Biochemical and molecular biomarkers and their association with anthropogenic chemicals in wintering Manx shearwaters (Puffinus puffinus).

Anthropogenic pollution poses a threat to marine conservation by causing chronic toxic effects. Seabirds have contact throughout their lives with pollutants like plastic, metals, polychlorinated biphenyls (PCBs), and organochlorine pesticides such as hexachlorocyclohexanes (HCHs). We assessed 155 Manx shearwaters (Puffinus puffinus) stranded along the Brazilian coast, analyzing associations between organic pollutants, plastic ingestion, biomarkers (transcript levels of aryl hydrocarbon receptor, cytochrome P450-1A-5 [CYP1A5], UDP-glucuronosyl-transferase [UGT1], estrogen receptor alpha-1 [ESR1], and heat shock protein-70 genes) and enzymes activity (ethoxy-resorufin O-deethylase and glutathione S-transferase [GST]). Plastic debris was found in 29 % of the birds. The transcription of UGT1 and CYP1A5 was significantly associated with hexachlorobenzene (HCB) and PCBs levels. ESR1 was associated with HCB and Mirex, and GST was associated with Drins and Mirex. While organic pollutants affected shearwaters more than plastic ingestion, reducing plastic availability remains relevant as xenobiotics are also potentially adsorbed onto plastics.

Highly Pathogenic Avian Influenza A(H5N1) Viruses from Multispecies Outbreak, Argentina, August 2023.

We report full-genome characterization of highly pathogenic avian influenza A(H5N1) clade 2.3.4.4b virus from an outbreak among sea lions (August 2023) in Argentina and possible spillover to fur seals and terns. Mammalian adaptation mutations in virus isolated from marine mammals and a human in Chile were detected in mammalian and avian hosts.

Exploring the relationship between environmental drivers and the manifestation of fibropapillomatosis in green turtles (Chelonia mydas) in eastern Brazil.

Fibropapillomatosis (FP) is a disease characterized by epithelial tumors that can impede life-sustaining activities of sea turtles, especially green turtles (Chelonia mydas). FP is caused by a herpesvirus, but environmental factors are also thought to play a role in triggering FP tumor growth. In this study, we evaluate the epidemiology of FP tumors in green turtles along the coast of Espírito Santo, Brazil, a region where juvenile green turtles are known to aggregate with high FP prevalence. A dataset comprising 2024 beach-cast green turtles recorded through daily beach surveys on 400 km of coastline from 2018 to 2021 (inclusive) was evaluated. FP tumors were recorded in 40.9% of the individuals in this dataset, and presence of FP tumors was predicted by individual variables (presence of marine leeches, stranding code, curved carapace length, body mass-size residual) and characteristics of the stranding site (distance to nearest metallurgical plant, mean sea surface salinity (SSS), annual range of sea surface temperature (SST)). Additionally, a second dataset comprising detailed information about the size and anatomical distribution of tumors in 271 green turtles with FP from the same region was evaluated. Hierarchical clustering analysis revealed these turtles could be classified in three groups according to the anatomical distribution of their tumors, and in turn the group to which each turtle was assigned could be predicted by the study period (2010-2014 vs. 2018-2022) and by characteristics of the stranding/capture site (green turtle stranding density, mean sea surface chlorophyll-a concentration, mean SSS, mean SST, annual range of SST). These results corroborate that individual and environmental factors play a significant role driving FP epidemiology. Furthermore, the results suggest that rather than behaving as a single entity, FP may be seen as a mosaic of distinct anatomical patterns that are not necessarily driven by the same environmental factors.

Haemoproteus jenniae (Haemoproteidae, Haemosporida) infects gulls (Larus spp.) in South Africa, with redescription of Haemoproteus skuae.

