Spheniscus Demersus Research Articles

Plasmodium matutinum Causing Avian Malaria in Lovebirds (Agapornis roseicollis) Hosted in an Italian Zoo.

Avian malaria is a worldwide distributed, vector-born disease of birds caused by parasites of the order Haemosporida. There is a lack of knowledge about the presence and pathogenetic role of Haemosporida in Psittacidae. Here we report a case of avian malaria infection in lovebirds (Agapornis roseicollis), with the genetic characterization of the Plasmodium species involved. The birds were hosted in a zoo located in Italy, where avian malaria cases in African penguins (Spheniscus demersus) were already reported. Animals (n = 11) were submitted for necropsy after sudden death and were subjected to further analyses including histopathology, bacteriology, and PCR for the research of haemosporidians. Clinical history, gross lesions and histopathological observation of schizonts, together with positive PCR results for Plasmodium spp., demonstrated that avian malaria was the cause of death for one animal and the possible cause of death for the other nine. The sequences obtained were compared using BLAST and analyzed for similarity to sequences available at the MalAvi database. Genetic analyses demonstrated a 100% nucleotide identity to Plasmodium matutinum LINN1 for all the obtained sequences. To our knowledge, this is the first report describing avian malaria in lovebirds.

Influence of ontogeny on stable isotope ratios and trophic discrimination factors of African penguin (Spheniscus demersus) tissues

Influence of ontogeny on stable isotope ratios and trophic discrimination factors of African penguin (Spheniscus demersus) tissues

Avian malaria affecting penguins in zoological gardens, aquariums and wildlife parks in the UK

Avian malaria has caused mortalities in captive penguins worldwide and it is a conservation threat for some wild penguins. The experience of staff working on penguins is highly valuable for the improvement of captivity conditions. An online questionnaire was designed to collect avian malaria information in penguins in the UK. Volunteers with diverse experience reviewed it for scientific value and clarity, and it was supported by the British and Irish Association of Zoos and Aquariums. Forty-two institutions were contacted by email, obtaining 27 complete and five partial responses. Eighteen institutions reported avian malaria in 38 suspected and 37 confirmed outbreaks in the past 20 years, mainly occurring in the summer and recent years. Proportional risk was higher in the south east region of England. Humboldt and African penguins were more commonly affected, and prevalence was mostly low or medium, but lethality was high. Different treatment protocols were reported with inconsistent outcomes. Penguins' susceptibility, their translocation across institutions, and the wide distribution of avian malaria make this disease a constant threat. Preventive and control measures are recommended, including testing for early diagnosis and treatment, avoiding prolonged stressful events in penguins, and monitoring and controlling mosquito vectors.

Passive acoustic monitoring of the endangered African Penguin (Spheniscus demersus) using autonomous recording units and ecoacoustic indices

African Penguins Spheniscus demersus are endangered and declining seabirds which make extensive use of vocal signals for intra‐specific vocal communication. Accordingly, passive acoustic monitoring tools could be developed as robust population monitoring methods that cause minimal disturbance to the birds. In this study, we collected soundscape recordings at the Stony Point penguin colony (Betty’s Bay, South Africa) during the breeding season in 2019 to document the circadian rhythms of vocal activity of this species and to investigate whether the magnitude of variation of three different ecoacoustic indices correlates with the number of ecstatic and mutual display songs counted in recordings, which might inform on the breeding activity of the colony. Indeed, whereas ecstatic display songs are produced by males during intersexual competition and territorial defence, mutual display songs are given by parents returning to the nest after foraging trips. We found that the vast majority of the display songs (> 80%) occurred between 04:00–08:00 and 17:30–21:30 h. We also found that the Acoustic Entropy Index was a good predictor of the number of penguins’ songs within a recording. Overall, our study shows that African Penguins vocalizations have the potential to assist the monitoring of this species while minimizing disturbance.

Long-term stability of vocal individuality cues in a territorial and monogamous seabird.

