFX1). While the majority of TF motifs enriched under sites of the nuclear receptor (NR) family (ARE, GRE, and PGR) in closed DARs at the incubation behavior stage. Footprint analysis indicated that the transcription factor RFX family exhibited higher binding on chromatin at the brooding stage. To further elucidate the effect of changes in chromatin accessibility on gene expression levels, a comparison of the transcriptome revealed 279 differentially expressed genes (DEGs). The transcriptome changes were associated with processes of steroid biosynthesis. By integrating ATAC-seq and RNA-seq, few DARs directly affect incubation behavior by regulating the transcription levels of genes. Five DAR-related DEGs were found to be closely related to maintaining the incubation behavior in geese. Footprinting analysis revealed a set of transcription factors (RFX1, RFX2, RFX3, RFX5, BHLHA15, SIX1, and DUX) which displayed the highest activity at the brooding stage. SREBF2 was predicted to be the unique differentially expressed transcription factor whose mRNA level was down-regulated and enriched in hyper-accessible regions of PRL in the broody stage. In the present study, we comprehensively profiled the transcriptome and chromatin accessibility in the pituitary related to incubation behavior. Our findings provided insight into the identification and analysis of regulatory elements in goose incubation behavior. The epigenetic alterations profiled here can help decipher the epigenetic mechanisms that contribute to the regulation of incubation behavior in birds. [ABSTRACT FROM AUTHOR]

Mortality rates are high for most avian species during early life stages, forming a critical source of natural selection that helps shape the diversity of avian life-history traits. We investigated hatching failure (i.e., non-predatory embryonic mortality, excluding abandoned or damaged eggs) and found significant variation among passerine species. Failure rates ranged from 1.0% to 12.7%, and species with cavity nests and larger clutches experienced greater rates of hatching failure. While past research has focused on the direct effects of predation on the nestling and fledgling stages, little is known about how predation may indirectly influence other sources of mortality such as hatching failure. We investigated the influence of nest predation risk and other factors on variation in hatching failure among 14 free-living grassland and shrubland songbird species. Across all species, 7.7% of 1,667 eggs failed to hatch. We found little evidence that variation in nest temperature influenced rates of hatching failure within and among species, although species with larger clutch sizes had more variable nest temperatures. Dissection of failed eggs revealed that most hatching failures occurred before or shortly after the onset of development; however, there was no difference between cavity and open-cup nesters in the rate of early-stage mortality. Our findings suggest there may be tradeoffs to having a large clutch, with a benefit of rearing more young at the cost of greater hatching failure, possibly due to delayed onset of incubation, poor incubation behavior, or inability to incubate large clutches. Additionally, as larger clutches are often laid in cavity nests, which have a relatively low predation risk, this may outweigh the costs of increased hatching failure. More experimental approaches, such as clutch size manipulations and egg-specific incubation behavior are needed to provide greater insight into factors driving variation in hatching failure across species. [ABSTRACT FROM AUTHOR]

Urbanization is currently considered one of the most rapid types of global environmental change. Urban habitats are biotically and abiotically different from their rural areas, i.e., the ambient temperature, predator, and food availability. These novel challenges create new selection pressures, which allow one to investigate eco-evolutionary responses to contemporary environmental change. A total of 118 breeding nests were monitored for nest predation in both urban and rural areas from 2018 to 2020. We used environmental factors from urban and rural areas and behavioral data from 439 Chinese Blackbird (Turdus mandarinus) valid incubation days to understand the impact of urbanization on the incubation behavior of blackbirds and its adaptation mechanism to the urban environment. Cities have warmer ambient temperatures and lower predation pressures than rural areas. Urban blackbirds chose the incubation strategy with shorter and more bouts, while rural blackbirds selected the incubation strategy with longer and fewer bouts. The plasticity of incubation behavior of urban blackbirds was higher than that of rural areas, and the range of egg temperature was also higher than that of rural areas. In addition, incubation temperature and the number of bouts per day were the key factors affecting the day survival rate of blackbirds, and the hatching rate of urban blackbirds was higher than that of rural blackbirds. Our results provide evidence for behavioral shifts in blackbirds during adaptation to urbanization and support the central role of behavioral adaptation in the successful colonization of new environments by wildlife. These help us understand the behavioral characteristics required for wildlife to live in cities and the urban adaptors faced environmental pressures.

Avian reproductive behavior is regulated through the neuroendocrine system. The transition from laying to brooding is strictly controlled by the hypothalamus-pituitary-gonadal (HPG) axis. Cross talk on the HPG axis relies on the circulatory system, where the dynamics of serum proteins can be observed during different reproductive phases. Some canonical hormones, such as prolactin and luteinizing hormone, play important roles in the transition through reproductive phases. However, little is known at the whole-proteome level. To discover novel serum proteins, we employed isobaric tags for relative and absolute quantification to assay the serum proteome during different reproductive phases in chicken. We identified a total of 1,235 proteins from chicken serum; 239 of these proteins showed differential expression between the laying and brooding stages, including a low concentration of steroid metabolism-related proteins and a high concentration of calcium signaling-related proteins (fold change ≥1.5 or ≤0.66; P

