Transcriptome analysis of pigeon pituitary gland: expression changes of genes encoding protein and peptide hormones at different breeding stages.

Unlike other poultry, parent pigeons produce "pigeon milk" in their crops to nurture their squabs, which is mainly controlled by prolactin (PRL). Exception for PRL, the pituitary gland may also release various other peptide and protein hormones. However, whether these hormones change during pigeon crop lactation and their potential physiological functions remain unclear. Here, to identify potential peptide or protein hormone genes that regulate crop lactation, we conducted transcriptome analysis of pigeon pituitary glands at 3 different breeding stages (the ceased stage-nonincubation and non-nurturing stage, the 11th d of the incubation, and the 1st d of the nurturing stage) using RNA sequencing (RNA-Seq). Our analysis identified a total of 15,191 mRNAs and screened out 297 differentially expressed genes (DEG), including PRL, VIP, etc. The expression abundance of PRL mRNA on the 1st d of the nurturing stage was respectively 4.93 and 3.62 folds higher when compared to the ceased stage and the 11th d of the incubation stage. Additionally, the expression abundance of VIP is higher in the 1st d of the nurturing stage than in the ceased stage. Protein-protein interaction (PPI) network and Molecular Complex Detection (MCODE) analysis identified several vital DEGs (e.g., GHRHR, VIP, etc.), being closely linked with hormone and enriched in neuropeptide signaling pathway and response to the hormone. Expression pattern analysis revealed that these DEGs exhibited 4 distinct expression patterns (profile 10, 16, 18, 19). Genes in profile 10 and 19 presented a trend with the highest expression level on 1st d of the nurturing stage, and functional enrichment analysis indicated that these genes are involved in neuropeptide hormone activity, receptor-ligand activity, and the extracellular matrix, etc. Taken together, being consistent with PRL, some genes encoding peptide and protein hormones (e.g., VIP) presented differentially expressed in different breeding stages. It suggests that these hormones may be involved in regulation of the crop lactation process or corresponding behavior in domestic pigeons. The results of this study help to gain new insights into the role of pituitary gland in regulating pigeon lactation.

Prolactin promotes crop epithelial proliferation of domestic pigeons (Columba livia) through the Hippo signaling pathway.

The purpose of this study was to investigate whether prolactin (PRL) regulates the proliferation of pigeon crop epithelium through the Hippo signaling pathway during the breeding cycle. Twenty-four pairs of adult pigeons were allotted to four groups by different breeding stages, and their crops and serum were sampled. Eighteen pairs of young pigeons were selected and divided into three groups for the injection experiments. The results showed that the serum PRL content and crop epithelial thickness of pigeons increased significantly at day 17 of incubation (I17) and day 1 of chick-rearing (R1). In males, the mRNA levels of yes-associated transcriptional regulator (YAP) and snail family transcriptional repressor 2 (SNAI2) were peaked at I17, and the gene levels of large tumor suppressor kinase 1 (LATS1), serine/threonine kinase 3 (STK3), TEA domain transcription factor 3 (TEAD3), connective tissue growth factor (CTGF), MYC proto-oncogene (c-Myc) and SRY-box transcription factor 2 (SOX2) reached the maximum value at R1. In females, the gene expression of YAP, STK3, TEAD3, and SOX2 reached the greatest level at I17, the expression profile of SAV1, CTGF, and c-Myc were maximized at R1. In males, the protein levels of LATS1 and YAP were maximized at R1 and the CTGF expression was upregulated at I17. In females, LATS1, YAP, and CTGF reached a maximum value at I17, and the expression level of phosphorylated YAP was minimized at I17 in males and females. Subcutaneous injection of prolactin (injected for 6 d, 10 µg per kg body weight every day) on the left crop of pigeons can promote the proliferation of crop epithelium by increasing the CTGF level and reducing the phosphorylation level of YAP. YAP-TEAD inhibitor verteporfin (injection for 6 d, 2.5 mg per kg body weight every day) can inhibit the proliferation of crop epithelium induced by prolactin by inhibiting YAP and CTGF expression. In conclusion, PRL can participate in crop cell proliferation of pigeons by promoting the expression of YAP and CTGF in Hippo pathway.

