Capillary Zone Electrophoresis in Humboldt Penguins (Spheniscus humboldti).

Electrophoresis is a useful diagnostic tool for detecting inflammation, including inflammation associated with infectious diseases (eg, aspergillosis in penguins). To our knowledge, reference intervals are not available for plasma proteins via electrophoresis in Humboldt penguins (Spheniscus humboldti). Therefore, preliminary reference intervals for blood plasma proteins measured by capillary zone electrophoresis were calculated for Humboldt penguins from a single zoological collection, and possible differences between the sexes and the ages of the birds were evaluated. Lithium heparinized plasma samples from 39 Humboldt penguins were analyzed. The following sex- and age-independent reference intervals were calculated: total protein 33.8-70.4 g/L, prealbumin 1.9-4.9 g/L, albumin 12.9-31.1 g/L, albumin: globulin ratio 0.7-1.7, α-globulins 4.5-11.6 g/L, ß-globulins 5.6-20.6 g/L, and γ-globulins 2.6-8.4 g/L. Male penguins had a significantly (P = 0.047) higher albumin: globulin ratio and lower percentage of ß-globulins (P = 0.015) in comparison with female penguins. Prealbumin (g/L) significantly (P = 0.021) decreased with increased age of the penguins. These results showed some differences between the sexes and ages of the penguins, which should be considered when interpreting the results. Further studies are needed to determine whether differences in other age groups or seasons exist, and also to evaluate which infectious diseases affect plasma proteins and how the reference values calculated here may deviate in ill penguins.

Vocal tract shape variation contributes to individual vocal identity in African penguins.

Variation in formant frequencies has been shown to affect social interactions and sexual competition in a range of avian species. Yet, the anatomical bases of this variation are poorly understood. Here, we investigated the morphological correlates of formants production in the vocal apparatus of African penguins. We modelled the geometry of the supra-syringeal vocal tract of 20 specimens to generate a population of virtual vocal tracts with varying dimensions. We then estimated the acoustic response of these virtual vocal tracts and extracted the centre frequency of the first four predicted formants. We demonstrate that: (i) variation in length and cross-sectional area of vocal tracts strongly affects the formant pattern, (ii) the tracheal region determines most of this variation, and (iii) the skeletal size of penguins does not correlate with the trachea length and consequently has relatively little effect on formants. We conclude that in African penguins, while the variation in vocal tract geometry generates variation in resonant frequencies supporting the discrimination of conspecifics, such variation does not provide information on the emitter's body size. Overall, our findings advance our understanding of the role of formant frequencies in bird vocal communication.

Splenic Small Cell Lymphoma with Presumed Secondary Immune-Mediated Hemolytic Anemia in an African Penguin (Spheniscus demersus).

A 2-year-old male African penguin (Spheniscus demersus) was presented to a veterinary teaching hospital for evaluation of a previously diagnosed subclinical, marked regenerative anemia. Physical examination at the zoological institution demonstrated biliverdinuria and pale oral mucous membranes. Diagnostic tests performed on the penguin since the diagnosis and prior to presentation to the veterinary teaching hospital included serial complete blood counts, plasma biochemistry panels, radiographic imaging, blood and plasma heavy metal testing, and infectious disease testing. The abnormal diagnostic test results were consistent with marked regenerative anemia and splenomegaly. At the veterinary teaching hospital, further diagnostic testing was ordered in an attempt to determine the cause of the biliverdinuria and pale oral mucous membranes. The diagnostic tests performed included a full-body contrast computed tomographic scan, upper gastrointestinal endoscopic procedure, bone marrow aspiration and evaluation, saline agglutination testing, blood Plasmodium species polymerase chain reaction screening, a vitamin profile panel, and repeat blood heavy metal testing. The complete blood count demonstrated a marked, regenerative anemia with the presence of dysplastic erythrocytes, and splenomegaly was found on the computed tomographic images without identifying a definitive cause. Primary disease differentials for the diagnosed regenerative anemia included a myelodysplastic syndrome and primary or secondary immune-mediated hemolytic anemia. The penguin was treated with oral prednisolone as an immunomodulatory agent; however, it did not result in a positive treatment response. The patient developed hyporexia, weight loss, and lethargy 2 months post presentation to the veterinary teaching hospital. Additional therapy with cyclophosphamide was initiated, and the penguin improved clinically, but then declined. The patient was euthanized due to a poor quality of life and prognosis 4 months after initial presentation and 1.5 years after the first complete blood count revealed the penguin to be anemic. Microscopic review of submitted postmortem tissue samples demonstrated a monomorphic population of neoplastic small lymphocytes infiltrating the spleen, consistent with splenic small cell lymphoma. The neoplastic cells did not label with the T-cell marker CD3 or B-cell markers CD20, CD79a, and Pax-5.

