Spatial distribution of vector mosquitoes plays a critical role in the dynamics of associated diseases' spread across diverse landscapes. In Botswana, six Districts are implicated as malaria endemic zones, one of which is the Central District comprising both malaria endemic and non-endemic sub-districts. Despite being the biggest in the country, mosquito diversity in this District is under-explored, more so in the malaria non-endemic sub-districts. Here, we thus sampled mosquito adults and larvae from the malaria endemic sub-district of Bobirwa and non-endemic sub-districts of Palapye and Serowe, to determine spatial mosquito abundance and diversity in the District. Overall, all the sub-districts had a representation of key mosquito taxa of medical and economic importance (Aedes, Culex and Anopheles), irrespective of malarial endemicity status. Bobirwa had the highest number of mosquitoes sampled (429) although the greatest species richness (0.8511) was observed from Palapye. Moreover, Palapye also recorded a species from another genus, Culiseta longiareolata, a known natural vector of avian malaria parasites. Given global climate shift projections for the region, there is a need for continuous area-wide surveillance for vector mosquitoes and associated parasites in curbing the risk of emerging and re-emerging infections. While the Anopheles-centric approach to mosquito control is still necessary, a holistic approach, incorporating other vector incriminated mosquito species is warranted, particularly given shifting climates and the presence of invasive disease associated vector mosquito species.

RESUMEN Los hemosporidios aviares forman un grupo diverso de parásitos transmitidos por vectores que pueden perjudicar a sus hospederos y amenazar la conservación de especies susceptibles. Evaluamos la prevalencia y parasitemia de hemosporidios en aves silvestres de la Reserva de la Biósfera Sierra de Huautla, Morelos, al sur de México. Las aves se capturaron usando redes de niebla en tres tipos de hábitat (conservado, perturbado y agrícola) y durante dos temporadas (lluvias y secas). Se realizaron frotis de capa fina para su análisis microscópico. Capturamos 142 aves pertenecientes a 17 especies. Los parásitos presentes fueron Haemoproteus spp., Plasmodium spp., y microfilarias. La prevalencia fue similar entre tipos de uso de suelo (conservado (26.3 %), perturbado (36.4 %) y agrícola (29.9 %)) y temporadas (lluvias (29.7 %) y secas (29.3 %)), pero varió por género y grupo de parásito (Haemoproteus spp. (28.2%), Plasmodium spp. (2.1 %), coinfecciones (5.6 %), y microfilarias (4.9 %)). La mayoría de las aves presentó una parasitemia baja (< 0.1% de eritrocitos infectados) y solo una, con apariencia malsana, presentó alta parasitemia (> 0.5 %). Reportamos por primera vez la infección de hemoparásitos en 12 especies de aves y 16 nuevas asociaciones parásito-hospedero. Este es el primer estudio de hemoparásitos en esta región y proporciona información fundamental para investigaciones futuras.

ABSTRACT Avian haemosporidians form a diverse group of vector-borne parasites that can cause detrimental effects on their hosts and threaten the conservation of susceptible species. We explored the prevalence and parasitemia of haemosporidians infecting wild birds from Sierra de Huautla Biosphere Reserve in Morelos, southern Mexico. Birds were caught using mist nets in three habitat conditions (conserved, disturbed, and agricultural) and during two seasons (rainy and dry). Thin blood smears were prepared from blood samples for microscopic analysis. We captured 142 birds belonging to 17 species. We identified Haemoproteus spp., Plasmodium spp., and microfilaria. Prevalence was similar among land-use types (conserved (26.3%), disturbed (36.4%) and agricultural (29.9%)), and between seasons (rainy (29.7%) and dry (29.3%)), but varied per parasite genus and group (Haemoproteus spp. (28.2%), Plasmodium spp. (2.1%), coinfections (5.6%), and microfilaria (4.9%)). Parasitemia was low in most birds (< 0.1% erythrocytes infected), and only one, with an unhealthy appearance, presented high parasitemia (> 0.5%). We reported for the first time 12 species of birds infected with haemosporidian parasites and 16 new host-parasite associations in total. This is the first study of blood parasites in this region and provides fundamental information for future research.

