Serological screening for Brucella spp. and Leptospira spp. antibodies in southern elephant seals Mirounga leonina from Elephant Island, Antarctica, in 2003 and 2004.

Brucella spp. and Leptospira spp. antibodies were surveyed in 35 southern elephant seals (SESs) Mirounga leonina at Elephant Island (South Shetland Islands), western Antarctic peninsula, in the Austral summer of 2003 and 2004. The rose Bengal test and a commercial competitive ELISA (c-ELISA) were used to detect Brucella spp. exposure, and the microscopic agglutination test (MAT) with 22 live serovars was used to determine anti-Leptospira spp. antibodies. We found evidence of Brucella spp. exposure in 3 of 35 (8.6%) SESs tested via the c-ELISA displaying high percentage inhibition (PI), similar to other studies in pinnipeds in which Brucella spp. antibodies have been determined. Two of the 3 positives were pups (PI = 70.4 and 86.6%), while the third was an adult female (PI = 48.8%). The 3 c-ELISA positive SESs were additionally tested via the serum agglutination test but were found to be negative. All individuals were negative for antibodies against 22 Leptospira spp. serovars by MAT. These results contribute to the knowledge and monitoring of zoonotic pathogens with epizootic potential in Southern Ocean pinnipeds. Given the potential impact that pathogens may have on the abundance of wild (sometimes threatened and endangered) populations, constant monitoring and surveillance are required to prevent pathogen spread, particularly under forecast climate change scenarios.

Toxoplasmosis and Sarcocystis spp. infection in wild pinnipeds of the Brazilian coast.

The protozoans Toxoplasma gondii and Sarcocystis spp. (Sarcocystidae: Apicomplexa) affect a wide variety of vertebrates. Both have been reported to infect pinnipeds, with impacts on health ranging from inapparent to fulminant disease and death. However, little is known regarding their infections and associated pathology in South American pinnipeds. We used histological techniques to survey for the presence of T. gondii and Sarcocystis spp. in 51 stranded pinnipeds from Brazil. Immunohistochemical and molecular assays were employed in those cases consistent with Sarcocystidae infection. T. gondii cysts were detected in the central nervous system and heart of a South American fur seal Arctocephalus australis, associated with meningoencephalitis, myocarditis and endocarditis, and confirmed by immunohistochemistry. Additionally, this animal presented Sarcocystis sp. cysts in brain and heart tissues. Four additional specimens-2 Subantarctic fur seals A. tropicalis, an Antarctic fur seal A. gazella and another South American fur seal-presented intrasarcoplasmic cysts compatible with Sarcocystis spp. in muscle samples. There was no inflammation associated with the Sarcocystis spp. tissue cysts and all cysts were negative for S. neurona immunohistochemistry. The B1 gene of T. gondii was amplified in the 5 pinnipeds infected by Sarcocystidae protozoans. To our knowledge, this is the first report of toxoplasmosis in wild South American pinnipeds and of Sarcocystis spp. in South American fur seals. Detection of terrestrial parasites in aquatic mammals could be an indicator of their presence in the marine environment.

Identification of a Novel Adélie Penguin Circovirus at Cape Crozier (Ross Island, Antarctica).

Understanding the causes of disease in Antarctic wildlife is crucial, as many of these species are already threatened by environmental changes brought about by climate change. In recent years, Antarctic penguins have been showing signs of an unknown pathology: a feather disorder characterised by missing feathers, resulting in exposed skin. During the 2018-2019 austral summer breeding season at Cape Crozier colony on Ross Island, Antarctica, we observed for the first time an Adélie penguin chick missing down over most of its body. A guano sample was collected from the nest of the featherless chick, and using high-throughput sequencing, we identified a novel circovirus. Using abutting primers, we amplified the full genome, which we cloned and Sanger-sequenced to determine the complete genome of the circovirus. The Adélie penguin guano-associated circovirus genome shares <67% genome-wide nucleotide identity with other circoviruses, representing a new species of circovirus; therefore, we named it penguin circovirus (PenCV). Using the same primer pair, we screened 25 previously collected cloacal swabs taken at Cape Crozier from known-age adult Adélie penguins during the 2014-2015 season, displaying no clinical signs of feather-loss disorder. Three of the 25 samples (12%) were positive for a PenCV, whose genome shared >99% pairwise identity with the one identified in 2018-2019. This is the first report of a circovirus associated with a penguin species. This circovirus could be an etiological agent of the feather-loss disorder in Antarctic penguins.