The stability of individual acoustic features is fundamental in social species, and more importantly in monogamous and territorial species, showing long-term fidelity both to the partner and the breeding site. In this study, the stability over time of two discrete vocal types was investigated in the African penguin (Spheniscus demersus), a monogamous and territorial seabird. Contact calls and ecstatic display songs were recorded from an ex situ colony in 2017 and in 2020. For each vocalisation, we measured 14 spectral and temporal acoustic parameters related to both source and filter components. Two separate leave-one-out cross-validated Discriminant Function Analyses (DFA) were then performed, generating the discriminant functions from the vocalisations collected in 2017 to classify those recorded in 2020. The DFA correctly classified 62% of the contact calls (10 subjects) and 80.9% of the ecstatic display songs (seven subjects) according to the correct emitter, showing that acoustic cues to individuality encoded in both vocal types remained unchanged over four consecutive breeding seasons. We suggest that, in this monogamous and territorial bird species, individual acoustic stability could be selected for to identify groupmates and neighbours over the years and to help couples to reunite in consecutive breeding seasons, increasing individual fitness.

Treatment with chloroquine is retinotoxic in captive African penguins (Speniscus demersus). Attenuation and recovery of electroretinographic responses.

To evaluate the effect of prophylactic anti-malarial chloroquine treatment, and its cessation, on electroretinographic (ERG) responses of captive African penguins. A brief ERG protocol ("QuickRetCheck") was recorded under mesopic conditions with manual restraint and no sedation or pupil dilation. Birds were recorded on two separate occasions, first while being treated with a daily chloroquine dose of 10mg/kg for 12days (n=15, treatment group) and second after 4months without chloroquine treatment (n=6, off-treatment group). Three birds were recorded on both occasions. Three other birds from the flock that died were studied histopathologically. Scotopic responses were unmeasurable in either recording and therefore were not analyzed. Mean a- and b-wave amplitudes of the mixed rod-cone responses to standard (3cd·s/m2 ) and high (10cd·s/m2 ) intensity flashes were higher in the off-treatment group. No difference in implicit times was observed. Sex, age, and number of previous chloroquine treatments did not affect ERG responses. Histopathology revealed Plasmodium spp.in the lungs, liver, and brain, but not in the eyes, of the necropsied birds, and there were no signs of retinitis or retinopathy. Daily chloroquine treatment was associated with attenuated ERG responses in penguins, which improve following cessation of treatment. Further work is warranted to establish a chloroquine dose that is efficacious yet has minimal adverse effects. Our results suggest that ERG responses of captive penguins undergoing ERG for any indication (such as prior to cataract surgery), must be evaluated in light of the birds' anti-malaria treatment status.

The Eyes of an African Penguin (Spheniscus demersus): General Morphology and Ophthalmopathology.

The Eyes of an African Penguin (Spheniscus demersus): General Morphology and Ophthalmopathology.

Plasmodium matutinum Transmitted by Culex pipiens as a Cause of Avian Malaria in Captive African Penguins (Spheniscus demersus) in Italy.

Avian malaria is a parasitic disease of birds caused by protozoa belonging to the genus Plasmodium, within the order Haemosporida. Penguins are considered particularly susceptible, and outbreaks in captive populations can lead to high mortality. We used a multidisciplinary approach to investigate the death due to avian malaria, occurred between 2015 and 2019, in eight African penguins (Spheniscus demersus) kept in two Italian zoos located in central Italy, and situated about 30 km apart. We also provided information about the presence and circulation of Plasmodium spp. in mosquitoes in central Italy by sampling mosquitoes in both zoos where penguin mortalities occurred. In the eight dead penguins, gross and histopathological lesions were consistent with those previously observed by other authors in avian malaria outbreaks. Organs from dead penguins and mosquitoes collected in both zoos were tested for avian malaria parasites by using a PCR assay targeting the partial mitochondrial conserved region of the cytochrome b gene. Identification at species level was performed by sequencing analysis. Plasmodium matutinum was detected in both dead penguins and in mosquitoes (Culex pipiens), while Plasmodium vaughani in Culex pipiens only. Parasites were not found in any of the PCR tested Aedes albopictus samples. Based on our phylogenetic analysis, we detected three previously characterized lineages: Plasmodium matutinum LINN1 and AFTRU5, P. vaughani SYAT05. In Culex pipiens we also identified two novel lineages, CXPIP32 (inferred morphospecies Plasmodium matutinum) and CXPIP33 (inferred morphospecies P. vaughani). Significantly, LINN1 and AFTRU5 were found to be associated to penguin deaths, although only LINN1 was detected both in penguins (along the years of the study) and in Culex pipiens, while AFTRU5 was detected in a single penguin dead in 2017. In conclusion, in our study Plasmodium matutinum was found to cause avian malaria in captive penguins kept in Europe, with Culex pipiens being its most probable vector. Our results are in agreement with previous studies suggesting that Culex pipiens is one of the main vectors of Plasmodium spp. in Europe and the Northern Hemisphere. Zoos maintaining captive penguins in temperate areas where Culex pipiens is abundant should be well aware of the risks of avian malaria, and should put every effort to prevent outbreaks, in particular during the periods when the number of vectors is higher.