This study evaluated whether prolactin (PRL) regulates the proliferation of pigeon crops through Hippo signaling pathway during the breeding cycle. Twenty-four pairs of adult pigeons were allotted to four groups by different breeding stages, and their crops and serum were sampled. Eighteen pairs of young pigeons were selected and divided into three groups for the injection experiments. The crop epithelial thickness and serum PRL content of pigeons increased significantly at day 17 of incubation (I17) and day 1 of chick-rearing (R1). In males and females, the mRNA and protein levels of yes-associated transcriptional regulator (YAP) reached the maximum value at R1 and I17, respectively, and phosphorylation level of YAP were minimized at I17. Subcutaneous injection of prolactin on pigeon crops can promote the proliferation of crop epithelium by increasing the connective tissue growth factor (CTGF) level and reducing the phosphorylation level of YAP. YAP-TEAD inhibitor verteporfin can inhibit the proliferation of crop epithelium induced by prolactin by inhibiting YAP and CTGF expression. In conclusion, PRL can participate in crop cell proliferation of pigeons by promoting the expression of YAP and CTGF in Hippo pathway.

Linoleic acid ameliorates intestinal mucosal barrier injury in early weaned pigeon squabs (Columba livia).

The study aimed to investigate whether linoleic acid could improve the intestinal barrier function of squabs under weaning stress conditions. Totally 320 7-d-old weaned squabs were randomly divided into four treatment groups, including control group (CON), 0.7% linoleic acid addition group (LA007), 1.4% linoleic acid addition group (LA014) and 2.1% linoleic acid addition group (LA021). At 21 d, eight squabs were randomly selected from each treatment group for sampling and determination. The results showed that adding linoleic acid could improve (P < 0.05) the body weight of weaned squabs, and LA014 had the best effect. With the increase of linoleic acid dosage, villi height and villi area increased linearly or quadratically (P < 0.05), and reached the maximum in LA021 or LA014, respectively. The linoleic acid supplementation could improve the intestinal tight junction of weaned squabs, and the LA014 was the most significant (P < 0.05). With the linoleic acid increasing, the levels of intestinal IL-6 and TNF-α decreased linearly (P < 0.05), while intestinal IL-10 increased quadratically (P < 0.05) and reached the maximum in LA014. Serum endotoxin and diamine oxidase levels decreased linearly (P < 0.05) and reached the lowest level in LA014. The ultrastructure of villi revealed that the length of ileal microvilli in LA014 was significantly increased (P < 0.05) and the microvilli became dense, and the mitochondria in epithelial cells returned to normal state. Further exploring the mechanism of linoleic acid alleviating intestinal injury caused by weaning stress in squabs, it was found that linoleic acid down-regulated (P < 0.05) the relative protein expression of TLR4, MyD88, phosphorylated JNK, and phosphorylated p38, reducing secretion of pro-inflammatory factors IL-6 and TNF-α. This study indicated that linoleic acid could alleviate intestinal barrier injury of early weaned squabs by down-regulating TLR4-MyD88-JNK/p38-IL6/TNF-α pathway.

Artificial feeding of early weaned squabs can reduce the burden of breeding pigeons and shorten the breeding cycle. However, similar to early weaned mammals, early weaned squabs would also inevitably undergo severe physiological and psychological stress responses in the early stage. The growth performance and immunity of early weaned squabs were inferior to those of the parent feeding squabs. Previous studies suggest that linoleic acid played an important role in the growth and development of squabs. Therefore, the study aimed to investigate whether linoleic acid could improve the intestinal barrier function of squabs under weaning stress conditions. Totally 320 7-d-old weaned squabs were randomly divided into four treatment groups, including control group and linoleic acid addition groups with three different doses. At 21 d, eight squabs were randomly selected from each treatment group for sampling and determination. The results indicated that under weaning stress conditions, linoleic acid could weaken the inflammatory response, and alleviate the intestinal epithelial barrier damage of weaned squabs, specifically by promoting the development of intestinal villi, strengthening the tight junction, reducing intestinal permeability, and promoting the secretion of anti-inflammatory factors.

Effects of a FCBP gene polymorphism, location, and sex on Young's modulus of the tenth primary feather in racing pigeons.