Suspected Mefloquine Toxicity in a Colony of Humboldt Penguins (Spheniscus humboldti).

Avian malaria is an important cause of mortality in captive penguins housed in outdoor exhibits. Mefloquine was used as a prophylaxis to treat a colony of 19 Humboldt penguins (Spheniscus humboldti) for avian malaria. A target dose of 30 mg/kg was obtained from anecdotal literature for sphenisciforms that was not based on pharmacokinetic or toxicity studies. For this reason, preliminary plasma concentrations of mefloquine were acquired after the first dose in some penguins to ensure that plasma concentrations reached human malaria prophylactic concentrations. Afterward, each penguin in the entire colony received mefloquine (26-31 mg/kg [125 mg in toto] PO q7d). Regurgitation was frequently observed starting after the fourth weekly administration. Plasma concentrations of mefloquine after the seventh dose showed elevated concentrations, and the treatment was immediately terminated. Eight penguins died during and after the treatment period. The first fatality occurred after the fifth weekly administration, and 7 birds died within 7-52 days after the seventh weekly administration. Three penguins were found dead without previous symptoms. The other five presented with marked lethargy, dyspnea, poor appetite, and vomiting, and all died despite medical care. The remaining 11 penguins of the colony survived without any supportive care; 5 did not exhibit any clinical disease signs, while the other 6 showed a mild apathy and decreased appetite. Mefloquine toxicity was highly suspected on the basis of clinical signs, the elevated mefloquine plasma concentrations, and no other underlying pathologic disease conditions identified through postmortem examinations. Nonspecific lesions, including pulmonary congestion and edema and hepatic perivascular hematopoiesis, were noted in the birds that died. Additionally, 1 case presented with myocarditis, and mycobacteria were observed within granulomas in the respiratory tract of 2 penguins. Caution is advised, and further studies are encouraged before administering mefloquine to penguins.

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.

The Effects of Critical Care Nutrition on Weight Gain in African Penguin (Spheniscus demersus) Chicks.

Nutritional support in malnourished animals is an essential aspect of wildlife rehabilitation; this support is especially relevant when providing lifesaving nutrition to endangered species such as the African penguin (Spheniscus demersus). This study investigated the short-term effects of a commercially available, semi-elemental, critical care diet compared with a hand-made fish formula. Twenty-one African penguin chicks were selected on admission to the Southern African Foundation for the Conservation of Costal Birds in Cape Town, South Africa, in November 2015. Initial assessment included body weight, a full clinical exam, white blood cell count, packed cell volume, and total plasma protein. Ten animals received the commercial critical care diet, whereas a control group of 11 animals were fed hand-made formula for the 2-week study period. All animals were weighed daily and blood sampling was repeated after 14 days. The median weight of both groups increased significantly over 14 days (critical care diet χ2 = 10.1, P = 0.002; control χ2 = 7.4, P = 0.006). The difference was not significant between the groups for start weight (χ2 = 0.1, P = 0.725) or end weight (χ2 = 0, P = 1.000) and was not significantly different in the change over time for either absolute numbers (χ2 = 1.7, P = 0.193) or percent gain (χ2 = 0.8, P = 0.36). The values for packed cell volume, total plasma protein, and white blood cell count increased in all animals after the 14-day study period was complete. On the basis of the results of this study, it was determined that the differing diets led to similar weight gain.

Long walk home: Magellanic penguins have strategies that lead them to areas where they can navigate most efficiently.