Frame‐by‐frame annotation of video recordings using deep neural networks

AbstractVideo data are widely collected in ecological studies, but manual annotation is a challenging and time‐consuming task, and has become a bottleneck for scientific research. Classification models based on convolutional neural networks (CNNs) have proved successful in annotating images, but few applications have extended these to video classification. We demonstrate an approach that combines a standard CNN summarizing each video frame with a recurrent neural network (RNN) that models the temporal component of video. The approach is illustrated using two datasets: one collected by static video cameras detecting seal activity inside coastal salmon nets and another collected by animal‐borne cameras deployed on African penguins, used to classify behavior. The combined RNN‐CNN led to a relative improvement in test set classification accuracy over an image‐only model of 25% for penguins (80% to 85%), and substantially improved classification precision or recall for four of six behavior classes (12–17%). Image‐only and video models classified seal activity with very similar accuracy (88 and 89%), and no seal visits were missed entirely by either model. Temporal patterns related to movement provide valuable information about animal behavior, and classifiers benefit from including these explicitly. We recommend the inclusion of temporal information whenever manual inspection suggests that movement is predictive of class membership.

Enhanced Virulence of Aeromonas hydrophila Is Induced by Stress and Serial Passaging in Mice.

Simple SummaryAeromonas hydrophila, which is an opportunistic zoonotic bacterium, has the ability to infect animals with injuries involving the condition of the aquatic environments. Factors including poor sanitation and water quality, stress, overcrowding, and rough handling can make animals more sensitive to infections and trigger outbreaks of A. hydrophila. A. hydrophila was previously isolated from an African black-footed penguin that died while in captivity at a zoo, following clinical signs of depression and anorexia, and in this study, we investigated the effect of stress and serial passaging in mice on A. hydrophila virulence. Serial passaging in mice enhanced the virulence of A. hydrophila, and A. hydrophila infection combined with administration of stress hormones or fasting increased mortality.Aeromonas hydrophila was isolated from an African black-footed penguin (Spheniscus demersus) that died while in zoo captivity. At necropsy, the virulence of A. hydrophila appeared to be enhanced by stress, so was assessed in the presence of in vitro and in vivo stressors and serial passaging in mice. Virulence genes from the isolate were amplified by PCR. In vitro assays were conducted to test the hemolytic activity, cytotoxicity, and effect of stress hormones on A. hydrophila virulence. In vivo assays were conducted to test the stress effect on mortality of A. hydrophila-infected mice and virulence in mice. Two virulence genes coding for hemolysin (ahh1) and aerolysin (aerA) were detected, and the cytotoxic potential of the isolate was demonstrated in baby hamster kidney and Vero cells. Some or all mice inoculated with A. hydrophila and exposed to stress hormones (epinephrine and norepinephrine) or low temperature died, while mice inoculated with A. hydrophila and exposed to fasting or agitation stressors or no stressors survived. We concluded that stress can be fatal in mice experimentally infected with A. hydrophila and that serial passaging in mice dramatically enhances the virulence of A. hydrophila.