Haemoproteus spp. are dipteran-borne protozoa that infect erythrocytes and reticulo-endothelial cells of birds. These parasites are not usually transmitted between birds belonging to different orders. The suborder Lari (order Charadriiformes) comprises ~170 avian species, the majority of which are aquatic, including gulls, terns, auklets, murres and skuas, among others. In spite of the diversity of this avian group, there is limited known diversity of haemosporidian parasites, with only 4 recorded Haemoproteus morphospecies thus far. We examined the blood smears of 21 kelp gulls (Larus dominicanus) captured at a breeding colony in South Africa, as well as Haemoproteus-positive archival blood smears of 15 kelp gulls and 1 Hartlaub's gull (Larus hartlaubii) sampled while under care at seabird rehabilitation facilities in South Africa. Haemoproteus sp. infection was detected in 19% of wild-caught kelp gulls. All parasites from the gulls were morphologically identified as Haemoproteus jenniae, a species previously recorded in Lari birds at the Galapagos Islands (Ecuador), Rocas Atoll (Brazil) and Poland. Gene sequencing uncovered a new cytochrome b lineage, LARDOM01, which was closely related to the previously reported H. jenniae lineage CREFUR01. Additionally, we evaluated a hapantotype blood smear of Haemoproteus skuae, which had been described infecting a brown skua (Catharacta antarctica) in South Africa. We provide a redescription of H. skuae and discuss the morphological characters distinguishing it from H. jenniae. Further research is necessary to improve our knowledge about the host and geographic distribution, health effects and phylogeny of H. jenniae and H. skuae.

Factors determining the number of seabirds impacted by oil spills and the success of their rehabilitation: Lessons learned from Namibia and South Africa.

The coastal waters of Namibia and South Africa have an extensive history of oil spills, with 71 recorded up to 2021. Thirty-nine spills reportedly affected 83,224 seabirds, with African penguins (Spheniscus demersus; 91.0 %) and Cape gannets (Morus capensis; 8.5 %) most affected. Spills affecting seabirds were caused by unknown sources (46 %), bulk/cargo carriers (43 %), tankers (38 %) and ship-to-ship transfers (14 %). The number of penguins oiled was predicted by the breeding population size within 25 to 75 km, but not the volume of oiled spilled, the month or the year. Rehabilitation records from penguins oiled in spills since 2001 reveal that the day of admission (relative to the start of the spill) was predictive of packed cell volume, body mass, and plasma total solids, with the latter two being predictive of rehabilitation success. Our results highlight the importance of rapid monitoring at colonies to locate oiled birds in the event of spills.

PHTHALATE ESTERS (PLASTICIZERS) IN THE UROPYGIAL GLAND AND THEIR RELATIONSHIP TO PLASTICS INGESTION IN SEABIRDS ALONG THE COAST OF ESPÍRITO SANTO, EASTERN BRAZIL.

Plastic ingestion is a problem for seabirds worldwide. In addition to direct health effects such as obstruction or perforation of the gastrointestinal tract, plastic ingestion can also lead to indirect health effects through the release of chemicals that may be absorbed and cause systemic and chronic toxicity. Among chemicals that can be released by plastics are phthalate esters, a group of chemicals widely used as plasticizers or additives to change the physical characteristics of plastics. In this study, three phthalate esters, dimethyl phthalate (DMP), dibuthyl phthalate (DBP), and diethylhexyl phthalate (DEHP), were quantified in the uropygial gland of 48 seabirds from 16 species collected ashore in a tropical region, the coast of Espírito Santo, Eastern Brazil. Including trace levels, DMP was detected in 16 birds (33%) from 10 species, with an average concentration of 0.014 ± 0.005 ng/µl (mean ± SD for individuals with concentrations above the practical level of detection of 0.01 ng/µl). DBP was detected in 15 birds (31%) from 11 species, with an average concentration of 0.049 ± 0.032 ng/µl. DEHP was detected in 21 birds (44%) from 11 species, with an average concentration of 0.115 ± 0.105 ng/µl. DMP concentration in the uropygial gland was positively associated with the presence, number, and mass of plastic items in the upper digestive tract. However, no such relationship was noted for DBP nor DEHP, suggesting the concentration of phthalate compounds in the uropygial gland might not always serve as a reliable proxy for plastic ingestion. In spite of relatively high frequencies of detection, the low concentrations of phthalates detected in this study suggest levels of exposure below known toxicity thresholds. Further studies on the potential adverse effects of phthalate exposure in seabirds are necessary, especially on the reproductive development of embryos and chicks.

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.

Sarcoptic mange outbreak decimates South American wild camelid populations in San Guillermo National Park, Argentina.

