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.

Haemosporidian parasites and incubation period influence plumage coloration in tanagers (Passeriformes: Thraupidae).

Birds are highly visually oriented and use plumage coloration as an important signalling trait in social communication. Hence, males and females may have different patterns of plumage coloration, a phenomenon known as sexual dichromatism. Because males tend to have more complex plumages, sexual dichromatism is usually attributed to female choice. However, plumage coloration is partly condition-dependent; therefore, other selective pressures affecting individuals' success may also drive the evolution of this trait. Here, we used tanagers as model organisms to study the relationships between dichromatism and plumage coloration complexity in tanagers with parasitism by haemosporidians, investment in reproduction and life-history traits. We screened blood samples from 2849 individual birds belonging to 52 tanager species to detect haemosporidian parasites. We used publicly available data for plumage coloration, bird phylogeny and life-history traits to run phylogenetic generalized least-square models of plumage dichromatism and complexity in male and female tanagers. We found that plumage dichromatism was more pronounced in bird species with a higher prevalence of haemosporidian parasites. Lastly, high plumage coloration complexity in female tanagers was associated with a longer incubation period. Our results indicate an association between haemosporidian parasites and plumage coloration suggesting that parasites impact mechanisms of sexual selection, increasing differences between the sexes, and social (non-sexual) selection, driving females to develop more complex coloration.

Migratory behaviour does not alter cophylogenetic congruence between avian hosts and their haemosporidian parasites.

Parasites display various degrees of host specificity, reflecting different coevolutionary histories with their hosts. Avian hosts follow multiple migration patterns representing short but also long distances. As parasites infecting migratory birds are subjected to multiple environmental and biotic changes through their flyways, migration may disrupt or strengthen cophylogenetic congruence between hosts and parasites. On the one hand, parasites might adapt to a single migratory host, evolving to cope with the specific challenges associated with the multiple habitats occupied by the host. On the other, as migrants can introduce parasites into new habitats, higher rates of host switching could also disrupt cophylogenetic patterns. We analysed whether migratory behaviour shapes avian haemosporidian parasite­host cophylogenetic congruence by testing if contributions of host­parasite links to overall congruence differ among resident and short-, variable- and long-distance migrants globally and within South America only. On both scales, we found significant overall cophylogenetic congruence by testing whether overall congruence differed between haemosporidian lineages and bird species. However, we found no difference in contribution towards congruence among links involving resident vs migratory hosts in both models. Thus, migratory behaviour neither weakens nor strengthens bird­haemosporidian cophylogenetic congruence, suggesting that other avian host traits are more influential in generating phylogenetic congruence in this host­parasite system.

Host life-history traits predict haemosporidian parasite prevalence in tanagers (Aves: Thraupidae).

Vector-borne parasites are important ecological drivers influencing life-history evolution in birds by increasing host mortality or susceptibility to new diseases. Therefore, understanding why vulnerability to infection varies within a host clade is a crucial task for conservation biology and for understanding macroecological life-history patterns. Here, we studied the relationship of avian life-history traits and climate on the prevalence of Plasmodium and Parahaemoproteus parasites. We sampled 3569 individual birds belonging to 53 species of the family Thraupidae. Individuals were captured from 2007 to 2018 at 92 locations. We created 2 phylogenetic generalized least-squares models with Plasmodium and Parahaemoproteus prevalence as our response variables, and with the following predictor variables: climate PC1, climate PC2, body size, mixed-species flock participation, incubation period, migration, nest height, foraging height, forest cover, and diet. We found that Parahaemoproteus and Plasmodium prevalence was higher in species inhabiting open habitats. Tanager species with longer incubation periods had higher Parahaemoproteus prevalence as well, and we hypothesize that these longer incubation periods overlap with maximum vector abundances, resulting in a higher probability of infection among adult hosts during their incubation period and among chicks. Lastly, we found that Plasmodium prevalence was higher in species without migratory behaviour, with mixed-species flock participation, and with an omnivorous or animal-derived diet. We discuss the consequences of higher infection prevalence in relation to life-history traits in tanagers.

Blood parasites of passerines in the Brazilian Pampas and their implications for a potential population supplementation program for the endangered Yellow Cardinal (Gubernatrix cristata).

