Diomedenema dinarctos n. sp. (Spiruromorpha: Desmidocercidae), a new species of lung nematode in Fiordland crested penguins (Eudyptes pachyrhynchus; Sphenisciformes), from South Island, New Zealand.

Fiordland crested penguins Eudyptes pachyrhynchus Gray are thick-billed, crested penguins endemic to New Zealand that breed in small colonies in inaccessible places. They are assessed as Vulnerable by the IUCN. This is the first report of helminth parasites from this penguin species. Herein a new species of Diomedenema is described (Spiruromorpha, Desmidocercidae) from the lungs of Fiordland crested penguins. Diomedenema dinarctos n. sp. has only two congeners and is closest morphologically to D. tavaresi. It differs from D. tavaresi in overall larger size, longer buccal cavity and oesophagus, in the distance of the excretory pore and nerve ring from the anterior end, and the ratio of spicule sizes. Eggs are twice the size reported for D. tavaresi. The only other species, D. diomedeae, is far larger in body size, with a shorter buccal cavity and oesophagus and a reduced body length to oesophagus ratio. The position in the body length of the vulva and the spicule ratio also differ strongly. We provide a DNA sequence for the 18S rDNA gene. Species of Diomedenema have been reported as causing death in albatross, so any deceased Fiordland crested penguins should be examined specifically for this worm to enable evaluation of this potentially damaging parasite in the population.

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.

Penguins are competent hosts of Haemoproteus parasites: the first detection of gametocytes, with molecular characterization of Haemoproteus larae.

BACKGROUND: The majority of penguins (Sphenisciformes) have evolved in areas with weak or absent transmission of haemosporidian parasites and are usually naïve to avian haemosporidian infections. Plasmodium parasites are transmitted by mosquitoes, and lethal avian malaria has been often reported in captive penguins in many countries. The related haemosporidian parasites belonging to Haemoproteus and Leucocytozoon have also been detected in penguins but less often than Plasmodium infections. The majority of Haemoproteus infection reports in penguins are based solely on PCR-based diagnostics. It remains unclear if haemoproteids can complete their life-cycle and produce infective stages (gametocytes) in penguins or whether these infections are abortive in penguins, and thus dead ends for transmission. In other words, it remains unknown if penguins are competent hosts for Haemoproteus parasites, which cause disease in non-adapted birds. METHODS: Two captive African penguins (Spheniscus demersus) and two Magellanic penguins (S. magellanicus) were found to be positive for Haemoproteus infection in two open-air aquariums in Japan, and the parasites were investigated using both PCR-based testing and microscopical examination of blood films. Samples from a black-tailed gull (Larus crassirostris) and previously tested gulls were used for comparison. RESULTS: The lineage hSPMAG12 was detected, and gametocytes of Haemoproteus sp. were seen in the examined penguins and gull. Observed gametocytes were indistinguishable from those of Haemoproteus larae, which naturally parasitize birds of the genus Larus (Laridae). The detected sequence information and Bayesian phylogenetic analysis supported this conclusion. Additionally, morphologically similar gametocytes and closely related DNA sequences were also found in other gull species in Japan. Phylogenetic analysis based on partial cytb sequences placed the lineage hSPMAG12 of H. larae within the clade of avian haemoproteids which belong to the subgenus Parahaemoproteus, indicating that Culicoides biting midges likely transmit the parasites between penguins and gulls. CONCLUSIONS: This study shows that some species of Haemoproteus parasites complete their development and produce gametocytes in penguins, which may be source of infection for biting midges transmitting haemoproteosis. To prevent haemosporidiosis in zoos, we call for control not only of mosquitoes, but also biting midges.

Atrophic Proventriculitis Associated with Gravid Females of Tetrameres sp. (Nematoda: Tetrameridae) in the Magellanic Penguin (Spheniscus magellanicus) on the Coast of Brazil.

The present study offers the first description of proventriculitis associated with the presence of gravid female nematodes of the genus Tetrameres (Nematoda: Tetrameridae) in 3 juvenile Magellanic penguins (Spheniscus magellanicus) found dead on the coast of Brazil. This study broadens knowledge on parasites associated with these hosts and the real impact of this association.