Toxoplasma gondii in cetaceans of Brazil: a histopathological and immunohistochemical survey.

Toxoplasmosis is a parasitic disease caused by the protozoan Toxoplasma gondii. In cetaceans, T. gondii infection is a significant cause of morbidity and mortality. Despite the worldwide range and broad cetacean host record of T. gondii infection, there is limited information on toxoplasmosis in cetaceans from the Southern hemisphere. We investigated the occurrence of T. gondii by histopathology and immunohistochemistry in tissue samples of 185 animals comprising 20 different cetacean species from Brazil. Three out of 185 (1.6%) animals presented T. gondii-associated lesions: a captive killer whale Orcinus orca, a free-ranging common bottlenose dolphin Tursiops truncatus and a free-ranging Guiana dolphin Sotalia guianensis. The main lesions observed in these animals were necrotizing hepatitis, adrenalitis and lymphadenitis associated with protozoal cysts or extracellular tachyzoites presenting immunolabeling with anti-T. gondii antibodies. This study widens the spectrum of species and the geographic range of this agent in Brazil, and provides the first reports of T. gondii infection in a captive killer whale and in a free-ranging common bottlenose dolphin in South America.

Toxoplasma gondii in cetaceans of Brazil: a histopathological and immunohistochemical survey / Toxoplasma gondii em cetáceos do Brasil: estudo histopatológico e imuno-histoquímico

Abstract Toxoplasmosis is a parasitic disease caused by the protozoan Toxoplasma gondii. In cetaceans, T. gondii infection is a significant cause of morbidity and mortality. Despite the worldwide range and broad cetacean host record of T. gondii infection, there is limited information on toxoplasmosis in cetaceans from the Southern hemisphere. We investigated the occurrence of T. gondii by histopathology and immunohistochemistry in tissue samples of 185 animals comprising 20 different cetacean species from Brazil. Three out of 185 (1.6%) animals presented T. gondii-associated lesions: a captive killer whale Orcinus orca, a free-ranging common bottlenose dolphin Tursiops truncatus and a free-ranging Guiana dolphin Sotalia guianensis. The main lesions observed in these animals were necrotizing hepatitis, adrenalitis and lymphadenitis associated with protozoal cysts or extracellular tachyzoites presenting immunolabeling with anti-T. gondii antibodies. This study widens the spectrum of species and the geographic range of this agent in Brazil, and provides the first reports of T. gondii infection in a captive killer whale and in a free-ranging common bottlenose dolphin in South America.

Resumo Toxoplasmose é uma doença parasitária causada pelo protozoário Toxoplasma gondii. A infecção por T. gondii é uma causa significativa de morbidade e mortalidade, nos cetáceos. Apesar da abrangência mundial e amplo registro de espécies de cetáceos infectadas por T. gondii, informações sobre toxoplasmose em cetáceos do hemisfério sul são limitadas. Neste estudo pesquisou-se por meio de histopatologia e imuno-histoquímica a ocorrência de T. gondii em amostras de tecido de 185 animais, compreendendo 20 diferentes espécies de cetáceos que ocorrem no Brasil. Três dos 185 (1,6%) animais apresentaram lesões associadas a T. gondii: uma orca Orcinus orca mantida em cativeiro, um golfinho-nariz-de-garrafa Tursiops truncatus e um boto-cinza Sotalia guianensis de vida livre. As principais lesões observadas nesses animais foram hepatite, adrenalite e linfadenite necrotizantes associadas a cistos protozoários ou taquizoítos extracelulares, marcados com anticorpos anti-T. gondii. O presente estudo amplia o espectro de espécies susceptíveis a esse agente e o seu alcance geográfico no Brasil, fornecendo o primeiro relato da infecção por T. gondii em uma orca mantida em cativeiro e em um golfinho-nariz-de-garrafa de vida livre na América do Sul.

Unique genome organization of non-mammalian papillomaviruses provides insights into the evolution of viral early proteins.

