Blood lead increases and haemoglobin decreases in urban birds along a soil contamination gradient in a mining city.

Lead contaminated soil is a persistent global threat to the health of animal populations. Nevertheless, links between soil lead and its adverse effects on exposed wildlife remain poorly understood. Here, we explore local geographic patterns of exposure in urban birds along a gradient of lead contamination in Broken Hill, an Australian mining city. Soil lead concentrations are linked to co-located blood lead measurements in rock pigeons (Columba livia), house sparrows (Passer domesticus), crested pigeons (Ocyphaps lophotes) and white-plumed honeyeaters (Lichenostomus ornatus). Median blood lead levels were highest in crested pigeons (59.6 µg/dL), followed by house sparrows (35.2 µg/dL), rock pigeons (35.1 µg/dL), and white-plumed honeyeaters (27.4 µg/dL). Blood lead levels in all species declined away from mining areas, the primary source of lead contamination in Broken Hill. Blood lead increased significantly and at the greatest rate relative to soil lead in the three ground foraging species (crested pigeons, house sparrows, rock pigeons). For these species, soil lead concentrations below 200 mg/kg and 900 mg/kg were needed to maintain a median blood lead concentration under the lower threshold of the subtoxic (20-50 µg/dL) and toxic (≥50 µg/dL) effect ranges previously identified for some bird species. We also investigated the effects of lead exposure on blood haemoglobin levels as a general measure of physiological condition in birds exposed to different levels of soil lead contamination. Overall, for every 1 µg/dL increase in blood lead, haemoglobin decreased by 0.11 g/L. The rate of this decrease was not significantly different between species, which supports the measurement of haemoglobin as a consistent though insensitive measure of physiological condition in chronically lead exposed birds. Our findings reflect the importance of lead contaminated soil as a widespread source of elevated blood lead and supressed haemoglobin levels in birds inhabiting urbanised and mining impacted environments.

PLASMA PROTEIN ELECTROPHORESIS IN THE WHITE STORK (CICONIA CICONIA): AGREEMENT BETWEEN AGAROSE GEL VERSUS CAPILLARY ZONE METHODS AND DEVELOPMENT OF REFERENCE INTERVALS.

The white stork (Ciconia ciconia) is a ciconiiform species widely represented in zoological institutions. Plasma protein electrophoresis is widely used in avian patients for assessment of inflammatory conditions, but reference intervals for this testing modality are lacking for the white stork. The two main electrophoretic methods are agarose gel electrophoresis (AGE) and capillary zone electrophoresis (CZE). This study assessed fresh plasma samples of healthy adult white storks (n = 30). Statistical analyses were performed to evaluate agreement between AGE and CZE. Typical electrophoretic fractions were obtained from both methods (prealbumin, albumin, α1, α2, ß, γ1, and γ2). The AGE and CZE methods were not equivalent for determining major electrophoretic fractions (except ß-globulins) and albumin:globulin ratio on plasma samples. An additional prealbumin fraction was seen with CZE. Reference intervals were established for each method as the smallest n group was 27 individuals for a given value; most values had normal distribution, and robust or parametric methods were used on the data.

Revisiting the question of nucleated versus enucleated erythrocytes in birds and mammals.

Erythrocyte enucleation is thought to have evolved in mammals to support their energetic cost of high metabolic activities. However, birds face similar selection pressure yet possess nucleated erythrocytes. Current hypotheses on the mammalian erythrocyte enucleation claim that the absence of cell organelles allows erythrocytes to 1) pack more hemoglobin into the cells to increase oxygen carrying capacity and 2) decrease erythrocyte size for increased surface area-to-volume ratio, and improved ability to traverse small capillaries. In this article, we first empirically tested current hypotheses using both conventional and phylogenetically informed analysis comparing literature values of mean cell hemoglobin concentration (MCHC) and mean cell volume (MCV) between 181 avian and 194 mammalian species. We found no difference in MCHC levels between birds and mammals using both conventional and phylogenetically corrected analysis. MCV was higher in birds than mammals according to conventional analysis, but the difference was lost when we controlled for phylogeny. These results suggested that avian and mammalian erythrocytes may employ different strategies to solve a common problem. To further investigate existing hypotheses or develop new hypothesis, we need to understand the functions of various organelles in avian erythrocytes. Consequently, we covered potential physiological functions of various cell organelles in avian erythrocytes based on current knowledge, while making explicit comparisons with their mammalian counterparts. Finally, we proposed by taking an integrative and comparative approach, using tools from molecular biology to evolutionary biology, would allow us to better understand the fundamental physiological functions of various components of avian and mammalian erythrocytes.