Determinants of external and blood parasite load in African penguins (Spheniscus demersus) admitted for rehabilitation.

We investigate the factors associated with the occurrence and abundance of external and blood parasites in African penguins (Spheniscus demersus), an endangered seabird that breeds exclusively on the coasts of Namibia and South Africa. External parasites were collected using the dust-ruffling method from 171 African Penguins admitted at a rehabilitation facility in the Western Cape, South Africa. Additionally, blood smears were obtained upon admission and weekly during rehabilitation and examined for blood parasites. Fleas Parapsyllus longicornis humboldti, ticks Ornithodoros capensis and lice Austrogoniodes demersus were recovered from 93, 63 and 40%, respectively, of the penguins upon admission to the centre. Rescue location and age group were identified as significant determinants of flea abundance, whereas month of admission was a significant determinant of tick abundance. Blood parasites were also common on admission, with Babesia being the most frequent (46% prevalence) whereas Borrelia was recorded sporadically (1.2%) and Plasmodium was recorded once. The prevalence and abundance of ticks on admission was positively associated with Babesia infection on admission. Our findings demonstrate the variability and contributing factor of parasite infections in an endangered species of penguin, and highlight the need for additional research on the parasite-host dynamics involving these potential disease vectors.

Phylogenetic Relationships of Cardiocephaloides spp. (Digenea, Diplostomoidea) and the Genetic Characterization of Cardiocephaloides physalis from Magellanic Penguin, Spheniscus magellanicus, in Chile.

PURPOSE: Cardiocephaloides is a small genus of strigeid digeneans with an essentially cosmopolitan distribution. Most members of Cardiocephaloides are found in larid birds, however, Cardiocephaloides physalis is an exception and parasitizes penguins in some coastal regions of South America and South Africa. No prior molecular phylogenetic studies have included DNA sequence data of C. physalis. Herein, we provide molecular phylogenetic analyses of Cardiocephaloides using DNA sequences from five species of these strigeids. METHODS: Adult Cardiocephaloides spp. were obtained from larid birds and penguins collected from 3 biogeographical realms (Palearctic, Nearctic and Neotropics). We have generated sequences of the complete ITS region and partial 28S gene of the nuclear ribosomal DNA, along with partial sequences of the mitochondrial CO1 gene for C. physalis, C. medioconiger and the type species of the genus, C. longicollis and used them for phylogenetic inference. RESULTS: Cardiocephaloides spp. appeared as a 100% supported clade in the phylogenetic tree based on 28S sequences. The position of C. physalis varied between the phylogenetic trees based on the relatively conservative 28S gene on one hand, and variable ITS1 and COI sequences on the other. Cardiocephaloides physalis was nested within the clade of Cardiocephaloides spp. in the 28S tree and appeared as the sister group to the remaining members of the genus in the ITS1 region and COI trees. We detected 0.4-1.6% interspecific divergence in 28S, 1.9-6.9% in the ITS region and 8.7-11.8% in CO1 sequences of Cardiocephaloides spp. Our 28S sequence of C. physalis from South America and a shorter sequence from Africa available in the GenBank were identical. CONCLUSION: Cardiocephaloides as represented in the currently available dataset is monophyletic with C. physalis parasitism in penguins likely resulting from a secondary host-switching event. Identical 28S sequences of C. physalis from South America and Africa cautiously confirm the broad distribution of this species, although comparison of faster mutating genes (e. g., CO1) is recommended for a better substantiated conclusion.

Epidemiology, hematology, and unusual morphological characteristics of Plasmodium during an avian malaria outbreak in penguins in Brazil.

