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.

H6N8 avian influenza virus in Antarctic seabirds demonstrates connectivity between South America and Antarctica.

Wild aquatic birds are the natural reservoirs of avian influenza viruses (AIVs). It is estimated that 100 million seabirds live in the Antarctic Peninsula and adjacent islands, regularly encountering migratory birds that use the islands to nest. Between 2010 and 2013, we collected samples from 865 seabirds in Elephant, King George and Livingston islands, around Antarctica Peninsula: chinstrap penguin (n = 143); gentoo penguin (n = 208); Adelie penguin (n = 46); brown skua (n = 90); Cape petrel (n = 115) and southern giant petrel (n = 263). Serum (n = 673) samples were analysed by competitive ELISA and swabs (n = 614) were tested by one step real-time RT-PCR for avian influenza virus (AIV). Sera from 30 chinstrap penguins, 76 brown skuas and a single Adelie penguin were seropositive for AIV. Thirteen swab samples were AIV positive by RT-PCR, and complete genome sequences of H6N8 AIVs isolated from brown skua and chinstrap penguin in 2011 were obtained. Phylogenetic analyses indicated that all gene segments of the H6N8 viruses were closely related to Argentinian and Chilean AIVs. The prevalence with which we identified evidence for AIVs infection in various Antarctic seabirds suggest viral circulation in Antarctic avifauna and interspecies viral transmission in the sub-Antarctic region.

A new haemosporidian parasite from the Red-legged Seriema Cariama cristata (Cariamiformes, Cariamidae).

Haemoproteids (Haemosporida, Haemoproteidae) are a diverse group of avian blood parasites that are transmitted by hematophagous dipterans. In this study, we describe Haemoproteus pulcher sp. nov. from a Red-legged Seriema (Cariama cristata) in southeast Brazil. Analysis of the mitochondrial cytb gene indicates this parasite is closely related to Haemoproteus catharti (from Turkey Vulture, Cathartes aura) and the unidentified haemosporidian lineages PSOOCH01 (from Pale-winged Trumpeter, Psophia leucoptera) and MYCAME08 (from Wood Stork, Mycteria americana). This group of parasites appears to represent an evolutionary lineage that is distinct from other Haemoproteus spp., being instead more closely related to Haemocystidium spp. (from reptiles), Plasmodium spp. (from reptiles, birds, and mammals) and other mammal-infecting haemosporidians (Nycteria, Polychromophilus, and Hepatocystis). Current evidence suggests that parasites of this newly discovered evolutionary lineage may be endemic to the Americas, but further studies are necessary to clarify their taxonomy, life cycle, vectors, hosts, geographic distribution and host health effects. Additionally, it should be borne in mind that some PCR protocols targeting the cytb gene might not reliably detect H. pulcher due to low primer affinity.

Successful Treatment of Capture Myopathy and Satellite Transmitter Injury in an Atlantic Yellow-nosed Albatross (Thalassarche chlororhynchos).

An Atlantic yellow-nosed albatross (Thalassarche chlororhynchos) was found on shore 3 days after having been captured at sea by researchers. It presented very lethargic, moderately dehydrated, and in poor body condition. It was mildly hypothermic, with moderate pediculosis, and dark malodorous feces with yellow urates. The bird had a 48-g satellite transmitter attached with a backpack-style chest harness, which caused an ulcerative lesion on the interscapular area. The bird was severely anemic (packed cell volume, 18%), and plasma chemistry results were suggestive of a severe rhabdomyolysis (aspartate transaminase, 3620 U/L; creatine kinase, 100 400 U/L). We hypothesized that capture myopathy occurred because of a combination of capture stress and prolonged physical restraint by researchers, stress associated with repeated attempts by the bird to remove the satellite-transmitter harness, and a lengthy road transport to the rehabilitation center. A treatment protocol, which relied on a combination of aggressive fluid therapy, selenium, vitamins E and B12, and multivitamin supplementation, was administered after the initial physical assessment of the albatross. Isoflurane inhalation anesthesia was used to minimize stress associated with the performed medical procedures (eg, physical examination, removal of the satellite transmitter harness, blood collection, and wound management). Measures were adopted while the bird was hospitalized to reduce stress (eg, quiet and comfortable environment with visual barriers and restricting handling of the patient to experienced staff). Clinical and hematological monitoring was used to assess the patient's condition as the bird gradually recovered while hospitalized. The albatross was successfully released 28 days after the initial presentation, suggesting that the medical protocol employed in this case may be useful for the treatment of capture myopathy in albatrosses and other birds.

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.

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.

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.

Empirical Primaquine Treatment of Avian Babesiosis in Seabirds.

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

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.

From incubation to release: Hand-rearing as a tool for the conservation of the endangered African penguin.

