Guard-hair microstructure of Brazilian felids: considerations for species identification

O exame da microestrutura da cutícula e medula dos pêlos é uma técnica simples e de baixo custo para identificar espécies de mamíferos para uma variedade de aplicações. Foram estudados pêlos-guarda de 66 indivíduos de oito espécies de felídeos brasileiros (Leopardus colocolo, L.geoffroyi, L.pardalis, L.tigrinus, L.wiedii, Panthera onca, Puma concolor, Puma yagouaroundi), através de amostras colhidas de animais anestesiados em zoológicos. A microestrutura dos pêlos-guarda foi analisada e descrita através de impressões cuticulares e preparações medulares, e posteriormente foi feito um teste cego para avaliar a acurácia da identificação específica. Embora tenham sido encontrados caracteres morfológicos distintos para cada espécie, a sutileza destes caracteres e sua sobreposição entre as diferentes espécies resultaram em uma acurácia relativamente baixa (75%). A identificação de pares ou trios de espécies cujos pêlos têm morfologia mais semelhante (Grupo A: L. pardalis, L. tigrinus, L. wiedii; Grupo B: L. colocolo, L. geoffroyi, P. yagouaroundi; Grupo C: P. concolor, P. onca) elevou a acurácia significativamente (91%). A identificação dos felídeos brasileiros através da microestrutura de seus pêlos é desafiadora e requer um exame cuidadoso de caracteres sutis, devendo ser apoiada por outras técnicas mais acuradas e/ou ser limitada principalmente às aplicações que não requerem identificação específica ou que trabalhem em escalas taxonômicas mais amplas.The analysis of cuticle and medulla hair microstructure is a simple and inexpensive technique to identify mammal species for a variety of applications. We studied the guard-hairs of 66 individuals of eight felid species occurring in Brazil (Leopardus colocolo, L.geoffroyi, L.pardalis, L.tigrinus, L.wiedii, Panthera onca, Puma concolor, Puma yagouaroundi), through hair samples collected from anesthetized zoo animals. The microstructure of the guard-hairs was analyzed and described through cuticle impressions and medulla preparations; a blind test was conducted to evaluate the accuracy of species identification. Although distinctive morphological characters could be identified for each species, the subtlety of these characters and the overlap of features among species resulted in a relatively poor accuracy (75%). The identification of pairs or trios of species whose hair has similar morphologies (Group A: L. pardalis, L. tigrinus, L. wiedii; Group B: L. colocolo, L. geoffroyi, P. yagouaroundi; Group C: P. concolor, P. onca) significantly improved accuracy (91%). The identification of Brazilian felids through the microstructure of their hair is challenging and requires careful examination of subtle features, and should be complemented by more accurate techniques and/or be limited mostly to applications where high accuracy is not essential or where a broader taxonomic scale is being evaluated

Ocurrencia de lepas pedunculadas (Conchoderma virgatum) (Cirripedia: thoracica) en pingüinos de Magallanes (Spheniscus magellanicus)

Magellanic Penguins (Spheniscus magellanicus) breed in Argentina, Chile and the Falkland (Malvinas) Islands, and seasonally migrate towards the coasts of Uruguay, Brazil and Peru. We report the finding of pedunculate barnacles Conchoderma virgatum (Cirripedia: Thoracica) on the body surface of seven out of 62 Magellanic Penguins. These birds were beach-cast along the coast of São Paulo state (23º57’S; 46º23’W), Brazil, on winter 2008 andwinter 2010. Barnacles were attached on the surface of the flippers, feet and lower chest, and in some cases the barnacle had over 40 grouped individuals, some measuring up to 4 cm. Affected penguins had low body mass (range = 1.6 – 2.05 kg) and were in generally poor health (cachexia, anemia and dehydration). The barnacles were considered to have occurred secondarily to the poor health status of the examined penguins. KEY WORDS.- Spheniscus, Conchoderma, penguin, parasite, BrazilPingüinos de Magallanes (Spheniscus magellanicus) son nativos de Argentina, Chile y de las Islas Malvinas (Falkland), y estacionalmente migran hacía las costas de Uruguay, Brasil y Perú. En este trabajo se reporta el hallazgo de percebes pedunculados Conchoderma virgatum (Cirripedia: Thoracica) sobre la superficie del cuerpo de siete pingüinos de Magallanes. Las aves fueron encontradas en las playas a lo largo de la costa del Estado de São Paulo(23º57’S, 46º23’O), Brasil, en los inviernos de 2008 y 2010. Los crustáceos estaban en lasuperficie de las aletas, pies y abdomen, y en algunos casos, los percebes tenían más de 40 individuos agrupados, algunos midiendo hasta 4 cm. Los pingüinos afectados tenían masa corporal baja (rango = 1.6 – 2.05 kg) y se encontraban en un mal estado de salud (caquexia, anemia y deshidratación). Se considera que los percebes observados ocurrieron posteriormente y de modo secundario al mal estado de salud de los pingüinos examinados. PALABRAS CLAVE.- Spheniscus, Conchoderma, pingüino, parásito, BrasilFAPESP 2009/53956-9CNPq 301517/2006-