Sarcoptic mange epidemics can devastate wildlife populations. In 2014, mange was first detected in vicuñas (Vicugna vicugna) and guanacos (Lama guanicoe) in San Guillermo National Park (SGNP), Argentina. This study describes the temporal dynamics of the outbreak, its effects on the park's wild camelid populations between 2017-2019, and investigates the potential source of the epidemic. From May 2017 to June 2018, transect surveys indicated a sharp decrease in the density of living vicuñas and guanacos by 68% and 77%, respectively. By April 2019 no vicuñas or guanacos were recorded on transect surveys, suggesting their near-extinction in the park. Clinical signs consistent with mange (e.g., intense scratching, hyperkeratosis, alopecia) were observed in 24% of living vicuñas (n = 478) and 33% of living guanacos (n = 12) during surveys, as well as in 94% of vicuña carcasses (n = 124) and 85% of guanaco carcasses (n = 20) examined. Sarcoptes scabiei was identified as the causal agent by skin scrapings, and the cutaneous lesions were characterized by histopathology (n = 15). Genetic characterization revealed that mites recovered from seven vicuñas (n = 13) and three guanacos (n = 11) shared the same genotype, which is consistent with a single source and recent origin of the epidemic. Tracing the potential source, we identified a governmental livestock incentive program which introduced llamas (Lama glama) in areas adjacent to SGNP in 2009, some of which had alopecic scaling consistent with sarcoptic mange. Though at the time of our study no llamas with mange were available for confirmatory sampling, we hypothesize that the introduction of mange-infected llamas may have triggered the outbreak in wild camelids. This unprecedented event in SGNP had devastating effects on dominating herbivores with potentially profound cascading effects at the community and ecosystem levels.

Risk assessment of SARS-CoV-2 in Antarctic wildlife.

The coronavirus disease 2019 (COVID-19) pandemic is caused by the severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2). This pathogen has spread rapidly across the world, causing high numbers of deaths and significant social and economic impacts. SARS-CoV-2 is a novel coronavirus with a suggested zoonotic origin with the potential for cross-species transmission among animals. Antarctica can be considered the only continent free of SARS-CoV-2. Therefore, concerns have been expressed regarding the potential human introduction of this virus to the continent through the activities of research or tourism to minimise the effects on human health, and the potential for virus transmission to Antarctic wildlife. We assess the reverse-zoonotic transmission risk to Antarctic wildlife by considering the available information on host susceptibility, dynamics of the infection in humans, and contact interactions between humans and Antarctic wildlife. The environmental conditions in Antarctica seem to be favourable for the virus stability. Indoor spaces such as those at research stations, research vessels or tourist cruise ships could allow for more transmission among humans and depending on their movements between different locations the virus could be spread across the continent. Among Antarctic wildlife previous in silico analyses suggested that cetaceans are at greater risk of infection whereas seals and birds appear to be at a low infection risk. However, caution needed until further research is carried out and consequently, the precautionary principle should be applied. Field researchers handling animals are identified as the human group posing the highest risk of transmission to animals while tourists and other personnel pose a significant risk only when in close proximity (< 5 m) to Antarctic fauna. We highlight measures to reduce the risk as well as identify of knowledge gaps related to this issue.

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.

Strandings of cetaceans on the Espírito Santo coast, southeast Brazil, 1975-2015.

Espírito Santo state is located on the eastern margin of Brazil, in a transitional tropical-subtropical area (18°S-21°S) dominated by oligotrophic waters. With the exception of humpback whales (Megaptera novaeangliae), the cetacean community of Espírito Santo has been understudied. In addition to the chronic impacts from fisheries, marine pollution, urban development, and coastal habitat degradation, in November 2015 the cetacean communities of Espírito Santo were challenged by the greatest environmental disaster in Brazil's history. The Mariana dam disaster caused 60 million cubic meters of mining waste to be washed into the Doce River, which ultimately flowed to the coastal waters of Espírito Santo, with a high concentration of heavy metals. This study reviews and updates information on cetacean strandings in the state of Espírito Santo (excluding humpback whales) prior to this disaster. From 1975 to September 2015, there were 461 recorded cetacean strandings, representing 20 species. An average 1.18 strandings per 100 km per month were recorded since a state-wide daily beach survey program was implemented in October 2010, contrasting with the 0.14 strandings per 100 km per month in previous years. Six species comprised the majority (94.7%) of stranding events: Guiana dolphin (Sotalia guianensis), Franciscana (Pontoporia blainvillei), rough-toothed dolphin (Steno bredanensis), bottlenose dolphin (Tursiops truncatus), sperm whale (Physeter macrocephalus), and melon-headed whale (Peponocephala electra). Oceanic cetaceans stranded most frequently on the southern portion of Espírito Santo, where the continental platform is narrower, whereas the strandings of coastal cetaceans such as Guiana dolphins and Franciscanas were concentrated near estuaries, especially the Doce River. This is particularly concerning in face of the Mariana dam disaster, which drastically altered the estuarine and coastal environment associated with the Doce River.