The incubation behavior of geese seriously affects their egg production performance. Studies on incubation behavior have identified functional genes, but the regulatory architecture relationship between functional genes and chromatin accessibility remains poorly understood. Here, we present an integrated analysis of open chromatin profiles and transcriptome to identify the cis-regulatory element and their potential transcription factors involved in regulating incubation behavior in goose pituitary. Assay for transposase-accessible chromatin sequencing (ATAC-seq) revealed that open chromatin regions increased in the pituitary during the transition from incubation behavior to laying. We identified 920 significant differential accessible regions (DARs) in the pituitary. Compared to the laying stage, most DARs had higher chromatin accessibility in the brooding stage. Motif analysis of open DARs showed that the most significant transcription factor (TF) occupied sites predominantly enriched in motifs binding to the RFX family (RFX5, RFX2, and RFX1). While the majority of TF motifs enriched under sites of the nuclear receptor (NR) family (ARE, GRE, and PGR) in closed DARs at the incubation behavior stage. Footprint analysis indicated that the transcription factor RFX family exhibited higher binding on chromatin at the brooding stage. To further elucidate the effect of changes in chromatin accessibility on gene expression levels, a comparison of the transcriptome revealed 279 differentially expressed genes (DEGs). The transcriptome changes were associated with processes of steroid biosynthesis. By integrating ATAC-seq and RNA-seq, few DARs directly affect incubation behavior by regulating the transcription levels of genes. Five DAR-related DEGs were found to be closely related to maintaining the incubation behavior in geese. Footprinting analysis revealed a set of transcription factors (RFX1, RFX2, RFX3, RFX5, BHLHA15, SIX1, and DUX) which displayed the highest activity at the brooding stage. SREBF2 was predicted to be the unique differentially expressed transcription factor whose mRNA level was down-regulated and enriched in hyper-accessible regions of PRL in the broody stage. In the present study, we comprehensively profiled the transcriptome and chromatin accessibility in the pituitary related to incubation behavior. Our findings provided insight into the identification and analysis of regulatory elements in goose incubation behavior. The epigenetic alterations profiled here can help decipher the epigenetic mechanisms that contribute to the regulation of incubation behavior in birds.

Abstract In ground nesting upland birds, reproductive activities contribute to elevated predation risk, so females presumably use multiple strategies to ensure nest success. Identification of drivers reducing predation risk has primarily focused on evaluating vegetative conditions at nest sites, but behavioral decisions manifested through movements during incubation may be additional drivers of nest survival. However, our understanding of how movements during incubation impact nest survival is limited for most ground nesting birds. Using GPS data collected from female Eastern Wild Turkeys (n = 206), we evaluated nest survival as it relates to movement behaviors during incubation, including recess frequency, distance traveled during recesses, and habitat selection during recess movements. We identified 9,361 movements off nests and 6,529 recess events based on approximately 62,065 hr of incubation data, and estimated mean nest attentiveness of 84.0%. The numbers of recesses taken daily were variable across females (range: 1‒7). Nest survival modeling indicated that increased cumulative distance moved during recesses each day was the primary driver of positive daily nest survival. Our results suggest behavioral decisions are influencing trade‐offs between nest survival and adult female survival during incubation to reduce predation risk, specifically through adjustments to distances traveled during recesses.

• Female' incubation behavior may regulate embryo exposure to maternal hormones. • Incubation temperature can affect testosterone metabolization into 5α-dihydrotestosterone or estradiol. • Incubation temperature at 36° C increased concentration of 5α-dihydrotestosterone, but did not affect estradiol titers. • Incubation temperature may constitute an additional pathway for hormone-mediated maternal effects. In birds, maternal hormones deposited into eggs in response to environmental stimuli can impact offspring phenotype. Although less studied, environmental conditions can also influence females' incubation behavior, which might play a role in regulating embryo exposure to maternal hormones through changes in incubation temperature that affect the activity of the enzymes responsible for converting testosterone (T) to 5α-dihydrotestosterone (DHT) or estradiol. Here, we tested the hypothesis that the initial T content of the yolk and incubation temperature determine exposure to T metabolites during early embryo development. In the Japanese quail (Coturnix japonica), we experimentally manipulated yolk T and incubation temperature (38° C versus 36° C) and analyzed DHT and estradiol titers on day four of incubation. We found that eggs with experimentally increased T and those incubated at 36° C showed higher DHT concentration in egg yolk (with no synergistic effect of the two treatments). Estradiol titers were not affected by T manipulation or incubation temperature. Our study suggests that incubation temperature influences DHT titers and may act as an understudied source of maternal influence on offspring phenotype. [ABSTRACT FROM AUTHOR]

• PRL molecules of 4 stages (premature non-laying, laying, trans, and incubation) were identified. • Different isoform composition between pituitary PRL and secreted PRL was identified. • Increasing isoform numbers and ratios to non-posttranslational modified PRL were observed with reproductive stages. • Presence of isoforms which differ pI at least in part originate phosphorylation to the amino acid residue of PRL molecule. Endocrine changes during bird reproduction are well documented. Prolactin (PRL) exhibits a strong relationship between incubation and broody behavior. The molecular forms of PRL in the anterior pituitary gland during the reproductive cycle have already been previously identified but not those in the secreted form. To identify the molecular forms of secreted PRL during the reproductive cycle, we thus monitored the physiological status and incubation behavior of 10 Silkie hens by a video recording system over 1–2 years. Nine out of ten mature hens exhibited incubation behavior multiple times during the experiment. Ten hens demonstrated two interesting features. In a typical clutch, hens spent 10–15 min in the nest to lay an egg. Once they spent over 1 h in the nest, the nest occupancy increased incrementally. This shift in the nest occupancy occurred 7–10 days before the incubation onset and was highly repeatable. Based on the behavior of the hens, we cultured the anterior pituitary gland during four stages (premature non-laying, laying, trans, and incubation) with physiological PRL-releasing factor, vasoactive intestinal peptide (VIP). Based on our two-dimensional protein analysis, glycosylated PRL (G-PRL) displayed several isoforms with varying isoelectric points (pI), whereas we could detect one primary signal for non-glycosylated PRL (NG-PRL). However, 3–4 NG-PRL isoforms were detected in the anterior pituitary gland. These results suggested that secreted PRL, especially from the trans and incubation stages, contains various isoforms and it is post-translationally glycosylated and phosphorylated. [ABSTRACT FROM AUTHOR]

