Antibiotic treatment increases yellowness of carotenoid feather coloration in male greenfinches (Chloris chloris).

Carotenoid plumage coloration is an important sexually selected trait in many bird species. However, the mechanisms ensuring the honesty of signals based on carotenoid pigments remain unclear. It has recently been suggested that intestinal integrity, which is affected by gut parasites and microbiota and influences nutrient absorption and acquisition, mediates the relationship between carotenoid ornamentation and individual quality. Here, we test whether carotenoid plumage coloration in greenfinches (Chloris chloris) is affected by the treatment of an antibiotic or an antiparasitic drug. We captured wild greenfinches (N = 71) and administered anticoccidial medication toltrazuril (TOLTRA) to one group, antibiotic metronidazole (METRO) to the second group to target trichomonosis, and the third group received no medication. In the METRO group, feathers grown during the experiment had significantly higher chroma of yellow parts, but there was no effect of TOLTRA on feather chroma. The results suggest that METRO increased the efficiency of carotenoid modification or deposition to the feathers rather than nutrient acquisition and/or freed energy resources that could be invested in coloration. Alternatively, though not measured, METRO might have affected microbial community and host physiology as microbial metabolites can modulate mitochondrial and immune function.

Dietary resistant potato starch improves growth performance and feather development in Pekin ducks fed a low phosphorus diet.

This study investigated whether dietary resistant potato starch (RPS) inclusion could ameliorate the negative impact of a low nonphytate phosphorus (nPP) diet on growth performance, feather growth, feather follicles (FF) development, and carcass traits by improving nutrient utilization and cecal microbiome fermentation capacity in Pekin ducks. The experiment was performed with a 2 × 2 randomized block design with 2 levels of RPS (0 or 12%) and 2 levels of nPP (low or normal, low: 0.22% at 1-14 d and 0.18% at 15-35 d of age; normal: 0.40% at 1-14 d and 0.35% at 15-35 d of age) for a total of 4 treatments, each with 8 replicate pens per treatment of 12 birds per pen. As regards growth performance and carcass traits, RPS inclusion markedly increased (P < 0.05) BW of 14 and 35 d, BWG and FI of 1-14 d, 15-35 d, and 1-35 d as well as abdominal fat and breast meat percentage of 35 d in ducks fed low nPP diets; moreover, RSP inclusion significantly reduced (P < 0.05) mortality in ducks fed low nPP diets. As regards feather growth and follicles development of 35 d, RPS inclusion significantly increased (P < 0.05) the fourth primary feather length, absolute feather weight, and the density of primary FF in the back skin in ducks fed low nPP diets. In regard to nutrition utilization, RPS supplementation significantly increased (P < 0.05) the availability of DM, CP, and energy, as well as dietary AME at 35 d of age in ducks fed low nPP diets. However, RPS supplementation had no effect (P > 0.05) on the concentration of cecal short-chain fatty acids and the activities of cecal phytase and cellulase in ducks fed low nPP diets. These results indicate that RPS can improve nutrient availability to ameliorate the negative effects on performance and feather development caused by a low nPP diet in Pekin ducks.

Estimated intestinal absorption of phosphorus and its deposition in chosen tissues, bones and feathers of chickens receiving chromium picolinate or chromium nanoparticles in diet.

The aim of the study was to determine whether the level and form of Cr in the diet of chickens influences its accumulation in tissues as well as intestinal absorption of P and its deposition in tissues. The experiment was carried out on 405 one-day-old male Ross 308 chickens that were randomly divided into five treatment groups. Control group was fed the diet without supplemental chromium; experimental groups were fed the diet with 3 or 6 mg/kg chromium picolinate (Cr-Pic) and with 3 or 6 mg/kg chromium nanoparticles (Cr-NP). Chromium was found to accumulate in the tissues of the ileum, liver, breast muscle, bones skin and in feathers of chickens. Chromium deposited in the ileum of chickens does not affect the ex vivo estimated intestinal absorption of P. The use of Cr in the diet of chickens carries the risk of lowering P levels in femur.

Local and systemic inflammatory responses to lipopolysaccharide in broilers: new insights using a two-window approach.