Viagem sem volta. [Depoimento a Rodrigo de Oliveira Andrade]

Fêmeas de pinguins-de-magalhães morrem mais que machos durante migração anual.1. Malária aviária e pinguins no Brasil: estudo epidemiológico e patológico de uma enfermidade com potencial risco à conservação da avifauna (nº 10/51801-5); Modalidade Projeto Temático; Coord. José Luiz Catão-Dias/ FMVZ-USP; Investimento R$665.198,08 (FAPESP) 2. Plasmodium spp. em aves silvestres da Fundação Parque Zoológico de São Paulo: identificação de espécie por microscopia e código de barras de DNA (nº 12/51427- 1); Modalidade Auxílio Regular a Projeto de Pesquisa; Coord. Karin Kirchgatter/Sucen-SES/SP; Investimento R$ 52.328,50 (FAPESP

Lead exposure in consumers of culled invasive alien mammals in El Palmar National Park, Argentina

Consumption of meat from animals hunted with Pb ammunition can cause toxic accumulation with consequent health risks, even if relatively small amounts are consumed in each exposure. In El Palmar National Park, Argentina, invasive alien mammals, wild boar (Sus scrofa) and axis deer (Axis axis), are culled with Pb ammunition and their meat is consumed. In this study, we evaluated blood Pb concentrations in 58 consumers of culled game and examined Pb exposure risk according to their demographics, duty, and consumption habits. Likewise, the likelihood of exposure was evaluated by quantifying Pb concentrations in meat samples of seven culled axis deer. Twenty-seven participants (46%) had detectable blood Pb levels (limit of detection = 3.3 μg/dL), with an average 4.75 ± 1.35 μg/dL (geometric mean ± geometric S.D.); the average for all participants was 3.25 ± 1.51 μg/dL. Blood Pb concentrations were significantly higher in hunters, in participants who reported consuming game meat more than 5 times per week, and in participants who reported frequently consuming cured game meat (compared to cooked or pickled). Pb concentration varied significantly along the trajectory of the bullet in deer muscle, being highest at mid-point but with detectable Pb levels even in distant tissue samples (control), suggesting potential for dietary intake by consumers. These findings provide evidence of Pb exposure risk in consumers and emphasize the relevance of replacing Pb ammunition with non-toxic alternatives. This change would reduce dietary exposure in frequent consumers and allow the use of game meat as safe food for people whilst eliminating collateral risks to wild animals and the environment.Fil: Tammone Santos, Agostina. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - Tandil. Centro de Investigación Veterinaria de Tandil. Universidad Nacional del Centro de la Provincia de Buenos Aires. Centro de Investigación Veterinaria de Tandil. Provincia de Buenos Aires. Gobernación. Comision de Investigaciones Científicas. Centro de Investigación Veterinaria de Tandil; ArgentinaFil: Caselli, Elsa Andrea. Universidad Nacional del Centro de la Provincia de Buenos Aires; ArgentinaFil: Condorí, Walter Ezequiel. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - Tandil. Centro de Investigación Veterinaria de Tandil. Universidad Nacional del Centro de la Provincia de Buenos Aires. Centro de Investigación Veterinaria de Tandil. Provincia de Buenos Aires. Gobernación. Comision de Investigaciones Científicas. Centro de Investigación Veterinaria de Tandil; ArgentinaFil: Fernández, Valentina. Universidad Nacional del Centro de la Provincia de Buenos Aires; Argentina. Universidad Nacional de San Martín. Instituto de Investigación e Ingeniería Ambiental. - Consejo Nacional de Investigaciones Científicas y Técnicas. Oficina de Coordinación Administrativa Parque Centenario. Instituto de Investigación e Ingeniería Ambiental; ArgentinaFil: Estein, Silvia Marcela. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - Tandil. Centro de Investigación Veterinaria de Tandil. Universidad Nacional del Centro de la Provincia de Buenos Aires. Centro de Investigación Veterinaria de Tandil. Provincia de Buenos Aires. Gobernación. Comision de Investigaciones Científicas. Centro de Investigación Veterinaria de Tandil; ArgentinaFil: Vanstreels, Ralph Eric Thijl. University of California at Davis; Estados UnidosFil: Sosa, Cristian. Administración de Parques Nacionales; ArgentinaFil: Delaloye, Aldo. Administración de Parques Nacionales; ArgentinaFil: Uhart, Marcela María. University of California at Davis; Estados Unido