Understanding how animals move in dense environments where vision is compromised is a major challenge. We used GPS and dead-reckoning to examine the movement of Magellanic penguins commuting through vegetation that precluded long-distance vision. Birds leaving the nest followed the shortest, quickest route to the sea (the 'ideal path', or 'I-path') but return tracks depended where the birds left the water. Penguins arriving at the beach departure spot mirrored the departure. Most of those landing at a distance from the departure spot travelled slowly, obliquely to the coast at a more acute angle than a beeline trajectory to the nest. On crossing their I-path, these birds then followed this route quickly to their nests. This movement strategy saves birds distance, time and energy compared to a route along the beach and the into the colony on the I-track and saves time and energy compared to a beeline trajectory which necessitates slow travel in unfamiliar areas. This suggests that some animals adopt tactics that take them to an area where their navigational capacities are enhanced for efficient travel in challenging environments.

Fatal Meningoencephalitis From a Rhizomucor pusillus Infection in a Juvenile Magellanic Penguin (Spheniscus magellanicus).

A 7-month-old, male Magellanic penguin (Spheniscus magellanicus), housed in an outdoor exhibit, developed acute neurologic signs that progressed to death over 2 days. On gross examination, the bird had congested, edematous lungs, and cerebellar hemorrhage. Histologic examination identified granulomatous pneumonia and encephalitis, with thrombosis and eosinophilic, branching fungal hyphae that had invaded the meningeal vessel walls. Polymerase chain reaction identified the fungus in the brain as Rhizomucor pusillus, an uncommon cause of mucormycosis. This organism has previously been reported in respiratory, skeletal, and sino-orbital lesions of avian species. This clinical report describes meningoencephalitis associated with Rhizomucor pusillus in a penguin.

Injectable Anesthesia With Medetomidine, Ketamine, and Butorphanol in Captive Humboldt Penguins (Spheniscus humboldti).

The effects of an injectable anesthesia with 0.05 mg/kg medetomidine, 5 mg/kg ketamine, and 0.5 mg/kg butorphanol administered together intramuscularly were evaluated in 22 captive Humboldt penguins (Spheniscus humboldti, 10 male and 12 female), with a mean age of 8.5 ± 8.23 years. The birds fasted for18-24 hours prior to the procedure. Induction was followed by 4 distinct progressive responses of the birds to the anesthetic effect, including onset of initial effects at 2.0 ± 1.7 minutes (x̄ ± SD), sternal recumbency with the head still elevated at 2.2 ± 1.6 minutes, lowering and placing the beak tip to the ground at 3.6 ± 3.4 minutes, and lateral positioning of the neck and head at 4.2 ± 3.4 minutes. A general state of sedation, muscle relaxation, and analgesia were noted 10.0 ± 2.8 minutes postinjection. However, according to an established scoring system for the assessment of anesthetic depth in avian patients, a surgical plane of anesthesia was not achieved. Muscle relaxation determined by the same scoring system lasted for 31.4 ± 17.1 minutes. The penguins' mean respiratory rate did not demonstrate significant change and spontaneous ventilation was present throughout the procedure. Relative peripheral arterial oxygen saturation decreased significantly from 92.83 ± 5.77% at 10 minutes to 90.91 ± 5.77% at 40 minutes following induction. The birds' heart rate also decreased significantly from 112.55 ± 23.97 beats/min at 10 minutes to 101.65 ± 25.42 beats/min at 40 minutes. The measured cloacal temperatures were maintained within normal range despite ambient temperatures of up to 28.3°C (82.9°F). Reversal of medetomidine with 0.25 mg/kg atipamezole was conducted after 45.1 ± 7.3 minutes. Recovery was smooth but of variable duration with patients being able or willing to stand steadily in an upright position after 50.1 ± 34.6 minutes. One penguin died during recovery from a ruptured left ventricle and consecutive pericardial tamponade, but no predisposing factors were identified. The anesthetic protocol proved to be effective for noninvasive and minor painful procedures (eg, diagnostic imaging, blood collection). Disadvantages to the administration of the combined anesthetic agents in the penguins included a short period of muscle relaxation and smooth but potentially prolonged recovery. The safety of the anesthetic protocol described for Humboldt penguins in this report has to be evaluated critically against the the death of 1 penguin during recovery.

Genetic evidence of hybridization between Magellanic (Sphensicus magellanicus) and Humboldt (Spheniscus humboldti) penguins in the wild.

