Evaluation of Metal Partitioning across Humboldt Penguin (Spheniscus humboldti) Egg Components.

Humboldt Penguin (Spheniscus humboldti) population declines are attributable to several multifaceted anthropogenic impacts. At present, the exposure of Humboldt Penguins to high concentrations of heavy metals in the marine environment is a preeminent concern, due to mining along the Peruvian coast near key rookery sites. Metal and selenium concentrations were determined in eggs collected from September 2020 to April 2021 from a managed-care penguin population at the Brookfield Zoo to establish reference values for health indices conducted on wild populations. Concentrations of 16 elements, with emphasis on those found in mine efflux-arsenic, cadmium, copper, lead, mercury, selenium, and zinc-were assessed via inductively coupled plasma mass spectrometry in yolk, albumen, and eggshell. Data analyses indicate a clear delineation between egg constituents, with lipid-rich yolk displaying notably higher concentrations (µg/g) of arsenic (0.20 ± 0.064), chromium (0.086 ± 0.03), cobalt (0.01 ± 0.003), iron (238.65 ± 54.72), lead (0.32 ± 0.97), manganese (2.71 ± 0.66), molybdenum (0.57 ± 0.14), tin (3.29 ± 0.99), and zinc (64.03 ± 13.01) than other components (albumen and eggshell). These data confirm that heavy metals are partitioned differently across Humboldt Penguin egg components, which provides insight into the potential connection between embryonic nutrient source contamination and subsequent chick viability.

Comparison of Plasma Trace Elements Between 2 Captive Humboldt Penguin (Spheniscus humboldti) Populations.

Environmental pollutants and their effect on wildlife health play an important role in the conservation of endangered species and can be clinically relevant in captive animals too. Data on relevant concentrations of trace elements in captive birds with no known exposures are rare. For this study, silver, arsenic, gold, barium, cadmium, cobalt, chromium, copper, iron, mercury, magnesium, manganese, molybdenum, thallium, selenium, and zinc were measured by inductively coupled plasma mass spectrometry in lithium heparinized plasma samples from 18 Humboldt penguins at 2 zoological collections in Germany. The results showed that the plasma concentrations of silver, arsenic, gold, cadmium, cobalt, chromium, mercury, and selenium differed significantly (P ≤ 0.05) between the 2 penguin collections. The results indicate that the location of the birds has a strong influence on the plasma trace element concentrations. Well water used in the enclosures was suspected to be associated with these significant differences. Trace elemental concentrations in feed (eg, marine fish) and contamination from enclosure construction materials may also play a role. This study could provide a basis for further comparative, biomonitoring, toxicity, and reference interval studies.

Evaluation of the Effects of Administering Ultradiluted Avena sativa and Echinacea angustifolia on the Hematological Parameters of Magellanic penguins (Spheniscus magellanicus) during the Reproductive Period.

BACKGROUND AND AIMS: Penguins are seabirds that manifest physiological and immunological alterations during the reproductive season. The objectives of this study were to evaluate the laboratory parameters of male and female Magellanic penguins and to determine the penguins' hematological response to homeopathic medicines during that reproductive period. METHODS: Penguins (N = 18), kept under human care in a zoological park setting, were evaluated during the reproductive period and were divided randomly into two groups: a group subjected to treatment with Echinacea angustifolia 6cH and Avena sativa 6cH (N = 8; four breeding couples), and a placebo control group (N = 10; five breeding couples). The investigators were blinded to treatment allocation. Two-way ANOVA was performed to determine whether the experimental group (control or verum) and the sex of the animal had any significant effect on the variation of each hematological parameter between the samples. One-way ANOVA was performed on hematological parameters for which the sex did not present a significant effect. The significance level was p ≤0.05. RESULTS: Significant effects were seen regarding the following: mean corpuscular volume (MCV), in which the verum group showed an increase (29.78 ± 52.95 fL) while the control group showed stability/reduction (-3.08 ± 46.36 fL) (p = 0.049); proportion of heterophils, in which the verum group showed a less marked increase (8.38 ± 12.53%) than that of the control group (18.00 ± 9.37%) (p = 0.010); lymphocyte concentration, in which the verum group showed less marked reduction (-4.39 ± 2.21 × 109 cells/L) than that of the control group (-1.56 ± 2.76 × 109 cells/L) (p = 0.001); and proportion of lymphocytes, in which the verum group showed a less marked reduction (-6.75 ± 10.35%) than that of the control group (-17.3 ± 8.73%) (p = 0.002). CONCLUSION: Comparison of samples collected before and during the reproductive period showed that, regardless of group allocation, there were differences in the effects on MCV, heterophils and lymphocytes. Treatment with Echinacea angustifolia and Avena sativa resulted in maintenance of lymphocyte levels in Magellanic penguins during the breeding period, thus aiding these birds' immunity.