Espinilho savanna ("seasonal steppe savanna") is a unique vegetation formation of the Pampas biome that is found near the tri-border of Brazil, Uruguay, and Argentina. The Yellow Cardinal (Gubernatrix cristata) is a flagship species of this ecosystem, but it is classified as "critically endangered" in Brazil due to habitat loss and poaching for the illegal trade. Population supplementation through the release of individuals that were captive-bred or apprehended by authorities from the illegal trade has been considered as a conservation strategy for this species; however, the risk of pathogen introduction is a critical concern. We used microscopy and molecular methods to investigate the occurrence of blood parasites in wild passerines (n = 64, including three Yellow Cardinals) at Espinilho State Park, Rio Grande do Sul, Brazil, and in captive Yellow Cardinals (n = 30) at three facilities in Brazil. Haemosporidian parasites were detected in the blood smears of 10.9% of the wild passerines, comprising the morphospecies Haemoproteus erythrogravidus in Rufous-collared Sparrow (Zonotrichia capensis), H. quiscalus in Grayish Baywing (Agelaioides badius), and H. tyranni in Great Kiskadee (Pitangus sulphuratus); these are the southernmost records for these morphospecies and their first record for the Pampas biome. No haemosporidian parasites were detected in the blood smears of the Yellow Cardinals, wild or captive. Microfilariae were detected in the blood smears of 14.1% of the wild passerines, including all wild Yellow Cardinals, and in 43.3% of captive Yellow Cardinals. Trypanosoma sp. was detected in the blood smear of one captive Yellow Cardinal. Nested PCR and gene sequencing of the cyt-b gene of Haemoproteus/Plasmodium was used to test a subset of wild passerines and captive Yellow Cardinals, allowing for the molecular barcoding of H. quiscalus lineage AGEBAD04 and H. tyranni lineage PITSUL01; additionally, DNA identical to that of lineage PITSUL01 was detected in the blood of one captive Yellow Cardinal. This study provides valuable data to support the conservation management of the Yellow Cardinal and other threatened passerines from the Pampas and highlights the need for further studies on the epidemiology and pathology of filarioid worms and trypanosomes in passerines from this biome.

Haemosporidian taxonomic composition, network centrality and partner fidelity between resident and migratory avian hosts.

Migration can modify interaction dynamics between parasites and their hosts with migrant hosts able to disperse parasites and impact local community transmission. Thus, studying the relationships among migratory hosts and their parasites is fundamental to elucidate how migration shapes host-parasite interactions. Avian haemosporidians are some of the most prevalent and diverse group of wildlife parasites and are also widely studied as models in ecological and evolutionary research. Here, we contrast partner fidelity, network centrality and parasite taxonomic composition among resident and non-resident avian hosts using presence/absence data on haemosporidians parasitic in South American birds as study model. We ran multilevel Bayesian models to assess the role of migration in determining partner fidelity (i.e., normalized degree) and centrality (i.e., weighted closeness) in host-parasite networks of avian hosts and their respective haemosporidian parasites. In addition, to evaluate parasite taxonomic composition, we performed permutational multivariate analyses of variance to quantify dissimilarity in haemosporidian lineages infecting different host migratory categories. We observed similar partner fidelity and parasite taxonomic composition among resident and migratory hosts. Conversely, we demonstrate that migratory hosts play a more central role in host-parasite networks than residents. However, when evaluating partially and fully migratory hosts separately, we observed that only partially migratory species presented higher network centrality when compared to resident birds. Therefore, migration does not lead to differences in both partner fidelity and parasite taxonomic composition. However, migratory behavior is positively associated with network centrality, indicating migratory hosts play more important roles in shaping host-parasite interactions and influence local transmission.

Plasmodium ouropretensis, n. sp., a new case of non-erythrocytic species within lizard malaria parasites.

Delimiting and describing Plasmodium species in reptiles remains a pressing problem in Haemosporida taxonomy. The few morphological characters used can overlap, and the significance of some life-history traits is not fully understood. Morphologically identical lizard Plasmodium forms have been reported infecting different cell types (red and white blood cells) in the same host and have been considered the same species. An example is Plasmodium tropiduri tropiduri, a species known to infect erythrocytes, thrombocytes and lymphocyte-like cells. Here, both forms of P. t. tropiduri were analysed using light microscope-based morphological characteristics and phylogenetic inferences based on almost complete mitochondrial genomes of parasites naturally infecting lizards in southeastern Brazil. Although morphologically similar, two distinct phylogenetic lineages infecting erythrocytes and non-erythrocytic cells were found. The lineage found in the erythrocytes forms a monophyletic group with species from Colombia. However, the non-erythrocytic lineage shares a recent common ancestor with Plasmodium leucocytica, which infects leucocytes in lizards from the Caribbean islands. Here, Plasmodium ouropretensis n. sp. is described as a species that infects thrombocytes and lymphocyte-like cells.