Young's modulus (E) is a measure for stiffness of a material and a higher E means a higher stiffness. The respective polymorphism of the feather corneous beta-protein gene causes the replacement of glycine by cysteine. We looked for possible effects of the three FCBP genotypes on E in the 10th primaries of racing pigeons. However, we did not find a statistically significant difference of E between the genotypes, even within the sexes and/or within different locations under our test conditions. Our findings do not preclude the possibility that under other conditions (temperature, moisture) an influence of the glycine/cysteine polymorphism on E may exist. Compared to the more proximal locations of the rachis (base and middle) we observed lower values for E in the distal region (tip). The 10th primary constitutes the leading edge of the pigeon wing and this special function may require higher stiffness in the proximal parts of the shaft. We observed significantly higher values of E in females than in males, which result only from statistically significantly higher values in the middle region. The higher stiffness of female primaries may also contribute to the better results of hens compared to cocks in pigeon races.

Uncovering the Sex-Specific Endocrine Responses to Reproduction and Parental Care.

Hormones mediate physiological and behavioral changes in adults as they transition into reproduction. In this study, we characterize the circulating levels of five key hormones involved in reproduction in rock doves (Columba livia): corticosterone, progesterone, estradiol, testosterone, and prolactin using univariate and multivariate approaches. We show similar patterns as previous studies in the overall patterns in circulating levels of these hormones, i.e., testosterone (males) and estradiol (females) high during nest-building or egg-laying, prolactin increasing at mid-incubation and peaking at hatching (both sexes), and elevated corticosterone levels in later incubation and early nestling development. In our investigation of hormone co-variation, we find a strong correlation between prolactin and corticosterone across sampling stages and similarities in earlier (early to mid-incubation) compared to later (late incubation to nestling d9) sampling stages in males and females. Finally, we utilized experimental manipulations to simulate nest loss or altered caregiving lengths to test whether external cues, internal timing, or a combination of these factors contributed most to hormone variation. Following nest loss, we found that both males and females responded to the external cue. Males generally responded quickly following nest loss by increasing circulating testosterone, but this response was muted when nest loss occurred early in reproduction. Similar treatment type, e.g., removal of eggs, clustered similarly in hormone space. These results suggest internal drivers limited male response early in reproduction to nest loss. In contrast, circulating levels of these hormones in females either did not change or decreased following nest manipulation suggesting responsiveness to external drivers, but unlike males, this result suggests that reproductive processes were decreasing.

Quantum magnetic imaging of iron organelles within the pigeon cochlea.

The ability of pigeons to sense geomagnetic fields has been conclusively established despite a notable lack of determination of the underlying biophysical mechanisms. Quasi-spherical iron organelles previously termed "cuticulosomes" in the cochlea of pigeons have potential relevance to magnetoreception due to their location and iron composition; however, data regarding the magnetic susceptibility of these structures are currently limited. Here quantum magnetic imaging techniques are applied to characterize the magnetic properties of individual iron cuticulosomes in situ. The stray magnetic fields emanating from cuticulosomes are mapped and compared to a detailed analytical model to provide an estimate of the magnetic susceptibility of the individual particles. The images reveal the presence of superparamagnetic and ferrimagnetic domains within individual cuticulosomes and magnetic susceptibilities within the range 0.029 to 0.22. These results provide insights into the elusive physiological roles of cuticulosomes. The susceptibilities measured are not consistent with a torque-based model of magnetoreception, placing iron storage and stereocilia stabilization as the two leading putative cuticulosome functions. This work establishes quantum magnetic imaging as an important tool to complement the existing array of techniques used to screen for potential magnetic particle-based magnetoreceptor candidates.

The mediation of pigeon egg production by regulating the steroid hormone biosynthesis of pigeon ovarian granulosa cells.