Chlamydiaceae-Like Bacterium in Wild Magellanic Penguins (Spheniscus magellanicus)

The family Chlamydiaceae is comprised of obligate intracellular bacteria, some of which are significant pathogens of humans and domestic animals. Magellanic penguins (Spheniscus magellanicus) are susceptible to Chlamydia psittaci outbreaks in captivity, and serological surveys detected antibodies against C. psittaci (or antigenically similar organisms) in wild populations of this species. To investigate the occurrence of Chlamydiaceae in wild Magellanic penguins, 167 cloacal swabs were collected from apparently healthy individuals at four breeding colonies along the coast of Chubut, Argentina. Real-time PCR revealed the presence of DNA from Chlamydiaceae-like bacteria in 61 cloacal swabs (36.5%). Detection did not correlate to nutritional status or to meaningful hematological abnormalities. Isolation in monkey kidney cells was unsuccessful, but DNA sequences for the 16S–23S rRNA region were obtained from one sample. Phylogenetic analysis showed a close relationship to unidentified Chlamydiaceae-like bacteria found in chinstrap penguins (Pygoscelis antarcticus) in Antarctica and seagulls in France and the Bering Sea, and to Chlamydiifrater spp. isolated from flamingos in France. Further studies are necessary to clarify the taxonomy and investigate the epidemiology, pathogenicity, and zoonotic potential of this group of Chlamydiaceae-like bacteria.Fil: Gallo Vaulet, Lucía. Universidad de Buenos Aires. Facultad de Farmacia y Bioquímica. Instituto de Fisiopatología y Bioquímica Clínica; ArgentinaFil: Vanstreels, Ralph Eric Thijl. University of California at Davis; Estados UnidosFil: Gallo, Luciana. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - Centro Nacional Patagónico. Instituto de Biología de Organismos Marinos; ArgentinaFil: Entrocassi, Andrea Carolina. Universidad de Buenos Aires. Facultad de Farmacia y Bioquímica. Instituto de Fisiopatología y Bioquímica Clínica; ArgentinaFil: Peker, Laura Alejandra. Universidad de Buenos Aires. Facultad de Farmacia y Bioquímica. Instituto de Fisiopatología y Bioquímica Clínica; ArgentinaFil: Blanco, Gabriela Silvina. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - Centro Nacional Patagónico. Centro para el Estudio de Sistemas Marinos; ArgentinaFil: Rago, María Virginia. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - Patagonia Norte. Instituto de Investigaciones en Biodiversidad y Medioambiente. Universidad Nacional del Comahue. Centro Regional Universidad Bariloche. Instituto de Investigaciones en Biodiversidad y Medioambiente; ArgentinaFil: Rodríguez Fermepin, Marcelo. Universidad de Buenos Aires. Facultad de Farmacia y Bioquímica. Instituto de Fisiopatología y Bioquímica Clínica; ArgentinaFil: Uhart, Marcela María. University of California at Davis; Estados Unido