The process of hybridization between closely related species plays an important role in defining the genetic integrity and overall genetic diversity of species. The distribution range of Magellanic (Spheniscus magellanicus) and Humboldt (Spheniscus humboldti) penguins is predominantly allopatric; however, the species share a region of sympatry where they may hybridize. We analyzed four types of genetic markers (including nuclear and mitochondrial markers) to assess their utility in detecting hybridization events between Magellanic and Humboldt penguins. Genetic assessment of non-introgressed reference samples allowed us to identify three types of informative markers (microsatellites, major histocompatibility complex, and mitochondrial DNA) and detect positive evidence of introgressive hybridization in the wild. Four out of six putative hybrids showed positive evidence of hybridization, revealed by the detection of Humboldt mitochondrial DNA and Magellanic species-specific alleles from nuclear markers. Bayesian Structure analysis, including samples from the sympatric region of the species in the southern Pacific Ocean, confirmed the use of nuclear markers for detecting hybridization and genetic admixture of putative hybrids, but revealed relatively low levels of genetic introgression at the population level. These findings provide insights into the role of hybridization in regions of species sympatry and its potential consequences on the levels of genetic introgression, genetic diversity, and conservation of these penguin species.

Compound-specific stable isotope analyses in Falkland Islands seabirds reveal seasonal changes in trophic positions.

BACKGROUND: While nitrogen and carbon stable isotope values can reflect ecological segregation, prey choice and spatial distribution in seabirds, the interpretation of bulk stable isotope values is frequently hampered by a lack of isotopic baseline data. In this study, we used compound-specific isotope analyses of amino acids (CSIA-AA) to overcome this constraint and to study interspecific differences, seasonal and historical changes in trophic positions of five seabird species, three penguins and two petrels, from a sub-Antarctic seabird community. RESULTS: CSIA-AA allowed comparing trophic positions of seabirds with temperate and polar distributions. Gentoo and Magellanic penguins had the highest trophic positions during the breeding season (3.7 and 3.9), but decreased these (2.9 and 3.3) during the feed-up for moult. Intra-specific differences were also detected in Thin-billed prions, where carbon isotope values clearly separated individuals with polar and temperate distributions, both in the breeding and interbreeding periods. Thin-billed prions that foraged in polar waters had lower trophic positions (3.2) than conspecifics foraging in temperate waters (3.8). We further investigated historical changes by comparing museum samples with samples collected recently. Our pilot study suggests that Rockhopper penguins, Magellanic penguins and Thin-billed prions with temperate non-breeding distributions had retained their trophic levels over a 90-100 year period, while Gentoo penguins and Thin-billed prions with polar non-breeding distributions had decreased trophic levels compared to historical samples. In contrast, Wilson's storm-petrels had slightly increased trophic levels compared to samples taken in 1924-1930. CONCLUSIONS: We applied compound-specific stable isotope analyses across a range of contexts, from intra-specific comparisons between stages of the breeding cycle to inter-specific seabird community analysis that would not have been possible using bulk stable isotope analyses alone due to differences in isotopic baselines.

A retrospective analysis of mortality in captive Magellanic penguins (Spheniscus magellanicus) in the United States, 2008-2018.

Mortality data for Magellanic penguins (Spheniscus magellanicus) housed in zoos and aquariums in the United States has not previously been published. Necropsy and histopathology records were examined for Magellanic penguins housed at 12 Association of Zoos and Aquariums institutions from 2008 through 2018. If birds lived through the first year, the mean longevity was found to be 18.9 years of age (standard deviation: 7.9). Prefledge chicks and geriatric penguins experienced the highest mortality rates. Aspergillosis was a major cause of death in this species. There was no significant difference in mortality between males and females. Based on these data, recommendations for the husbandry and veterinary care of captive Magellanic penguins can be made.

THE RADIOGRAPHIC AND ENDOSCOPIC ANATOMY AND DIGESTIVE MECHANISMS OF CAPTIVE AFRICAN PENGUINS (SPHENISCUS DEMERSUS).