The inflammatory response involves a complex interplay of local tissue activities designed to recruit leukocytes and proteins from the blood to the infected tissue. For egg-type chickens, we established the growing feather (GF) as an accessible tissue test site to monitor tissue responses to injected test-material. For commercial broilers, whose health depends to a large extent on innate immune system functions, the GF test system offers an important novel window to directly assess their natural defenses. This study was conducted to adapt the GF test system for use in broilers, and use it to simultaneously examine local (GF) and systemic (blood) inflammatory responses initiated by GF pulp injection of lipopolysaccharide (LPS). Specifically, GF of 12 male and 12 female, 5-week-old broilers were injected with LPS (16 GF/chicken; 1 µg LPS/GF). Blood and GF were collected at 0 (before), 6, and 24 h after GF injection. GF pulp was used to determine leukocyte-infiltration and gene-expression profiles, reactive-oxygen-species generation, and superoxide dismutase (SOD) activity. Blood was used to determine blood cell profiles and SOD activity. A time effect (P ≤ 0.05) was observed for most aspects examined. In GF, LPS injection resulted in heterophil and monocyte infiltration reaching maximal levels at 6 and 24 h, respectively. Reactive-oxygen-species generation, SOD activity, and mRNA levels of IL-1ß, IL-8, IL-6, IL-10, and cathelicidin B1 were elevated, whereas those of TNF-α, LITAF, SOD1, and SOD2 decreased after LPS injection. In blood, levels of heterophils and monocytes were elevated at 6 h, lymphocytes and RBC decreased at 6 h, and thrombocytes and SOD activity increased at 24 h. Assessment of LPS-induced activities at the site of inflammation (GF) provided novel and more relevant insights into temporal, qualitative, and quantitative aspects of inflammatory responses than blood. Knowledge generated from this dual-window approach may find direct application in identification of individuals with robust, balanced innate defenses and provide a platform for studying the effects of exogenous treatments (e.g., nutrients, probiotics, immunomodulators, etc.) on inflammatory responses taking place in a complex tissue.

Alteration of dietary cysteine affects activities of genes of the transsulfuration and glutathione pathways, and development of skin tissues and feather follicles in chickens.

The dietary requirement for cysteine is not determined in poultry since it is not an essential amino acid. The cysteine need is expected to be met through the transsulfuration pathway where homocysteine, a precursor of methionine, is converted to cysteine. Cysteine is a major component of plumage, and the degree to which cysteine is involved in plumage and other keratized proteins are unknown. We randomly assigned chicks to control and treatment (deficient in cysteine) diets for 49 d. The thickness of the skin layers, feather follicle length, and thickness were measured at days 10, 24, 34, and 49. We also measured the hepatic mRNA expressions of cystathionine beta synthase (CBS), cystathionine γ-lyase (CTL), cysteine dioxygenase (CDO), and glutathione synthetase (GSS). Chickens fed the treatment diet had reduced epidermis thickness and shorter feather follicles compared with the controls. The chicken fed the treatment diet also had increased mRNA expression of CBS and CTL indicating a disruption of the transsulfuration pathway. The treatment chickens also had a decreased hepatic CDO and increased GSS mRNA expressions which are in concordance with the homeostatic regulation of cysteine. Compromised cysteine metabolism could affect thermoregulation and subsequently affect feed efficiency and welfare of the birds.

Do glucocorticoids or carotenoids mediate plumage coloration in parrots? An experiment in Platycercus elegans.