Lycopodium clavatum as a homeopathic treatment in Magellanic penguins (Sphenicus magellanicus)

Uric acid is the main form of excretion of nitrogenous components in birds. The assessment verification samples, control, check of uric acid concentrations is widely used to detect kidney disease. High concentrations (up to five times) of uric acid in plasma can lead to the precipitation of this acid in the form of crystals, which accumulate in the tissues, mainly in the synovial joints and the viscera surface. Aims: report the evolution of homeopathic treatment in increased uric acid in Magellanic penguins (Spheniscus magellanicus) from Santo André Municipal Zoo. Methodology: 12 animals showed an increase in uric acid (reference values 0,3-16,1mg/dL) and received homeopathic treatment with Lycopodium clavatum 6cH. The protocol established was the administration of two globules of Lycopodium clavatum 6cH, once a day, inside fishes offered in the animal's beak. Treatment started on 25/09/2019. On 30/10/2019 was collected a new blood sample, those who were not discharged were kept on treatment and new collections were made on 27/11/2019 and 17/12/2019. Results: 8 animals had an average uric acid of 20.45mg /dl and responded to the 35-days treatment, normalizing the biochemical values with an average of 12.24mg /dl. Two animals with 18.27 mg /dl were discharged in 62 days with 11.70 mg /dl and another two penguins with 19.85 mg /dl were discharged in 82 days later with 9.57 mg /dl of uric acid. Conclusion: Homeopathy proved to be successful in the treatment of increased uric acid in Magellanic penguins, easy to administer, affordable and without side effects.

Fraction distribution and dynamic cycling of phosphorus in lacustrine sediment at Inexpressible Island, Antarctica.

Phosphorus (P) chemistry and its dynamic cycling are essential for understanding aquatic primary productivity and ecosystem structure. However, there is a lack of knowledge on P chemistry in pristine aquatic ecosystems, such as in Antarctica. Here, we applied the Standards, Measurements and Testing Program (SMT) procedure and nuclear magnetic resonance spectroscopy (NMR) to reveal P speciation in two types of lacustrine sediment cores collected from Inexpressible Island, Ross Sea, East Antarctica. The Positive Matrix Factorization Model and Generalized Additive Models were applied to quantitatively identify the P sources and estimate relative effects of various environmental factors on the speciation. Our results demonstrate that orthophosphate, mainly as Ca-P, is the major component and the ortho-monoesters are the predominant organic phosphorus (OP) form in lacustrine sediments. Ornithogenic lacustrine sediments have a higher content of P as Ca-P than sediments with little or no penguin influence. Our model further suggests that penguin guano is the most important source for Ca-P, accounting for 80%, while detrital input is the predominant source for Fe/Al-P (up to 90%). The content of ortho-monoesters, as revealed by NMR, declines with depth, reflecting mineralization process of OP in the sediments. Moreover, we observed higher relative proportions of organic P in the sediments with little guano influence and the deposition of organic P are likely facilitated by microbial mats. Overall, our data suggest that burial of P in Antarctic lakes is sensitive to different P sources and sedimentary environments. The relatively higher bioavailable phosphorus in lacustrine sediments largely controls growth of aquatic microbial mats in oligotrophic lakes and ponds in Antarctica. The sediment profile data also indicate that P burial increased during the Medieval Climate Anomaly period, and climate warming is more conducive to P burial through the expansion of penguin populations and productivity of microbial mats. Our findings represent the first systematic understanding of natural P cycling dynamics and its main controlling factors in pristine ponds with different organic sources in Antarctica.