First report of avian malaria in a Manx shearwater (Puffinus puffinus).

This study reports the case of a Manx shearwater (Puffinus puffinus) that died from avian malaria while under care at a rehabilitation center in Espírito Santo, Brazil. The bird was rescued on October 2018, and remained under care until it died suddenly on January 2019. A blood smear produced 8 days before death was negative for parasites, whereas a blood smear produced post-mortem revealed a high parasitemia by a parasite resembling Plasmodium cathemerium. The sequence of a 412 bp segment of the cyt-b gene was identical to that of lineage PADOM09, and phylogenetic analysis corroborated that this parasite was closely-related to known lineages of P. cathemerium. The acuteness and severity of the infection documented in this case suggest that seabirds of the order Procellariiformes might be highly susceptible to Plasmodium infections, raising the concern that avian malaria may present a significant threat to their conservation.

Morphological and molecular characterization of Plasmodium cathemerium (lineage PADOM02) from the sparrow Passer domesticus with complete sporogony in Culex pipiens complex.

Avian malaria is a mosquito-borne disease caused by Plasmodium spp. protozoa. Although these parasites have been extensively studied in North America and Eurasia, knowledge on the diversity of Plasmodium, its vectors and avian hosts in Africa is scarce. In this study, we report on natural malarial infections in free-ranging sparrows (Passer domesticus) sampled at Giza Governorate, Egypt. Parasites were morphologically characterized as Plasmodium cathemerium based on the examination of thin blood smears from the avian host. Sequencing a fragment of the mitochondrial cytochrome b gene showed that the parasite corresponded to lineage PADOM02. Phylogenetic analysis showed that this parasite is closely related to the lineages SERAU01 and PADOM09, both of which are attributed to P. cathemerium. Experimental infection of Culex pipiens complex was successful, with ookinetes first detected at 1-day post infection (dpi), oocysts at 4 dpi and sporozoites at 6 dpi. The massive infection of the salivary glands by sporozoites corroborates that Cx. pipiens complex is a competent vector of PADOM02. Our findings confirm that Plasmodium lineage PADOM02 infects sparrows in urban areas along the Nile River, Egypt, and corroborate that Cx. pipiens complex is a highly competent vector for these parasites. Furthermore, our results demonstrate that this lineage corresponds to the morphospecies P. cathemerium and not P. relictum as previously believed.

PROGNOSTIC INDICATORS OF IMMATURE REHABILITATED AFRICAN PENGUINS (SPHENISCUS DEMERSUS) IN SOUTH AFRICA.

The Southern African Foundation for the Conservation of Coastal Birds (Cape Town, South Africa) regularly receives African Penguins (Spheniscus demersus) for rehabilitation. The impact of life history and clinical parameters at admission of 3,975 chicks, 1,687 blues (chicks without downy plumage that are about to fledge or have recently fledged), and 850 fledged juveniles admitted over a 12 yr period (2002-13) were assessed in relation to rehabilitation outcomes using multivariate analysis. Younger chicks were more likely to die during rehabilitation compared with older chicks, and chicks admitted because they had been abandoned by their parents were more likely to die than those admitted preemptively (chicks that were removed when rangers considered their nests were in high-risk areas). Blues admitted because of injury and juveniles admitted because of debilitation, injury, and molt were more likely to die or be euthanized relative to the reference category oiling. Plasmodium infection contributed to natural death for all age groups and to the decision to euthanize chicks, whereas Borrelia infection contributed to both natural death and the decision to euthanize blues. Babesia infection was associated with decreased odds of euthanasia among juveniles. Low hematocrit at admission contributed to natural death of chicks and blues and euthanasia of blues. Low total plasma protein, on the other hand, contributed to natural death in chicks, blues, and juveniles, whereas high total plasma protein had a protective effect against natural death in chicks. These results indicate a need to focus on prevention and early intervention among compromised immature penguins in order to increase the rehabilitation success for this endangered species.

Molecular characterization and lesions associated with Diomedenema diomedeae (Aproctoidea: Desmidocercidae) from grey-headed albatrosses (Thalassarche chrysostoma) on Subantarctic Marion Island.