Conspicuous coloration can indicate phenotypic quality, and may reflect exposure or vulnerability to stress, or access to essential nutrients such as pigments. Although the production of pigmented colours is well understood, much less is known about how structural colours are affected by physiological state. In this study, we tested whether glucocorticoids (corticosterone) predicted expression of plumage coloration in an Australian parrot, the crimson rosella (Platycercus elegans). Parrots provide an interesting and unique test, as they possess conspicuous coloration produced by distinctive pigments known as psittacofulvins, in addition to structural coloration. We have previously documented that coloration in P. elegans is condition-dependent and responds to dietary manipulation. Here, n = 21 P. elegans underwent a dietary manipulation (including food restriction or carotenoid supplementation) during which they moulted, and the change in reflectance was measured for three structural and three pigmentary plumage patches. Stress-induced corticosterone (10 min after handling) measured at the start of the experiment predicted change in coloration in two pigmentary patches (crown and front). We also found that change in stress-induced corticosterone during the experiment was associated with the change in coloration of the crown and two structural patches (cheek and epaulette). Baseline corticosterone (<3 min after handling) was not associated with any measure of coloration. We found no effects of dietary manipulation on baseline or stress-induced corticosterone, but carotenoid supplementation was associated with an increase in a measure of chronic stress (heterophil/lymphocyte ratio), and the corticosterone response to handling decreased over the course of the study. Our results suggest that corticosterone may be linked to colour expression more broadly than previously recognised, including psittacofulvin and structural coloration in parrots, and they confirm the independence of plumage pigmentation in parrots from carotenoid accumulation. Moreover, our study provides new insight into the stress responses of Psittaciformes, one of the most highly threatened avian orders.

Leaves of Moringa, Rosemary and Olive as a Phytogenic Feed Additives in Muscovy Duck diets.

BACKGROUND AND OBJECTIVE: Alternative sources of natural ingredients such as herbs or phytogenic plants have beneficial effects which are used as feed supplements or medicines in poultry. The current research objective was to investigate the influence of using phytogenic plants as feed additives on Muscovy ducklings performance. MATERIALS AND METHODS: A total number of 315 of 14 days old Muscovy ducklings were divided randomly into 7 experimental groups, each group was allocated into three replicates. The experimental treatments were arranged in one way classification by using three sources of phytogenic plants being Moringa oleifera leaves meal (MLM), Rosmarinus officinalis leaves meal (RLM) and Olea europaea leaves meal (OLM) with two levels of plant addition (1 or 2%) compared to the commercial diet (control). The experimental diets were formulated to be iso-caloric and iso-nitrogenous. Feed and water were offered ad libitum from 14 up to 70 days old. RESULTS: Data showed that phytogenic plants did not affect negatively growth performance of ducklings but improved gain compared to the control group. The highest value of live body weights (4185 g) was recorded for diet that supplemented with 1% MLM at 70 days old while, the lowest weight recorded for diet supplemented with 2% OLM being 3507 g compared to the control. A significant decrease in FI (g/bird) was noticed for treatment groups during the interval and the whole experimental periods compared with control group. CONCLUSION: Ducklings fed diet supplemented with phytogenic plants recorded the highest values of feather, carcass, heart, liver and gizzard (%) compared to the control group.

The influence of petroleum oil films on the feather structure of tropical and temperate seabird species.

Feather fouling is a primary cause of seabird mortality during marine hydrocarbon oil spills. Understanding how oils interact with feathers is an important step in mitigating this threat. Seabird feathers from 12 taxa, representing most seabird families from the tropics and southern latitudes, were exposed to crude and condensate oil films under laboratory settings. Feathers were measured for changes in mass proportional to feather size, and for barbule clumping. Seabird feathers from six distinct families exposed to very thin oil sheens (<0.3 µm) showed no significant change in proportional mass relative to control treatments, and 10 of the 12 species exposed to these films revealed no significant difference in barbule clumping. By contrast, exposure to both crude and condensate oil films ≥3 µm resulted in significant increases in feather mass and clumping. Our findings highlight the importance of considering the influence of oil on feather structure when compiling threat assessments involving seabirds.

Inhibition of the Transsulfuration Pathway Affects Growth and Feather Follicle Development in Meat-Type Chickens.