Isolation, Phylogenetic and Gephyromycin Metabolites Characterization of New Exopolysaccharides-Bearing Antarctic Actinobacterium from Feces of Emperor Penguin.

A new versatile actinobacterium designated as strain NJES-13 was isolated from the feces of the Antarctic emperor penguin. This new isolate was found to produce two active gephyromycin analogues and bioflocculanting exopolysaccharides (EPS) metabolites. Phylogenetic analysis based on pairwise comparison of 16S rRNA gene sequences showed that strain NJES-13 was closely related to Mobilicoccus pelagius Aji5-31T with a gene similarity of 95.9%, which was lower than the threshold value (98.65%) for novel species delineation. Additional phylogenomic calculations of the average nucleotide identity (ANI, 75.9-79.1%), average amino acid identity (AAI, 52.4-66.9%) and digital DNA-DNA hybridization (dDDH, 18.6-21.9%), along with the constructed phylogenomic tree based on the up-to-date bacterial core gene (UBCG) set from the bacterial genomes, unequivocally separated strain NJES-13 from its close relatives within the family Dermatophilaceae. Hence, it clearly indicated that strain NJES-13 represented a putative new actinobacterial species isolated from the gut microbiota of mammals inhabiting the Antarctic. The obtained complete genome of strain NJES-13 consisted of a circular 3.45 Mb chromosome with a DNA G+C content of 67.0 mol%. Furthering genome mining of strain NJES-13 showed the presence of five biosynthetic gene clusters (BGCs) including one type III PKS responsible for the biosynthesis of the core of gephyromycins, and a series of genes encoding for bacterial EPS biosynthesis. Thus, based on the combined phylogenetic and active metabolites characterization presented in this study, we confidently conclude that strain NJES-13 is a novel, fresh actinobacterial candidate to produce active gephyromycins and microbial bioflocculanting EPS, with potential pharmaceutical, environmental and biotechnological implications.

Nutritional Secondary Hyperparathyroidism and Fibrous Osteodystrophy in a Captive African Penguin (Spheniscus demersus) Similar to Osteomalacia in Poultry.

A 9-yr-old female black-footed African penguin (Spheniscus demersus) was presented for necropsy after a history of reproductive abnormalities, paresis of limbs, weakness, and sudden death. Postmortem examination revealed soft keel, collapsed rib cage with beading of the ribs, and bilateral parathyroid enlargement. Classic histologic lesions of fibrous osteodystrophy with osteomalacia were observed in the ribs, vertebrae, and to a lesser extent in the femur and tibiotarsus associated with hyperplasia of parathyroid glands. This represents the first report of nutritional secondary hyperparathyroidism in birds of the order Spheniciformes, most likely caused by low levels of calcium supplementation during egg laying. The reproductive abnormalities observed in this penguin and others from the same group (asynchronous egg-laying cycles, abnormal breeding behavior) were most likely exacerbated by the lack of an adequate photoperiod mimicking the natural daylight pattern.

Reporte de caso­Hiperparatiroidismo secundario nutricional y osteodistrofia fibrosa en un pingüino africano (Spheniscus demersus) en cautiverio similar a la osteomalacia observada en de aves de corral. Una hembra de pingüino africano de patas negras (Spheniscus demersus) de nueve años fue sometida a necropsia después de un historial de anomalías reproductivas, paresia de extremidades, debilidad y muerte súbita. El examen post mortem reveló que la quilla del esternón estaba blanda, la caja torácica colapsada, se observaron "perlas raquíticas" en las costillas y agrandamiento bilateral de las paratiroides. Se observaron lesiones histológicas clásicas de osteodistrofia fibrosa con osteomalacia en las costillas, vértebras y en menor medida, en el fémur y tibiotarsus asociadas con hiperplasia de glándulas paratiroides. Esto representa el primer informe de hiperparatiroidismo secundario nutricional en un ave del orden Spheniciformes, muy probablemente causado por un bajo nivel de suplementos de calcio durante la producción de huevos. Las anomalías reproductivas observadas en este pingüino y otros del mismo grupo (ciclos de puesta de huevos asincrónicos, comportamiento de reproducción anormal) probablemente se vieron exacerbadas por la falta de un fotoperíodo adecuado que imitara el patrón de luz natural.