Abstract Aims. The goal of the current study was to determine the prevalence of Plasmodium in the population of Phoenicopterus ruber kept under human care at Zoofari Conservation Center, Morelos, Mexico Settings and Design. Avian malaria vectors belonging to the family Culicidae has been reported at different Mexican States. Malaria infections compromise different host ranges including Phoenicopteriformes (Phoeniconaias minor, Phoenicopterus chilensis). The singled population housed in Zoofari, Conservation Center may harbor subclinical malaria (Plasmodium) infections. Methods and Material A descriptive crosssectional study in a singled population of American flamingos (Phoenicopterus ruber). We compared and contrasted two diagnostic techniques, microscopy and polymerase chain reaction (PCR) for avian haemosporidia. In this study 80 blood smears stained in a Romanowsky stain were examined by microscopy, also molecular analysis were conducted, PCR was performed with the use of primers that amplifies parasite mtDNA from species of Haemoproteus, Plasmodium, Leucocytozoon. Results. A total of 80 Phoenicopterus ruber birds were examined for malarial parasites using both microscopy and PCR diagnostic methods, none of them were positives. Conclusions. Our study corresponds to the first of its kind in the region. After implementing two different diagnostic methods for avian malaria, no evidence was obtained of the presence of hemoparasites in the Phoenicopterus ruber population. This type of research allows us to understand the dynamics and identify emerging diseases that can affect animals and man in a certain geographic region.

Resumen Objetivos. Establecer la prevalencia de Plasmodium en la población de Phoenicopterus ruber mantenidos bajo cuidado humano en el Centro de Conservación Zoofari, Morelos, México. Configuración y Diseño Se han presentado reportes de vectores de malaria aviar pertenecientes a la familia Culicidae en estado de Morelos, México. La malaria aviar puede afectar diversas especies de aves incluyendo Phoenicopteriformes (Phoeniconaias minor, Phoenicopterus chilensis) la población de interés podría ser portadora subclínica de Plasmodium. Materiales y Métodos se realizó un estudio exploratorio en el total de la población de Phoenicopterus ruber. En la investigación se implementaron dos técnicas de diagnóstico de hemoparásitos, la primera corresponde a análisis de frotis sanguíneos teñidos con una tinción rápida tipo Romanowsky los cuales fueron evaluados mediante microscopía óptica convencional y la segunda concierne la realización de la PCR utilizando iniciadores para amplificar ADN mitocondrial de tres especies de hemoparásitos (Haemoproteus, Plasmodium, Leucocytozoon). Análisis Estadístico utilizado. los resultados negativos del estudio no permiten realizar análisis estadístico Resultados. En la evaluación mediante microscopía convencional no se evidenció en ningún frotis sanguíneo, presencia de merogonias o gránulos de hemozoína, obteniendo una totalidad de 80 muestras negativas. Las PCR realizadas bajo las condiciones utilizadas no demostraron amplificación exitosa de ADN parasitario. Conclusiones. Nuestro estudio corresponde al primero de este tipo en la región. Después de implementar dos técnicas diferentes de diagnóstico para malaria aviar no se obtiene evidencia de la presencia de hemoparásitos en la población de Phoenicopterus ruber. Este tipo de investigaciones permite entender la dinámica e identificar enfermedades emergentes que puedan afectar a los animales y al hombre en una determinada región geográfica.

Resumo O objetivo deste estudo foi verificar a presença e a identificação espécies de parasitas hemosporídeos em pombos (Zenaida auriculata) no Brasil. Duzentos e onze pombos machos e fêmeas foram capturados em quatro regiões diferentes de Londrina, sul do Brasil. Amostra de sangue foi coletada em tubos contendo EDTA por meio de punção cardíaca, após eutanásia em câmara de CO2. Uma nested PCR com alvo no gene mitocondrial citocromo b (cyt b) de Haemoproteus spp./Plasmodium spp. foi realizada, seguida de digestão enzimática para identificar o gênero. A árvore filogenética foi construída para determinar a relação com outras espécies. Das 211 pombas, 209 (99,05%) foram positivas para Haemoproteus spp./Plasmodium spp. A análise RFLP demonstrou que 72,72% (152/209) das pombas foram positivas somente para Haemoproteus spp.; 6,22% (13/209) foram positivas somente para Plasmodium e 21,05% (44/209) das pombas tiveram infecções mistas. A análise genética mostrou quatro amostras homólogas com H. multipigmentatus e uma com Plasmodium spp. Este é o primeiro estudo molecular de hemoparasitas em pombos no Brasil. E é também a primeira descrição da infecção por H. multipigmentatus e Plasmodium spp. em pombos Z. auriculata no Brasil.

