Leucocytozoon cariamae n. sp. and Haemoproteus pulcher coinfection in Cariama cristata (Aves: Cariamiformes): first mitochondrial genome analysis and morphological description of a leucocytozoid in Brazil.

The distribution of avian haemosporidians of the genus Leucocytozoon in the Neotropics remains poorly understood. Recent studies confirmed their presence in the region using molecular techniques alone, but evidence for gametocytes and data on putative competent hosts for Leucocytozoon are still lacking outside highland areas. We combined morphological and molecular data to characterize a new Leucocytozoon species infecting a non-migratory red-legged seriema (Cariama cristata), the first report of a competent host for Leucocytozoon in Brazil. Leucocytozoon cariamae n. sp. is distinguished from the Leucocytozoon fringillinarum group by its microgametocytes that are not strongly appressed to the host cell nucleus. The bird studied was coinfected with Haemoproteus pulcher, and we present a Bayesian phylogenetic analysis based on nearly complete mitochondrial genomes of these 2 parasites. Leucocytozoon cariamae n. sp. morphology is consistent with our phylogenetic analysis indicating that it does not share a recent common ancestor with the L. fringillinarum group. Haemoproteus pulcher and Haemoproteus catharti form a monophyletic group with Haemocystidium parasites of Reptilia, supporting the polyphyly of the genus Haemoproteus. We also discussed the hypothesis that H. pulcher and H. catharti may be avian Haemocystidium, highlighting the need to study non-passerine parasites to untangle the systematics of Haemosporida.

Preliminary assessment of anti-α-Gal IgG and IgM levels in patients with patent Plasmodium vivax infection

Anti-α-Gal responses may exert a protective effect in falciparum malaria. However, the biological role of such antibodies is still unknown during Plasmodium vivax infections. We investigated IgG and IgM responses to α-Gal in individuals with vivax malaria. Anti-α-Gal IgG and IgM levels were higher in these patients than in controls, but no significant correlation was found between parasitaemia and anti-α-Gal response, nor between this response and ABO blood group status. This is the first study to investigate anti-α-Gal antibodies in P. vivax-infected patients; a larger survey is necessary to achieve a better understanding of host immune response during vivax malaria.

Patterns of avian malaria in tropical and temperate environments: testing the "The enemy release hypothesis" / Padrões de malária aviária em região tropical e temperada: testando a "hipótese da liberação do inimigo"

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.

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.

Cytokine modulation of human blood viscosity from vivax malaria patients.

Malaria is a major infectious disease in several countries and is caused by protozoa of the genus Plasmodium. In vivax malaria patients, inflammatory processes occur, as well as changes in cytokines and blood flow. The present study analyzed the cytokine modulation of blood viscosity from patients infected with Plasmodium vivax (P. vivax). Blood samples were collected from 42 non-infected individuals (control group) and 37 individuals infected with P. vivax. The IL-2, IL-4, IL-6, IL-10, TNFα, TGF-ß and IL-17 cytokine concentrations in the serum were assessed, and the blood rheological properties were determined. The analysis of blood viscosity for shear rates revealed that the blood viscosity of the infected patients was significantly greater than that of the non-infected individuals. The viscosity of the blood was greater in the infected individuals than in the non-infected subjects. The serum from individuals with P. vivax infections exhibited higher IFN-γ and IL-17 concentrations and lower TGF-ß levels. Incubation of the blood from infected individuals with IL-17 or IL-17 associated with IFN-γ reduced the viscosity to rates equivalent to the blood from non-infected individuals. Independently of cytokine modulation, no correlation was found between the parasitemia and blood viscosity of the infected patients. These data suggest that the alterations of blood viscosity are relevant as an auxiliary tool for the clinical diagnosis of disease. In malaria, erythrocytes are more sensitive to osmotic shock, and the reduction of viscosity by IL-17 may be related to a possible immunomodulator agent during infection.

Blood parasites in passerine birds from the Brazilian Atlantic Forest / Hemoparasitos em passeriformes da Mata Atlântica Brasileira

Parasites may lead bird species to extinction, affect host temporal and spatial population dynamics, alter community structure and alter individuals’ social status. We evaluated blood parasite prevalence and intensity according to bird families and species, among 925 birds that were caught in 2000 and 2001, in the Atlantic Forest in the State of Minas Gerais, Brazil. We applied Giemsa staining to thin blood smears, to detect blood parasites. The birds (n = 15.8%) in 11 families, were infected by at least one parasite genus, especially Muscicapidae (28.3%) and Conopophagidae (25%). Among the 146 infected birds, Plasmodium was detected in all bird families and had the highest prevalence (54.8%). Trypanosoma, Haemoproteus and microfilaria had lower prevalence rates (23.3, 23.3 and 2.1%, respectively). Birds caught during the rainy season were more infected than birds caught during the dry season. The overall low prevalence of blood parasites in birds is similar to the patterns found elsewhere in the Neotropical region.