Homeopathic Treatment of Pododermatitis in Magellanic Penguins (Spheniscus magellanicus).

BACKGROUND: Problems with the feet are common in penguins, with a particular predisposition to pododermatitis. This condition usually occurs due to the changes in normal activity that result from being held captive, but its precise pathogenesis is still undetermined. METHODS/PATIENTS: This veterinary case study reports the use of oral homeopathic treatment on acute and chronic pododermatitis in five Magellanic penguins in a zoological park setting. During treatment, the patients remained in the penguins' living area, and the effect of the treatment on the progression of their lesions was assessed visually once weekly. The treatment consisted of a combination of Arnica montana and Calcarea carbonica. RESULTS: After treatment, the appearance of the lesions had noticeably improved: in the majority of penguins there was no longer evidence of infection or edema in the feet. The rate of recovery depended on the initial severity of the lesion. Those penguins that still showed signs of infection nevertheless exhibited a clear diminution of the size and thickness of the lesions. Homeopathic treatment did not cause any side effects. CONCLUSION: Homeopathy offers a useful treatment option for pododermatitis in captive penguins, with easy administration and without side effects.

Male Antarctic fur seals: neglected food competitors of bioindicator species in the context of an increasing Antarctic krill fishery.

The fishery for Antarctic krill is currently managed using a precautionary, ecosystem-based approach to limiting catch, with performance indices from a long-term monitoring program focused on several krill-dependent predators that are used to track ecosystem health. Concerns over increased fishing in concentrated areas and ongoing efforts to establish a Marine Protected Area along the Peninsula, a key fishing region, is driving the development of an adaptive management system for the fishery. The cumulative effects of fishing effort and interactions among krill-dependent predators and their performance is at present neglected in the CCAMLR Ecosystem Monitoring Program. However, we show considerable overlap between male Antarctic fur seals and the krill fishery in a complex mosaic, suggesting potential for cumulative impacts on other krill dependent predators. A holistic view is required as part of future efforts to manage the krill fishery that incorporates various sources of potential impacts on the performance of bioindicator species, including the fishery and its interactions with various krill dependent predators.

Nest-type associated microclimatic conditions as potential drivers of ectoparasite infestations in African penguin nests.

Nest design and characteristics can influence the microclimatic conditions in the nest. Nest-dwelling ectoparasites are sensitive to temperature and moisture and as such the conditions in the nest can influence parasite infestations. The endangered African penguin (Spheniscus demersus) breeds in different nest types and as yet little is known with regard to the microclimate and parasite infestation within these nests. This study characterized the microclimatic conditions in natural open, natural covered (with vegetation) and artificial nests, and assessed the relationship between nest characteristics (type, age, distance from the coast, orientation and entrance opening) and in-nest ectoparasite infestations and the health of African penguins in Stony Point, South Africa. Penguins (50 adults and 192 chicks) and their nests (n = 308) were sampled in 2016 and 2017. Soil temperature was higher in artificial than in natural nests, and soil and nest material moisture was lower in artificial and natural covered nests than natural open. Ectoparasite infestations were higher under warmer and drier conditions, in artificial nests and nests near the coastline. Penguin (adult and chick) body mass and chick body condition were lower in warmer nests and total plasma protein (in adults and checks) was lower in drier nests. Given the potential adverse effects of ectoparasites on host species, it is recommended that conservation agencies implement a monitoring programme to assess the ectoparasite infestation in artificial nests across multiple colonies. This information will facilitate a more holistic penguin conservation management plan that may prevent further detrimental effects on this endangered penguin species.

HEMATOLOGY AND PLASMA CHEMISTRY VALUES IN BEACHED MAGELLANIC PENGUIN (SPHENISCUS MAGELLANICUS) IN NORTHERN ARGENTINA DURING THE NONBREEDING SEASON.