Abstract The aim of this study was to verify the presence and identify the species of haemosporidian parasites in eared doves (Zenaida auriculata) in Brazil. Two hundred and eleven male and female eared doves were trap-captured in four different regions of Londrina city, in southern Brazil. Whole blood was collected in EDTA tubes through heart puncture after euthanasia in a CO2 chamber. A nested PCR targeting the mitochondrial cytochrome b gene (cyt b) of Haemoproteus spp./Plasmodium spp. was performed, followed by an enzymatic digestion to identify the genus. Phylogenetic trees were constructed to determine the closely related species. Out of 211 eared doves, 209 (99.05%) were positive for Haemoproteus spp. and/or Plasmodium spp. RFLP analysis showed that 72.72% (152/209) of eared doves were positive only for Haemoproteus spp., 6.22% (13/209) were positive only for Plasmodium spp., and 21.05% (44/209) of eared doves had mixed infections. Genetic analysis found four samples that were homologous with Haemoproteus multipigmentatus and one that was homologous with Plasmodium sp. This is the first molecular study of hemoparasites from eared doves in Brazil, and it is also the first description of H. multipigmentatus and Plasmodium spp. infection in eared doves in Brazil.

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.

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: De acordo com a hipótese da liberação do inimigo (HLI), a disseminação de espécies invasoras será facilitada pela liberação de seus inimigos ao ocuparem novas áreas. No entanto, a HLI raramente é testada comparando-se as populações de espécies nativas (não invasivas, estabelecidas há muito tempo) que apresentam expansão ou alteração de habitats, com populações das mesmas espécies em habitats que foram invadidos. Testamos a HLI com relação aos níveis de parasitas no sangue (prevalência e intensidade de Plasmodium spp. e Haemoproteus spp.). De (a) duas espécies estreitamente relacionadas e amplamente distribuídas de Turdus (Turdus leucomelas e T. merula), e (b) um pardal invasor (Passer domesticus) cujo alcance se expandiu do Velho Mundo para o Novo Mundo desde o século 18. Um total de 158 aves foram amostradas em Portugal e 99 no Brasil. Todas as espécies foram parasitadas e 55% dos indivíduos foram parasitados, sendo que a intensidade média da infecção foi de 28 parasitas por 10.000 eritrócitos. Avaliamos se as diferenças nos níveis de infecção (prevalência e intensidade) foram devidas ao local (tropical/Novo Mundo e temperado/Velho Mundo) ou espécies hospedeiras. A HLI foi corroborada: Passer domesticus e Turdus merula apresentaram valores mais elevados de parasitismo no Velho Mundo do que no Novo Mundo. Assim, P. domesticus parece estar se beneficiando de sua expansão "recente" em comparação com T. leucomelas, através da liberação ecológica de seus parasitas nativos porque os parasitas da área recentemente invadida parecem infestar espécies nativas.

Abstract: According to the enemy release hypothesis (ERH) the spread of invasive species will be facilitated by release from their enemies as they occupy new areas. However, the ERH has rarely been tested by comparing populations of native (non-invasive, long established) species with expanding or shifting ranges, to the same species as invasive in another area. We tested the ERH with respect to blood parasite levels (prevalence and intensity of Plasmodium spp. and Haemoproteus spp.) of (a) two closely related, widely distributed species of thrush (Turdus leucomelas and T. merula), and (b) an invasive sparrow (Passer domesticus) whose range has expanded from the Old World to the New World since the 18th century. A total of 158 birds were sampled in Portugal and 99 in Brazil. All bird species were parasitized, and 55% of the individuals collected were parasitized, and the mean intensity of infection was of 28 parasites per 10,000 erythrocytes. We assessed whether differences in levels of infection (prevalence and intensity) were due to site (tropical/New World and temperate/Old World) or host species. The ERH was supported: Passer domesticus and Turdus merula had higher levels of parasitism in the Old World than in the New World. Thus, P. domesticus seems to be benefitting from its "recent" range expansion, compared to T. leucomelas, through ecological release from its native parasites and because the parasites of the recently invaded area seem to be infesting native species instead.