Parasitos podem levar espécies de aves à extinção, afetar as dinâmicas temporais e espaciais dos hospedeiros, alterar a estrutura de comunidades e o status social de indivíduos. Avaliou-se a prevalência e a intensidade de parasitos em famílias e espécies de 925 aves capturadas, entre 2000 e 2001, na Mata Atlântica de Minas Gerais. Foram coradas com Giemsa extensões de sangue para detectar parasitos hematozoários. As aves (n= 15,8%) 11 famílias estavam infectadas por pelo menos um gênero de parasito, especialmente Muscicapidae (28,3%) e Conopophagidae (25%). Entre as 146 aves infectadas, Plasmodium foi detectado em todas as famílias e possuiu a maior prevalência (54,8%). Trypanosoma,Haemoproteus e microfilaria possuíram baixas prevalências (23,3, 23,3 e 2,1%, respectivamente). Aves capturadasdurante a estação chuvosa estavam mais infectadas do que aves capturadas durante a estação seca. A baixa prevalência geral de parasitos do sangue das aves é semelhante aos padrões encontrados em outras localidades da região Neotropical.

Recent advances in the study of avian malaria: an overview with an emphasis on the distribution of Plasmodium spp in Brazil

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.

Long-term humoral and cellular immune responses elicited by a heterologous Plasmodium vivax apical membrane antigen 1 protein prime/adenovirus boost immunization protocol.

Apical membrane antigen 1 (AMA-1) is an invasion-related Plasmodium antigen that is expressed during both intracellular and extracellular asexual stages of the parasite's life cycle, making it an ideal target for induction of humoral and cellular immune responses that can protect against malaria. We show here that when it is administered as a recombinant protein (P) in Montanide ISA720 adjuvant, followed by a recombinant human type 5 adenovirus (Ad), intense and long-lasting Plasmodium vivax AMA-1-specific antibody responses (including both IgG1 and IgG2a), as well as proliferative memory T cell responses, can be detected in immunized mice. Memory T cells displayed both central (CD44(hi) CD62L(hi)) and effector (CD44(hi) CD62L(lo)) phenotypes, with the central memory phenotype prevailing (56% of AMA-1-specific proliferating cells). Considering the main traits of the memory immune responses induced against AMA-1, this particular sequence of immunogens (P followed by Ad), but no others (Ad/Ad, Ad/P, or P/P), displayed an optimal synergistic effect. These results give further support to the need for preclinical studies of P. vivax vaccine candidate AMA-1 administered in prime/boost protocols that include recombinant proteins and adenoviral vectors.

Identification of a highly antigenic linear B cell epitope within Plasmodium vivax apical membrane antigen 1 (AMA-1).

Apical membrane antigen 1 (AMA-1) is considered to be a major candidate antigen for a malaria vaccine. Previous immunoepidemiological studies of naturally acquired immunity to Plasmodium vivax AMA-1 (PvAMA-1) have shown a higher prevalence of specific antibodies to domain II (DII) of AMA-1. In the present study, we confirmed that specific antibody responses from naturally infected individuals were highly reactive to both full-length AMA-1 and DII. Also, we demonstrated a strong association between AMA-1 and DII IgG and IgG subclass responses. We analyzed the primary sequence of PvAMA-1 for B cell linear epitopes co-occurring with intrinsically unstructured/disordered regions (IURs). The B cell epitope comprising the amino acid sequence 290-307 of PvAMA-1 (SASDQPTQYEEEMTDYQK), with the highest prediction scores, was identified in domain II and further selected for chemical synthesis and immunological testing. The antigenicity of the synthetic peptide was identified by serological analysis using sera from P. vivax-infected individuals who were knowingly reactive to the PvAMA-1 ectodomain only, domain II only, or reactive to both antigens. Although the synthetic peptide was recognized by all serum samples specific to domain II, serum with reactivity only to the full-length protein presented 58.3% positivity. Moreover, IgG reactivity against PvAMA-1 and domain II after depletion of specific synthetic peptide antibodies was reduced by 18% and 33% (P = 0.0001 for both), respectively. These results suggest that the linear epitope SASDQPTQYEEEMTDYQK is highly antigenic during natural human infections and is an important antigenic region of the domain II of PvAMA-1, suggesting its possible future use in pre-clinical studies.

Naturally acquired antibodies to merozoite surface protein (MSP)-1(19) and cumulative exposure to Plasmodium falciparum and Plasmodium vivax in remote populations of the Amazon Basin of Brazil

To infer recent patterns of malaria transmission, we measured naturally acquired IgG antibodies to the conserved 19-kDa C-terminal region of the merozoite surface protein (MSP)-1 of both Plasmodium vivax (PvMSP-1(19)) and Plasmodium falciparum (PfMSP-1(19)) in remote malaria-exposed populations of the Amazon Basin. Community-based cross-sectional surveys were carried out between 2002 and 2003 in subjects of all age groups living along the margins of the Unini and Jaú rivers, Northwestern Brazil. We found high prevalence rates of IgG antibodies to PvMSP-1(19) (64.0 - 69.6 percent) and PfMSP-1(19) (51.6 - 52.0 percent), with significant differences in the proportion of subjects with antibodies to PvMSP-1(19) according to age, place of residence and habitual involvement in high-risk activities, defining some groups of highly exposed people who might be preferential targets of malaria control measures. In contrast, no risk factor other than age was significantly associated with seropositivity to PfMSP-1(19). Only 14.1 percent and 19.3 percent of the subjects tested for antibodies to PvMSP-1(19) and PfMSP-1(19) in consecutive surveys (142 - 203 days apart) seroconverted or had a three fold or higher increase in the levels of antibodies to these antigens. We discuss the extent to which serological data correlated with the classical malariometric indices and morbidity indicators measured in the studied population at the time of the seroprevalence surveys and highlight some limitations of serological data for epidemiological inference.