Changes in malaria patterns in Brazil over 28 years (1990-2017): results from the Global Burden of Disease Study 2017.

BACKGROUND: This study presents the malaria burden in Brazil from 1990 to 2017 using data from the Global Burden of Diseases, Injuries, and Risk Factors Study 2017 (GBD 2017), by analyzing disease burden indicators in federated units of the Legal Amazon and Extra-Amazon regions, as well as describing malaria cases according to Plasmodium species occurring in the country. METHODS: We used estimates from the GBD 2017 to report years of life lost due to premature death (YLLs), years lived with disability (YLDs), and disability-adjusted life years (DALYs) for malaria in Brazil, grouped by gender, age group, and Brazilian federated unit, from 1990 to 2017. Results are presented as absolute numbers and age-standardized rates (per 100,000 inhabitants) with 95% uncertainty intervals (UI). RESULTS: At the national level, the age-standardized DALYs rate due to malaria decreased by 92.0%, from 42.5 DALYs per 100,000 inhabitants (95% UI 16.6-56.9) in 1990 to 3.4 DALYs per 100,000 inhabitants (95% UI 2.7-4.7) in 2017. The YLLs were the main component of the total DALYs rate for malaria in 1990 (67.3%), and the YLDs were the main component of the metric in 2017 (61.8%). In 2017, the highest sex-age DALYs rate was found among females in the "< 1-year-old" age group, with a 6.4 DALYs per 100,000 inhabitants (95% UI 1.8-14.7) and among males in the age group of "20 to 24 years old", with a 4.7 DALYs per 100,000 inhabitants (95% UI 3.3-9.9). Within the Brazilian Amazon region, the three federated units with the highest age-standardized DALYs rates in 2017 were Acre [28.4 (95% UI 14.2-39.1)], Roraima [28.3 (95% UI 13.5-40.2)], and Rondônia [24.7 (95% UI 11.4-34.8)]. Concerning the parasite species that caused malaria, 73.5% of the total of cases registered in the period had Plasmodium vivax as the etiological agent. CONCLUSIONS: The results of the GBD 2017 show that despite the considerable reduction in the DALYs rates between 1990 and 2017, malaria remains a relevant and preventable disease, which in recent years has generated more years of life lost due to disability than deaths. The states endemic for malaria in the Amazon region require constant evaluation of preventive and control measures. The present study will contribute to the direction of current health policies aimed at reducing the burden of malaria in Brazil, as knowing the geographical and temporal distribution of the risk of death and disability of this disease can facilitate the planning, implementation, and improvement of control strategies aimed at eliminating the disease.

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.

Allele-specific antibodies to Plasmodium vivax merozoite surface protein-1: prevalence and inverse relationship to haemoglobin levels during infection.

BACKGROUND: Antigenic polymorphisms are considered as one of the main strategies employed by malaria parasites to escape from the host immune responses after infections. Merozoite surface protein-1 (MSP-1) of Plasmodium vivax, a promising vaccine candidate, is a highly polymorphic protein whose immune recognition is not well understood. METHODS AND RESULTS: The IgG responses to conserved (MSP-119) and polymorphic (block 2 and block 10) epitopes of PvMSP-1 were evaluated in 141 P. vivax infected patients. Ten recombinant proteins corresponding to block 2 (variants BR07, BP29, BP39, BP30, BEL) and block 10 (BR07, BP29, BP39, BP01, BP13) often observed in Brazilian P. vivax isolates were assessed by ELISA in order to determine levels of specific antibodies and their respective seroprevalence. The magnitude and the frequency of variant-specific responses were very low, except for BR07 variant (>40%), which was the predominant haplotype as revealed by block 10 PvMSP-1 gene sequencing. By contrast, 89% of patients had IgG against the C-terminal conserved domain (PvMSP-119), confirming the high antigenicity of this protein. Using multiple linear and logistic regression models, there was evidence for a negative association between levels of haemoglobin and several IgG antibodies against block 2 variant antigens, with the strongest association being observed for BP39 allelic version. This variant was also found to increase the odds of anaemia in these patients. CONCLUSIONS: These findings may have implications for vaccine development and represent an important step towards a better understanding of the polymorphic PvMSP-1 domain as potential targets of vaccine development. These data highlight the importance of extending the study of these polymorphic epitopes of PvMSP-1 to different epidemiological settings.

Anti-erythrocyte antibodies may contribute to anaemia in Plasmodium vivax malaria by decreasing red blood cell deformability and increasing erythrophagocytosis.