Blood parasites of bird communities in Sri Lanka and their mosquito vectors.

Avian blood parasites have been shown to have significant health effects on avifauna worldwide. Sri Lanka, a tropical island rich with resident and migratory birds, has not been properly evaluated for avian blood parasites or their vectors. We investigated the presence of avian haemoparasites in Sri Lankan birds and the potential mosquito vectors of those pathogens. Blood samples were collected from local/migratory birds captured by standard mist nets from Anawilundawa bird sanctuary, Hanthana mountain range, and the University of Peradeniya park. Mosquitoes were collected from Halgolla forest reserve and the forest patches in Kurunegala and Gampola areas in addition to the above mist-netting localities. Part of the mitochondrial cytochrome b (cytb) gene was amplified and sequenced to detect the presence of haemoparasites from avian blood samples (86) and mosquito samples (480). Blood parasites of the two genera, i.e., Haemoproteus (4 species; Haemoproteus sp. 1-4) and Plasmodium (5 species; Plasmodium sp. 1-5) were identified from seven bird species (four resident and three migratory). Among these, three bird species (Red-vented bulbul (3/16), Asian Brown flycatcher (1/1), and India pitta (1/1)) were positive for Plasmodium spp., while four (Yellow-browed bulbul (1/4), oriental white-eye (1/4), brown-headed Barbet (1/4), and Indian blue robin (1/1)) were positive for Haemoproteus spp. Two mosquito species were also positive for Plasmodium (3) and Haemoproteus (1) species. Phylogenetic analysis and haplotype networks created using positive sequences of haemoparasites showed that a Plasmodium clade was shared by Cx nigropunctatus mosquitoes and the migratory bird, Indian pitta. The majority (85%) of the Plasmodium and Haemoproteus sequences of this study were not linked to the well-characterized species suggesting the distinct nature of the lineages. Associations between mosquito species and blood parasites of birds suggest the possible vector status of these mosquitoes.

HEALTH STATUS OF NAZCA BOOBIES (SULA GRANTI) ON DAPHNE MAJOR ISLAND IN THE GALÁPAGOS DETERMINED BY HEMATOLOGY, BIOCHEMISTRY, AND PHYSICAL EXAMINATION.

Island species are particularly vulnerable to environmental disturbances and introduced pathogens. Conducting health assessments of wild populations in the Galápagos improves the ability of wildlife managers and veterinarians to detect deteriorations in health status. Seabirds in particular are useful species to monitor due to their colonial breeding and wide migration range. Nazca boobies (Sula granti) in a breeding colony at Daphne Major (n = 30) were given physical examinations, and blood samples were collected for hematology and biochemistry using an iSTAT Portable Clinical Analyzer. Female boobies had longer wing length than males, as well as lower blood urea nitrogen, creatinine, and white blood cell counts. This could be attributed to sexual dimorphism or differences in foraging and mating strategies between the sexes. The time between capture and blood collection had a significant inverse relationship on plasma sodium, potassium, hemoglobin, anion gaps, and lymphocyte counts, suggesting that blood sampling in Nazca boobies should be done in less than 5 m to avoid the impacts of stress on hematological parameters. This is the first health assessment on the breeding colony of Nazca boobies at Daphne Major, and the results can inform future monitoring in this species as well as other sulids.

CLINICAL CHEMISTRY, VITAMIN, ELECTROPHORESIS, AND HEMATOLOGIC ANALYTES OF BLACK-HEADED IBIS (THRESKIORNIS MELANOCEPHALUS).