Avian malaria is a mosquito-borne disease caused by Plasmodium spp. protozoa, and penguins are considered particularly susceptible to this disease, developing rapid outbreaks with potentially high mortality. We report on an outbreak of avian malaria in Magellanic penguins (Spheniscus magellanicus) at a rehabilitation center in Espírito Santo, southeast Brazil. In August and September 2015, a total of 89 Magellanic penguins (87 juveniles and 2 adults) received care at Institute of Research and Rehabilitation of Marine Animals. Over a period of 2 weeks, Plasmodium infections were identified in eight individuals (9.0%), four of which died (mortality = 4.5%, lethality = 50%). Blood smears and sequencing of the mitochondrial cytochrome b gene revealed the presence of Plasmodium lutzi SPMAG06, Plasmodium elongatum GRW06, Plasmodium sp. PHPAT01, Plasmodium sp. SPMAG10, and Plasmodium cathemerium (sequencing not successful). Two unusual morphological features were observed in individuals infected with lineage SPMAG06: (a) lack of clumping of pigment granules and (b) presence of circulating exoerythrocytic meronts. Hematological results (packed cell volume, plasma total solids, complete blood cell counts) of positive individuals showed differences from those of negative individuals depending on the lineages, but there was no overarching pattern consistently observed for all Plasmodium spp. The epidemiology of the outbreak and the phylogeography of the parasite lineages detected in this study support the notion that malarial infections in penguins undergoing rehabilitation in Brazil are the result of the spillover inoculation by plasmodia that circulate in the local avifauna, especially Passeriformes.

Evaluation of antibodies against Toxoplasma gondii and Leptospira spp. in Magellanic penguins (Spheniscus magellanicus) on Magdalena Island, Chile.

Toxoplasmosis has been reported in many avian species, but little information is available from wild penguin populations. Leptospira can infects domestic and wild animals. Spheniscus magellanicus belong to the order Sphenisciformes, family Spheniscidae, and are colonial birds. These seabirds live in temperate waters along the Atlantic shores of South America, and their total population has been estimated to be 1,300,000 breeding pairs. Magdalena Island (Chile) hosts an important breeding colony but, over recent decades, a marked decline in the number of birds has been seen. The objective of this study was to determine occurrences of antibodies against T. gondii and Leptospira spp. in penguins (Spheniscus magellanicus) on Magdalena Island, from where no previous data on these agents were available. Serum samples were collected from 132 penguins on Magdalena Island. Antibodies against Toxoplasma gondii were detected using the modified agglutination test (Titer ≥20), and anti-Leptospira spp. antibodies were detected using the microscopic agglutination test (Titer ≥100). T. gondii antibodies were detected in 57 (43.18%) of the 132 serum samples, with titers that ranged from 20 to 320. None of the penguins in this study was reactive to anti-Leptospira spp. antibodies. This is the first report of T. gondii seropositivity in free-living Magellanic penguins in Chile.

Species of Contracaecum Parasitizing the Magellanic Penguin Spheniscus magellanicus (Spheniscidae) from the Argentinean Coast.

Anisakid nematodes have a worldwide distribution and are associated with fishes, birds, and marine mammals from freshwater, brackish, and marine systems. The aims of this work are to report for the first time Contracaecum mirounga parasitizing the Magellanic penguin Spheniscus magellanicus, to report another Contracaecum species in the same host species, and to discuss the validity of Contracaeceum spheniscus. Several dead chicks, juveniles, and adults of S. magellanicus were collected along the Argentinean coast from 2002 to 2009. Nematodes were removed from digestive tracts and studied using both light and scanning electron microscopy. Nematode prevalences were 2.38% for C. mirounga and 12.5% for Contracaecum sp. Contracaecum mirounga was found in 1 penguin from Península Valdés, Chubut. This species is known as a specific parasite of marine mammals such as Pinnipedia, thereby suggesting that this nematode is not as specific as believed. Another species of Contracaecum sp. was found parasitizing 1 penguin from the Río de la Plata coast. It possessed an unusual interlabial morphology and arrangement of male caudal papillae. Despite the low prevalence, the distinct morphological features are convincing and support the presence of a new Contracaecum species. However, a formal description is not presented because sufficient male specimens are lacking. Finally, C. spheniscus is considered a junior synonym of Contracaecum pelagicum. Future molecular studies might be helpful to determine the real diversity of Contracaecum species parasitizing S. magellanicus considering the number of sibling species recognized among the anisakids.