The African penguin (Spheniscus demersus) population is estimated at 25,000 breeding pairs, approximately 5% of that at the start of the 20th century, and the species is currently classified as Endangered. In the last two decades, the hand-rearing of penguin chicks that were abandoned by their parents due to oil spills or other circumstances has become a valuable conservation tool to limit mortality and to bolster the population at specific colonies. We summarize and evaluate the techniques employed by the Southern African Foundation for the Conservation of Coastal Birds (SANCCOB) to incubate and hand-rear African penguin eggs and chicks. From 2012 to 2016, a total of 694 eggs and 2819 chicks were received by SANCCOB's Chick Rearing Unit. It was estimated that 13% of the eggs were infertile, and 81% of the fertile eggs hatched successfully. The overall release rate for chicks was 77%, with a higher release rate for chicks that were pre-emptively removed (93%) followed by chicks that had been abandoned by their parents (78%), chicks admitted due to avian pox lesions (61%), chicks that hatched from artificially-incubated eggs (57%), and chicks admitted due to injuries or deformities (25%). Rescuing and hand-rearing eggs and chicks has been a successful strategy for African penguins, and might be also applicable for the conservation of other threatened seabird species whose population are critically low or during natural or anthropogenic events that could have disastrous population impacts (e.g. oil spills, disease outbreaks, catastrophic weather events, strong El Niño years, etc.).

First report of a hatched, hand-reared, and released African oystercatcher.

The African oystercatcher Haematopus moquini is a near-threatened wader that is endemic to southern Africa. In the past, the species suffered a drastic decrease in nesting success due to human disturbance. We present the case report of an African oystercatcher that was hatched, hand-reared, and released in the Western Cape, South Africa. African oystercatchers are semi-altricial birds that tend to be highly sensitive to stress; as a result, strategies to minimize stress and the employment of surrogate parents and pre-release acclimatization are important to ensure post-release survival of hand-reared chicks. Considering the lack of literature on the incubation and hand-rearing of oystercatchers, this case report provides a basis for the development of hand-rearing techniques that might be useful for the protection of this and other threatened wader species.

Lack of Detection of Avian Influenza, Newcastle Disease, and West Nile Viruses in Wild Birds of Northeastern Brazil.

We tested 529 wild birds captured in northeastern Brazil for infection by avian influenza, Newcastle disease, and West Nile. Viruses were not detected by real-time PCR with the exception of one Tropical Gnatcatcher ( Polioptila plumbea) positive for influenza virus, but this could not be confirmed by viral isolation or gene sequencing.

Antibodies Against Henipa-Like Viruses in Brazilian Bats.

Bats are reservoir hosts for many paramyxoviruses, some of which cause human and zoonotic diseases of public health importance. We developed a Nipah virus nucleoprotein enzyme-linked immunosorbent assay to detect cross-reactive antibodies in serum samples from several bat species in Brazil. Our results warrant further investigation of henipa-like virus reservoirs in the Western hemisphere.

Do blood parasites infect Magellanic penguins (Spheniscus magellanicus) in the wild? Prospective investigation and climatogeographic considerations.

Magellanic penguins (Spheniscus magellanicus) are native to Argentina, Chile and the Falkland Islands. Magellanic penguins are highly susceptible to blood parasites such as the mosquito-borne Plasmodium spp., which have been documented causing high morbidity and mortality in zoos and rehabilitation centres. However, to date no blood parasites have been detected in wild Magellanic penguins, and it is not clear whether this is reflective of their true absence or is instead related to an insufficiency in sampling effort or a failure of the diagnostic methods. We examined blood smears of 284 Magellanic penguins from the Argentinean coast and tested their blood samples with nested polymerase chain reaction tests targeting Haemoproteus, Plasmodium, Leucocytozoon and Babesia. No blood parasites were detected. Analysing the sampling effort of previous studies and the climatogeography of the region, we found there is strong basis to conclude that haemosporidians do not infect wild Magellanic penguins on the Argentinean coast. However, at present it is not possible to determine whether such parasites occur on the Chilean coast and at the Falkland Islands. Furthermore, it is troubling that the northward distribution expansion of Magellanic penguins and the poleward distribution shift of vectors may lead to novel opportunities for the transmission of blood parasites.

Lesions associated with drowning in bycaught penguins.