Natural gas compressor stations emit loud, low-frequency noise that travels hundreds of meters into undisturbed habitat. We used experimental playback of natural gas compressor noise to determine whether and how noise influenced settlement decisions and reproductive output as well as when in the nesting cycle birds were most affected by compressor noise. We established 80 nest boxes to attract Eastern Bluebirds (Sialia sialis) and Tree Swallows (Tachycineta bicolor) to locations where they had not previously nested and experimentally introduced shale gas compressor noise to half the boxes while the other 40 boxes served as controls. Our experimental design allowed us to control for the confounding effects of both physical changes to the environment associated with compressor stations as well as site tenacity or the tendency for birds to return to the specific locations where they had previously bred. We incorporated behavioral observations with video cameras placed within boxes to determine how changes in behavior might lead to any noted changes in fitness. Neither species demonstrated a preference for box type (quiet or noisy), and there was no difference in clutch size between box types. In both species, we observed a reduction in incubation time, hatching success, and fledging success (proportion of all eggs that fledged) between quiet and noisy boxes but no difference in provisioning rates. Nest success (probability of fledging at least one young; calculated from all nests that were initiated) was not affected by noise in either species suggesting that noise did not increase rates of either depredation or abandonment but instead negatively impacted fitness through reduced hatching and fledging success. Compressor noise caused behavioral changes that led to reduced reproductive success; for Eastern Bluebirds and Tree Swallows, gas infrastructure can create an equal-preference ecological trap where birds do not distinguish between lower and higher quality territories even when they incur fitness costs. [ABSTRACT FROM AUTHOR]

Avian incubation provides an opportunity to test how parental behavior and ecological conditions interact to shape variation in offspring traits along geographic gradients. In particular, the duration of the incubation period is shorter at higher latitudes, but the degree to which this pattern arises from genetic divergence in rates of growth and development versus from parentally‐mediated variation in egg temperatures is controversial. At higher latitudes parents have higher daytime incubation attentiveness, i.e. they spend a greater proportion of the day on the nest. However, interpreting latitudinal variation in behavior is complicated by latitudinal patterns in ambient temperature and day length. Here, we use 24‐h video recordings to compare the incubation behavior of orange‐crowned warblers Leiothlypis celata in California and Alaska and test how attentiveness varies between populations and as a function of temperature. Birds in Alaska had higher nest attentiveness during the day, despite experiencing similar ambient temperatures. However, when analyzed over 24‐h, the longer nighttime period in California almost entirely canceled out daytime attentiveness differences between populations, and differences in 24‐h attentiveness were small. Our work highlights how incorporating nighttime incubation behavior qualitatively alters latitudinal patterns of attentiveness, and how lower ambient temperatures cannot account for the higher attentiveness in a high latitude population. These populations differ in their incubation period lengths, and differentiating between evolved versus environmentally‐induced variation in offspring growth and development will help understand the fitness consequences of variation in developmental periods. [ABSTRACT FROM AUTHOR]

In ground nesting upland birds, reproductive activities contribute to elevated predation risk, so females presumably use multiple strategies to ensure nest success. Identification of drivers reducing predation risk has primarily focused on evaluating vegetative conditions at nest sites, but behavioral decisions manifested through movements during incubation may be additional drivers of nest survival. However, our understanding of how movements during incubation impact nest survival is limited for most ground nesting birds. Using GPS data collected from female Eastern Wild Turkeys (n = 206), we evaluated nest survival as it relates to movement behaviors during incubation, including recess frequency, distance traveled during recesses, and habitat selection during recess movements. We identified 9,361 movements off nests and 6,529 recess events based on approximately 62,065 hr of incubation data, and estimated mean nest attentiveness of 84.0%. The numbers of recesses taken daily were variable across females (range: 1‒7). Nest survival modeling indicated that increased cumulative distance moved during recesses each day was the primary driver of positive daily nest survival. Our results suggest behavioral decisions are influencing trade‐offs between nest survival and adult female survival during incubation to reduce predation risk, specifically through adjustments to distances traveled during recesses. [ABSTRACT FROM AUTHOR]

The incubation behavior of the Japanese Nagoya chicken breed is a commercial issue because it often causes a sudden and sharp drop in egg production. In this study, whether the incidence of incubation behavior in Nagoya laying hens was associated with calls and the presence of roosters in the same laying house was investigated. Four experiments were conducted using commercial layer-type Nagoya hens where the hatching time of the experimental birds and the treatment order in the presence of males were changed . In Experiment 1, the proportion of incubation behavior in the presence of roosters kept in another pen located between pen-rearing hens (51.3%) was higher than that in their absence (15.9%) or with only rooster calls (23.8%). In Experiments 2, 3, and 4, the proportion of incubation behavior in the presence of roosters (47.3%, 33.3%, and 37.9%, respectively) was higher than that in their absence (33.3%, 17.4%, and 25.6%, respectively). In all experiments, approximately 70% of the incubating hens observed in the absence of roosters exhibited incubation behavior, even in the presence of roosters. Therefore, the presence of roosters may enhance egg incubation behavior in Nagoya laying hens., Competing Interests: Conflicts of Interest: The authors declare no conflict of interest., (2023 Japan Poultry Science Association.)