The aim of this study was to determine the molecular mechanism of miR-205b targeting 11ß-hydroxysteroid dehydrogenase type 1 (HSD11B1) on the apoptosis and proliferation of granulosa cells (GC) of pigeons. Our previous studies suggested that HSD11B1 was the target gene of miR-205b and played a key role in steroid hormone biosynthesis and GC development. The adenovirus-miR-205b recombinant virus and adenovirus-cli-miR-205b-sh recombinant virus were generated, verified, and their characteristics determined. The recombinant viruses were used to infect the GC of pigeons, with real time quantitative PCR used to examine the expressions of HSD11B1 and related genes. The HSD11B1 antibody was obtained and verified, and Western blotting was used to detect the protein level of HSD11B1. The Cell Counting Kit-8 assay kit was used to detect cell viability, and the Annexin V-FITC/PI kit was used for the apoptosis assays. The expression of HSD11B1 was significantly lower in the overexpression (OE) than in OE negative control (OE-NC) treatments and significantly higher in short hairpin (SH) than in SH negative control (SH-NC) treatments. The expression levels of cytochrome P4503A5 was significantly higher in SH and lower in OE treatments, and the rhythms of cytochrome P450 aromatase mRNA levels were similar. The mRNA level of cytochrome P450scc in OE was lower than in OE-NC treatments and higher in SH than in SH-NC treatments. The protein expressions of HSD11B1 were decreased in the GC of OE, whereas increased in the SH group. The Cell Counting Kit-8 assay revealed that overexpression of miR-205b significantly suppressed proliferation of the GC of pigeons, whereas interference of miR-205b significantly induced the proliferation of the GC. The overexpression and the interference of miR-205b did not have a significant effect on cell cycle. The overexpression of miR-205b significantly increased the number of apoptotic cells, whereas the interference of miR-205b decreased the number of apoptotic cells. These findings indicated that miR-205b mediated pigeon egg production by regulating the steroid hormone biosynthesis of the pigeon ovarian GC by targeting HSD11B1, which may be useful in increasing pigeon egg production.

c-Fos revealed lower hippocampal participation in older homing pigeons when challenged with a spatial memory task.

Homing pigeons experience age-related spatial-cognitive decline similar to that seen in mammals. In contrast to mammals, however, previous studies have shown the hippocampal formation (HF) of old, cognitively impaired pigeons to be greater in volume and neuron number compared with young pigeons. As a partial explanation of the cognitive decline in older birds, it was hypothesized that older pigeons have reduced HF activation during spatial learning. The present study compared HF activation (via the activity-dependent expression of the immediate early gene c-Fos) between younger and older pigeons during learning of a spatial, delayed nonmatch-to-sample task. On the last day of training, c-Fos activation significantly correlated with behavioral performance in the young, but not old, pigeons suggesting more HF engagement by the young pigeons in solving the task. The behavioral correlation was additionally associated with consistently higher, but insignificant c-Fos activation across practically every HF subdivision in the young compared with the old pigeons. In sum, the results of the present study are consistent with the hypothesis that age-related decline in the spatial cognitive ability of homing pigeons is in part a result of an older HF being less responsive to the processing of spatial information. However, alternative interpretations of the data are discussed.

Intraocular Pressure Measurement by Rebound Tonometry (TonoVet) in Normal Pigeons (Columba livia).

We evaluated the applicability of a rebound tonometer (TonoVet) in pigeon eyes and established normal reference intraocular pressure (IOP) values in healthy pigeons; 20 eyes of euthanized pigeons were used for calibration of the TonoVet and 48 eyes of 24 adult pigeons were used for measurement of reference IOP. First, IOP of pigeon eyes ex vivo were measured using the 'd' and the 'p' modes of the TonoVet and compared to manometric IOP values from 5 to 80 mm Hg. Then, to establish normal reference values, IOP was measured from clinically normal pigeons in vivo. The 'd' and the 'p' modes of the TonoVet showed a strong linear correlation with the manometric IOP (R2 = .996 and .991, respectively). The obtained regression formulas were: y1 = 0.439x + 2.059 and y2 = 0.330x - 0.673, respectively (y1, 'd' mode of TonoVet; y2, 'p' mode of TonoVet; x, manometric IOP). The 'd' and the 'p' modes consistently measured one-half and one-third of the actual IOP, respectively. Therefore, the formula obtained through the 'd' mode was applied to obtain reference values. The calibrated IOP of normal pigeon eyes was 19.5 ± 4.4 mm Hg. The actual IOP could be calculated using the presented formula. Considering the limitations of the 'p' mode, use of the 'd' mode is more appropriate. Therefore, the TonoVet rebound tonometry under the 'd' mode is a reliable method for measuring IOP in pigeons.

3D functional ultrasound imaging of pigeons.

Recent advances in ultrasound Doppler imaging have facilitated the technique of functional ultrasound (fUS) which enables visualization of brain-activity due to neurovascular coupling. As of yet, this technique has been applied to rodents as well as to human subjects during awake craniotomy surgery and human newborns. Here we demonstrate the first successful fUS studies on awake pigeons subjected to auditory and visual stimulation. To allow successful fUS on pigeons we improved the temporal resolution of fUS up to 20,000 frames per second with real-time visualization and continuous recording. We show that this gain in temporal resolution significantly increases the sensitivity for detecting small fluctuations in cerebral blood flow and volume which may reflect increased local neural activity. Through this increased sensitivity we were able to capture the elaborate 3D neural activity pattern evoked by a complex stimulation pattern, such as a moving light source. By pushing the limits of fUS further, we have reaffirmed the enormous potential of this technique as a new standard in functional brain imaging with the capacity to unravel unknown, stimulus related hemodynamics with excellent spatiotemporal resolution with a wide field of view.