The evaluation of laboratory blood analytes is an important tool for health assessment in avian medicine. Unfortunately, there is a lack of suitable reference values for many zoo bird species. The goal of the present study was to establish reference intervals for a wide range of blood analytes in healthy black-headed ibis (Threskiornis melanocephalus) kept in a zoologic garden. Lithium heparinized blood samples from 15 individuals were collected, and 18 different clinical chemistry analytes were measured. New are especially the minimum and maximum values for bile acid (2.38-49.4 µmol/L), cholinesterase (439-1260 U/L), vitamin A (0.65-1.70 mg/L), vitamin E (26.3-52.5 mg/L), and capillary zone electrophoresis (prealbumin, 10.2-23.4%; albumin, 35.9-44.1%; A/G ratio, 1.07-1.69; α-, 15.7-20.0%; ß-, 13.5-19.1%; γ-globulin, 6.08-11.3%). The measured values for clinical chemistry, vitamin, electrophoresis, and hematologic analytes are a basis for further studies and for diagnostics and clinical treatment in this ibis species.

Veterinary utility of dried blood spots for detailed analysis of chlorinated pesticides and polychlorinated biphenyls by gas chromatography tandem mass spectrometry.

Persistent organic pollutants (POPs) are organic compounds of anthropogenic origin that resist atmospheric and microbial degradation and thus persist in the environment and in food chains for exceptionally long periods of time. Veterinarians and wildlife researchers need simple methodologies for monitoring and measuring such compounds including two large and diverse categories, organochlorine pesticides (OCs) and polychlorinated biphenyls (PCBs), compounds that have been largely banned from production and use except for specific exceptions. We present development of methodologies for detection and quantitation of 22 OCs and 10 PCB congeners by tandem quadrupole gas chromatography-mass spectrometric analysis of Dried Blood Spots (DBS). Development was enabled by (1) optimization of suspension and extraction methodologies for DBS; (2) strategic streamlining and condensation of Multiple Reaction Monitoring (MRM) settings on GC/MS/MS; and (3) improvement of GC settings to accommodate all 32 compounds in a single chromatographic run per sample. The method was validated for parameters of linearity, limits of detection and quantitation, recovery and precision, and results from blood were shown to correlate well with those from DBS despite both being only 50 uL in volume. The method was applied successfully to blood samples from nine avian specimens submitted to the MSU Veterinary Diagnostic Lab, and all were shown to bear the burden of varying levels of OCs and/or PCB compounds.

PLASMA BIOCHEMISTRY AND PROTEIN ELECTROPHORESIS REFERENCE INTERVALS OF THE COMMON LOON (GAVIA IMMER).

There are no published plasma biochemistry reference intervals for any species within the order Gaviiformes, which includes the common loon (Gavia immer). Because of their unique classification and lack of close taxonomic relatives, species-specific values for clinical data in loons are needed. This study determined reference intervals for plasma biochemical values in adult common loons, and reference intervals for protein electrophoresis values in both adult and juvenile common loons. Healthy, wild adult (n = 148, age >3 yr) and juvenile (n = 31, age 4-12 wk) common loons were sampled on freshwater summer breeding territories at study sites across North America. Plasma biochemical analytes included glucose (Glu), total calcium, phosphorus, sodium, potassium, chloride, blood urea nitrogen (BUN), uric acid, cholesterol, triglycerides, creatine kinase, γ-glutamyl transferase, alkaline phosphatase (ALP), lactate dehydrogenase, aspartate aminotransferase, amylase, and bile acids. Protein electrophoresis data included albumin to globulin ratio (A: G), prealbumin, albumin, α1-globulin, α2-globulin, ß-globulin, and γ-globulin. Adult females had significantly higher Glu, ALP, and BUN than adult males. Juvenile loons had higher ß-globulins than adults, whereas adults had higher α1-globulins. Establishment of complete reference intervals will improve clinical assessment of captive loons, and allow researchers to better understand the health of wild loons in response to the multiple environmental stressors faced by these species.

HEALTH STATUS AND BASELINE HEMATOLOGY, BIOCHEMISTRY, AND BLOOD GAS VALUES OF GALAPAGOS SHEARWATERS (PUFFINUS SUBALARIS).