The molecular mechanism underlying in the onset and maintenance of incubation behavior are not fully understood, and it is still unknown the reason why White Leghorn, a layer strain, hens never display incubation behavior. Therefore, to explore specific hypothalamic genes regulating incubation behavior, cap analysis of gene expression (CAGE) were applied to comparison between incubating Silkie and laying White Leghorn hens. In addition, mRNA expression of some differentially expressed genes (DEGs) and melanocortinergic appetite genes including agouti‐related peptide (AgRP) and pro‐opiomelanocortin (POMC) was also analyzed on Silkie hens under natural anorexia and starvation. The CAGE identified 217 hypothalamic DEGs in incubating Silkie hens, and that of two, transthyretin (TTR) and prolactin‐releasing peptide (PrRP), suggested as appetite gene, were markedly up– and down‐regulated in incubating hens, respectively. In addition, AgRP and POMC expression also increased in incubating bird. mRNA expression of TTR, PrRP, and appetite genes were not differed significantly by starvation, although TTR mRNA expression was relatively high in fasting hens. Consequently, transcriptome by CAGE identified a number of hypothalamic genes differentially expressed by incubation behavior in Silkie hens. Of these, it is suggested that TTR and PrRP may, at least in part, be related to adaptation to natural anorexia in incubating Silkie chickens. [ABSTRACT FROM AUTHOR]

Our study was conducted to examine differences in incubation behavior among breeding sites and the relationship between factor affecting environmental change and incubation behavior of the Blackfaced Spoonbill (Platalea minor). We set up the remote sensor cameras at three breeding sites (Mae-do, Namdongji, Guji-do) to observe incubation behavior in Gyeonggi Bay, South Korea from 2015 to 2018. We analyzed effects of breeding year, day of incubation started, day of incubation, the time of incubation exchanges and sex on incubation bout length. Mean incubation bout length of females (Mae-do: 7.19 ± 0.23 hours, Namdongji: 6.08 ± 0.23 hours, Guji-do: 7.96 ± 0.30 hours) was longer than males (Mae-do: 6.14 ± 0.21 hours, Namdongji: 5.45 ± 0.28 hours, Guji-do: 7.38 ± 0.29 hours). Mean incubation bout length was longer in Guji-do than other study sites. Incubation bout length tended to increase with the clutch initiation date. Males incubated their eggs at day time while female did at night time, these tendencies were observed more clearly in Guji-do. The proportion of time spent incubating of females was higher than males. Males' proportion increased as incubation progressed and increased rate in Guji-do was higher than other study sites. Our results showed that incubation rhythm of the Black-faced Spoonbill differed among breeding sites and varied with the environmental cycle. [ABSTRACT FROM AUTHOR]

The genus Lanio is distributed from southern Mexico to northern Brazil. Until now, nesting information has been limited and scattered among Lanio species. Nests descriptions are available for 3 of the 4 species, and only L. versicolor has nestling descriptions. No information is available on incubation and feeding behavior or incubation and nestling periods for any Lanio species. We present detailed nesting biology information for L. versicolor, such as incubation and nestling behavior, eggs and nest thermal dynamics, and nestling feeding behavior, and also present the first evidence of cooperative breeding for the genus Lanio based on 11 nests monitored in the buffer zone of Manu National Park, Cusco, Peru. Nests were cup-shaped, and all had a leaf located above the nest. The clutch size was 2 eggs, white with maroon or wine-colored spots covering the shell surface. Incubation and nestling periods were 15 and 11 d, respectively, and daytime nest attentiveness was 78%. Nest architecture, clutch size, and nestling period were consistent with previous reports for the genus Lanio. We found similarities in the nesting biology of Lanio spp. with other species of the Tachyphoninae subfamily. Our study significantly increases the information available for the genus Lanio and provides novel and important information of cooperative breeding for a Neotropical mixed-species flock "leader" species. Yet, more studies are needed to fill the large information gap on nesting of tropical birds such as the species in the genus Lanio. [ABSTRACT FROM AUTHOR]

Behavioral adjustments and parental decisions during reproduction can influence the thermal environment at nests, yet our understanding into how environmental factors (i.e., temperature and precipitation) constrain an adult's ability to balance self-maintenance and incubation demands is limited. To expand our understanding of how species respond to environmental factors, we investigated the reproductive ecology of two ground-nesting species (northern bobwhite [ Colinus virginianus ] and scaled quail [ Callipepla squamata ]) in a region (i.e., the Southern Great Plains) prone to thermal variability (i.e., extreme hot and cold temperatures). Specifically, our objective was to examine how temperature and precipitation directly influenced behavioral adjustments (i.e., off-bout duration, frequency, and nest attentiveness) and parental decisions (i.e., nest site selection), and indirectly influenced nest fate. Overall, we found that parents chose to nest in sites that were significantly cooler in temperature than randomly selected sites, and parents further altered the thermal environment experienced by embryos through incubation behavior. Daily precipitation and average ambient temperature and/or their interaction best predicted incubation behaviors, yet each species differed in the timing (i.e., morning vs. evening), frequency, and duration of off-bouts. Furthermore, successful nests were associated with cooler nest site temperatures for bobwhite and warmer nest site temperatures for scaled quail. Our finding of relatively stable (35.5 °C) incubation temperature for developing embryos of both species suggests that ground-nesting birds are able to regulate microclimate through behavioral adjustments and parental decisions even under extreme temperature fluctuations. Nevertheless, the ability for a ground-nesting species to effectively modify behavioral adjustments and decisions may be altered during long periods of enhanced physiological and environmental stress. [ABSTRACT FROM AUTHOR]