LDHA gene is associated with pigeon survivability during racing competitions.

Pigeon racing is a popular sport worldwide. Pigeons are under continuous selection to improve speed, spatial orientation, and endurance during long flights. However, numerous genetic and non-genetic factors affect survivability and homing ability, making such traits difficult for breeders to control. Polymorphisms in the lactate dehydrogenase A gene (LDHA) likely affects pigeon racing and homing abilities, due to its role in physical and mental performance. Additionally, the adenylate cyclase activating polypeptide 1 gene (ADCYAP1) has been associated with physiological and behavioral shifts that occur during avian migration. In this study, we examined the association between LDHA and ADCYAP1 genotypes with pigeon survivability during racing competitions. Survivability was evaluated through the estimated breeding value (EBV) of each individual's total race distances during its athletic life. ADCYAP1 was not polymorphic among our samples, while LDHA genotypes were significantly associated with deviated EBV values of longer total race distance; individuals carrying the S+ genotype had higher EBV (i.e., greater survivability). Thus, the LDHA locus might be useful for marker-assisted selection, empowering breeders and trainers to maximize pigeon quality. Moreover, data obtained from breeding will also improve our understanding of the genetic mechanism underlying navigation and flight for wild migrating bird species.

Changes in hormone profiles, growth factors, and mRNA expression of the related receptors in crop tissue, relative organ weight, and serum biochemical parameters in the domestic pigeon (Columba livia) during incubation and chick-rearing periods under artificial farming conditions.

The present study was conducted to determine the changes in concentrations of hormones and growth factors and their related receptor gene expressions in crop tissue, relative organ weight, and serum biochemical parameters in male and female pigeons during incubation and chick-rearing periods under artificial farming conditions. Seventy-eight pairs of 60-week-old White King pigeons with 2 fertile eggs per pair were randomly divided into 13 groups by different breeding stages. Serum prolactin and insulin-like growth factor-1 (IGF-1) concentrations in crop tissue homogenates were the highest in both male and female pigeons at 1 d of chick-rearing (R1), while epidermal growth factor (EGF) in female pigeons peaked at d 17 of incubation (I17) (P < 0.05). mRNA expression of the prolactin and EGF receptors in the crop tissue increased at the end of incubation and the early chick-rearing stage in both sexes. However, estrogen, progesterone, and growth hormone receptor expression each decreased during the early chick-rearing stage (P < 0.05). In male pigeons, IGF-1 receptor gene expression reached its peak at R7, while in female pigeons, it increased at the end of incubation. The relative weight of breast and abdominal fat in both sexes and thighs in the males was lowest at R7, and then gradually increased to the incubation period level. Serum total protein, albumin, and globulin concentrations increased to the highest levels at I17 (P < 0.05). Total cholesterol, triglyceride, and low-density lipoprotein reached their highest values at I17 in male pigeons and R25 in female pigeons (P < 0.05). In conclusion, hormones, growth factors, and their receptors potentially underlie pigeon crop tissue development. Changes in organs and serum biochemical profiles suggested their different breeding-cycle patterns with sexual effects.

Sex-biased transcriptomic response of the reproductive axis to stress.

Stress is a well-known cause of reproductive dysfunction in many species, including birds, rodents, and humans, though males and females may respond differently. A powerful way to investigate how stress affects reproduction is by examining its effects on a biological system essential for regulating reproduction, the hypothalamic-pituitary-gonadal (HPG) axis. Often this is done by observing how a stressor affects the amount of glucocorticoids, such as cortisol or corticosterone, circulating in the blood and their relationship with a handful of known HPG-producing reproductive hormones, like testosterone and estradiol. Until now, we have lacked a full understanding of how stress affects all genomic activity of the HPG axis and how this might differ between the sexes. We leveraged a highly replicated and sex-balanced experimental approach to test how male and female rock doves (Columba livia) respond to restraint stress at the level of their transcriptome. Females exhibit increased genomic responsiveness to stress at all levels of their HPG axis as compared to males, and these responsive genes are mostly unique to females. Reasons for this may be due to fluctuations in the female endocrine environment over the reproductive cycle and/or their evolutionary history, including parental investment and the potential for maternal effects. Direct links between genome to phenome cause and effect cannot be ascertained at this stage; however, the data we report provide a vital genomic foundation on which sex-specific reproductive dysfunction and adaptation in the face of stress can be further experimentally studied, as well as novel gene targets for genetic intervention and therapy investigations.