The African penguin (Spheniscus demersus) is the only penguin species that breeds on the African continent and it is currently classified as endangered. Its conservation is assisted by the Southern African Foundation for the Conservation of Coastal Birds (SANCCOB) which is a seabird rehabilitation facility based at the Rietvlei Wetland Reserve in Tableview, Cape Town. Despite the success of SANCCOB in rehabilitating diseased, injured or oiled penguins, significant mortalities have occurred at the facility as a result of avian malaria. Avian malaria can be contracted during rehabilitation during which penguins are inadvertently exposed to additional threats. An enzyme-linked immunosorbent assay (ELISA) was used to assess the anti-Plasmodium antibody levels of penguins to avian malaria on entry into the SANCCOB facility from 2001 to 2004 and during their rehabilitation process. Using blood smear data, avian malaria prevalence and malaria-related deaths were also monitored from 2002 to 2013. Significant increases in anti-Plasmodium antibody levels after admission were found during summer months. New infection and not parasite recrudescence was concluded to be the cause of this increase. This source was confirmed by a dramatic drop in penguin mortalities upon exclusion of mosquito vectors in 2008. Mortalities did not depend on the birds' abilities to produce an anti-Plasmodium antibody response and oiling had no influence on immunity or prevalence of avian malaria infections. This study highlights the importance of mosquito vector control to control pathogen exposure in wild bird rehabilitation centres. SIGNIFICANCE: •Efforts to assist with the conservation of endangered species can unintentionally add to the conservation burden. •Rehabilitation influences exposure of African penguins to avian malaria. •Avian malaria prevalence and mortality are not influenced by oiling or anti-Plasmodium antibody responses. •Vector control can limit avian malaria exposure in wild bird rehabilitation centres.

Nos últimos anos infecção por protozoários hemosporídeos dos gêneros Plasmodium e Haemoproteus, tem sido considerada um dos fatores mais importantes relacionados com a extinção e / ou declínio da população de várias espécies de aves em todo o mundo. No Brasil, apesar da grande biodiversidade aviária, poucos estudos foram desenvolvidos para detectar a infecção, especialmente entre as aves silvestres mantidas em cativeiro. Assim, o objetivo deste estudo foi analisar a prevalência de infecção por Plasmodium spp. e Haemoproteus spp. em aves silvestres em cativeiro na Mata Atlântica do sudeste do Brasil, utilizando microscopia convencional e reação em cadeia da polimerase. Amostras de sangue de 119 aves mantidas em cativeiro no Ibama durante o período de julho de 2011 a julho de 2012, foram coletadas. A densidade parasitária foi determinada com base apenas em leituras de esfregaços de sangue por microscopia fotônica. A prevalência média de infecção por Plasmodium spp. e Haemoproteus spp. obtida por exame microscópico de esfregaços sanguíneos e PCR foi semelhante (83,19% e 81,3%, respectivamente), com Caracara plancus e Saltator similis sendo as espécies mais parasitadas. A parasitemia média determinada pela contagem microscópica de formas evolutivas de Plasmodium spp. e Haemoproteus spp. foi de 1,51%. Os resultados obtidos neste estudo reforçam a importância do manejo de aves em cativeiro, especialmente quando serão reintroduzidas na natureza.