Understanding impacts of zoo visitors: Quantifying behavioural changes of two popular zoo species during COVID-19 closures

Visitors are normally a prominent and constant feature in a zoo animals’ environment with more than 700 million people visiting zoos and aquariums worldwide, annually. Animal-visitor interactions can be enriching and stimulating and are now considered within the Five Domains of animal welfare assessment. Zoo closures as a result of COVID-19 provided a unique opportunity to monitor the impact of abrupt and prolonged removal of visitors on two popular zoo species. Data were collected at four facilities (n = 3 slender-tailed meerkats, n = 1 African penguin) during COVID-19 zoo closures and up to one month following reopening to the public. Meerkats showed increased positive social interactions, increased alert behaviours, and reduced environmental interactions in the first month post-opening, as compared to closure periods. They also used more of their enclosures during periods of closure and spent longer than would be expected in zones furthest from visitor viewing areas when facilities reopened. African penguins showed no behavioural change between open and closure periods. Enclosure usage during both observation periods was relatively even and no differences were observed in enclosure use between open and closure periods. These results will enable an advanced understanding of the impact that people have on the behaviour of zoo animals, which has ramifications for animals used in close encounters and other ‘visitor experiences’ in the future. Understanding relationships between animals and people is applicable in all managed animal settings. The results from this study are of practical use in managing visitor access to animals moving forwards, including enclosure location and design, to ensure a positive visitor experience that does not negatively impact animal behaviour.

Genomic Characterisation of a Novel Avipoxvirus Isolated from an Endangered Yellow-Eyed Penguin (Megadyptes antipodes).

Emerging viral diseases have become a significant concern due to their potential consequences for animal and environmental health. Over the past few decades, it has become clear that viruses emerging in wildlife may pose a major threat to vulnerable or endangered species. Diphtheritic stomatitis, likely to be caused by an avipoxvirus, has been recognised as a significant cause of mortality for the endangered yellow-eyed penguin (Megadyptes antipodes) in New Zealand. However, the avipoxvirus that infects yellow-eyed penguins has remained uncharacterised. Here, we report the complete genome of a novel avipoxvirus, penguinpox virus 2 (PEPV2), which was derived from a virus isolate obtained from a skin lesion of a yellow-eyed penguin. The PEPV2 genome is 349.8 kbp in length and contains 327 predicted genes; five of these genes were found to be unique, while a further two genes were absent compared to shearwaterpox virus 2 (SWPV2). In comparison with penguinpox virus (PEPV) isolated from an African penguin, there was a lack of conservation within the central region of the genome. Subsequent phylogenetic analyses of the PEPV2 genome positioned it within a distinct subclade comprising the recently isolated avipoxvirus genome sequences from shearwater, canary, and magpie bird species, and demonstrated a high degree of sequence similarity with SWPV2 (96.27%). This is the first reported genome sequence of PEPV2 from a yellow-eyed penguin and will help to track the evolution of avipoxvirus infections in this rare and endangered species.

Non-invasive hormone monitoring as a robust method for determining adrenocortical activity in injured, emaciated and oil-contaminated African penguins undergoing rehabilitation

Anthropogenic activity is a major driver of seabird injury and mortality in the 21st century. Although most seabirds perish within the natural environment as a result of human activities, some are rescued and admitted to rehabilitation centres. Despite the considerable number of admissions, little is known regarding the physiological response seabirds have to specific admission reasons and the rehabilitation process. In this study, we aimed to determine the effect of injury, emaciation, oiling, individual removal from the natural environment and the rehabilitation process on the physiological stress response of the African penguin (Spheniscus demersus). Urofaecal samples were collected from African penguins throughout a three-stage rehabilitation process and quantified for glucocorticoid metabolites (ufGCM). The three stages included an initial ICU crate stage (Stage 1), an ICU pen stage (Stage 2) and a rehabilitation stage (Stage 3). Data were analysed using a generalised linear model in order to determine the effect of admission reason, age and rehabilitation stage (Stage 1, 2 and 3) on ufGCM levels. Although the model indicated that only Stage 1 was a significant driver of adrenocortical activity in the study population, ufGCM levels of injured and emaciated animals within Stage 1 were considerably higher than those of birds with any other admission reason across all stages. This is the first study examining the causes and effect of rehabilitation on the physiological stress response in African penguins. Enhanced care and attention should be given to rescued individuals, especially during the first stage of rehabilitation, to reduce perception of additional stressors and thus increase the chance of full recovery.

Urofaecal glucocorticoid metabolite concentrations in African penguin (Spheniscus demersus) chick populations experiencing different levels of human disturbance.