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

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

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

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.

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

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

Can a Terrestrial Ectoparasite Disperse with Its Marine Host?

One of the most extreme examples of parasite adaptation comes from terrestrial ectoparasites exploiting marine hosts. Despite the ubiquity of such ectoparasitism and its ecological and evolutionary importance, investigations of the responses of ectoparasites to conditions encountered on their hosts are rare. In the case of penguins and their ticks, current understanding suggests that ticks freely parasitize their hosts on land but are incapable of surviving extended oceanic journeys. We examined this conjecture by assessing the physiological capacity of little penguin ticks to endure at-sea foraging and dispersal events of their hosts. Survival in penguins ticks was not significantly compromised by exposure to depths commonly associated with host dives (40 and 60 m), repeated seawater exposure relevant to the most common (30 s) and longest (120 s) recorded host dives, or extended (48 h) exposure to seawater. Mean (±SD) closed-phase durations in adult and nymphal ticks exhibiting discontinuous gas exchange ( 339±237 and 240±295 s, respectively) exceeded that of the maximum recorded host dive duration (120 s). Normoxic-anoxic-normoxic respirometry also confirmed spiracle closure. Mean metabolic rates ( 0.354±0.220 and 4.853±4.930 µL/h at 25°C for unfed and fed adult females, respectively) were significantly influenced by temperature; optimal and LT50 temperatures for adult ticks and fed nymphal ticks were typically higher than swimming penguin body temperatures. These findings suggest that marine host dispersal is unlikely to present an insurmountable barrier to long-distance tick dispersal. Such dispersal has important implications for evolutionary theory, conservation, and epidemiology.

Nasal mites (Mesostigmata: Rhinonyssidae) in African penguins (Spheniscus demersus).

Rhinonyssids are obligate haematophagous mites that parasitize the nasal cavity of vertebrates, and occur in a wide range of birds worldwide. Two species of nasal mites are known to occur in penguins: Rhinonyssus sphenisci, which has been recorded from Humboldt and Magellanic penguins (Spheniscus humboldti and S. magellanicus, respectively), and Rhinonyssus schelli, which has been recorded in Adélie and Gentoo penguins (Pygoscelis adeliae and P. papua, respectively). We examined the nasal cavity of African penguins (Spheniscus demersus) that died while under care at a rehabilitation centre (N = 40) or that were found dead at breeding colonies (N = 67). Nasal mites were found in the nasal cavity and/or paranasal of sinuses of 21 penguins, some of which had signs of mild-to-moderate sinusitis. Prevalence was higher in juveniles (29%) and adults (27%) than in chicks (10%). Mean intensity was 5.9 ± 12.9 mites per infected host (range 1-60). The mites presented morphological characteristics that were at times consistent with either R. sphenisci or R. schelli, and therefore we conservatively classified them as 'R. sphenisci sensu lato'. Our morphometric results raise the question of whether the specific status of R. schelli is justified.

Pathological and molecular characterization of avian malaria in captive Magellanic penguins (Spheniscus magellanicus) in South America.

Avian malaria is a mosquito-borne disease that affects multiple avian species and is caused by protozoans of the genus Plasmodium. An avian malaria infection caused by Plasmodium sp. in Magellanic penguins (Spheniscus magellanicus) with high mortality is described in a zoo in Southern Brazil. Clinically, three birds presented signs of inappetence, anorexia, pale mucosa, dyspnea, and opisthotonus, with death in a clinical course of 5-8 h. At the necropsy, all birds exhibited pale mucosa, marked splenomegaly and hepatomegaly, in addition to moderate leptomeningeal blood vessels ingurgitation in the brain. Microscopically, multiple exoerythrocytic meronts were observed in the cytoplasm of endothelial cells in the spleen, liver, heart, lungs, brain, kidneys, and pancreas. The spleen had a multifocal perivascular inflammatory infiltrate of lymphocytes, plasma cells, and macrophages, which also exhibited hemosiderosis and erythrophagocytosis. The liver had a multifocal periportal inflammatory infiltrate of lymphocytes, macrophages, and plasma cells, in addition to marked hemosiderosis in the hepatic sinusoids. Fragments of spleen, liver, brain, skeletal muscle, and lung were tested by the polymerase chain reaction technique for the detection of a fragment of the cytochrome B gene from haemosporidians, which resulted positive for Plasmodium spp. After sequencing, the samples were phylogenetically associated to Plasmodium sp. detected in Turdus albicollis (KU562808) in Brazil and matched to the lineage TURALB01 previously detected in T. albicollis. Avian malaria infections caused by Plasmodium sp. of lineage TURALB01 may occur in S. magellanicus with high mortality, and, thus, it is essential to detect and characterize the agent involved to obtain the differential diagnosis of the condition.