Homing pigeons externally exposed to Deepwater Horizon crude oil change flight performance and behavior.

The Deepwater Horizon oil spill was the largest in U.S. history, contaminating thousands of miles of coastal habitat and affecting the lives of many avian species. The Gulf of Mexico is a critical bird migration route area and migrants that were oiled but did not suffer mortality as a direct result of the spill faced unpredictable fates. This study utilized homing pigeons as a surrogate species for migratory birds to investigate the effects a single low level external oiling event has on the flight performance and behavior of birds flying repeated 161 km flights. Data from GPS data loggers showed that lightly oiled pigeons changed their flight paths, increased their flight durations by 2.6 fold, increased their flight distances by 28 km and subsequently decreased their route efficiencies. Oiled birds also exhibited reduced rate of weight gain between flights. Our data suggest that contaminated birds surviving the oil spill may have experienced flight impairment and reduced refueling abilities, likely reducing overall migration speed. Our findings contribute new information on how oil spills affect avian species, as the effects of oil on the flight behavior of long distance free-flying birds have not been previously described.

Effect of selenium supplementation on pigeon reproductive performance, selenium concentration and antioxidant status.

The effects of dietary supplementation of sodium selenite (SS) on the reproductive performance and the concentration of selenium, glutathione peroxidase (GSH-Px), superoxide dismutase (SOD), and malondialdehyde (MDA) were determined, and expression of glutathione peroxidase 4 (GPx4) and bone morphogenic protein 15 (BMP15) was evaluated. Paired pigeons (n = 864) were fed: T1 received no SS, while T2, T3, and T4 received 0.5, 1.0, and 1.5 mg of SS/kg of dry matter (DM), respectively. Treatments were performed in triplicate with 72 pairs in each replicate. The results showed that selenium supplementation significantly affected pigeon reproductive performance. Birds fed 1.0 mg of SS/kg displayed higher egg production (P > 0.05), higher birth rate, and lower dead sperm rate than the control group (P < 0.05). Selenium and biochemical analyses revealed a higher selenium concentration in the 1.5 mg of SS/kg group than in the control group (P < 0.05), while GSH-Px was higher in the 0.5 mg of SS/kg group than in the control group (P < 0.05). Neither the MDA nor the SOD content were affected significantly in liver, chest muscle, or leg muscle (P > 0.05); however, in plasma, MDA was lower in the control group (P < 0.05), while SOD was higher in the control group (P < 0.05). qRT-PCR results revealed up-regulation of GPx4 in hypothalamus, pituitary and testis tissues in supplemented groups (P < 0.05). However, expression in ovary differed; GPx4 mRNA levels were lower in the 1.5 mg of SS/kg and control groups than in the 1.0 or 0.5 mg of SS/kg groups (P < 0.05). Expression of BMP15 in the hypothalamus, pituitary, and testis tissues was unaffected (P > 0.05), while in ovary, BMP15 was down-regulated in the 1.5 mg of SS/kg group (P < 0.05). These results suggest pigeons supplemented with SS up-regulated GPx4, 1.0 mg of SS/kg exhibited superior reproductive performance, while 1.5 mg of SS/kg increased the selenium concentration, and 0.5 mg of SS/kg up-regulated GSH-Px activity.

Low level exposure to crude oil impacts avian flight performance: The Deepwater Horizon oil spill effect on migratory birds.

In 2010, the Deepwater Horizon oil spill released 134 million gallons of crude oil into the Gulf of Mexico making it the largest oil spill in US history. The three month oil spill left tens of thousands of birds dead; however, the fate of tens of thousands of other migratory birds that were affected but did not immediately die is unknown. We used the homing pigeon as a surrogate species for migratory birds to investigate the effects of a single external oiling event on the flight performance of birds. Data from GPS data loggers revealed that lightly oiled pigeons took significantly longer to return home and spent more time stopped en route than unoiled birds. This suggests that migratory birds affected by the oil spill could have experienced long term flight impairment and delayed arrival to breeding, wintering, or crucial stopover sites and subsequently suffered reductions in survival and reproductive success.