Seabirds have been widely used for monitoring the health of the oceans in diverse marine regions. Among low-cost survey strategies, systematic surveys of seabirds beached on coasts have been developed since the 20th century. However, these studies do not always address blood aspects. The assessment of the health status of birds based on the analysis of hematological and plasma chemistry is crucial to evaluate the overall health status profile of live organisms. Here, the authors study the variability of blood parameters by sex, age class, and year of beached Magellanic Penguin during the nonreproductive period in northern Argentina. Of 44 penguins, 77% were categorized as younger juveniles and the rest as older juveniles, and were captured and studied in coastal areas of Buenos Aires Province during the summers of 2017 and 2018. The mean body weight of beached penguins was affected by the age class of the individuals; most of the younger juveniles showed poor condition in terms of body mass (1,761 ± 235 g). No significant differences were observed in body weight between years and sex. Still, there were significant differences between years for alkaline phosphatase (ALP) and creatine phosphokinase (CPK) values. Twelve of the 20 blood parameters analyzed differ significantly with the age class of the beached penguins; younger juveniles were in a state of inanition. Our results may serve as a necessary first step in improving the conservation status of the Magellanic Penguin in nonbreeding grounds of Argentina, and call for a better knowledge of the health status of the species along its annual cycle.

Transformation of sulfur species in lake sediments at Ardley Island and Fildes Peninsula, King George Island, Antarctic Peninsula.

In Antarctica, penguins transport sulfur and other bio-elements in the form of guano from marine to terrestrial environments where they become deposited in ornithogenic soils and sediments, including lake or pond systems. Transformation of sulfur species in these terrestrial and aquatic deposits, however, has rarely been studied. Here, we used the cold diffusion method to analyze various sulfur species in a lake deposit of ornithogenic sediment core (C1) and a pristine lake sediment core (C2), collected from Ardley Island and Fildes Peninsula, Antarctic Peninsula, respectively. The total organic carbon, total phosphorus, total nitrogen and various sulfur species in C1 were more fluctuant and much higher than those in C2, indicating a primary source from penguin guano. In core C1, organic sulfur (Org-S) was the main form of sulfur, and sulfate (SO42-) was the main form of inorganic sulfur. The acid volatile sulfur (AVS) in C1 was much higher than pyrite sulfur (CRS). In the pristine lake sediment core C2, Org-S and SO42- were the main sulfur species. CRS was the primary form of reduced inorganic sulfur in C2 sediments in contrast to the AVS in C1, indicating that AVS had been effectively transformed into CRS in C1. Our results demonstrate that the high levels of organic matter in C1 likely limited the transformation of AVS to CRS.

Comparative morphometric evaluation of hepatic hemosiderosis in wild Magellanic penguins (Spheniscus magellanicus) infected with different Plasmodium spp. subgenera / Avaliação morfométrica comparativa da hemossiderose hepática em pinguins-de-Magalhães (Spheniscus magellanicus) de vida livre infectados por diferentes subgêneros de Plasmodium spp

Abstract Avian malaria is one of the most important diseases of captive penguins. We employed morphometric techniques to evaluate hepatic hemosiderosis in rehabilitating wild Magellanic penguins (Spheniscus magellanicus) that were negative (n = 9) or naturally infected by different subgenera of Plasmodium spp. (n = 24), according with: Plasmodium subgenera (Haemamoeba, Huffia, Other lineages, and Unidentified lineages), severity of Plasmodium histopathological lesions, and concurrent diseases, age class (juvenile or adult plumage), sex (male, female or not determined), body score (emaciated, thin, good, excellent, not available), molt, presence or absence of oil contamination upon admission, iron supplementation, and rehabilitation center. The percentage of the area occupied by hemosiderin was called 'Index of Hepatic Hemosiderosis (IHH)'. Plasmodium-positive females presented significantly higher IHH values (17.53 ± 12.95%) than males (7.20 ± 4.25%; p = 0.041). We observed higher levels of congestion (p = 0.0182) and pneumonia (p = 0.0250) severity between Unidentified lineages vs. Huffia. We believe that the hepatic hemosiderosis observed in this study was multifactorial, the result of pathological processes caused by malaria, molting, hemoglobin and myoglobin catabolism during migration, anemia, concomitant diseases, and iron supplementation, all possibly potentiated by decreased liver mass. Further studies are needed to clarify the mechanisms of these hypotheses.