Fisheries bycatch, the incidental mortality that occurs as a result of entanglement in fishing gear, is an important conservation threat to penguins and other seabirds. Identification of entanglement and drowning in beach-cast carcasses of seabirds remains a challenge, as it is still unclear what lesions are to be expected in a bycaught seabird. We necropsied 2 Magellanic penguins Spheniscus magellanicus that were entangled and drowned in gillnets. Marked distension of the lungs with foamy red fluid and marked oedema of the dorsal visceral pleura were prominent lesions consistent with those described in cases of 'wet drowning' in humans. On the other hand, the air sacs contained very small quantities of liquid, suggesting that absence of water in the air sacs might not be a reliable sign to exclude drowning. Other relevant findings included cutaneous lacerations and bruising in one bird and cervical and pectoral rhabdomyolysis in both birds. While cutaneous or subcutaneous hematomas may be an indication of bycatch, especially if linear or cross-linear patterns consistent with fishing nets are present, these lesions might not always be discernible and their absence does not suffice to exclude the possibility of entanglement in fishing nets. Additionally, our findings suggest that the histological examination of skeletal muscles, particularly of the neck, may provide additional clues to corroborate the diagnosis of drowning in penguins.

Surveillance of Avian Influenza Virus in Aquatic Birds on the Brazilian Amazon Coast.

The occurrence of avian influenza viruses (AIV) has been extensively studied in aquatic birds in the Northern hemisphere; however, much less information is available for the South American region. In 2009-2010, we sampled 1006 wild aquatic birds (90% Charadriiformes, 9% Anseriformes, and 1% other groups) at three locations on the Brazilian Amazon coast, a region that serves as a major stop-over and wintering site along the Atlantic Americas flyway. Real-time RT-PCR identified five samples as positive; however, no AIV isolates could be obtained and Illumina sequencing did not produce gene sequences that would allow further characterization of the virus.

Serological Survey of Hantavirus in Inhabitants from Tropical and Subtropical Areas of Brazil.

Brazil has reported more than 1,600 cases of hantavirus cardiopulmonary syndrome (HPS) since 1993, with a 39% rate of reported fatalities. Using a recombinant nucleocapsid protein of Araraquara virus, we performed ELISA to detect IgG antibodies against hantavirus in human sera. The aim of this study was to analyze hantavirus antibody levels in inhabitants from a tropical area (Amazon region) in Rondônia state and a subtropical (Atlantic Rain Forest) region in São Paulo state, Brazil. A total of 1,310 serum samples were obtained between 2003 and 2008 and tested by IgG-ELISA, and 82 samples (6.2%), of which 62 were from the tropical area (5.8%) and 20 from the subtropical area (8.3%), tested positive. Higher levels of hantavirus antibody were observed in inhabitants of the populous subtropical areas compared with those from the tropical areas in Brazil.

Molecular Characterization of Subtype H11N9 Avian Influenza Virus Isolated from Shorebirds in Brazil.

Migratory aquatic birds play an important role in the maintenance and spread of avian influenza viruses (AIV). Many species of aquatic migratory birds tend to use similar migration routes, also known as flyways, which serve as important circuits for the dissemination of AIV. In recent years there has been extensive surveillance of the virus in aquatic birds in the Northern Hemisphere; however in contrast only a few studies have been attempted to detect AIV in wild birds in South America. There are major flyways connecting South America to Central and North America, whereas avian migration routes between South America and the remaining continents are uncommon. As a result, it has been hypothesized that South American AIV strains would be most closely related to the strains from North America than to those from other regions in the world. We characterized the full genome of three AIV subtype H11N9 isolates obtained from ruddy turnstones (Arenaria interpres) on the Amazon coast of Brazil. For all gene segments, all three strains consistently clustered together within evolutionary lineages of AIV that had been previously described from aquatic birds in North America. In particular, the H11N9 isolates were remarkably closely related to AIV strains from shorebirds sampled at the Delaware Bay region, on the Northeastern coast of the USA, more than 5000 km away from where the isolates were retrieved. Additionally, there was also evidence of genetic similarity to AIV strains from ducks and teals from interior USA and Canada. These findings corroborate that migratory flyways of aquatic birds play an important role in determining the genetic structure of AIV in the Western hemisphere, with a strong epidemiological connectivity between North and South America.

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.

Avian influenza virus (H11N9) in migratory shorebirds wintering in the Amazon Region, Brazil.

Aquatic birds are the natural reservoir for avian influenza viruses (AIV). Habitats in Brazil provide stopover and wintering sites for water birds that migrate between North and South America. The current study was conducted to elucidate the possibility of the transport of influenza A viruses by birds that migrate annually between the Northern and Southern Hemispheres. In total, 556 orotracheal/cloacal swab samples were collected for influenza A virus screening using real-time RT-PCR (rRT-PCR). The influenza A virus-positive samples were subjected to viral isolation. Four samples were positive for the influenza A matrix gene by rRT-PCR. From these samples, three viruses were isolated, sequenced and characterized. All positive samples originated from a single bird species, the ruddy turnstone (Arenaria interpres), that was caught in the Amazon region at Caeté Bay, Northeast Pará, at Ilha de Canelas. To our knowledge, this is the first isolation of H11N9 in the ruddy turnstone in South America.