Body mass change in flying homing pigeons externally exposed to Deepwater Horizon crude oil.

The Deepwater Horizon oil spill contaminated thousands of miles of habitat valuable to hundreds of species of migratory and resident birds of the Gulf of Mexico. Many birds died as a direct result of the oil spill; however, the indirect effects of oil exposure on the flight ability and body condition of birds are difficult to assess in situ. This study utilizes the homing pigeon as a surrogate species for migratory birds to investigate the effect of multiple external oil exposures on the flight performance and body mass change of birds over a series of repeated flights from 136.8km flight distance. Oiled pigeons took significantly longer to return home, lost more weight during flight, and were unable to recover their weight, resulting in reduction of body weight overtime. Based on our data, migratory birds that were oiled, even partially, by the Deepwater Horizon oil spill likely took longer to complete migration and were likely in poor body condition, increasing their risk of mortality and reproductive failure.

The effect of endoscopic sterilization on reproductive behavior and pair bond maintenance of feral pigeons (Columba livia).

OBJECTIVE: Problems related to feral pigeons (Columba livia) in cities mainly result from their large numbers due to uncontrolled population growth. The aim of this study was to evaluate whether endoscopic guided sterilization affects the reproductive behavior of feral pigeons under experimental conditions, with the intention of assessing this technique as a potential method for feral pigeon population control. MATERIALS AND METHODS: Five groups of four pairs of feral pigeons each were studied from 8 weeks before, to 7 weeks after sterilization. Both the male and female of the first pair of each group were sterilized, in the second pair only the female and in the third pair only the male was sterilized. The fourth pair acted as a control. All eggs laid were candled to assess fertility. RESULTS: Surgical sterilization had minimal effects on behavior and therefore seems not to have impact on possible field application for population control. All pairs maintained their pair bonds and continued to defend their nesting sites against other pigeons. Only one female copulated with a foreign fertile male while her primary partner was debilitated due to surgery, but returned to him as soon as he recovered. All eggs laid more than 5 days after male sterilization were infertile, whereas all control pairs had fertile eggs. Only one fertile clutch was produced, 5 days after the male's sterilization. Therefore it is assumed that males remain fertile for a limited period of time. CONCLUSION: Endoscopic sterilization seems to be a promising method for field control of feral pigeon populations and sterilization of the male only seems sufficient.

BLOOD GAS, LACTATE, AND HEMATOLOGY EFFECTS OF VENIPUNCTURE TIMING AND LOCATION AFTER MIST-NET CAPTURE OF MOURNING DOVES (ZENAIDA MACROURA), BOAT-TAILED GRACKLES (QUISCALUS MAJOR), AND HOUSE SPARROWS (PASSER DOMESTICUS).

Venous blood gas partial pressures, pH, bicarbonate and lactate concentrations, packed cell volume, white blood cell differential counts, and heterophil/lymphocyte ratios were measured from Mourning Doves (Zenaida macroura), Boat-tailed Grackles (Quiscalus major), and House Sparrows (Passer domesticus). Birds were bled promptly after mist-net capture and banding or following a targeted delay of 45-60 min, in order to assess the impacts of a brief holding period commonly practiced in large-scale bird banding operations. Additionally, effects of venipuncture location (basilic [=ulnar] vein versus jugular vein) were evaluated in male Boat-tailed Grackles sampled promptly after capture and banding. All comparisons were with unpaired samples; no birds were subjected to more than one venipuncture. All three species exhibited moderate improvements in blood gas and acid-base status after the delay, with reductions in lactate concentrations with or without concurrent increases in pH and bicarbonate. Boat-tailed Grackles exhibited an increased proportion of heterophils in the differential white blood cell count following a delay in sampling, suggestive of a stress leukogram. There were no significant differences between basilic and jugular venipuncture results from male Boat-tailed Grackles. Most metabolic, respiratory, and acid-base alterations were minor, but a small number of birds exhibited values (e.g., temperature-corrected pH <7.3, lactate >10 mmol/L) that could be of concern if combined with other adverse conditions. For such birds, a short delay between capture and processing could benefit their blood gas and acid-base status, although loss of time foraging or feeding young and greater activation of the hypophyseal-pituitary-adrenal axis are additional considerations.