Resumo Malária aviária é uma das mais relevantes doenças em pinguins cativos. Foram aplicadas técnicas morfométricas para avaliar a hemossiderose hepática em pinguins-de-Magalhães (Spheniscus magellanicus ) de vida livre em reabilitação negativos (n = 9) e naturalmente infectados por diferentes subgêneros de Plasmodium spp. (n = 24), quanto a: subgênero de Plasmodium (Haemamoeba , Huffia, Outras Linhagens, e Linhagens não identificadas), severidade das lesões histopatológicas causadas por Plasmodium e doenças concomitantes, faixa etária (plumagem juvenil ou adulta), sexo (macho, fêmea, indeterminado), condição corporal (emaciado, magro, bom, excelente, indisponível), muda, presença/ausência de óleo a admissão, suplementação de ferro, e centro de reabilitação. A porcentagem da área ocupada por hemossiderina foi denominada "Índice de Hemossiderose Hepática (IHH)". Fêmeas Plasmodium -positivas apresentaram IHH significativamente mais elevado que machos, respectivamente, 17,53 ± 12,95% e 7,20 ± 4,25% (p = 0,041). Níveis mais elevados de congestão (p = 0,0182) e pneumonia (p = 0,0250) foram observados entre Linhagens não identificadas vs. Huffia. Possivelmente, a hemossiderose hepática observada nesse estudo seja multifatorial, resultado de processos patológicos causados por malária, muda, catabolismo de hemoglobina e mioglobina durante a migração, anemia, doenças concomitantes e suplementação de ferro, potencialmente intensificados por massa hepática reduzida. Estudos complementares são necessários para esclarecer os mecanismos de tais hipóteses.

Comparative morphometric evaluation of hepatic hemosiderosis in wild Magellanic penguins (Spheniscus magellanicus) infected with different Plasmodium spp. subgenera.

Avian malaria is one of the most important diseases of captive penguins. We employed morphometric techniques to evaluate hepatic hemosiderosis in rehabilitating wild Magellanic penguins (Spheniscus magellanicus) that were negative (n = 9) or naturally infected by different subgenera of Plasmodium spp. (n = 24), according with: Plasmodium subgenera (Haemamoeba, Huffia, Other lineages, and Unidentified lineages), severity of Plasmodium histopathological lesions, and concurrent diseases, age class (juvenile or adult plumage), sex (male, female or not determined), body score (emaciated, thin, good, excellent, not available), molt, presence or absence of oil contamination upon admission, iron supplementation, and rehabilitation center. The percentage of the area occupied by hemosiderin was called 'Index of Hepatic Hemosiderosis (IHH)'. Plasmodium-positive females presented significantly higher IHH values (17.53 ± 12.95%) than males (7.20 ± 4.25%; p = 0.041). We observed higher levels of congestion (p = 0.0182) and pneumonia (p = 0.0250) severity between Unidentified lineages vs. Huffia. We believe that the hepatic hemosiderosis observed in this study was multifactorial, the result of pathological processes caused by malaria, molting, hemoglobin and myoglobin catabolism during migration, anemia, concomitant diseases, and iron supplementation, all possibly potentiated by decreased liver mass. Further studies are needed to clarify the mechanisms of these hypotheses.

PLASMA CONCENTRATIONS OF VITAMIN A1, B1, D3, AND E IN HUMBOLDT PENGUINS ( SPHENISCUS HUMBOLDTI) BEFORE AND AFTER DIETARY VITAMIN SUPPLEMENTATION OF THEIR FISH DIET.