The Galápagos shearwater, Puffinus subalaris, is a seabird endemic to the Galápagos archipelago. Hematology, blood chemistry, and general health parameters have not been published for this species. Analyses were run on blood samples drawn from 20 clinically healthy Galápagos shearwaters captured by hand at their nests at Islote Pitt on San Cristóbal Island in July 2016. A portable blood analyzer (iSTAT) was used to obtain near immediate field results for pH, pO2, pCO2, TCO2, HCO3 -, hematocrit, hemoglobin, sodium, potassium, chloride, ionized calcium, creatinine, urea nitrogen, anion gap, and glucose. Blood lactate was measured using a portable Lactate Plus analyzer. The reported results provide baseline data that can be used for comparisons among populations and in detecting changes in health status among Galápagos shearwaters.

Organ Histopathology and Hematological Changes Associated With Heat Exposure in Australian Desert Birds.

An inability of the body to appropriately respond to extreme temperatures will result in pathological changes to vital organs and adverse hematological changes. Mild heat exposure of a bird to a temperature above the zone of thermoneutrality can induce subclinical heat stress, which may be a precursor to illness. The ability to identify subtle changes that may be associated with subclinical heat stress can be important in early diagnosis and treatment of heat stress in birds. Pathological changes to internal body organs, post-heat exposure, were microscopically examined in 13 budgerigars (Melopsittacus undulatus), 15 zebra finches (Taeniopygia guttata), and 8 diamond doves (Geopelia cuneata) as model species for the bird orders Psittaciformes, Passeriformes, and Columbiformes, respectively. There was mild to moderate congestion of the lungs of 28/36 birds examined, including all of the budgerigars and diamond doves. In 8/15 zebra finches no significant lung congestion was noted. Interstitial and pulmonary hemorrhage was in observed in one diamond dove. The most common hepatic pathologic change identified was micro- and macro-vesicular hepatocellular vacuolation (4/15 zebra finches, 5/13 budgerigars, and 8/8 diamond doves). There was mild to moderate congestion in the kidneys of 1/15 zebra finch, 2/ 13 budgerigars, and 4/8 diamond doves, as well as in the gastrointestinal tract of 1/15 zebra finch and 7/8 budgerigars. Budgerigars showed a decrease in hematocrit and a significant change in the numbers of heterophils and lymphocytes following heat exposure. The basophil population of cells remained relatively stable in both budgerigars and diamond doves. These findings indicate avian species differences in body organ and hematological changes following exposure to similar elevations in environmental temperatures.

Prognostic Indicators of Avian Survival.

The objective of this study was to evaluate whether blood glucose, packed cell volume, and total protein are effective diagnostic tests to predict short-term (48 h following admittance to a veterinary hospital) prognosis for wild birds with traumatic injuries. The cohort study conducted for this investigation took place at the Jivdaya Charitable Trust in Ahmedabad, India, over a 7-d period, at the time of the International Kite Flying Festival of Uttarayan. A small blood sample was collected from every avian patient prior to surgical treatment and immediately analyzed. Patient outcome was reviewed and recorded at the end of a 48-h period following admittance to the veterinary hospital. Statistical analysis revealed no significant correlations between blood glucose levels (P = .24), packed cell volume (P = .80), or total protein levels (P = .47) outside the reference intervals and short-term outcome of the patients. There is a lack of evidence regarding the use of these diagnostic tests to ascertain short-term prognosis in avian species. While research has been conducted on avian wound healing, studies fail to identify any correlation between duration of injury and patient outcome. Despite a lack of statistically significant results from this study, the findings should not be dismissed and could be used as a basis for future studies on this subject.

Metal and metalloid levels in blood of semipalmated sandpipers (Calidris pusilla) from Brazil, Suriname, and Delaware Bay: Sentinels of exposure to themselves, their prey, and predators that eat them.