BACKGROUND: Plasmodium vivax accounts for the majority of human malaria infections outside Africa and is being increasingly associated in fatal outcomes with anaemia as one of the major complications. One of the causes of malarial anaemia is the augmented removal of circulating non-infected red blood cells (nRBCs), an issue not yet fully understood. High levels of auto-antibodies against RBCs have been associated with severe anaemia and reduced survival of nRBCs in patients with falciparum malaria. Since there are no substantial data about the role of those antibodies in vivax malaria, this study was designed to determine whether or not auto-antibodies against erythrocytes are involved in nRBC clearance. Moreover, the possible immune mechanisms elicited by them that may be associated to induce anaemia in P. vivax infection was investigated. METHODS: Concentrations of total IgG were determined by sandwich ELISA in sera from clinically well-defined groups of P. vivax-infected patients with or without anaemia and in healthy controls never exposed to malaria, whereas the levels of specific IgG to nRBCs were determined by cell-ELISA. Erythrophagocytosis assay was used to investigate the ability of IgGs purified from each studied pooled sera in enhancing nRBC in vitro clearance by THP-1 macrophages. Defocusing microscopy was employed to measure the biomechanical modifications of individual nRBCs opsonized by IgGs purified from each group. RESULTS: Anaemic patients had higher levels of total and specific anti-RBC antibodies in comparison to the non-anaemic ones. Opsonization with purified IgG from anaemic patients significantly enhanced RBCs in vitro phagocytosis by THP-1 macrophages. Auto-antibodies purified from anaemic patients decreased the nRBC dynamic membrane fluctuations suggesting a possible participation of such antibodies in the perturbation of erythrocyte flexibility and morphology integrity maintenance. CONCLUSIONS: These findings revealed that vivax-infected patients with anaemia have increased levels of IgG auto-antibodies against nRBCs and that their deposition on the surface of non-infected erythrocytes decreases their deformability, which, in turn, may enhance nRBC clearance by phagocytes, contributing to the anaemic outcome. These data provide insights into the immune mechanisms associated with vivax malaria anaemia and may be important to the development of new therapy and vaccine strategies.

Blood parasites of penguins: a critical review.

Blood parasites are considered some of the most significant pathogens for the conservation of penguins, due to the considerable morbidity and mortality they have been shown to produce in captive and wild populations of these birds. Parasites known to occur in the blood of penguins include haemosporidian protozoans (Plasmodium, Leucocytozoon, Haemoproteus), piroplamid protozoans (Babesia), kinetoplastid protozoans (Trypanosoma), spirochete bacteria (Borrelia) and nematode microfilariae. This review provides a critical and comprehensive assessment of the current knowledge on these parasites, providing an overview of their biology, host and geographic distribution, epidemiology, pathology and implications for public health and conservation.

Epidemiology and pathology of avian malaria in penguins undergoing rehabilitation in Brazil.

Seabird rehabilitation is a valuable strategy to mitigate the impacts of oil pollution and other anthropogenic factors, and can significantly contribute to the conservation of penguins. However, infectious diseases such as avian malaria (Plasmodium spp.) can hamper the success of rehabilitation efforts. We combined morphological and molecular diagnostic methods to investigate the epidemiology and pathology of Plasmodium in Magellanic penguins (Spheniscus magellanicus) at rehabilitation centers along 2500 km of the coastline of Brazil. True prevalence of malarial parasites was estimated between 6.6% and 13.5%. We identified five species, three of which had not been described infecting penguins (P. cathemerium, P. nucleophilum, P. unalis); an additional five distinct Plasmodium lineages were also distinguished, and albeit unidentified these clearly correspond to species that also have not yet been reported in penguins. Our results indicate that the diversity of plasmodia that may infect these birds is greater than previously recognised. Considering the well-defined seasonality observed in this study, it is clear that rehabilitation centers could benefit by narrowing their preventative efforts on penguins maintained or admitted during the Austral spring-summer, particularly by preventing mosquitoes from coming into contact with penguins.

Evaluating anti-Orthopoxvirus antibodies in individuals from Brazilian rural areas prior to the bovine vaccinia era.

Vaccinia virus naturally circulates in Brazil and is the causative agent of a zoonotic disease known as bovine vaccinia (BV). We retrospectively evaluated two populations from the Amazon and Southeast Regions. BV outbreaks had not been reported in these regions before sample collection. Neutralising antibodies were found in 13 individuals (n = 132) with titres ranging from 100 ≥ 6,400 neutralising units/mL. Univariate analysis identified age and vaccination as statistically significant risk factors in individuals from the Southeast Region. The absence of detectable antibodies in vaccinated individuals raises questions about the protection of smallpox vaccine years after vaccination and reinforces the need for surveillance of Orthopoxvirus in Brazilian populations without evidence of previous outbreaks.