In a practical feeding trial at Ouwehand Zoo, plasma concentrations of vitamin A1, calcidiol (D3), α-tocopherol (E), and B1 in 17 Humboldt penguins ( Spheniscus humboldti) were measured before and after supplementation to gain insight into the effect of supplementing these vitamins in animals being fed thawed frozen-fish diets. None of the penguins received vitamin supplements for at least 6 mo before the supplementation trial, which was conducted prior to their normal nesting and molting period. During the trial period, eight penguins received daily vitamin A1, D3, tocopheryl acetate, and B1 supplementation placed in their fish immediately prior to feeding and nine control penguins received no supplementation. Concentrations of vitamins A1, D3, α-tocopherol, and B1 were also measured in the thawed ready-to-feed fish. Concentrations of vitamins B1 and α-tocopherol were below the Association of Zoos and Aquariums (AZA) recommendations for penguin diets, while concentrations of vitamins A1 and D3 were far above AZA recommendations. At the start of the study and after 70 days of supplementation, plasma concentrations were determined for these vitamins. Vitamin B1 concentrations in plasma increased significantly ( P < 0.05) between Day 0 (mean 39.9 µg/L) and day 70 (mean 160.5 µg/L) in the supplemented group. Plasma vitamin D3 and α-tocopherol did not show a significant change. Vitamin A1 levels in the supplemented group decreased significantly from 1.65 mg/L on day 0 to 1.4 mg/L on day 70. In the control group no significant changes were observed. The results of the study support the necessity of supplementing vitamin B1 in penguins fed thawed frozen fish. Depletion of vitamin A and E concentrations in frozen food fish over time support recommendations to regularly measure vitamin concentrations in different batches of frozen fish.

UV-light and dietary vitamin D and their effects on ionized calcium and 25-OH-D plasma concentrations in captive gentoo penguins (Pygoscelis papua).

In this study, the effect of ultraviolet (UV) light and dietary vitamin D on calcium metabolism in permanently indoor-housed gentoo penguins (Pygoscelis papua) was investigated. The study consisted of three periods, each completed with blood samples to analyse plasma concentrations of 25-OH-D, 1,25-(OH)2 -D, ionized (iCa) and total calcium (tCa). During the first study period (D), animals were housed under routine conditions without UV-light and fed a diet of different fish species, supplemented with 1,000 IU vitamin D per animal and day. The following study period (Baseline) of 28-day duration consisted of the same diet without any vitamin D supplementation and without UV-light. During the study period (UVB) artificial UV-light was added for 3 weeks. The vitamin D content of fish was measured by high-performance liquid chromatography. It varied between fish species and between facilities, ranging from no measurable content in capelin (Mallotus villosus) to 7,340 IU vitamin D/kg original matter (OM) in herring (Clupea spp). The average dietary vitamin D content was 311 IU/kg OM at facility 1 and 6,325 IU/kg OM at facility 2, resulting in a vitamin D intake per animal and day without supplementation of 130 IU (25.5 IU/kg body weight BW) and 2,454 IU (438.2 IU/kg BW) respectively. The supplementation of vitamin D elevated significantly the plasma concentrations of 25-OH-D by an intraindividual difference of 15 (range -2 to 59) nmol/L and tCa by 0.1 (0.0-0.3) mmol/L only at facility 2. The exposure to UV-light raised the blood concentrations of tCa at facility 2 by 0.15 (0.1-0.2) mmol/L, and of iCa and tCa for females at facility 1 by 0.23 (0.13-0.41) mmol/L and 1.8 (1.1-2.5) mmol/L respectively. No significant influence of the study periods (D) and (UVB) was found for the concentrations of 1,25-(OH)2 -D at both facilities.

SUSPECTED HYPOVITAMINOSIS A-ASSOCIATED SALT GLAND ADENITIS IN NORTHERN ROCKHOPPER PENGUINS ( EUDYPTES MOSELEYI).

Supraorbital salt-excreting glands are present in at least 10 avian orders and are largest in marine species, including penguins. Diseases of the avian salt gland have been described infrequently. From September 2015, five captive northern rockhopper penguins ( Eudyptes moseleyi) were presented over a 6-wk period for unilateral or bilateral supraorbital swellings. In September 2016, two cases recurred and two additional cases were identified. Histopathology demonstrated salt gland adenitis with extensive squamous metaplasia. Blood plasma testing demonstrated marked vitamin A and E deficiencies within the colony. Prolonged frozen storage of feed-fish was implicated as a cause of vitamin depletion; reducing storage times and addition of dietary supplementation prevented recurrence.