In recent years haemosporidian infection by protozoa of the genus Plasmodium and Haemoproteus, has been considered one of the most important factors related to the extinction and/or population decline of several species of birds worldwide. In Brazil, despite the large avian biodiversity, few studies have been designed to detect this infection, especially among wild birds in captivity. Thus, the objective of this study was to analyze the prevalence of Plasmodium spp. and Haemoproteus spp. infection in wild birds in captivity in the Atlantic Forest of southeastern Brazil using microscopy and the polymerase chain reaction. Blood samples of 119 different species of birds kept in captivity at IBAMA during the period of July 2011 to July 2012 were collected. The parasite density was determined based only on readings of blood smears by light microscopy. The mean prevalence of Plasmodium spp. and Haemoproteus spp. infection obtained through the microscopic examination of blood smears and PCR were similar (83.19% and 81.3%, respectively), with Caracara plancus and Saltator similis being the most parasitized. The mean parasitemia determined by the microscopic counting of evolutionary forms of Plasmodium spp. and Haemoproteus spp. was 1.51%. The results obtained from this study reinforce the importance of the handling of captive birds, especially when they will be reintroduced into the wild.

O pinguim-de-Magalhães (Spheniscus magellanicus) migra das suas colônias reprodutivas até o extremo sul do Brasil. Esses pinguins frequentemente são acometidos por doenças, notavelmente a malária aviária, que é relatada no Brasil e no mundo. A detecção de Plasmodium spp. no esfregaço sanguíneo é o método de rotina mas apresenta baixa sensibilidade quando comparado aos métodos moleculares. Considerando o impacto negativo da malária aviária nos pinguins, o objetivo deste estudo foi detectar a presença de Plasmodium spp. em pinguins-de-Magalhães usando a Reação em Cadeia da Polimerase (PCR), verificar as alterações clínicas, hematológicas e bioquímicas e o provável prognóstico em resposta à infecção. Amostras de sangue foram obtidas de 75 pinguins para determinar o hematócrito (Ht), contagens totais de eritrócitos e leucócitos, volume globular médio (VGM), concentração de ácido úrico, proteínas totais, albumina, globulinas e atividade da aspartato aminotransferase (AST). O sangue total foi usado para ensaios de PCR. A detecção de Plasmodium spp. foi obtida em 32,0% dos indivíduos pela PCR e em 29,3% pela análise microscópica. Anorexia, diarreia e alterações neurológicas foram mais frequentes nos pinguins com malária, e uma diferença significativa no peso entre pinguins infetados e não infectados foi detectada. Ht e VGM não mostraram diferença significativa. A contagem de eritrócitos e leucócitos foi menor nos animais com a malária aviária e leucopenia esteve presente em alguns pinguins. Contagens de basófilos e linfócitos foram mais baixas nos pinguins infectados, bem como elevadas contagens de monócitos estavam presentes. Não houve diferença significativa para a atividade da AST entre os animais infectados e não infectados. Houve um aumento significativo nos valores de ácido úrico, entretanto houve redução nos valores da albumina entre os pinguins infectados avaliados. Concluiu-se que Plasmodium spp. ocorre em pinguins-de-Magalhães de centros de reabilitação no sudeste brasileiro, comprometendo o peso dos animais infectados e com alterações clínicas aparecendo em casos graves da doença. Percebeu-se que embora alterações hematológicas possam não ser conclusivas, leucopenia, monocitose e diminuição de basófilos e linfócitos revelaram prognóstico desfavorável. A infecção por Plasmodium spp. pode cursar com aumento da concentração de ácido úrico e baixos valores de albumina.

Magellanic penguins (Spheniscus magellanicus) routinely migrate from their breeding colonies to Southern Brazil often contracting diseases during this migration, notably avian malaria, which has been already reported in Brazil and throughout the world. Detection of Plasmodium spp. in blood smears is the routine diagnostic method of avian malaria, however it has a low sensitivity rate when compared to molecular methods. Considering the negative impact of avian malaria on penguins, the aim of this study was to detect the presence of Plasmodium spp. in Magellanic penguins using Polymerase Chain Reaction (PCR) and by verifying clinical, hematological, and biochemical alterations in blood samples as well as to verify the likely prognosis in response to infection. Blood samples were obtained from 75 penguins to determine packed cell volume (PCV), red blood cell (RBC) and white blood cell (WBC) counts, mean corpuscular volume (MCV), uric acid, total protein, albumin, globulin and aspartate aminotransferase (AST) activity levels. Whole blood samples were used for PCR assays. Plasmodium spp. was detected in 32.0% of the specimens using PCR and in 29.3% using microscopic analyses. Anorexia, diarrhea and neurological disorders were more frequent in penguins with malaria and a significant weight difference between infected and non-infected penguins was detected. PCV and MCV rates showed no significant difference. RBC and WBC counts were lower in animals with avian malaria and leukopenia was present in some penguins. Basophil and lymphocyte counts were lower in infected penguins along with high monocyte counts. There was no significant difference in AST activities between infected and non-infected animals. There was a significant increase in uric acid values, however a decrease in albumin values was observed in infected penguins. Based on this study, we concluded that Plasmodium spp. occurs in Magellanic penguins of rehabilitation centers in Southeastern Brazil, compromising the weight of infected animals with clinical alterations appearing in severe cases of this disease. It was also noted that, although the hematological abnormalities presented by these animals may not have been conclusive, leukopenia, monocytosis and the decrease of basophils and lymphocytes revealed an unfavorable prognosis, and Plasmodium spp. infections may progress with elevated uric acid concentration and low albumin levels.