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

Outbreak of avian malaria associated to multiple species of plasmodium in magellanic penguins undergoing rehabilitation in Southern Brazil

Avian malaria is a mosquito-borne disease caused by Plasmodium spp. Avian plasmodia are recognized conservationthreatening pathogens due to their potential to cause severe epizootics when introduced to bird populations with which they did not co-evolve. Penguins are considered particularly susceptible, as outbreaks in captive populations will often lead to high morbidity and rapid mortality. We used a multidisciplinary approach to investigate an outbreak of avian malaria in 28 Magellanic penguins (Spheniscus magellanicus) at a rehabilitation center during summer 2009 in Florianópolis, Brazil. Hemosporidian infections were identified by microscopic and molecular characterization in 64% (18/28) of the penguins, including Plasmodium (Haemamoeba) tejerai, Plasmodium (Huffia) elongatum, a Plasmodium (Haemamoeba) sp. Lineage closely related to Plasmodium cathemerium, and a Haemoproteus (Parahaemoproteus) sp. lineage closely related to Haemoproteus syrnii. P. tejerai played a predominant role in the studied outbreak and was identified in 72% (13/18) of the hemosporidian-infected penguins, and in 89% (8/9) of the penguins that died, suggesting that this is a highly pathogenic parasite for penguins; a detailed description of tissue meronts and lesions is provided. Mixed infections were identified in three penguins, and involved P. elongatum and either P. tejerai or P. (Haemamoeba) sp. that were compatible with P. tejerai but could not be confirmed. In total, 32% (9/28) penguins died over the course of 16 days despite oral treatment with chloroquine followed by sulfadiazine-trimethoprim. Hemosporidian infections were considered likely to have occurred during rehabilitation, probably from mosquitoes infected while feeding on local native birds, whereas penguin-mosquitopenguin transmission may have played a role in later stages of the outbreak. Considering the seasonality of the infection, rehabilitation centers would benefit from narrowing their efforts to preventavian malaria outbreaks to the penguins that are maintained throughout summerFAPESP 2009/53956-9, 2010/51801-5FAPEMIGCAPESCNP

Investigation of avian malaria and other blood parasites in penguins along the Atlantic coast of South America