There is limited information on the breeding biology of members of the speciose genus Hemitriccus (Aves: Tyrannidae), published nest descriptions exist for only six of the 22 species. The Black-throated Tody-Tyrant (Hemitriccus granadensis) is one of these species that lacks information regarding its nesting biology. We provide the first nest, egg, nestling, and incubation behavior descriptions for the species with notes on its courtship behavior. We found three nests and observed one courtship display at three field sites, two in Colombia and one in Peru. The nest of H. granadensis is an enclosed "purse-like" pendant pouch inside of a moss ball suspended from a small branch with a side entrance and obscured by vegetation. Two nests had a single egg, which averaged 18.1 x 12.9 mm and 1.55 g. Egg appearance varied between nests, the egg at the Colombian site was tan and unmarked, whereas the egg at the Peruvian site was bright white with a few scattered, red specks. We cannot describe the full incubation period or nestling period because of egg infertility and nestling death. However, one nest was discovered with an egg that was incubated for 19 days. Nest attentiveness was 77.5% and average inner nest temperature differed between the Colombian (±SD) (23.79 ± 1.5°C) and Peruvian (17.27 ± 1.54°C) nests. We provide observations and video footage of the courtship display that match previous observations of this species. Overall, the nest shape resembles those of other Hemitriccus species, but clutch size differs from information published for the genus. [ABSTRACT FROM AUTHOR]

For organisms living in seasonal environments, synchronizing the peak energetic demands of reproduction with peak food availability is a key challenge. Understanding the extent to which animals can adjust behavior to optimize reproductive timing, and the cues they use to do this, is essential for predicting how they will respond to future climate change. In birds, the timing of peak energetic demand is largely determined by the timing of clutch initiation; however, considerable alterations can still occur once egg laying has begun. Here, we use a wild population of great tits ( Parus major) to quantify individual variation in different aspects of incubation behavior (onset, duration, and daily intensity) and conduct a comprehensive assessment of the causes and consequences of this variation. Using a 54-year dataset, we demonstrate that timing of hatching relative to peak prey abundance (synchrony) is a better predictor of reproductive success than clutch initiation or clutch completion timing, suggesting adjustments to reproductive timing via incubation are adaptive in this species. Using detailed in-nest temperature recordings, we found that postlaying, birds improved their synchrony with the food peak primarily by varying the onset of incubation, with duration changes playing a lesser role. We then used a sliding time window approach to explore which spring temperature cues best predict variance in each aspect of incubation behavior. Variation in the onset of incubation correlated with mean temperatures just prior to laying; however, incubation duration could not be explained by any of our temperature variables. Daily incubation intensity varied in response to daily maximum temperatures throughout incubation, suggesting female great tits respond to temperature cues even in late stages of incubation. Our results suggest that multiple aspects of the breeding cycle influence the final timing of peak energetic demand. Such adjustments could compensate, in part, for poor initial timing, which has significant fitness impacts. [ABSTRACT FROM AUTHOR]

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Avian mesotocin (MT) is homologous to oxytocin in mammals. Native Thai chickens ( Gallus domesticus ) strongly express maternal behaviors including incubation and rearing. However, the role of MT during incubation behavior has never been studied. The objective of this study was to determine the physiological function(s) of the MTergic system in incubation behavior in native Thai chickens. The brains were collected from incubating (INC) and nest-deprived (ND) hens at different time points (days 3, 6, 8, 10, 14, 18, and 21; n = 6). Immunohistochemistry technique was used to compare the numbers of MT-immunoreactive (-ir) neurons between the INC and ND hens within the Nucleus supraopticus, pars ventralis (SOv), Nucleus preopticus medialis (POM), and Nucleus paraventricularis magnocellularis (PVN). The results revealed that the numbers of MT-ir neurons within the SOv, POM, and PVN remained high during the incubating stage. The number of MT-ir neurons in the SOv was lower than that of the POM and PVN. Disruption of incubation behavior by nest deprivation caused the numbers of MT-ir neurons within the SOv, POM, and PVN to decrease throughout the observation periods. For the first time, this study demonstrates that the MTergic system within the SOv, POM, and PVN may be involved with incubation behavior. In addition, these results further suggest that the MTergic neurons in these nuclei are not only regulated by rearing behavior but also might have a role in the initiation and maintenance of incubation behavior in this tropical species. [ABSTRACT FROM AUTHOR]

The Sooty Antbird ( Hafferia fortis) inhabits Andean foothills and lowland humid forest from Colombia to the western Brazilian Amazon. Despite its wide distribution, many natural history aspects are poorly known. The nest, eggs, and nestlings are described, with no additional nesting information available. Our study provides detailed behavioral information on the nesting biology of H. fortis based on seven nests that were monitored in lowland humid forest in southeastern Peru. Nests were large domes found mostly on slopes or in ravines. The clutch size was always two eggs. Both sexes share daytime incubation and feed the nestlings, but the female exclusively conducted nighttime incubation and brooding. Daytime nest attentiveness was 83 ± 9.5% (SD; n = 13 days), and was similar in both sexes. Nestling period was 13 days, and adults conducted 3.86 feeding trips per day, which increased as nestling developed. Nesting characteristics observed in H. fortis are similar to other Hafferia species, although there is little information about incubation and brooding behavior for the longipes clade, and in general for the Thamnophilidae. Additionally, we described an unusual nest defense behavior during a nocturnal attack by a snake ( Oxyrhopus petola). [ABSTRACT FROM AUTHOR]

Brood parasitism is an uncommon and understudied strategy in Amazonian bird communities, within which only 5 species are known to be brood parasites. We present data on the brood-parasitic behavior of the Pavonine Cuckoo ( Dromococcyx pavoninus) in 3 host species of small-bodied flycatchers in the Peruvian Amazon that construct hanging globular nests with side entrances. During the 7 yr of the study, we encountered 74 nests of these 3 hosts, but parasitism occurred only in 9 nests (12.2%) in 2 yr. Only 1 Pavonine Cuckoo egg was deposited in each host nest ( n = 7), and eggs were markedly dissimilar in size and coloration between hosts and parasite. Incubation investment per day was slightly higher (4%) for 1 parasitized nest than for nonparasitized nests. Overall, 33% of parasitic eggs ( n = 6) hatched; cuckoo nestlings apparently removed host eggs and killed host nestlings. The nestling period lasted 24 days, and the growth-rate constant based on nestling mass ( k = 0.23) was slower for parasite nestlings than for their hosts ( k = 0.27 and 0.31). Food provisioning rates were greater in 1 parasitized nest (2.1 ± 0.7 feedings hr−1 nestling−1) than in nonparasitized nests (1.1 ± 0.4). Nestling cuckoos may further mimic the plumage of their host nestlings. Our results suggest that Pavonine Cuckoos negatively affect their hosts' breeding success and are engaged in a coevolutionary arms race with hosts that have defenses against parasitism. [ABSTRACT FROM AUTHOR]