Despite the importance of ecotourism in species conservation, little is known about the industry’s effects on wildlife. In South Africa, some African penguin (Spheniscus demersus) colonies have become tourist attractions. The species is globally endangered, with population sizes decreasing over the past 40 years. As African penguin chicks are altricial and unable to move away from anthropogenic stressors, it is important to evaluate the effect of tourist activities on baseline glucocorticoid levels as a measure of potential disturbance. Chicks at three study sites within two breeding colonies (Robben Island, Stony Point), with varying levels of exposure to tourism (low/moderate/high) were monitored. Urofaecal samples were collected to determine urofaecal glucocorticoid metabolite (ufGCM) concentrations as an indication of baseline stress physiology. Morphometric measurements were taken to compare body condition between sites. Penguin chicks experiencing low, infrequent human presence had significantly higher mean (± standard deviation) ufGCM levels [1.34 ± 1.70 μg/g dry weight (DW)] compared to chicks experiencing both medium (0.50 ± 0.40 μg/g DW, P = 0.001) and high levels of human presence (0.57 ± 0.47 μg/g DW, P = 0.003). There was no difference in chick body condition across sites. These results suggest that exposure to frequent human activity may induce habituation/desensitization in African penguin chicks. Acute, infrequent human presence was likely an important driver for comparatively higher ufGCM levels in chicks, though several other environmental stressors may also play an important role in driving adrenocortical activity. Nevertheless, as unhabituated chicks experiencing infrequent anthropogenic presence showed significantly higher ufGCM levels, managers and legislation should attempt to minimize all forms of activity around important breeding colonies that are not already exposed to regular tourism. Although the results of this study are crucial for developing enhanced conservation and management protocols, additional research on the long-term effect of anthropogenic activities on African penguin physiology is required.

Capillary Electrophoresis Assessment of Plasma Protein Changes in an African Penguin (Spheniscus demersus) With Aspergillosis

A decrease of avian biodiversity in the African continent has been the result of anthropogenic pressure in the region. This has resulted in the African penguin (Spheniscus demersus) being placed on the endangered species list and requires conservation efforts to maintain its free-ranging population and placement under managed care. In the latter environment, infection by Aspergillus fumigatus can be common. The diagnosis and treatment of this fungal disease in birds has presented with many difficulties, largely due to the diversity and limited knowledge that exists about this species. In this study, we implement a high-resolution capillary electrophoresis system for the fractionation of African penguin plasma, followed by mass spectrometry analysis for the identification of proteins associated with aspergillosis. Several protein differences were revealed, including changes in acute phase proteins and lipid metabolism. In addition, our results demonstrated that fibrinogen β chain is a protein largely present during the inflammatory process in an African penguin infected with A. fumigatus. These findings present a new avenue for the measurement of plasma proteins as a potential method for identifying important biomarkers to aid in monitoring African penguin health.

Seasonal changes in reproductive anatomy and gonadal hormone concentrations of African penguins (Spheniscus demersus)

Several standard descriptions of the avian male and female reproductive tract have been reported, including effects of age, stage of reproductive maturity and gonadal hormone concentrations. Limited information on penguin reproductive biology and a lack of information on the African penguin (Spheniscus demersus) necessitated a detailed description of salient structural features of this species and provided an opportunity to evaluate seasonal changes in gonadal steroid hormone concentrations. Tissues from 36 males (adults and juveniles) and 29 females (adults and juveniles) were used for macro-anatomical descriptions and histology of the testes and ovaries. In addition, concentrations of gonadal steroid hormones for eight captive African penguins (four females and four males) were quantified during two breeding and one non-breeding season. The testes were asymmetrical, with the right testis having smaller dimensions compared to the left testis. Marked spermatogenic cellular associations and spermatid developmental stages were present in adult testes only during the breeding season. There was variation in the dimensions of the single ovary during follicular development related to the age and breeding status of the females. Testosterone, dihydrotestosterone, and estradiol concentrations fluctuated during the breeding and non-breeding periods, with males and females having similar steroid concentrations. The results from this study confirm that the breeding status in African penguins can be deduced based on testicular and ovarian histological structures. The results from the present study focused on African penguin reproductive biology should be considered in management strategies for the conservation of the species.