Increased mitochondrial energy efficiency in skeletal muscle after long-term fasting: its relevance to animal performance.

In the final stage of fasting, skeletal muscle mass and protein content drastically decrease when the maintenance of efficient locomotor activity becomes crucial for animals to reactivate feeding behaviour and survive a very long period of starvation. As mitochondrial metabolism represents the main physiological link between the endogenous energy store and animal performance, the aim of this study was to determine how a very long, natural period of fasting affected skeletal muscle mitochondrial bioenergetics in king penguin (Aptenodytes patagonicus) chicks. Rates of mitochondrial oxidative phosphorylation were measured in pectoralis permeabilized fibres and isolated mitochondria. Mitochondrial ATP synthesis efficiency and the activities of respiratory chain complexes were measured in mitochondria isolated from pectoralis muscle. Results from long-term (4-5 months) naturally fasted chicks were compared with those from short-term (10 day) fasted birds. The respiratory activities of muscle fibres and isolated mitochondria were reduced by 60% and 45%, respectively, on average in long-term fasted chicks compared with short-term fasted birds. Oxidative capacity and mitochondrial content of pectoralis muscle were lowered by long-term fasting. Bioenergetic analysis of pectoralis muscle also revealed that mitochondria were, on average, 25% more energy efficient in the final stage of fasting (4-5 months) than after 10 days of fasting (short-term fasted birds). These results suggest that the strong reduction in respiratory capacity of pectoralis muscle was partly alleviated by increased mitochondrial ATP synthesis efficiency. Such oxidative phosphorylation optimization can impact animal performance, e.g. the metabolic cost of locomotion or the foraging efficiency.

Metals and metalloids in Little Penguin (Eudyptula minor) prey, blood and faeces.

Piscivorous species like the Little Penguin (Eudyptula minor) are particularly at risk of being negatively impacted by pollution due to their heightened exposure through aquatic food chains. Therefore, determining the concentration of heavy metals in the fish prey of seabirds is an essential component of assessing such risk. In this study, we report on arsenic, cadmium, mercury, lead and selenium concentrations in three fish species, which are known to comprise a substantial part of the diet of Little Penguins at the urban colony of St Kilda, Melbourne, Australia. Metal concentrations in the fish sampled were generally within the expected limits, however, arsenic and mercury were higher than reported elsewhere. Anchovy (Engraulis australis) and sandy sprat (Hyperlophus vittatus) contained higher Hg concentrations than pilchard (Sardinops sagax), while sandy sprat and pilchard contained more selenium. We present these findings together with metal concentrations in Little Penguin blood and faeces, sampled within weeks of the fish collection. Mercury concentrations were highest in the blood, while faeces and fish prey species contained similar concentrations of arsenic and lead, suggesting faeces as a primary route of detoxification for these elements. We also investigated paired blood - faecal samples and found a correlation for selenium only. Preliminary data from stable isotope ratios in penguin blood indicate that changes in penguin blood mercury concentrations cannot be explained by trophic changes in their diet alone, suggesting a variation of bioavailable Hg within this semi-enclosed bay.

Trace-elements, methylmercury and metallothionein levels in Magellanic penguin (Spheniscus magellanicus) found stranded on the Southern Brazilian coast.

Magellanic penguins have been reported as good biomonitors for several types of pollutants, including trace-elements. In this context, selenium (Se), total mercury, methylmercury, inorganic mercury (Hg(inorg)), cadmium (Cd) and lead (Pb), as well as metallothionein (MT) levels, were evaluated in the feathers, liver and kidney of juvenile Magellanic penguins found stranded along the coast of Southern Brazil. The highest concentrations of all trace-elements and methylmercury were found in internal organs. Concentrations of Cd and Se in feathers were extremely low in comparison with their concentrations in soft tissues. The results showed that both Se and MT are involved in the detoxification of trace-elements (Cd, Pb and Hg(inorg)) since statistically significant relationships were found in liver. Conversely, hepatic Se was shown to be the only detoxifying agent for methylmercury.