Identifying animals as sentinels for humans and other animal species is an excellent method for understanding exposure to environmental contamination at different times and places. Shorebirds are useful sentinels because they have a world-wide distribution, eat a range of prey, and are eaten by a range of other species, including humans. We collected blood from semipalmated sandpipers (Calidris pusilla) wintering in northern (Suriname N = 71) and northeastern (Brazil N = 61) South America to examine levels of heavy metals and metalloids (arsenic, selenium), and compare them to blood levels in sandpipers at a heavily used stopover site in New Jersey (N = 30; Delaware Bay, NJ). Since blood represents relatively recent exposure, it can provide information on where and when the birds were exposed. Levels were highest in Brazil for arsenic and particularly selenium; highest in Suriname for cadmium and lead; and highest in New Jersey for chromium. Samples from Brazil and Suriname presented higher levels of mercury than did those from New Jersey. There were no geographic differences for cobalt. Levels of all metals were generally within an order of magnitude. The significant geographic difference for selenium was interesting because it is regulated in the body. Selenium levels in the NJ sample were directly proportional to levels found in their principle food at this migration stopover site (eggs of horseshoe crab, Limulus polyphemus). Mean selenium level was almost an order of magnitude higher in the semipalmated sandpiper blood samples from Brazil (mean of 27,500 µg/L= ppb) compared to the other sampling locations (mean > 5330 µg/L). This is a toxic level and cause for concern and further investigation, alerting us to look for other evidence of excess selenium exposure. Otherwise the levels of other metals are generally not high enough to cause harm to the sandpipers themselves or to predators that eat them. We discuss the implications for these birds and their exposure to contaminants at different stopover sites.

White blood cell count in birds: evaluation of a commercially available method.

BACKGROUND: To conduct a hematological analysis of avian blood samples, standard automated cell counting is unreliable because all avian blood cells are nucleated. Therefore, quantitative white blood cell counting in birds is still performed manually, whereby the Natt-Herrick method is widely used in veterinary laboratories. The aim of this study was to evaluate a new commercially available single test system for avian white blood cell counting, the Natt-Herricks-Tic®, which would allow easy in-house analysis by clinicians or technicians. A total of 40 avian ethylenediaminetetraacetic acid (EDTA) blood samples from 24 different species were included in the study. To assess method agreement, each blood sample was analyzed for total white blood cell count with the test method and the Natt-Herrick reference method. To determine the imprecision of the reference method and the Natt-Herricks-Tic® method, the noncorrected white blood cell count was determined ten consecutive times from one avian EDTA blood sample for each method. RESULTS: The Natt-Herricks-Tic® method performed well concerning staining quality and countability of the granulocytes by the hemocytometer. In the agreement study, the Natt-Herricks-Tic® method showed a small proportional systematic error with a small positive mean bias of 282 white blood cells/µL but had wide 95% limits of agreement (- 4683 cells/µL to 5227 cells/µL), indicating random error. The precision study resulted in a coefficient of variation of 16% for the Natt-Herricks-Tic® method (the mean ± standard deviation: 9.7 ×  103/µL ± 1.5 × 103/µL) and 23% (the mean ± standard deviation: 7.9 × 103/µL ± 1.8 × 103/µL) for the reference method. CONCLUSIONS: The Natt-Herricks-Tic® method showed acceptable precision for a manual method and demonstrated good agreement with the reference method. It can be recommended as a reliable and suitable method for determining white blood cell counts in avian EDTA blood if nonstatistical quality control measures are used in the daily routine. The application of individual reference intervals for the interpretation of white blood cell counts in birds may improve the diagnostic performance of this important analyte in a clinical setting.

Detecting total immunoglobulins in diverse animal species with a novel split enzymatic assay.

BACKGROUND: Total immunolobulin G concentration is a useful, albeit underutilized, diagnostic parameter for health assessments of non-domestic animal species, due to a lack of functional diagnostic tools. Traditional assays, including enzyme-linked immunosorbent assay or radial immunodiffusion, require development of specific reagents (e.g., polyclonal antisera and appropriate protocols) for each animal species, precluding wide and easy adoption in wildlife welfare. As an alternative, bacterial virulence factors able to bind IgGs in antigen-independent manner can be used. To further simplify the diagnostic procedure and increase the number of species recognized by an assay, in this study a recently developed Split Trehalase immunoglobulin assay (STIGA) with bIBPs as a sensing elements was used to detect antibodies in 29 species from 9 orders. Three bacterial immunoglobulin binding proteins (protein G, protein A and protein L) were incorporated into STIGA reagents to increase the number of species recognized. RESULTS: IgG concentrations were detected through glucose production and produced signals were categorized in 4 categories, from not active to strong signal. Activation was detected in almost all tested animal species, apart from birds. Incorporation of Protein G, Protein A and Protein L allowed detection of IgGs in 62, 15.5 and 6.9% of species with a strong signal, respectively. Assays combining 2 bacterial immunoglobulin binding proteins as sensing element generally gave poorer performance than assays with the same bacterial immunoglobulin binding proteins fused to both trehalase fragments. CONCLUSIONS: STIGA assays have potential to be further developed into an easily adoptable diagnostic test for total amount of IgGs in almost any serum sample, independent of species.