Intercolony health evaluation of wild African penguins Spheniscus demersus, in relation to parasites, along the southwest coast of South Africa

Clinical parameters of African penguins Spheniscus demersus have been recorded mostly from birds admitted to rehabilitation centres and are typically presented as mean values across a region. It is uncertain whether these values are representative of wild penguins and whether they are influenced by parasite infestations. We assessed general clinical parameters from 793 African penguins (210 adults and 583 chicks) at five South African colonies, in 2016 and 2017, and then established the relationship between those values and parasite infestations. Penguins at Dassen Island had indications of a different health status: lower body condition and total plasma protein, and higher haematocrit and ectoparasite richness. Overall, haematocrit values were lower for the mainland colonies than for the island colonies. All clinical parameters were significantly lower in spring than in autumn/winter. Both ectoparasite and haemoparasite taxon richness were negatively related to haematocrit values, while helminth taxon richness was positively related to body mass and haematocrit. At the Stony Point colony, tick abundance and ectoparasite and haemoparasite taxon richness were associated with lower haematocrit in adults and chicks. This study highlights intercolony variations in the health status of African penguins and potential factors (such as parasite infestations and/or food-resource accessibility) that might affect specific colonies, thereby helping to focus conservation efforts.

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.

Mosquito identification and haemosporidian parasites detection in the enclosure of the African penguins (Spheniscus demersus) at the SANBI zoological garden.

The National Zoological Gardens (NZG) is a facility of the South African National Biodiversity Institute (SANBI) and the largest zoo in southern Africa. Among the 9000 captive animals kept by the NZG, is the endangered African penguin (Spheniscus demersus). There have been several post-mortem reports on deaths of penguins in the NZG due to haemosporidian infections, however, the haemosporidian lineages involved and possible insect vector are unknown. Haemosporidians are apicomplexan parasites that infect vertebrates through blood-sucking dipteran insects. Therefore, the current study aimed to identify mosquitoes that are potential vectors found within the African penguin enclosure as well as to detect the haemosporidian parasites from these insects using nested-PCR and real-time PCR (qPCR) analyses. Mosquito samples were collected using an overnight UV-light trap setup for 3 months. From the 65 pooled samples representing 325 mosquitoes, morphological and molecular analysis showed that Culex pipiens (52.31%) was the dominant species followed by Cx.theileri (30.77%) and Cx. quinquefasciatus (16.92%). Nested-PCR detected parasite DNA of Leucocytozoon sp. and Plasmodium sp. The Cx. pipiens had the highest minimum infection rate (MIR) of 5.88% by nested-PCR and 9.41% by qPCR whilst Cx. quinquefasciatus had MIR of 3.64% in both assays and no haemosporidian parasites were detected from Cx.theileri. One Cx. pipiens sample had a co-infection of both Plasmodium sp. and Leucocytozoon sp. detected by nested-PCR. These findings suggest that effective control measures for blood-sucking dipteran insects is required at the NZG and more studies should be conducted to determine the actual prevalence of these haemosporidian parasites among other bird species within NZG.

Outbreak of cryptosporidiosis in African penguins Spheniscus demersus.

Cryptosporidium spp. are parasitic intracellular protozoa that infect the digestive, respiratory, and urinary tracts of vertebrates. The disease affects many different avian species across all continents, and >25 species and genotypes of Cryptosporidium have been documented infecting birds. We report on an outbreak of cryptosporidiosis in African penguin Spheniscus demersus chicks admitted to a rehabilitation center in South Africa from February 2012 to October 2013. Eighteen cases were confirmed through histopathology. The most frequent clinical signs were regurgitation (78%), dyspnea (72%), decreased weight gain or weight loss (72%), and lethargy (50%). Clinical signs began 8-46 d after hatching or admission (median: 13 d), and death followed 1-41 d after the onset of clinical signs (median: 13.5 d). The most frequent necropsy findings were stomach distended with undigested food or gas (78%), mildly reddened lungs (56%), spleen petechial hemorrhage (44%), and kidney congestion (39%). The most frequent histopathological findings were necrotic bursitis (89%), necrotic enteritis (83%), and bursal atrophy (67%). Small round or oval basophilic bodies (3-5 µm diameter) consistent with Cryptosporidium sp. were closely associated with the surface of the epithelial cells or in the lumen of the bursa (89%), large intestine (61%), small intestine (44%), trachea (22%), and ventriculus (6%). Transmission electron microscopy of 1 case confirmed that these organisms were Cryptosporidium sp. To our knowledge, this is the first report of cryptosporidiosis in penguins, raising concern of the potential implications for the conservation of these species.