From an ecological point of view, parasites are considered important selective forces that influence all aspects of a host's life. The purpose of this study was to assess the health state of Turdus leucomelas Vieillot, 1818 (Turdidae: Passeriformes) inhabiting a Brazilian Cerrado in the breeding and molting seasons through a hematological parameter analysis and an evaluation of infection by blood parasites. The birds were collected with mist-nets, ringed and blood sampled to assess hematological parameters (hematocrit, hemoglobin concentration and white blood cells) and blood parasites. We detected blood parasites through optical microscopy and subsequently used PCR (amplification of the 18sRNA gene) to verify the presence of Plasmodium spp. (avian malaria). During the breeding season, the hemoglobin level and CHGM percentage were greater. Global leukocyte count was positively related to hemoglobin level, CHGM percentage and body weight. Of the total 31 T. leucomelas individuals examined, 18 presented Plasmodium parasites (58% of prevalence). There was a significant relationship between the presence of Plasmodium spp. and decreased CHGM. These results suggest a connection between the health parameters of wild birds and the physiological stress associated with the breeding season.

Avian malaria parasites (Plasmodium) have a worldwide distribution except for Antarctica. They are transmitted exclusively by mosquito vectors (Diptera: Culicidae) and are of particular interest to health care research due to their phylogenetic relationship with human plasmodia and their ability to cause avian malaria, which is frequently lethal in non-adapted avian hosts. However, different features of avian Plasmodium spp, including their taxonomy and aspects of their life-history traits, need to be examined in more detail. Over the last 10 years, ecologists, evolutionary biologists and wildlife researchers have recognized the importance of studying avian malaria parasites and other related haemosporidians, which are the largest group of the order Haemosporida by number of species. These studies have included understanding the ecological, behavioral and evolutionary aspects that arise in this wildlife host-parasite system. Molecular tools have provided new and exiting opportunities for such research. This review discusses several emerging topics related to the current research of avian Plasmodium spp and some related avian haemosporidians. We also summarize some important discoveries in this field and emphasize the value of using both polymerase chain reaction-based and microscopy-based methods in parallel for wildlife studies. We will focus on the genus Plasmodium, with an emphasis on the distribution and pathogenicity of these parasites in wild birds in Brazil.

Aborda a contribuição de cientistas brasileiros aos estudos sobre o protozoário causador da malária. Ao colocar em foco os trabalhos de Henrique Aragão e Wladimir Lobato Paraense, destaca a importância da malária aviária para o entendimento da malária humana e sua terapêutica, a rede de relações científicas estabelecidas, as agendas comuns de pesquisa, as trocas de informações entre pesquisadores, assim como o papel por eles desempenhado no contexto internacional das descobertas científicas.

The article explores Brazilian investigators' contributions to research on the protozoan causative agent of malaria. Focusing on the work of Henrique Aragão and Wladimir Lobato Paraense, it underscores the importance of avian malaria in elucidating human malaria and treatment options, and also examines the network of scientific relations forged by these researchers, their shared research agendas, exchange of information with other researchers, and role within the international context of scientific discoveries.