Urbanization is currently considered one of the most rapid types of global environmental change. Urban habitats are biotically and abiotically different from their rural areas, i.e., the ambient temperature, predator, and food availability. These novel challenges create new selection pressures, which allow one to investigate eco-evolutionary responses to contemporary environmental change. A total of 118 breeding nests were monitored for nest predation in both urban and rural areas from 2018 to 2020. We used environmental factors from urban and rural areas and behavioral data from 439 Chinese Blackbird (Turdus mandarinus) valid incubation days to understand the impact of urbanization on the incubation behavior of blackbirds and its adaptation mechanism to the urban environment. Cities have warmer ambient temperatures and lower predation pressures than rural areas. Urban blackbirds chose the incubation strategy with shorter and more bouts, while rural blackbirds selected the incubation strategy with longer and fewer bouts. The plasticity of incubation behavior of urban blackbirds was higher than that of rural areas, and the range of egg temperature was also higher than that of rural areas. In addition, incubation temperature and the number of bouts per day were the key factors affecting the day survival rate of blackbirds, and the hatching rate of urban blackbirds was higher than that of rural blackbirds. Our results provide evidence for behavioral shifts in blackbirds during adaptation to urbanization and support the central role of behavioral adaptation in the successful colonization of new environments by wildlife. These help us understand the behavioral characteristics required for wildlife to live in cities and the urban adaptors faced environmental pressures. [ABSTRACT FROM AUTHOR]

The genus Chlorospingus is distributed from central Mexico to northern Argentina. Despite its wide distribution, natural history information is sparse. We present the first detailed breeding biology information for C. flavigularis, plus descriptions of the nest, eggs, and other natural history information for C. parvirostris and C. flavopectus based on 26 nests monitored (22 from C. flavigularis, two from C. parvirostris, and two from C. flavopectus) in the buffer zone of Manu National Park, Cusco, Peru. Nests of C. flavigularis were bulky open-cups placed on a variety of substrates (e.g., ferns, moss). The clutch size of C. flavigularis varied between two to three eggs, but clutch size for C. flavopectus and C. parvirostris was consistently two eggs. The eggs of all three species were white with small light-brown speckles. Incubation and nestling period for C. flavigularis was 17 and 14 days, respectively, and daytime nest attentiveness was 60%. Nest architecture, clutch size, and eggs encountered for these species were consistent with previous reports for the genus Chlorospingus. More nesting behavior studies are needed to fill the knowledge gap regarding all species of Chlorospingus to help us understand nesting requirements for this cloud forest genus. [ABSTRACT FROM AUTHOR]

Nest attendance behavior is a critical component of avian ecology that influences nest survival and population productivity. Birds that provide uniparental care during incubation and brood-rearing must balance the benefit of reproductive success with the costs of physiological needs and predation risk. We used miniature nest cameras to record 5904 h of video footage at 33 nests of Greater Prairie-Chickens ( Tympanuchus cupido) during 2010 and 2011 in northcentral Kansas. We quantified the timing and duration of incubation bouts to address alternative hypotheses about physiological requirements and predation risk as drivers of incubation behavior. We also identified nest predators and determined timing of predation events, and tested for effects of nest attendance and monitoring technique on nest survival (video vs. telemetry). Female prairie chickens exhibited high incubation constancy per day (~95 %) and typically took two ~40-min recesses per day: one after sunrise and one before sunset. Mesocarnivores were responsible for 75 % (18 of 24) of nest losses, and most nest predation events occurred during crepuscular or overnight hours. Controlled comparisons provided no evidence that video surveillance attracted predators to nests. Variation in nest attendance had a minimal effect on nest survival compared to height of vegetative cover at the nest site. Timing of recesses did not indicate avoidance of predator activity in our study system. The bimodal pattern of incubation breaks observed in most grouse species is likely driven by physiological requirements of the female rather than predation pressure. Female Greater Prairie-Chickens appear to prioritize their metabolic needs and future reproductive potential over current nest survival. [ABSTRACT FROM AUTHOR]

The Lesser Crested Tern (Thalasseus bengalensis) is a ground-nesting seabird that breeds only on subtropical and tropical islands from northern Africa to northern Australia. Despite their wide distribution, few studies have been conducted on the breeding ecology of this species. On Jana Island, Saudi Arabia, which is northeast of the Saudi Arabia mainland, the midday ground temperature may reach 60 °C during the summer. In this study, two cameras were used to record the incubation behavior of Lesser Crested Terns to evaluate their coping ability in this extremely hot environment. The behavioral mechanism that the seabirds used to maintain optimal egg temperatures was also evaluated. The results show that Lesser Crested Terns attend their eggs continuously during a 24-hr period without leaving the nests, except when a disturbance occurs. This behavior prevents the eggs from reaching lethal temperatures. In addition, position of the sun influenced the incubation behavior. Most incubating Lesser Crested Terns faced west in the morning, and began rotating clockwise until they faced east in the evening. This behavior may play a vital role in preventing both eggs and incubating adults from overheating. Received 16 November 2015, accepted 31 January 2016. [ABSTRACT FROM AUTHOR]

Copyright of Journal of Field Ornithology is the property of Resilience Alliance and its content may not be copied or emailed to multiple sites or posted to a listserv without the copyright holder's express written permission. However, users may print, download, or email articles for individual use. This abstract may be abridged. No warranty is given about the accuracy of the copy. Users should refer to the original published version of the material for the full abstract. (Copyright applies to all Abstracts.)