Cysteine is a nonessential amino acid in poultry nutrition. Poultry diets are deficient in cysteine, but the bird's cysteine need is met through the transsulfuration pathway (TSP) where homocysteine is converted to cysteine: a process catalyzed by cystathionine ß-synthase (CBS) and cystathionine γ-lyase (CTH). Cysteine is also a major component of keratinized protein found in feathers, but the extent to which cysteine is involved in feather and skin development in poultry is unknown. We randomly assigned chicks to control and treatment (control diet plus 100 mg/kg body weight of propargylglycine which is an inhibitor of CTH) diets. The thickness of skin layers, primary feather follicle parameters, growth, and mRNA expression of CBS and CTH were measured. Inhibition of TSP corresponded with the upregulation of liver mRNA of both CBS and CTH and reduction in growth from 35 to 40 days of age. The epidermis thickness, feather follicle length, and diameter were reduced from 10 to 40 days of age. Incorporation of cysteine into keratinized protein may be more sensitive to the level of available cysteine than into nonkeratinized proteins. Thus, disruption of the TSP could affect the thermoregulatory ability of the bird.

How chemotherapy and radiotherapy damage the tissue: Comparative biology lessons from feather and hair models.

Chemotherapy and radiotherapy are common modalities for cancer treatment. While targeting rapidly growing cancer cells, they also damage normal tissues and cause adverse effects. From the initial insult such as DNA double-strand break, production of reactive oxygen species (ROS) and a general stress response, there are complex regulatory mechanisms that control the actual tissue damage process. Besides apoptosis, a range of outcomes for the damaged cells are possible including cell cycle arrest, senescence, mitotic catastrophe, and inflammatory responses and fibrosis at the tissue level. Feather and hair are among the most actively proliferating (mini-)organs and are highly susceptible to both chemotherapy and radiotherapy damage, thus provide excellent, experimentally tractable model systems for dissecting how normal tissues respond to such injuries. Taking a comparative biology approach to investigate this has turned out to be particularly productive. Started in chicken feather and then extended to murine hair follicles, it was revealed that in addition to p53-mediated apoptosis, several other previously overlooked mechanisms are involved. Specifically, Shh, Wnt, mTOR, cytokine signalling and ROS-mediated degradation of adherens junctions have been implicated in the damage and/or reparative regeneration process. Moreover, we show here that inflammatory responses, which can be prominent upon histological examination of chemo- or radiotherapy-damaged hair follicle, may not be essential for the hair loss phenotype. These studies point to fundamental, evolutionarily conserved mechanisms in controlling tissue responses in vivo, and suggest novel strategies for the prevention and management of adverse effects that arise from chemo- or radiotherapy.

Effect of dietary amino acid density on broiler breeder reproductive performance.

The effects of 3 dietary amino acid (AA) profiles during lay on reproductive performance of broiler breeders were investigated. A flock of 2,376 Ross 708 female broiler breeders were randomly divided into 3 treatments with 12 replicates of 66 females per replicate, and fed 2 breeder feed phases (breeder 1: 25 to 36 wk; breeder 2: 37 to 60 wk) which were formulated to 3 different AA densities (Aviagen advice = control; 10% increased AA in Aviagen advice = high-AA; and estimated AA needs for maintenance, growth, and egg mass = estimated-AA). No differences in total or hatching egg production due to dietary treatments were observed. Hens fed high-AA had a higher incidence of double-yolk eggs. During the second phase of lay, hens fed estimated-AA had lower livability compared to the other treatments. Hens fed estimated-AA also laid smaller eggs compared to control and high-AA fed birds. Back feather cover deteriorated as hens grew older, and hens fed the estimated-AA diets had poorer feather cover from 41 wk onward. Overall, these results suggest that feeding higher AA levels than Aviagen recommendations presented no benefit in reproductive performance or feather quality. The estimated-AA feeding approach supported laying performance similar as other treatments, although affected egg weight and back feather cover.

Efficacy of 2-hydroxy-4-methylthiobutanoic acid compared to DL-Methionine on growth performance, carcass traits, feather growth, and redox status of Cherry Valley ducks.