The family Papillomaviridae contains more than 320 papillomavirus types, with most having been identified as infecting skin and mucosal epithelium in mammalian hosts. To date, only nine non-mammalian papillomaviruses have been described from birds (n = 5), a fish (n = 1), a snake (n = 1), and turtles (n = 2). The identification of papillomaviruses in sauropsids and a sparid fish suggests that early ancestors of papillomaviruses were already infecting the earliest Euteleostomi. The Euteleostomi clade includes more than 90 per cent of the living vertebrate species, and progeny virus could have been passed on to all members of this clade, inhabiting virtually every habitat on the planet. As part of this study, we isolated a novel papillomavirus from a 16-year-old female Adélie penguin (Pygoscelis adeliae) from Cape Crozier, Ross Island (Antarctica). The new papillomavirus shares ∼64 per cent genome-wide identity to a previously described Adélie penguin papillomavirus. Phylogenetic analyses show that the non-mammalian viruses (expect the python, Morelia spilota, associated papillomavirus) cluster near the base of the papillomavirus evolutionary tree. A papillomavirus isolated from an avian host (Northern fulmar; Fulmarus glacialis), like the two turtle papillomaviruses, lacks a putative E9 protein that is found in all other avian papillomaviruses. Furthermore, the Northern fulmar papillomavirus has an E7 more similar to the mammalian viruses than the other avian papillomaviruses. Typical E6 proteins of mammalian papillomaviruses have two Zinc finger motifs, whereas the sauropsid papillomaviruses only have one such motif. Furthermore, this motif is absent in the fish papillomavirus. Thus, it is highly likely that the most recent common ancestor of the mammalian and sauropsid papillomaviruses had a single motif E6. It appears that a motif duplication resulted in mammalian papillomaviruses having a double Zinc finger motif in E6. We estimated the divergence time between Northern fulmar-associated papillomavirus and the other Sauropsid papillomaviruses be to around 250 million years ago, during the Paleozoic-Mesozoic transition and our analysis dates the root of the papillomavirus tree between 400 and 600 million years ago. Our analysis shows evidence for niche adaptation and that these non-mammalian viruses have highly divergent E6 and E7 proteins, providing insights into the evolution of the early viral (onco-)proteins.

IDENTIFICATION OF RESPIRATORY AND GASTROINTESTINAL PARASITES OF THREE SPECIES OF PINNIPEDS (ARCTOCEPHALUS AUSTRALIS, ARCTOCEPHALUS GAZELLA, AND OTARIA FLAVESCENS) IN SOUTHERN BRAZIL.

In order to improve understanding of parasitism in South American pinnipeds, respiratory and gastrointestinal samples were collected from 12 Arctocephalus australis (South American fur seal), one Arctocephalus gazella (Antarctic fur seal), and one Otaria flavescens (South American sea lion). Ova and larvae were microscopically identified from fecal samples and respiratory secretions collected from live A. australis undergoing rehabilitation at Centro de Recuperação de Animais Marinhos (CRAM-FURG) in Rio Grande, Rio Grande do Sul, Brazil during June-July 2012. Adult parasites were collected from the lungs and gastrointestinal tracts of animals that died while undergoing treatment or were found dead along the southern Brazil coast. Parasites were identified by polymerase chain reaction and DNA sequencing, microscopic examination, comparison with keys, and histologic examination of tissues. Lung parasites of the Parafilaroides genus (Metastrongyloidea, Filaroididae) were identified at necropsy in both A. australis and A. gazella and gastrointestinal parasites were found in all three species of pinniped studied. Gastrointestinal parasites identified in A. australis included the nematodes Contracaecum sp. and Pseudoterranova cattani, the cestodes Adenocephalus pacificus (previously Diphyllobothrium pacificum), one from the Tetrabothridae family and one undetermined, and the acanthocephalans Corynosoma sp. and Bolbosoma sp.; from A. gazella the nematode Contracaecum sp. and the acanthocephalan Corynosoma sp.; and from O. flavescens the acanthocephalan Corynosoma sp. Ova from fecal samples from A. australis represent ascarid nematodes, Parafilaroides sp., Adenocephalus pacificus, acanthocephalans, and an egg determined either to be a trematode or pseuophyllidean cestode. With limited information surrounding parasitism, these findings are an important contribution to knowledge of the health of Southern Hemisphere pinnipeds.

INCIDENCE DENSITY, PROPORTIONATE MORTALITY, AND RISK FACTORS OF ASPERGILLOSIS IN MAGELLANIC PENGUINS IN A REHABILITATION CENTER FROM BRAZIL.