In birds, the duration of egg incubation (the time from incubation onset to hatching) can affect multiple components of nest success, but what affects incubation duration? Previous studies suggest that incubation duration is affected by both parental behavior and components of the egg, which have yet to be determined. One egg component that may be related to incubation behavior and the time until hatching is eggshell porosity, which affects the exchange of metabolic gasses and water vapor across the shell and, thus, the speed of embryonic development and incubation duration. We tested whether eggshell porosity was associated with the timing of incubation onset by female House Wrens ( Troglodytes aedon Vieillot, 1809), and whether porosity varied within clutches in a manner that might be associated with incubation periods and hatching patterns (i.e., synchronous vs. asynchronous hatching). Eggshell porosity was unrelated to the onset of maternal incubation and did not differ between early- and later-laid eggs within clutches, but differed significantly among females and covaried with egg size. We conclude that producing all eggshells of similar porosity within clutches, while adjusting incubation onset once most or all eggs are laid, provide facultative maternal control over variation in hatching patterns. [ABSTRACT FROM AUTHOR]

The Black Antbird ( Cercomacroides serva) inhabits the east Andean foothills and Amazonian lowlands from Colombia to north Bolivia and western Brazil. It is uncommon throughout its distribution. We provide the first nest and egg description for C. serva with detailed information on bi-parental incubation and nestling feeding behavior. We found four nests at two elevations in the buffer zone of Manu National Park in southeastern Peru. The C. serva nest is a pensile hanging cup, much higher at the back that at the front, made with dark fibers, dry leaves, sticks and fresh moss. The entrance was strongly oblique in three of the four nests. Egg coloration varied between and within the two eggs clutch size. On average, parents made 5.7 ± 2.08 (±SD) foraging trips per day that lasted 39.2 ± 36.17 mins, resulting in 84.6 ± 9.2% nest attentiveness. Both sexes share daytime incubation, but night incubation was exclusively conducted by the female. Nest attentiveness did not differ between males and females, but daytime incubation bouts did vary between the sexes. Afternoon incubation was mainly conducted by the female (77.7%); in contrast, morning incubation was conducted primarily by the male (66.1%). The nestling period was 12-14 days, and the female spent more time feeding and brooding the nestling, a behavior that differs from that of other antbirds. Overall, nesting characteristics of C. serva resemble previous reproductive information published for other Cercomacroides species [ABSTRACT FROM AUTHOR]

Abstract: Hormones regulate many aspects of an individual's phenotype, including various physiological and behavioral traits. Two hormones have been described as important players in the regulation of parental investment in birds: the glucocorticoid hormone corticosterone and prolactin, a pituitary hormone, widely involved in mediating parental behavior. In comparison with corticosterone, the role of prolactin on parental investment remains poorly documented, and most studies so far have been correlative. In this study, the effects of an experimental decrease of prolactin levels on the incubation behavior of a long-lived seabird species were assessed. Male Adélie penguins were treated with self-degradable bromocriptine pellets, inhibiting prolactin secretion. Filming and subsequent video analysis allowed the determination of a behavioral time budget for birds and their position on the nest, while dummy eggs recorded incubation parameters. Incubation duration and breeding success at hatching were also monitored. As expected, bromocriptine-treatment significantly decreased plasma prolactin levels, but did not affect corticosterone levels. The behavioral time budget of penguins was not affected by the treatment. However, treated birds spent significantly more time in an upright position on the nest. These birds also incubated their eggs at lower temperatures and turned their eggs more frequently than controls, resulting in a lengthened incubation period. Despite this, the treatment was insufficient to trigger nest desertion and eggs of treated birds still hatched, indicating that several endocrine signals are required for the induction of nest abandonment. We suggest that the decreased prolactin levels in treated birds offset their timeline of breeding, so that birds displayed behavior typical of early incubation. [Copyright &y& Elsevier]

Copyright of Journal of Field Ornithology is the property of Resilience Alliance and its content may not be copied or emailed to multiple sites or posted to a listserv without the copyright holder's express written permission. However, users may print, download, or email articles for individual use. This abstract may be abridged. No warranty is given about the accuracy of the copy. Users should refer to the original published version of the material for the full abstract. (Copyright applies to all Abstracts.)

We provide the first detailed nesting biology information for the Yellow-breasted Warbling-Antbird (Hypocnemis subflava), a member of the recently split Warbling Antbird complex. We found seven pouch-shaped nests with two eggs or nestlings within Manu National Park, SE Peru. Long incubation sessions (93.7 ± 7.78 minutes, range = 4-480 minutes, n = 18 days) were untaken by both parents and high diurnal nest attentiveness (% of time on the nest) averaged 85.8 ± 1.4% (n = 16) as a result of taking fewer short foraging trips (6.3 ± 0.6 trips/day, range = 3-11; n = 14) that lasted 16.19 ± 1.78 min (range = 1-91; n = 18). Incubation behavior produced an average inner nest temperature of 32.3 ± 0.24 °C (n = 15 days) and average egg temperature of 36.4 ± 0.09 °C (n = 3). Individual nests at different elevations exhibit differences in incubation behavior. The nestling period was 11 days (n = 1) and both parents brooded and fed the nestlings. Nest shape, location, and composition were similar to other species in the complex, but egg coloration was variable among species. [ABSTRACT FROM AUTHOR]

Copyright of Journal of Field Ornithology is the property of Resilience Alliance and its content may not be copied or emailed to multiple sites or posted to a listserv without the copyright holder's express written permission. However, users may print, download, or email articles for individual use. This abstract may be abridged. No warranty is given about the accuracy of the copy. Users should refer to the original published version of the material for the full abstract. (Copyright applies to all Abstracts.)