The objective of this study was to compare the bio-efficacy of 2-hydroxy-4-methylthiobutanoic acid (DL-HMTBA) with that of DL-methionine (DLM) as sources of methionine in terms of the growth performance, carcass traits, feather growth, and redox statuses of Cherry Valley ducks. Six hundred and thirty male ducks were randomly allotted to 9 dietary treatment groups with 7 replicates of 10 birds each. The first group received a basal diet (BD) without methionine addition that was deficient in the total number of sulfur amino acids. In Groups 2 to 5 and Groups 6 to 9, the BD was supplemented with 4 increasing doses of methionine as either DLM or DL-HMTBA. The trial was run from ages 1 to 42 d. Dietary supplementation with DLM and DL-HMTBA improved body weight gain and feed intake as well as weights of carcasses, breast meat, and feathers compared with the BD. No significant difference was observed between the 2 methionine sources on growth performance, carcass traits, and feather growth. Concentrations of some redox markers in the pectoralis major muscle were improved by addition of methionine to the BD. However, a significant difference was observed between DLM and DL-HMTBA in this respect, as the supplementation of DL-HMTBA significantly increased the total antioxidant capacity, the activities of glutathione peroxidase, and the concentration of reduced glutathione in the pectoralis major muscle, compared with DLM. No significant difference between methionine sources was found with regard to the concentrations of oxidized glutathione and malondialdehyde in the pectoralis major muscle. Both DLM and DL-HMTBA increased malondialdehyde concentrations in the pectoralis major muscle compared with the BD. In conclusion, these results indicated that DLM and DL-HMTBA have equal biological value for the growth performance, carcass traits, and feather growth of Cherry Valley duck. Moreover, the improved antioxidant capacity observed with DL-HMTBA makes this a better candidate than DLM for lowering the oxidation process in the meat during post-mortem storage and thereby contributes to a better duck meat quality.

Effects of an early-life paraquat exposure on adult resistance to oxidative stress, plumage colour and sperm performance in a wild bird.

Early-life stressful conditions can shape individual phenotypes and ultimately influence fitness. Oxidative stress is a pervasive threat that affects many fitness-related traits and can modulate life-history trade-offs. Yet, the extent to which exposure to oxidative stress during early life can have long-lasting effects on key fitness-related traits remains to be elucidated, particularly in natural populations of vertebrates. Using a wild population of great tits Parus major, we experimentally dosed 11-day-old birds with paraquat, a pro-oxidant molecule, aiming at increasing oxidative stress. One year later, we recaptured 39 of them as adult recruiting breeders and quantified effects of the paraquat exposure on their resistance to oxidative stress, carotenoid-based plumage coloration and male sperm performance. Despite the absence of a short-term effect of paraquat on oxidative stress measured two days later, the pre-fledging exposure to paraquat induced a reduction in individual oxidative damage measured at adulthood. Paraquat-dosed individuals also had brighter plumage, but no effect was observed on male sperm performance. For the first time in a natural population of vertebrates, we experimentally show that an early-life acute exposure to a pro-oxidant has long-lasting effects on individual resistance to oxidative stress at adulthood. Our results are in line with the environmental matching and the hormesis hypotheses but may also reflect selective disappearance of individuals with lower resistance to oxidative stress.

Spotlight on Ibrutinib in PCNSL: Adding Another Feather to Its Cap.

In this issue Grommes and colleagues elegantly show that the irreversible inhibitor of Bruton tyrosine kinase, ibrutinib, promotes a high proportion of durable responses in primary central nervous system lymphoma, a type of diffuse large B-cell lymphoma (DLBCL), and also in secondary DLBCL relapsing to the central nervous system. Mutations in the B-cell antigen receptor-associated protein CD79B with upregulation of the MTOR pathway were associated with diminished response, but preclinical combination of PIK3CA and PIK3CD inhibitors synergized with ibrutinib to overcome this resistance mechanism, providing opportunity for further targeted therapy of this difficult-to-treat disease. Cancer Discov; 7(9); 940-2. ©2017 AACRSee related article by Grommes et al., p. 1018.

Omega-3 polyunsaturated fatty acids provided during embryonic development improve the growth performance and welfare of Muscovy ducks (Cairina moschata).