Population pharmacokinetics of itraconazole solution after a single oral administration in captive lesser flamingos (Phoeniconaias minor).

Aspergillosis is an infectious, non-contagious fungal disease of clinical importance in flamingo collections. Itraconazole is an antifungal drug commonly used in the treatment and prophylaxis of avian aspergillosis. Studies have shown that dosage regimes in birds vary based on different itraconazole presentation and administration methods. This investigation used a population pharmacokinetic approach to study itraconazole in lesser flamingos. Itraconazole was administered orally at 10 mg/kg to 17 flamingos. A sparse blood sampling was performed on the subjects, and samples were collected at 1, 2, 3, 5, 8, 12, 16, 21, and 24 hr post-drug administration. Twelve flamingos were sampled three times, three birds bled twice and two sampled once. Itraconazole in plasma was quantified using high-pressure liquid chromatography (HPLC). A one-compartment pharmacokinetic model with first order absorption was fitted to the data using nonlinear mixed effects modeling (NLME) to determine values for population parameters. We identified a long half-life (T½) of more than 75 hr and a maximum plasma concentration (CMAX ) of 1.69 µg/ml, which is above the minimal inhibitory concentrations for different aspergillus isolates. We concluded that plasma drug concentrations of itraconazole were maintained in a population of flamingos above 0.5 ug/ml for at least 24 hr after a single oral dose of 10 mg/kg of itraconazole solution.

HEMATOLOGY, BIOCHEMISTRY, AND METAL CONTAMINANT LEVELS IN WILD-CAUGHT CLAPPER RAIL (RALLUS CREPITANS) IN NORTHEASTERN FLORIDA.

The clapper rail (Rallus crepitans) is native to salt marshes along the eastern United States. Populations are likely stable, but may be at risk due to the degradation of wetland habitat by contaminants. Contaminants can cause adverse effects in birds such as alteration of immune and reproductive function, and previous studies have used this species as a sentinel for estuarine health. Blood samples were collected from clapper rails in Florida and hematology counts, plasma biochemistry panels, and metal assessments using inductively coupled plasma-mass spectrometry were performed. Biochemical and hematology data were too limited to determine if contaminants were adversely affecting clapper rails in this study, but cadmium, lead, and zinc were increased for several birds. Although contaminant levels were not consistently elevated for all birds, additional research is needed to assess if clapper rails in this region are at risk of contaminant exposure due to increasing urbanization and development pressures.

PAIRED BIOCHEMICAL ANALYSIS OF PIGMENTED PLASMA SAMPLES FROM ZOO-KEPT AMERICAN FLAMINGOS (PHOENICOPTERUS RUBER) USING A POINT-OF-CARE AND A STANDARD WET CHEMISTRY ANALYZER.

American flamingos (Phoenicopterus ruber) are commonly kept in zoological collections, making health monitoring essential. Use of point-of-care (POC) blood analyzers that require small volumes of whole blood samples produces prompt results allowing for rapid clinical decision-making. To evaluate and compare blood biochemistry analysis results analyzed by a POC biochemistry analyzer and a laboratory wet biochemistry analyzer, blood was collected from 17 apparently healthy zoo-kept American flamingos. Analyzer agreement was investigated using the Passing-Bablock regression analysis and Spearman correlation coefficients. Plasma samples from all birds were bright yellow in color. The results from the POC analyzer used in this study were found to be outside acceptance and clinical allowable error limits when compared with the laboratory analyzer for phosphorus (Phos), total protein (TP), albumin (Alb), glucose (Glu), creatine kinase (CK), and potassium (K). For aspartate aminotransferase (AST), results were within clinical allowable error but outside the acceptance limits, and for calcium (Ca) and sodium (Na), results were within both limits. The POC analyzer failed to measure the uric acid (UA) concentrations of all the samples, and reported all bile acids (BA) concentrations as below its minimal measurable limit. The use of analyzer-specific reference intervals is recommended for most analytes tested. The POC analyzer used in this study cannot be recommended for measuring UA concentrations in brightly colored samples from American flamingos.