The Prince Edward Islands are Subantarctic islands in the southwest Indian Ocean that are of global importance as seabird nesting sites, and are breeding grounds for five species of albatrosses (Procellariiformes: Diomedeidae). In March-April 2016 numerous chicks of one of these species, the grey-headed albatross (Thalassarche chrysostoma), were found dead at colonies on Marion Island (46°57'S 37°42'E), the larger of the two Prince Edward Islands. Affected chicks were weak, prostrated, apathetic, had drooping wings, and many eventually died while sitting on the nest. Five carcasses were necropsied, and samples were obtained for pathological and parasitological analysis. Four chicks appeared to have died from starvation, and one died due to air-sac helminthiasis, with extensive hemorrhage in the air sacs and multifocal pyogranulomatous air-sacculitis. The air sac parasites were identified as Diomedenema diomedeae (Aproctoidea: Desmidocercidae). Phylogenetic analysis of the nuclear 18S rRNA gene and mitochondrial COI gene confirmed that D. diomedeae belongs to the suborder Spirurina and showed that it is most closely related to the Diplotriaenidae (superfamily Diplotriaenoidea), a family of parasites that infect the air sacs and subcutaneous tissues of a variety of bird species. To our knowledge this is the first record of the occurrence of a nematode in the respiratory tract of an albatross and the first study to provide DNA sequences for a species of the superfamily Aproctoidea.

Parasitaemia data and molecular characterization of Haemoproteus catharti from New World vultures (Cathartidae) reveals a novel clade of Haemosporida.

BACKGROUND: New World vultures (Cathartiformes: Cathartidae) are obligate scavengers comprised of seven species in five genera throughout the Americas. Of these, turkey vultures (Cathartes aura) and black vultures (Coragyps atratus) are the most widespread and, although ecologically similar, have evolved differences in morphology, physiology, and behaviour. Three species of haemosporidians have been reported in New World vultures to date: Haemoproteus catharti, Leucocytozoon toddi and Plasmodium elongatum, although few studies have investigated haemosporidian parasites in this important group of species. In this study, morphological and molecular methods were used to investigate the epidemiology and molecular biology of haemosporidian parasites of New World vultures in North America. METHODS: Blood and/or tissue samples were obtained from 162 turkey vultures and 95 black vultures in six states of the USA. Parasites were identified based on their morphology in blood smears, and sequences of the mitochondrial cytochrome b and nuclear adenylosuccinate lyase genes were obtained for molecular characterization. RESULTS: No parasites were detected in black vultures, whereas 24% of turkey vultures across all sampling locations were positive for H. catharti by blood smear analysis and/or PCR testing. The phylogenetic analysis of cytochrome b gene sequences revealed that H. catharti is closely related to MYCAMH1, a yet unidentified haemosporidian from wood storks (Mycteria americana) in southeastern USA and northern Brazil. Haemoproteus catharti and MYCAMH1 represent a clade that is unmistakably separate from all other Haemoproteus spp., being most closely related to Haemocystidium spp. from reptiles and to Plasmodium spp. from birds and reptiles. CONCLUSIONS: Haemoproteus catharti is a widely-distributed parasite of turkey vultures in North America that is evolutionarily distinct from other haemosporidian parasites. These results reveal that the genetic diversity and evolutionary relationships of avian haemosporidians are still being uncovered, and future studies combining a comprehensive evaluation of morphological and life cycle characteristics with the analysis of multiple nuclear and mitochondrial genes will be useful to redefine the genus boundaries of these parasites and to re-evaluate the relationships amongst haemosporidians of birds, reptiles and mammals.

PROGNOSTIC INDICATORS OF REHABILITATION OUTCOMES FOR ADULT AFRICAN PENGUINS ( SPHENISCUS DEMERSUS).

: The Southern African Foundation for the Conservation of Coastal Birds facility near Cape Town, South Africa, receives ∼900 African Penguins ( Spheniscus demersus) for rehabilitation every year. Data were analyzed from 3,657 adult African Penguins over a 12-yr period (2002-13), and multivariate logistic regression analysis was used to evaluate whether individual history and clinical parameters upon admission could predict the outcome of rehabilitation. Penguins admitted due to molt or debilitation were more likely to die during rehabilitation than those admitted due to oiling. Individuals admitted during summer and spring were more likely to die during rehabilitation than those admitted during winter. Penguins diagnosed with Plasmodium infection at some point during rehabilitation were more likely to die than those that were consistently negative, and no significant effect was found for other blood parasite infections. Penguins admitted with low body mass, low total plasma protein, or low hematocrit were more likely to die during rehabilitation than those with normal values. With regard to euthanasia, penguins admitted due to molt, debilitation, injury, or other causes and those admitted during spring or with low plasma protein were more likely to be euthanized.