The anatomy of the avian gastrointestinal (GI) tract is uniquely suited to each species' dietary requirements. African penguins (Spheniscus demersus) are charismatic and popular exhibit animals. As their prevalence grows, there is a need to understand their unique digestive tract to diagnose abnormalities. Reference material specific to the digestive tract of piscivores is scant, and knowledge of the GI tract of a healthy penguin is based on information from other birds. The purpose of this study is to determine the normal gross anatomy, transit time, and histopathologic structures of the penguin GI tract. Twelve clinically healthy penguins were selected for this study from the colony at the Maryland Zoo in Baltimore, which, at the time of this study, consisted of 55 birds. All penguins underwent a barium contrast study, and radiographic images were obtained until the entire GI tract was empty. Approximately 2 wk later, each penguin was anesthetized, and an endoscopic evaluation of the anterior GI tract was performed. Time from barium administration to defecation ranged from 17 to 70 min, and on average, barium clearance was 17.6 hr (range, 5-36 hr). Fluid from the ventriculus had an average pH of 2.75 and contained a mixed bacterial population. Koilin presence and thickness appreciated on endoscopy did not correspond with the thickness determined on histopathology. The results of this study provide a comparative baseline to use during diagnostic workups and help guide treatment decisions.

OBJECTIVE GAIT ANALYSIS IN HUMBOLDT PENGUINS (SPHENISCUS HUMBOLDTI) USING A PRESSURE-SENSITIVE WALKWAY.

Assessment of pododermatitis, osteoarthritis, and other causes of lameness in penguins can be challenging. Subjective gait analysis using visual observation and response to analgesic therapy can be affected by observer variation and caregiver placebo bias. A pressure-sensitive walkway (PSW), however, allows for objective gait analysis and assessment of analgesic therapeutic response. In this study, a 3-m-long PSW was used to analyze gait in 21 adult Humboldt penguins (Spheniscus humboldti). Medical record reviews and comprehensive examinations were performed on all penguins; five penguins were considered abnormal, with either right-sided (n = 3) or bilateral historical lameness-causing disease (n = 2) and were analyzed separately from the normal data set. All penguins walked across the PSW four times and gait parameters (step and stride distances and velocities, maximum force, impulse, and peak pressure) were calculated for each foot in each penguin. Statistical comparisons were made between right and left feet, sexes, and normal and abnormal penguins for each gait parameter. Among normal penguins, there were no significant differences between feet or sex. Left step width was shorter in abnormal penguins than that of normal penguins. Study results established baseline values for Humboldt penguins. This will allow objective monitoring of progression and response to therapy in penguin lameness cases, both current and future. The data also provide a foundation to compare gait parameters with other penguin populations and species.

FACTORS AFFECTING ABNORMAL MOLTING IN THE MANAGED AFRICAN PENGUIN (SPHENISCUS DEMERSUS) POPULATION IN NORTH AMERICA.

Abnormal molting, including partial or incomplete molt, arrested molt cycle, or inappropriate frequency of molt, is a primary concern for the managed African penguin (Spheniscus demersus) population and is documented across institutions. To identify factors associated with increased odds of abnormal molts and characterize intervention opportunities, a comprehensive survey evaluating numerous husbandry and medical parameters was created. Survey results represent 45 North American African penguin holding facilities and 736 unique animals. Of these individuals, 135 (18.3%) demonstrated an abnormal molt over the 5-yr study period (2012-2017). Increased odds ratios for abnormal molt included biologic (age, sex, etc.), geographic (elevation, latitude), and husbandry (exhibit design, diet, etc.) variables. The mean age of affected animals was 15.2 yr (1-45 yr, n = 135) compared with 9.92 yr (4 mo-38 yr, n = 601) for unaffected animals. In addition, although statistically insignificant, males were overrepresented in the affected cohort compared with a near even distribution among unaffected animals. Identified factors with increased odds for abnormal molting included advanced age and facilities using freshwater pools. Normally molting penguins were more commonly found with saltwater pool access and natural lighting exposure. Anecdotal medical intervention attempts are discussed, although further research is needed to define their use. Of attempted interventions, subcutaneous 5.4-mg melatonin implants placed in anticipation of environmental molting cues showed the most promise at inducing catastrophic molt, with 14 of 17 (82.3%) of affected individuals molting normally following this treatment. Survey analysis indicated that abnormal molt is a complex, multifactorial process, and modifiable factors that may predispose animals to abnormally molt exist. Addressing these factors in future exhibit design may mitigate the prevalence of this condition. Despite these efforts, it is likely that medical interventions will be required to aid in the treatment of abnormal molting in this 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.