Comparative mtDNA phylogeographic patterns reveal marked differences in population genetic structure between generalist and specialist ectoparasites of the African penguin (Spheniscus demersus).

To address factors affecting genetic diversity and dispersal of ectoparasites, we compared mitochondrial DNA (mtDNA) population genetic structures of the generalist soft tick Ornithodoros capensis to the more host-specific nest flea Parapsyllus humboldti. A total of 103 ticks and 92 fleas were sampled at five distinct South African island/mainland African penguin (Spheniscus demersus) colonies. With its wide host range, O. capensis showed no evidence of significant cytochrome c oxidase subunit I (COI) mtDNA population differentiation among the five sampling sites (φst = 0.00 ± 0.004; p = 0.80), as well as a higher level of genetic diversity (π = 0.8% ± 0.06%) when compared to P. humboldti. In contrast, the flea showed significant population structure among most of the same sampling sites (φst = 0.22 ± 0.11; p ≤ 0.05) and a lower level of genetic diversity (π = 0.2% ± 0.01%). Our findings suggest that despite both parasites being mostly nest bound, O. capensis have few barriers to dispersal among island and mainland colonies. However, P. humboldti are more dependent on the African penguin for dispersal and thus have more impediments to gene flow among the same colonies. These findings broadly support the SGVH (specialist generalist variation hypothesis) and provide the first evidence for this hypothesis in parasites restricted to seabird colonies.

Occurrence of tissue cyst forming coccidia in Magellanic penguins (Spheniscus magellanicus) rescued on the coast of Brazil.

The main motivation for this study was to determine the occurrence of Toxoplasma gondii, a cosmopolitan widespread zoonotic parasite distribution that can infect a wide variety of mammals and birds, in Magellanic penguins (Spheniscus magellanicus) in Brazil. In recent decades there has been a significant increase in the number of penguins originating from Argentinian and Chilean Patagonia, where these birds are born, that arrive on the Brazilian coast, where many of them are stranded and rescued. Tissue samples were collected from 330 individuals surveyed from 2012-2015 at the Institute for Marine Animal Research and Rehabilitation (IPRAM) located in Cariacica, state of Espirito Santo, Brazil. Serum were collected from 145 animals surveyed in 2015 for the detection of anti-T. gondii antibodies using the Modified Agglutination Test (MAT ≥20) and 18 birds were positive, with titers of 20 (7 birds), 40 (9 birds) and 80 (2 birds). Mouse bioassay for the isolation of T. gondii was performed using tissues from 54 penguins that were also surveyed in 2015, but no isolates were obtained. DNA from tissue samples of 330 individuals was PCR amplified and sequenced to detect tissue cyst forming coccidians by using pan sarcocystids-directed primers (based on 18S rDNA). These samples were from animals surveyed in 2015 and from frozen stocked tissues from animals surveyed in the years 2012 and 2013. The positives were PCR amplified and sequenced with genus Sarcocystis-specific primers (based on internal transcribed spacer 1, RNA polymerase beta subunit coding gene, and cytochrome B coding gene) and with Sarcocystis falcatula/Sarcocystis neurona- specific primers (based on surface antigens SAG2, SAG3 and SAG4). Sixteen (3.0%) of pectoral muscle samples were positive by all the seven molecular markers and all the samples were identical to each other. Organisms close related to Sarcocystis falcatula were confirmed in all cases. This is the first report on molecular detection of infection by S. falcatula-related organisms and the first report of seropositivity for T. gondii in free-living Magellanic penguins in Brazil. Felids and didephid opossums are definitive hosts of T. gondii and S. falcatula, respectively. Where the penguins acquire the infective forms of the parasites shed by the terrestrial mammals remains to be elucidated.