The welfare of ducks can be affected by unwanted behaviors such as excessive reactivity and feather pecking. Providing long-chain n-3 polyunsaturated fatty acids (LC n-3 PUFA) during gestation and early life has been shown to improve the brain development and function of human and rodent offspring. The aim of this study was to test whether the pecking behavior of Muscovy ducks during rearing could be reduced by providing LC n-3 PUFA during embryonic and/or post-hatching development of ducklings. Enrichment of eggs, and consequently embryos, with LC n-3 PUFA was achieved by feeding female ducks (n-3F) a diet containing docosahexaenoic (DHA) and linolenic acids (microalgae and linseed oil). A control group of female ducks (CF) was fed a diet containing linoleic acid (soybean oil). Offspring from both groups were fed starter and grower diets enriched with DHA and linolenic acid or only linoleic acid, resulting in four treatment groups with 48 ducklings in each. Several behavioral tests were performed between 1 and 3 weeks of age to analyze the adaptation ability of ducklings. The growth performance, time budget, social interactions, feather growth, and pecking behavior of ducklings were recorded regularly during the rearing period. No significant interaction between maternal and duckling feeding was found. Ducklings from n-3F ducks had a higher body weight at day 0, 28, and 56, a lower feed conversion ratio during the growth period, and lower reactivity to stress than ducklings from CF ducks. Ducklings from n-3F ducks also exhibited a significantly reduced feather pecking frequency at 49 and 56 days of age and for the whole rearing period. Moreover, consumption of diets enriched with n-3 PUFA during the starter and grower post-hatching periods significantly improved the tibia mineralization of ducklings and the fatty acid composition of thigh muscles at 84 days of age by increasing the n-3 FA content.

The Feather Model for Chemo- and Radiation Therapy-Induced Tissue Damage.

Chemo- and radiation therapy are the main modalities for cancer treatment. A major limiting factor is their toxicity to normal tissue, thus reducing the dose and duration of the therapy. The hair follicle, gastrointestinal tract, and hematopoietic system are among the target organs that often show side effects in cancer therapy . Although these organs are highly mitotic in common, the molecular mechanism of the damage remains unclear. The feather follicle is a fast-growing mini-organ, which allows observation and manipulation on each follicle individually. As a model system, the feather follicle is advantageous because of the following reasons: (1) its complex structure is regulated by a set of evolutionarily conserved molecular pathways, thus facilitating the effort to dissect the specific signaling events involved; (2) its morphology allows the continuity of normal-perturbed-normal structure in a single feather, thus "recording" the damaging effect of chemo- and radiation therapy; (3) further histological and molecular analysis of the damage response can be performed on each plucked feather; thus, it is not necessary to sacrifice the experimental animal. Here, we describe methods of applying the feather model to study the molecular mechanism of chemo- and radiation therapy-induced tissue damage.

T lymphocytes dominate local leukocyte infiltration in response to intradermal injection of functionalized graphene-based nanomaterial.

Graphene-based nanomaterials (GBN) have many potential biomedical applications. However, information regarding their biological properties and interactions with cells and/or soluble factors within a complex tissue is limited. The objective of this study was to use the growing feather (GF) of chickens as a minimally invasive cutaneous test-site to assess and monitor leukocyte recruitment in response to intradermal GBN injection. Specifically, the dermis of 20 GFs per chicken was injected with 10 µl of phosphate-buffered saline (PBS)-vehicle or 10 µl of 300 µg ml-1 oxygen-functionalized (f) GBN (6 chickens/treatment). GFs were collected before- (0) and at 0.25, 1, 2, 3, 4, 5, and 7 days post-injection and used for leukocyte-population analysis of immunofluorescently stained pulp cell suspensions or histological examination. Based on flow-cytometric cell population analysis, lymphocytes and macrophages were the major leukocyte-populations infiltrating GFs in response to f-GBN presence. Compared with PBS-controls, levels of T cells (γδ-, αß-, CD4- and CD8-T cells) were greatly elevated in f-GBN-injected GFs within 6 h and remained elevated throughout the 7-day examination period. f-GBN's effects on local tissue leukocyte recruitment were not reflected in the blood, except for a higher percentage of lymphocytes on 7 days. These observations together with a visual examination of f-GBN-injected GF tissue-sections suggest a delayed-type hypersensitivity-like, inflammatory cell-mediated response to the non-biodegradable f-GBN. The GF 'in vivo test-tube'system together with blood sampling provided unique insight into the time-course, qualitative, and quantitative aspects of immune system activities initiated by the presence of f-GBN in a complex tissue of a living animal. Copyright © 2017 John Wiley & Sons, Ltd.