Aspergillosis, an opportunistic mycosis caused by the Aspergillus genus, affects mainly the respiratory system and is considered one of the most significant causes of mortality in captive penguins. This study aimed to examine a 6-yr period of cases of aspergillosis in penguins at the Centro de Recuperação de Animais Marinhos (CRAM-FURG), Rio Grande, Brazil. A retrospective cohort study was conducted using the institution's records of penguins received from January 2004 to December 2009. Animals were categorized according to the outcome "aspergillosis," and analyzed by age group, sex, oil fouling, origin, prophylactic administration of itraconazole, period in captivity, body mass, hematocrit, and total plasma proteins. A total of 327 Magellanic penguins (Spheniscus magellanicus) was studied, 66 of which died of aspergillosis. Proportionate mortality by aspergillosis was 48.5%, and incidence density was 7.3 lethal aspergillosis cases per 100 penguins/mo. Approximately 75% of the aspergillosis cases occurred in penguins that had been transferred from other rehabilitation centers, and this was considered a significant risk factor for the disease. Significant differences were also observed between the groups in regard to the period of time spent in captivity until death, hematocrit and total plasma proteins upon admission to the center, and body mass gain during the period in captivity. The findings demonstrate the negative impacts of aspergillosis on the rehabilitation of Magellanic penguins, with a high incidence density and substantial mortality.

Epidemiology and pathology of avian malaria in penguins undergoing rehabilitation in Brazil.

Seabird rehabilitation is a valuable strategy to mitigate the impacts of oil pollution and other anthropogenic factors, and can significantly contribute to the conservation of penguins. However, infectious diseases such as avian malaria (Plasmodium spp.) can hamper the success of rehabilitation efforts. We combined morphological and molecular diagnostic methods to investigate the epidemiology and pathology of Plasmodium in Magellanic penguins (Spheniscus magellanicus) at rehabilitation centers along 2500 km of the coastline of Brazil. True prevalence of malarial parasites was estimated between 6.6% and 13.5%. We identified five species, three of which had not been described infecting penguins (P. cathemerium, P. nucleophilum, P. unalis); an additional five distinct Plasmodium lineages were also distinguished, and albeit unidentified these clearly correspond to species that also have not yet been reported in penguins. Our results indicate that the diversity of plasmodia that may infect these birds is greater than previously recognised. Considering the well-defined seasonality observed in this study, it is clear that rehabilitation centers could benefit by narrowing their preventative efforts on penguins maintained or admitted during the Austral spring-summer, particularly by preventing mosquitoes from coming into contact with penguins.

Epidemiology and molecular phylogeny of Babesia sp. in Little Penguins Eudyptula minor in Australia.

Blood parasites are potential threats to the health of penguins and to their conservation and management. Little penguins Eudyptula minor are native to Australia and New Zealand, and are susceptible to piroplasmids (Babesia), hemosporidians (Haemoproteus, Leucocytozoon, Plasmodium) and kinetoplastids (Trypanosoma). We studied a total of 263 wild little penguins at 20 sites along the Australian southeastern coast, in addition to 16 captive-bred little penguins. Babesia sp. was identified in seven wild little penguins, with positive individuals recorded in New South Wales, Victoria and Tasmania. True prevalence was estimated between 3.4% and 4.5%. Only round forms of the parasite were observed, and gene sequencing confirmed the identity of the parasite and demonstrated it is closely related to Babesia poelea from boobies (Sula spp.) and B. uriae from murres (Uria aalge). None of the Babesia-positive penguins presented signs of disease, confirming earlier suggestions that chronic infections by these parasites are not substantially problematic to otherwise healthy little penguins. We searched also for kinetoplastids, and despite targeted sampling of little penguins near the location where Trypanosoma eudyptulae was originally reported, this parasite was not detected.

Phenological changes in the southern hemisphere.

Current evidence of phenological responses to recent climate change is substantially biased towards northern hemisphere temperate regions. Given regional differences in climate change, shifts in phenology will not be uniform across the globe, and conclusions drawn from temperate systems in the northern hemisphere might not be applicable to other regions on the planet. We conduct the largest meta-analysis to date of phenological drivers and trends among southern hemisphere species, assessing 1208 long-term datasets from 89 studies on 347 species. Data were mostly from Australasia (Australia and New Zealand), South America and the Antarctic/subantarctic, and focused primarily on plants and birds. This meta-analysis shows an advance in the timing of spring events (with a strong Australian data bias), although substantial differences in trends were apparent among taxonomic groups and regions. When only statistically significant trends were considered, 82% of terrestrial datasets and 42% of marine datasets demonstrated an advance in phenology. Temperature was most frequently identified as the primary driver of phenological changes; however, in many studies it was the only climate variable considered. When precipitation was examined, it often played a key role but, in contrast with temperature, the direction of phenological shifts in response to precipitation variation was difficult to predict a priori. We discuss how phenological information can inform the adaptive capacity of species, their resilience, and constraints on autonomous adaptation. We also highlight serious weaknesses in past and current data collection and analyses at large regional scales (with very few studies in the tropics or from Africa) and dramatic taxonomic biases. If accurate predictions regarding the general effects of climate change on the biology of organisms are to be made, data collection policies focussing on targeting data-deficient regions and taxa need to be financially and logistically supported.