TNF-α is an important human cytokine that imparts dualism in malaria pathogenicity. At high dosages, TNF-α is believed to provoke pathogenicity in cerebral malaria; while at lower dosages TNF-α is protective against severe human malaria. In order to understand the human TNF-α gene and to ascertain evolutionary aspects of its dualistic nature for malaria pathogenicity, we characterized this gene in detail in six different mammalian taxa. The avian taxon, Gallus gallus was included in our study, as TNF-α is not present in birds; therefore, a tandemly placed duplicate of TNF-α (LT-α or TNF-β) was included. A comparative study was made of nucleotide length variations, intron and exon sizes and number variations, differential compositions of coding to non-coding bases, etc., to look for similarities/dissimilarities in the TNF-α gene across all seven taxa. A phylogenetic analysis revealed the pattern found in other genes, as humans, chimpanzees and rhesus monkeys were placed in a single clade, and rats and mice in another; the chicken was in a clearly separate branch. We further focused on these three taxa and aligned the amino acid sequences; there were small differences between humans and chimpanzees; both were more different from the rhesus monkey. Further, comparison of coding and non-coding nucleotide length variations and coding to non-coding nucleotide ratio between TNF-α and TNF-β among these three mammalian taxa provided a first-hand indication of the role of the TNF-α gene, but not of TNF-β in the dualistic nature of TNF-α in malaria pathogenicity.

Neste trabalho, três isolados de Plasmodium juxtanucleare foram analisados com base na morfologia, morfometria e parâmetros parasitológicos. Cada isolado foi coletado de aves (Gallus gallus) adultas infectadas naturalmente de áreas rurais de três municípios brasileiros: Seropédica (22º 48' S; 43º 41' W), no estado do Rio de Janeiro; Cruzeiro (22º 33' S; 44º 57' W), no estado de São Paulo; e Santa Bárbara do Tugúrio (21º 15' S; 43º 27' W), no estado de Minas Gerais. As amostras de sangue coletadas de cada ave infectada foram inoculadas em três grupos de dez aves (21 dias de idade). Esfregaços sangüíneos das aves infectadas experimentalmente foram realizados de dois em dois dias durante um período de 69 dias para avaliar a parasitemia. Para análises morfofisiológica e morfométrica, foram mensurados 30 indivíduos de cada estágio intraeritrocítico. Foram tomadas as medidas do diâmetro maior (DM) e diâmetro menor (dm), com os quais foi estimado o índice morfométrico (Mi=md/MD) e o tamanho (T=pab, a= md/ 2; b=MD/2). Os resultados indicaram uma parasitemia homogênea entre os três isolados, havendo diferenças apenas nas formas e tamanhos dos estádios parasitários.

In this work, three isolates of Plasmodium juxtanucleare have been analyzed based on morphological, morphometric and parasitic parameters. Each isolate was sampled from naturally infected adult chicken (Gallus gallus) from rural areas of three Brazilian municipalities: Seropédica (22º 48' S; 43º 41'W), in the state of Rio de Janeiro; Cruzeiro (22º 33' S; 44º 57'W), in the state of São Paulo; and Santa Bárbara do Tugúrio (21º 15' S; 43º 27' W), in the state of Minas Gerais. The blood samples taken from each infected chicken were inoculated in three groups of ten young chicken (21 days old). Blood smears of the experimentally infected chicken were sampled every two days until the 69th day in order to evaluate the parasitemia. For the morphological-descriptive and morphometric analyses, we measured 30 individuals from each of the intraerythocytic states, measures of the major (MD) and minor diameters (md), the estimation of morphometric index (Mi=md/MD) and size (T=pab, a= md/2; b=MD/2). The results indicated low and homogeneous parasitemia rates in the three strains, which showed differences among shape and size of the parasitic stadia displayed.

The technique to generate transgenic mosquitoes requires adaptation for each target species because of aspects related to species biology, sensitivity to manipulation and rearing conditions. Here we tested different parameters on the microinjection procedure in order to obtain a transgenic Neotropical mosquito species. By using a transposon-based strategy we were able to successfully transform Aedes fluviatilis (Lutz), which can be used as an avian malaria model. These results demonstrate the usefulness of the piggyBac transposable element as a transformation vector for Neotropical mosquito species and opens up new research frontiers for South American mosquito vectors.