Molecular characterization of Babesia peircei and Babesia ugwidiensis provides insight into the evolution and host specificity of avian piroplasmids.

There are 16 recognized species of avian-infecting Babesia spp. (Piroplasmida: Babesiidae). While the classification of piroplasmids has been historically based on morphological differences, geographic isolation and presumed host and/or vector specificities, recent studies employing gene sequence analysis have provided insight into their phylogenetic relationships and host distribution and specificity. In this study, we analyzed the sequences of the 18S rRNA gene and ITS-1 and ITS-2 regions of two Babesia species from South African seabirds: Babesia peircei from African penguins (Spheniscus demersus) and Babesia ugwidiensis from Bank and Cape cormorants (Phalacrocorax neglectus and P. capensis, respectively). Our results show that avian Babesia spp. are not monophyletic, with at least three distinct phylogenetic groups. B. peircei and B. ugwidiensis are closely related, and fall within the same phylogenetic group as B. ardeae (from herons Ardea cinerea), B. poelea (from boobies Sula spp.) and B. uriae (from murres Uria aalge). The validity of B. peircei and B. ugwidiensis as separate species is corroborated by both morphological and genetic evidence. On the other hand, our results indicate that B. poelea might be a synonym of B. peircei, which in turn would be a host generalist that infects seabirds from multiple orders. Further studies combining morphological and molecular methods are warranted to clarify the taxonomy, phylogeny and host distribution of avian piroplasmids.

Health evaluation of African penguins (Spheniscus demersus) in southern Africa.

The African penguin (Spheniscus demersus) is an endangered seabird that breeds along the coast of Namibia and South Africa, and disease surveillance was identified as a priority for its conservation. Aiming for the establishment of baseline data on the presence of potential pathogens in this species, a comprehensive health assessment (blood smear examination, haematology, biochemistry and serology) was conducted on samples obtained from 578 African penguins at 11 breeding colonies and a rehabilitation centre. There were 68 penguins that were seropositive for at least one of seven pathogens tested: avian encephalomyelitis virus, avian infectious bronchitis virus, avian reovirus, infectious bursal disease virus, Newcastle disease virus, Mycoplasma gallisepticum and Mycoplasma synoviae. All samples were seronegative for avian influenza virus subtypes H5 and H7 and infectious laryngotracheitis virus. The apparent prevalence of Babesia sp. and Borrelia sp. in blood smears was consistent with previous studies. Babesia-infected individuals had a regenerative response of the erythrocytic lineage, an active inflammatory response and hepatic function impairment. These findings indicate that African penguins may be exposed to conservation-significant pathogens in the wild and encourage further studies aiming for the direct detection and/or isolation of these microorganisms.

Isolation and Characterization of IgM and IgY Antibodies from Plasma of Magellanic Penguins (Spheniscus magellanicus).

Infectious diseases such as aspergillosis, avian malaria, and viral infections are significant threats to the conservation of penguins, leading to morbidity and mortality of these birds both in captivity and in the wild. The immune response to such infectious diseases is dependent on different mechanisms mediated by cells and soluble components such as antibodies. Antibodies or immunoglobulins are glycoproteins that have many structural and functional features that mediate distinct effector immune functions. Three distinct classes of antibodies have been identified in birds: immunoglobulin A (IgA), immunoglobulin M (IgM), and immunoglobulin Y (IgY). In this study we aim to establish an efficient laboratory method to obtain IgM and IgY antibodies from plasma samples of healthy adult Magellanic penguins (Spheniscus magellanicus). The protocol was developed combining plasma delipidation, sequential precipitation with caprylic acid and ammonium sulfate, and size-exclusion chromatography. The efficiency of the protocol and the identity of the purified IgM and IgY antibodies were confirmed through enzyme-linked immunosorbent assay, Western blotting, one-dimensional and two-dimensional polyacrylamide gel electrophoresis, and lectin binding assay. Structural and physicochemical properties of IgM and IgY from Magellanic penguins were consistent with those of other avian species. This purification protocol will allow for more detailed studies on the humoral immunity of penguins and for the development of high specificity serologic assays to test Magellanic penguins for infectious pathogens.