Survival and movements of Magellanic penguins rehabilitated from oil fouling along the coast of South America, 2000-2010.

Oil pollution is a significant conservation concern. We examined data from six institutions along the coast of South America: Emergency Relief Team of the International Fund for Animal Welfare, Fundación Mundo Marino, Centro de Recuperação de Animais Marinhos, Natura Patagonia, Associação R3 Animal, and Mar del Plata Aquarium and data from resightings in Argentina, Brazil, Chile and Falkland/Malvinas Islands. From 2000 to 2010, 2183 oiled Magellanic penguins were rehabilitated as part of the routine activities of these institutions or during emergency responses to eight oil spills in which they were involved; all rehabilitated penguins were flipper banded and released. Since their release, 41 penguins were resighted until 31 December 2011. The results demonstrate that, when combined with other prevention strategies, the rehabilitation of Magellanic penguins is a strategy that contributes to the mitigation of adverse effects of oil spills and chronic pollution to the species.

Unusual incidence of chronic pneumonia associated with cholesterol deposits in stranded and bycaught franciscanas Pontoporia blainvillei.

Franciscanas Pontoporia blainvillei are small dolphins endemic to Brazil, Uruguay and Argentina. During routine pathologic examinations, chronic pneumonia associated with cholesterol deposits was found in 16.7% of stranded and incidentally bycaught franciscanas (n = 60), and was more frequent in dolphins from the Brazilian state of São Paulo (Franciscana Management Area II) and frequently accompanied by splenic lymphoid hyperplasia. It is unclear why these otherwise uncommon lesions were relatively frequent in the studied dolphins, and further research is advised.

Magellanic penguin mortality in 2008 along the SW Atlantic coast.

Magellanic penguins migrate from Patagonia reaching northern Argentina, Uruguay, and southern Brazil on their winter migration, in parallel with the seasonal pulse of anchovy spawning. In 2008, Magellanic penguins went further north than usual. Many died and a few swam nearly to the Equator. Twelve groups surveyed 5000 km of coastline encountering 3371 penguins along the coast. Most penguins arrived in northern Brazil (68.4%) without petroleum (2933, 87%). Almost all penguins without petroleum were juveniles (2915, 99%) and 55% were alive when found. Penguins were dehydrated, anemic, hypothermic, and emaciated. Of the penguins with petroleum, 13% arrived in the southern half of Brazil, showing that petroleum pollution remains a problem along the SW Atlantic coast. The mortality occurred in the winter of 2008 when sea surface temperature were unusually cold perhaps reducing the prey for penguins.

Chronic oil pollution harms Magellanic penguins in the Southwest Atlantic.

Petroleum pollution is a problem for seabirds along the Southwest Atlantic coast. Twenty-five groups from Salvador, Brazil (12 degrees 58'S) to San Antonio Oeste, Argentina (40 degrees 43'S) survey or rehabilitate sick or oiled seabirds. Four groups, one each in Brazil and Uruguay, and two in Argentina, kept counts of birds found alive and in need of rehabilitation. An average of 63.7% of the seabirds found were Magellanic penguins (Spheniscus magellanicus), with 3869 reported since 1987. Mainly adult penguins were found in Argentina (1605 of 2102 penguins of known age class) and Uruguay (158 of 197). Juveniles were most common in Brazil (234 of 325). Oil fouling was the most frequent cause of injury or sickness. The number of oiled penguins reported in their wintering range has greatly increased since the early 1990s and is strongly correlated with petroleum exports from Argentina. Our results show that chronic petroleum pollution is a problem for wildlife from Southern Brazil through Northern Argentina, and regulations and enforcement are failing to protect living resources.

Corneal endothelium of the Magellanic penguin (Spheniscus magellanicus) by scanning electron microscopy.

The corneal endothelium is essential for the maintenance of the corneal transparency. The aim of this study was to examine the morphology of the endothelial surface and perform morphometric analysis of the normal corneal endothelial cells of the Magellanic penguin (Spheniscus magellanicus) using scanning electron microscopy. The present work demonstrates that the corneal endothelium of the Magellanic penguin is similar to those described in other vertebrates.