The authors discuss the study by G. A. Rebstock and P. D. Boersma which explores the variation in the temperature and egg incubation period within clutches of the Magellanic Penguin (Spheniscus magellanicus). Authors of the study include the period before the delayed onset of incubation that leads to confusion. The authors argue that the conclusions of the study are based on an unconventional definition of incubation period.

Abstract: Gonadotropin releasing hormone-I (GnRH-I) is known to regulate the avian reproductive system. We investigated the roles of GnRH-I in the regulation of the reproductive system of the native Thai chicken. The distribution of GnRH-I neurons and changes in GnRH-I-immunoreactive (-ir) neurons throughout the reproductive stages and between incubating and nest-deprived hens were analyzed utilizing immunohistochemical techniques. The results revealed that GnRH-I-ir neurons were distributed in a discrete region lying close to the third ventricle from the level of preoptic area through the anterior hypothalamus, with the greatest abundance found within the nucleus commissurae pallii (nCPa). The number of GnRH-I-ir neurons in the nCPa was highest in laying hens when compared with that in the other reproductive stages. Nest deprivation caused an increase in the number of GnRH-I-ir neurons in the nCPa of nest-deprived hens when compared with incubating hens. These results indicate that GnRH-I expression is correlated with the reproductive state in the native Thai chicken and may be, in part, regulated by it. This study also confirms a pivotal role of GnRH-I in controlling avian reproduction of this non-seasonal breeding, equatorial species. [Copyright &y& Elsevier]

The article discusses a study which examined the natural and human influences that affect incubation patterns and sex role in Black Oystercatcher (Haematopus bachmani) in Alaska. Researchers used 24-hour-per-day video monitoring of breeding pairs to examine incubation patterns. The study found that female and male shorebirds have different but complementary incubation patterns, which facilitates an efficient egg care, and are affected by disturbance and nest predation threats.

Abstract: We investigated the neuroendocrine changes involved in the transition from incubating eggs to brooding of the young in turkeys. Numbers of mesotocin (MT; the avian analog of mammalian oxytocin) immunoreactive (ir) neurons were higher in the nucleus paraventricularis magnocellularis (PVN) and nucleus supraopticus, pars ventralis (SOv) of late stage incubating hens compared to the layers. When incubating and laying hens were presented with poults, all incubating hens displayed brooding behavior. c-fos mRNA expression was found in several brain areas in brooding hens. The majority of c-fos mRNA expression by MT-ir neurons was observed in the PVN and SOv while the majority of c-fos mRNA expression in dopaminergic (DAergic) neurons was observed in the ventral part of the nucleus preopticus medialis (POM). Following intracerebroventricular injection of DA or oxytocin (OT) receptor antagonists, hens incubating eggs were introduced to poults. Over 80% of those injected with vehicle or the D1 DA receptor antagonist brooded poults, while over 80% of those receiving the D2 DA receptor antagonist or the OT receptor antagonist failed to brood the poults. The D2 DA/OT antagonist groups also displayed less c-fos mRNA in the dorsal part of POM and the medial part of the bed nucleus of the stria terminalis (BSTM) areas than did the D1 DA/vehicle groups. These data indicate that numerous brain areas are activated when incubating hens initially transition to poult brooding behavior. They also indicate that DAergic, through its D2 receptor, and MTergic systems may play a role in regulating brooding behaviors in birds. [Copyright &y& Elsevier]

The article presents a study concerning parental behavior controls during incubation stage and asynchrony of hatching in Magellanic Penguins. It states that two eggs were tested whether its temperatures differ and the delayed onset of incubation affects the pattern. Moreover, it notes that parental incubation behavior define why the incubation period of second eggs was shorter that of first eggs.

The article discusses a 2010 study to determine what factors during incubation have an effect on Wood Ducks, Aix sponsa in Latin. The researchers manipulated the incubation of females to find out how nest temperature, female body mass and incubation behavior affected their reproduction. They found no relationship between how long females incubated and how much body mass they lost after, but that the earlier they incubated, the more weight they lost, while the later they incubated, the more mass they maintained after.

Abstract: Hyperprolactinemia is associated with incubation behavior and ovarian regression in birds. To investigate the association of prolactin (PRL), vasoactive intestinal peptide (VIP), and dopamine (DA) with the neuroendocrine regulation of incubation behavior, changes in the number of visible VIP-immunoreactive (VIP-ir) neurons in the nucleus inferioris hypothalami (IH) and nucleus infundibuli hypothalami (IN) and tyrosine hydroxylase-immunoreactive (TH-ir) neurons in the nucleus intramedialis (nI) and nucleus mamillaris lateralis (ML) of incubating native Thai hens were compared with those of nest-deprived hens. TH was used as a marker for dopaminergic (DAergic) neurons. Blood samples were collected to determine PRL levels. The localization and the number of visible VIP-ir and TH-ir neurons were determined by immunohistochemistry. Disruption of incubation behavior was accompanied by a precipitous decline in plasma PRL levels. The number of visible VIP-ir neurons in the IH–IN and TH-ir neurons in the nI and ML were high during incubation and decreased when hens were deprived of their nests. This study indicated an association between VIP neurons in the IH–IN and DA neurons in the nI and ML with the degree of hyperprolactinemia, suggesting that the expression of incubation behavior in birds might be, in part, regulated by the DAergic input from the nI and ML to VIP neurons in the IH–IN and subsequent PRL release. [Copyright &y& Elsevier]