Molecular detection of Leucocytozoon in red-legged seriemas (Cariama cristata), a non-migratory bird species in the Brazilian Cerrado.

Avian Haemosporidian parasites - Plasmodium, Haemoproteus, Leucocytozoon and Fallisia - have a wide distribution except for Antarctica. Leucocytozoon sp. has been poorly described in Brazil, and few studies have indicated infections in birds from the Atlantic Forest, Pantanal, Pampa and Amazon biomes. This study describes, for the first time, the occurrence of Leucocytozoon infection in red-legged seriemas (Cariama cristata) in the Brazilian savanna (Cerrado biome) using molecular diagnosis. Leucocytozoon spp. lineage CARCRI01 was detected in three C. cristata, a non-migratory bird, confirming transmission in mid-elevation areas in central Brazil. Further studies are needed to certify whether infections in red-legged seriemas were not abortive and to elucidate Leucocytozoon infection at low altitudes in the Brazilian lands.

Polyester Nanocapsules for Intravenous Delivery of Artemether: Formulation Development, Antimalarial Efficacy, and Cardioprotective Effects In Vivo.

Artemether (ATM) is an effective antimalarial drug that also has a short half-life in the blood. Furthermore, ATM is also cardiotoxic and is associated with pro-arrhythmogenic risks. We aimed to develop a delivery system enabling the prolonged release of ATM into the blood coupled with reduced cardiotoxicity. To achieve this, we prepared polymeric nanocapsules (NCs) from different biodegradable polyesters, namely poly(D,L-lactide) (PLA), poly-ε-caprolactone (PCL), and surface-modified NCs, using a monomethoxi-polyethylene glycol-block-poly(D,L-lactide) (PEG5kDa-PLA45kDa) polymer. Using this approach, we were able to encapsulate high yields of ATM (>85%, 0−4 mg/mL) within the oily core of the NCs. The PCL-NCs exhibited the highest percentage of ATM loading as well as a slow release rate. Atomic force microscopy showed nanometric and spherical particles with a narrow size dispersion. We used the PCL NCs loaded with ATM for biological evaluation following IV administration. As with free-ATM, the ATM-PCL-NCs formulation exhibited potent antimalarial efficacy using either the "Four-day test" protocol (ATM total at the end of the 4 daily doses: 40 and 80 mg/kg) in Swiss mice infected with P. berghei or a single low dose (20 mg/kg) of ATM in mice with higher parasitemia (15%). In healthy rats, IV administration of single doses of free-ATM (40 or 80 mg/kg) prolonged cardiac QT and QTc intervals and induced both bradycardia and hypotension. Repeated IV administration of free-ATM (four IV doses at 20 mg/kg every 12 h for 48 h) also prolonged the QT and QTc intervals but, paradoxically, induced tachycardia and hypertension. Remarkably, the incorporation of ATM in ATM-PCL-NCs reduced all adverse effects. In conclusion, the encapsulation of ATM in biodegradable polyester NCs reduces its cardiovascular toxicity without affecting its antimalarial efficacy.

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.

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.

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.

Hepatozoon spp. (Apicomplexa: Hepatozoidae) infection and selected hematological values of the neotropical rattlesnake, Crotalus durissus collilineatus (Linnaeus, 1758) (Serpentes: Viperidae), from Brazil.

This study aims to establish the hematological values of Crotalus durissus collilineatus snakes captured in Brazil as well as to verify the effects of hematozoan infection on these snakes. Eighty-three blood samples were drawn from C. d. collilineatus specimens for analysis. The sample set was composed of 30 males and 30 females, recently caught from the wild, and 11 males and 12 females bred in captivity. Blood samples were used to determine red blood cell counts, white blood cell counts, thrombocyte counts, hematocrit values, hemoglobin concentration, and total plasma protein. Blood smears were used to diagnose Hepatozoon spp. infection and to calculate the parasitic load in the sample as well as the percentage of immature red cells. Results obtained for the wild-caught animals, with and without parasites, were compared among themselves and with the values obtained for the captive-bred animals. Hematological values for C. durissus were established. Wild-caught snakes had an infection rate of 38.3%, while no Hepatozoon sp. infection was detected in the captive-bred animals. The snakes which were not infected by the Hepatozoon sp. exhibited average weight, length, and weight-length ratios higher than those of the infected animals. An increase in immature red cells was noted in the Hepatozoon-infected snakes.