Embora não existam colônias reprodutivas de pinguins na costa do Brasil, o país é uma importante área de invernada para o pinguim-de-Magalhães (Spheniscus magellanicus), uma espécie nativa do sul da América do Sul. Quando encontradas debilitadas em praias brasileiras, estas aves são comumente levadas a centros de reabilitação especializados para receber cuidados veterinários e, posteriormente, serem liberadas à natureza. Durante esta permanência em reabilitação, no entanto, enfermidades infecciosas como a malária aviária podem ser importantes limitantes à recuperação destas aves. A malária aviária é uma enfermidade causada por protozoários do gênero Plasmodium (Apicomplexa: Haemosporida), veiculados às aves por meio da picada de mosquitos. Enquanto estes parasitas são relativamente pouco patogênicos para a maioria das espécies aviárias, algumas aves como os pinguins são excepcionalmente mais suscetíveis a estes patógenos, podendo constituir uma significativa ameaça à sua conservação. O presente estudo investiga a ocorrência de Plasmodium spp. e outros hemoparasitas em pinguins-de-Magalhães em centros de reabilitação na costa brasileira, assim como outras espécies de pinguins em vida livre nas Ilhas South Shetland. Métodos diagnósticos morfológicos (esfregaços sanguíneos, histopatologia) e moleculares (reação em cadeia de polimerase aninhada, sequenciamento genético) foram utilizados para estudar pinguins reabilitados em diferentes instituições em seis estados do Brasil entre 1999 e 2013. Um surto de malária aviária particularmente relevante foi estudado em detalhes em um centro de reabilitação em Florianópolis, SC, tendo sido demonstrado o envolvimento de três diferentes linhagens de Plasmodium spp. em um único evento epizoótico, com elevada morbidade e mortalidade. Além disto, a ocorrência de Plasmodium spp. foi documentada em pinguins-de-Magalhães em centros de reabilitação ao longo de grande parte da costa brasileira, do extremo sul do Rio Grande do Sul à Bahia, com uma prevalência estimada entre 6.6% e 13.5%. Estas infecções apresentam marcante sazonalidade, incidindo exclusivamente nos meses mais quentes do ano (outubro a abril), e podem envolver uma grande variedade de linhagens de Plasmodium spp., algumas das quais nunca haviam sido reportadas em pinguins como P. cathemerium, P. nucleophilum e P. tejerai. Em contraste não foram encontrados hemoparasitas em pinguins amostrados nas Ilhas South Shetland, um achado consistente com estudos anteriores. À parte destas investigações epidemiológicas, foi realizada uma extensa revisão e compilação dos aspectos da literatura científica acerca desta enfermidade e outras hemoparasitoses em pinguins buscando estabelecer um panorama mais claro acerca da sua distribuição geográfica e implicações epidemiológicas e para a conservação. Com base nisto, torna-se possível desenvolver uma discussão crítica do atual estado da arte e apontar as atuais lacunas de conhecimento que possam direcionar estudos futuros. Em suma, a malária aviária é uma enfermidade relevante para as mais diversas espécies de pinguins em todo o mundo, e possui particular importância para a reabilitação e conservação destas aves na América do Sul.Despite no penguin breeding colonies along the Brazilian coast, the country is an important wintering area for the Magellanic penguin (Spheniscus magellanicus), a species native to the south of South America. When Magellanic penguins are found alive ashore on Brazilian beaches, they are taken to rehabilitation centers to receive veterinary care and then are released back into the wild. However, while in rehabilitation, infectious diseases such as avian malaria may become important limiting factors for the recovery of these birds. Avian malaria is a disease caused by protozoa of the genus Plasmodium (Apicomplexa: Haemosporida), which are transmitted to birds through mosquitoes. While these parasites are relatively non-pathogenic for most avian species, some birds such as penguins are exceptionally susceptible, such that Plasmodium poses a significant conservation threat. This study investigates the occurrence of Plasmodium spp. and other blood parasites in Magellanic penguins at rehabilitation centers along the coast of Brazil, and in other species of penguins at the South Shetland Islands, South Atlantic Ocean. In Brazil, a combination of morphological (blood smears, histopathology) and molecular (nested polymerase chain reaction, gene sequencing) diagnostic methods were employed to investigate the presence of haemoparasites in penguins undergoing rehabilitation in six states between 1999 and 2013. A particularly significant avian malaria outbreak was studied in detail at a rehabilitation center in Florianópolis, SC, where the involvement of three distinct Plasmodium spp. in a single epizootic event was demonstrated, with resultant high morbidity and mortality. The occurrence of Plasmodium spp. was documented in Magellanic penguins at rehabilitation centers along most of the Brazilian coast, from southernmost Rio Grande do Sul to Bahia, with an estimated prevalence between 6.6% and 13.5% of captive penguins. These infections were markedly seasonal, with the incidences exclusively restricted to the warmer months of the year (October to April), and involving a broad variety of Plasmodium spp. lineages, some of which had not yet been reported in penguins, such as P. cathemerium, P. nucleophilum and P. tejerai. In contrast, no blood parasites were detected in the penguins sampled at the South Shetland Islands, a finding that is consistent with previous studies. Aside from these epidemiological investigations, an extensive revision and compilation of the scientific literature was conducted for this disease and other penguins haemosporidioses, aiming to establish an integrated understanding of their geographic distribution and epidemiological and conservation implications. On this basis, it is possible to critically examine the state of the art and identify knowledge gaps that can be addressed in future studies. Avian malaria is a significant disease and conservation threat for most penguin species throughout the world, and has particular importance for the rehabilitation and conservation of penguins in South America