Not one hormone or another: Aggression differentially affects progesterone and testosterone in a South American ovenbird.

Behaviors such as territorial interactions among individuals can modulate vertebrate physiology and vice versa. Testosterone has been pointed out as a key hormone that can be rapidly affected by aggressive interactions. However, experimental evidence for such a link is mixed. In addition, behaviors can elicit changes in multiple hormones, which in turn have the potential to synergistically feedback to behavior. For example testosterone and progesterone can act interdependently in modulating male behavior. However, if aggression can affect progesterone levels in males remain unknown and - to the best of our knowledge - no one has yet tackled if and how aggressive behavior simultaneously affects testosterone and progesterone in free-living animals. We addressed these questions by performing simulated territorial intrusion experiments measuring both hormones and their ratio in male rufous horneros (Aves, Furnarius rufus) during the mating and parental care periods. Aggression affected testosterone and progesterone differentially depending on the period of testing: challenged birds had higher levels of progesterone during the mating period and lower levels of testosterone during parental care compared to controls. Challenged individuals had similar progesterone to testosterone ratios during both periods and these ratios were higher than those of control birds. In summary, territorial aggression triggered hormonal pathways differentially depending on the stage of the breeding cycle, but equally altered their ratio independent of it. Our results indicate that multiple related hormones could be playing a role rather than each hormone alone in response to social interactions.

Carotenoid-based skin ornaments reflect foraging propensity in a seabird, Sula leucogaster.

Carotenoid-based ornaments are common signalling features in animals. It has long been proposed that such ornaments communicate information about foraging abilities to potential mates. However, evidence linking foraging with ornamentation is largely missing from unmanipulated, free-ranging populations. To investigate this relationship, we studied a coastal population of brown booby (Sula leucogaster brewsteri), a seabird with a carotenoid-based gular skin ornament. δ13C values from both feathers and blood plasma were negatively correlated with male gular colour, indicating birds that consumed more pelagic prey in offshore locations had more ornamented skin than those that fed on nearshore, benthic prey. This relationship was supported by our GPS tracking results, which revealed longer, more offshore foraging trips among highly ornamented males. Our data show that brown booby ornaments are honest indicators of foraging propensity; a link consistent with the rarity hypothesis and potentially driven by the concentration of carotenoids found in phytoplankton versus benthic algae. Carotenoid-based ornaments may reflect foraging tendencies in animals such as coastal predators that use food webs with distinct carotenoid profiles.

Haematospirillum and insect Wolbachia DNA in avian blood.

In this study, blood samples of 259 Acrocephalus sp. warblers were molecularly analysed for Anaplasmataceae and Rhodospirillaceae based on PCR amplification of 16S rRNA gene fragments. One bird blood sample (from Reed Warbler, Acrocephalus scirpaceus) yielded a sequence with 99.8% identity to Haematospirillum jordaniae. This is the first molecular evidence for the occurrence of this species in the blood of any vertebrate other than human. Another bird blood sample (from Marsh Warbler: Acrocephalus palustris) yielded a Wolbachia sequence, closely related to a moth endosymbiont with 99.8% identity. A nematode origin of Wolbachia DNA detected here in avian blood can be excluded, because results of phylogenetic analysis showed its closest alignment with insect wolbachiae. This is the first finding of insect Wolbachia DNA in the circulatory system of birds, which can be explained either by the inoculation of wolbachiae by blood-sucking vectors, or passing of Wolbachia DNA from the gut into the blood of this insectivorous bird species.