Schistosomes and Microfilarial Parasites in Magellanic Penguins.

The Magellanic Penguin ( Spheniscus magellanicus) is native to Argentina, Chile, and the Falkland/Malvinas Islands, and is a regular winter migrant in Uruguayan and Brazilian coastal waters. The species is known to be susceptible to a variety of gastrointestinal nematodes, cestodes, trematodes, and acanthocephalans, as well as renal trematodes and pulmonary nematodes. Schistosomes (Platyhelminthes, Trematoda, Schistosomatidae) and microfilariae (Nematoda, Secernentea, Onchocercidae) were histologically identified in Magellanic Penguins ( Spheniscus magellanicus) that died while under care at rehabilitation centers in southern Brazil. Phylogenetic analysis of the COI gene, ITS-1 region, 5.8S rRNA gene, ITS-2 region, and 28S rRNA gene sequences of the schistosome revealed that it is closely related to, but distinct from, a schistosome reported from the African Penguin ( Spheniscus demersus). The schistosomes from Magellanic and African Penguins were grouped with Gigantobilharzia huronensis, Gigantobilharzia melanoidis, and Dendritobilharzia pulvurenta; however, the lack of a clearly monophyletic origin precludes determining their genus. The incidental discovery of novel parasites during a study that did not specifically aim to investigate the occurrence of helminths underscores the value of histopathological examination as an exploratory diagnostic approach.

Sappinia sp. (Amoebozoa: Thecamoebida) and Rosculus sp. (SAR: Cercozoa) Isolated From King Penguin Guano Collected in the Subantarctic (South Georgia, Salisbury Plain) and their Coexistence in Culture.

Two amoeboid organisms of the genera Sappinia Dangeard, 1896 and Rosculus Hawes, 1963 were identified in a sample containing king penguin guano. This sample, collected in the Subantarctic, enlarges the list of fecal habitats known for the presence of coprophilic amoebae. The two organisms were co-isolated and subcultured for over 6 mo, with continuous efforts being invested to separate each one from the mixed culture. In the mixed culture, Rosculus cells were fast growing, tolerated changes in culturing conditions, formed cysts, and evidently were attracted by Sappinia trophozoites. The separation of the Rosculus strain was accomplished, whereas the Sappinia strain remained intermixed with inseparable Rosculus cells. Sappinia cell populations were sensitive to changes in culturing conditions; they improved with reduction of Rosculus cells in the mixed culture. Thick-walled cysts, reportedly formed by Sappinia species, were not seen. The ultrastructure of both organisms was congruent with the currently accepted generic characteristics; however, some details were remarkable at the species level. Combined with the results of phylogenetic analyses, our findings indicate that the ultrastructure of the glycocalyx and the presence/absence of the Golgi apparatus in differential diagnoses of Sappinia species require a critical re-evaluation.

A new species of Diomedenema (Nematoda, Rhabditida, Spiruromorpha) from Spheniscus magellanicus (Aves, Sphenisciformes) found on the southern coast of Brazil.

A new species of the genus Diomedenema, a spiruromorph nematode, collected from the lung of Spheniscus magellanicus (Sphenisciformes) found on the southern coast of the state of Rio Grande do Sul, Brazil, is described. The new species is differentiated from the only previously described species of the genus, D. diomedeae Johston & Mawson, 1952, by males possessing a set of caudal papillae with three pairs of precloacal, two pairs of adcloacal and one pair of postcloacal papillae; precloacal papillae with the papillae of the first two pairs being closer to each other than those of the third pair; a longer and pointed tail in males; and females with the vulva at mid-body. This is the first report of a nematode infecting the lung of a sphenisciforme host.