Trouble on takeoff: Crude oil on feathers reduces escape performance of shorebirds.

The ability to takeoff quickly and accelerate away from predators is crucial to bird survival. Crude oil can disrupt the fine structure and function of feathers, and here we tested for the first time how small amounts of oil on the trailing edges of the wings and tail of Western sandpipers (Calidris mauri) affected takeoff flight performance. In oiled birds, the distance travelled during the first 0.4s after takeoff was reduced by 29%, and takeoff angle was decreased by 10° compared to unoiled birds. Three-axis accelerometry indicated that oiled sandpipers produced less mechanical power output per wingbeat during the initial phase of flight. Slower and lower takeoff would make oiled birds more likely to be targeted and captured by predators, reducing survival and facilitating the exposure of predators to oil. Whereas the direct mortality of heavily-oiled birds is often obvious and can be quantified, our results show that there are significant sub-lethal effects of small amounts crude oil on feathers, which must be considered in natural resource injury assessments for birds.

Development of methods for avian oil toxicity studies using the double crested cormorant (Phalacrocorax auritus).

Oral and external dosing methods replicating field exposure were developed using the double crested cormorant (DCCO) to test the toxicity of artificially weathered Deepwater Horizon Mississippi Canyon 252 oil. The majority of previous oil dosing studies conducted on wild-caught birds used gavage methods to dose birds with oil and determine toxicity. However, rapid gut transit time of gavaged oil likely reduces oil absorption. In the present studies, dosing relied on injection of oil into live feeder fish for oral dosing of these piscivorous birds, or applying oil to body contour feathers resulting in transdermal oil exposure and oral exposure through preening. Both oral and external oil dosing studies identified oil-related toxicity endpoints associated with oxidative stress such as hemolytic anemia, liver and kidney damage, and immuno-modulation or compromise. External oil application allowed for controlled study of thermoregulatory stress as well. Infrared thermal images indicated significantly greater surface temperatures and heat loss in treated birds following external oil applications; however, measurements collected by coelomically implanted temperature transmitters showed that internal body temperatures were stable over the course of the study period. Birds exposed to oil externally consumed more fish than control birds, indicating metabolic compensation for thermal stress. Conversely, birds orally dosed with oil experienced hypothermia and consumed less fish compared to control birds.

The Importance of Carotenoid Dose in Supplementation Studies with Songbirds.

Carotenoid coloration is the one of the most frequently studied ornamental traits in animals. Many studies of carotenoid coloration test the associations between carotenoid coloration and measures of performance, such as immunocompetence and oxidative state, proceeding from the premise that carotenoids are limited resources. Such studies commonly involve supplementing the diets of captive birds with carotenoids. In many cases, however, the amount of carotenoid administered is poorly justified, and even supposedly carotenoid-limited diets may saturate birds' systems. To quantify the relationships among the amount of carotenoids administered in experiments, levels of circulating carotenoids, and quantities of carotenoids deposited into colored ornaments, we performed a meta-analysis of 15 published studies that supplemented carotenoids to one of seven songbird species. We used allometric scaling equations to estimate the per-gram carotenoid consumption of each study's subjects, and we used meta-regression to evaluate the effects of this carotenoid dose on differences in coloration and plasma carotenoid levels between supplemented and control groups of birds. After accounting for supplementation duration and species, we observed a significant positive correlation between carotenoid intake and response of plasma carotenoid level to supplementation. The presence of supplemental carotenoids also tended to increase the expression of ornamental coloration, but the magnitude of the carotenoid dose did not significantly affect how strongly coloration changed with supplementation. Further, coloration effect sizes had no significant relationship with plasma carotenoid effect sizes. We also found significant heterogeneity in responses among studies and species, and the parameters used to measure color significantly affected response to supplementation. Our results emphasize the importance of performing dosage trials to determine what supplementation levels provide limited versus surplus carotenoids and of measuring the natural level of carotenoid intake by the study species to validate the appropriateness of supplementation levels for a particular study species and experimental design.