Parasite diversity associated with African penguins (Spheniscus demersus) and the effect of host and environmental factors.

The African penguin (Spheniscus demersus) is a critically endangered species endemic to southern Africa. Limited information is available on the parasite diversity associated with the species in natural settings. This study explores the diversity and incidence of parasites associated with African penguins and their nests, and records the effect of host and environmental factors on parasite infestation. Ecto-, haemo- and helminth parasites were recorded from 210 adult birds, 583 chicks and 628 nests across five colonies (two mainland and three islands) along the south-western coast of South Africa, in 2016 and 2017. Mean nest density (total and active nests) and climate variables (temperature and precipitation) were obtained for each colony. Parapsyllus humboldti was the most abundant and prevalent ectoparasite on penguins and in nests (69.10 and 57.80%, respectively), while Piroplasmorida/Haemospororida (33.51%) and Cardiocephaloides spp. (56.17%) were the most prevalent haemo- and helminth parasites of penguins, respectively. In general parasite abundance and prevalence was significantly affected by penguin age (chicks vs adults), location (mainland vs islands), nest density (total and active nests) and season (spring vs autumn/winter). It is concluded that parasite infestations are structured and that penguin chicks at mainland colonies are more susceptible to parasite infestations during spring.

Empirical Primaquine Treatment of Avian Babesiosis in Seabirds.

Babesia species are tickborne hemoprotozoans. Although experiments have shown that primaquine is highly effective in the treatment of Babesia species infections in mammals, this drug has not been widely used for the treatment of avian babesiosis. Consequently, the treatment of this disease for avian patients has traditionally relied on an empirically established imidocarb treatment. In this study, the authors examined the efficacy of primaquine as a treatment alternative for avian babesiosis (Babesia peircei and Babesia ugwidiensis) in seabirds. Retrospective analysis was performed on the medical records and blood smears of 446 B peircei-positive African penguins (Spheniscus demersus) and 41 B ugwidiensis-positive Cape cormorants (Phalacrocorax capensis) admitted for rehabilitation at the Southern African Foundation for the Conservation of Coastal Birds (SANCCOB, Cape Town, South Africa). Treatment with primaquine (1 mg/kg PO q24h for 10 days) was effective in rapidly and markedly decreasing the proportion of Babesia-positive blood smears in African penguins and Cape cormorants. No regurgitation, loss of appetite, or any other signs after administration of primaquine were observed during the study period. The use of primaquine can be a particularly advantageous treatment alternative for avian babesiosis in circumstances in which it is not possible to determine confidently whether the intraerythrocytic inclusions seen in blood smears correspond to Babesia or Plasmodium or in cases in which there is a coinfection by Babesia and Plasmodium.

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.

ENVIRONMENTAL FUNGAL LOADS IN AN INDOOR-OUTDOOR AFRICAN PENGUIN ( SPHENISCUS DEMERSUS) EXHIBIT.

Aspergillosis continues to be one of the most important causes of disease in captive penguins. As such, designing exhibits and holding areas that minimize the risk of aspergillosis is of great interest; however, very little has been published regarding this topic. The goal of this study was to assess total fungal spore loads as well as the loads of Aspergillus spp. encountered in multiple indoor and outdoor microenvironments around the exhibit for a large colony of African penguins ( Spheniscus demersus). Air samples were collected via impaction at the microenvironments on a monthly basis over a 1-yr period. Results of this study indicated seasonal trends in both total fungal spore loads as well as Aspergillus spp. loads, with the lowest levels encountered during January through April. During the warmer, more humid spring, summer, and fall months when outdoor microenvironments experienced the highest fungal loads, the air-handling system and the pleated filters used indoors are thought to have reduced the fungal loads in the indoor microenvironments compared with the outdoor microenvironments. Additionally, surrounding planting beds were thought to contribute to the higher total fungal loads and Aspergillus spp. loads in the outdoor microenvironments. Results of this study are useful in understanding the factors that contribute to Aspergillus spp. loads in areas that house penguins, and can be used in guiding design, construction, and landscaping of penguin enclosures.