Related Haemosporidian Parasites Research Articles

RNAscope in situ hybridization reveals microvascular sequestration of Plasmodium relictum pSGS1 blood stages but absence of exo-erythrocytic dormant stages during latent infection of Serinus canaria

BackgroundBirds chronically infected with avian malaria parasites often show relapses of parasitaemia after latent stages marked by absence of parasites in the peripheral circulation. These relapses are assumed to result from the activation of dormant exo-erythrocytic stages produced during secondary (post-erythrocytic) merogony of avian Plasmodium spp. Yet, there is no morphological proof of persistent or dormant tissue stages in the avian host during latent infections. This study investigated persistence of Plasmodium relictum pSGS1 in birds with latent infections during winter, with the goal to detect presumed persisting tissue stages using a highly sensitive RNAscope® in situ hybridization technology.MethodsFourteen domestic canaries were infected with P. relictum pSGS1 by blood-inoculation in spring, and blood films examined during the first 4 months post infection, and during winter and spring of the following year. After parasitaemia was no longer detectable, half of the birds were dissected, and tissue samples investigated for persisting tissue stages using RNAscope ISH and histology. The remaining birds were blood-checked and dissected after re-appearance of parasitaemia, and their tissues equally examined.ResultsSystematic examination of tissues showed no exo-erythrocytic stages in birds exhibiting latent infections by blood-film microscopy, indicating absence of dormant tissue stages in P. relictum pSGS1-infected canaries. Instead, RNAscope ISH revealed rare P. relictum blood stages in capillaries of various tissues and organs, demonstrating persistence of the parasites in the microvasculature. Birds examined after re-appearance of parasitemia showed higher numbers of P. relictum blood stages in both capillaries and larger blood vessels, indicating replication during early spring and re-appearance in the peripheral circulation.ConclusionsThe findings suggest that persistence of P. relictum pSGS1 during latent infection is mediated by continuous low-level erythrocytic merogony and possibly tissue sequestration of infected blood cells. Re-appearance of parasitaemia in spring seems to result from increased erythrocytic merogony, therefore representing recrudescence and not relapse in blood-inoculated canaries. Further, the study highlights strengths and limitations of the RNAscope ISH technology for the detection of rare parasite stages in tissues, providing directions for future research on persistence and tissue sequestration of avian malaria and related haemosporidian parasites.

Mitochondrial and apicoplast genome copy abundances of haemosporidian parasites are explained by host species and parasitic lineage

Endosymbiotic organelles, such as mitochondria and plastids, contain own remnant genomes (nucleoids), whose variable abundance in cells may be adaptive to the physiological necessities and functions of the cells. Unicellular apicomplexan parasites contain one mitochondrium and one apicoplast with variable genome copy numbers. We measured the abundance of mitochondrial, apicoplast and nuclear genome copies in a set of avian blood samples infected with haemosporidian blood parasites, belonging to the three main genera Plasmodium, Haemoproteus and Leucocytozoon. We designed general primers suitable for qPCR, amplifying fragments of the mitogenome, plastome and nuclear genomes of avian malaria and related haemosporidian parasites. We measured the amplification of these fragments in 153 samples of 23 avian host species and infected with 33 parasitic lineages. We estimate on average several hundred mitochondrial genome copies and several tens of apicoplast copies per haploid gametocyte cell with substantial variation among samples. Host species appeared to differ in their mitogenome abundance while parasitic lineages differed in plastome abundance per cell (per nuclear copy signal). We did not find consistent differences between parasite genera or higher avian taxa. Parasite lineages and host bird species did not differ consistently in infection intensity, estimated from parasite to host nuclear signals, which may indicate that samples were taken at different stages of infection. However, this and similar results remain to be cross-validated with in-situ imaging techniques. The novel molecular tools introduced here offer avenues for the characterization of nucleoid abundance of haemosporidian parasites over environmental conditions and parasitic developmental stages. Such measures will improve our understanding of parasite physiology, ecology, the coadaptation and coevolution with hosts and suggest possible augmentations to standard methods in the research field.

Unravelling the mosquito-haemosporidian parasite-bird host network in the southwestern Iberian Peninsula: insights into malaria infections, mosquito community and feeding preferences.

Vector-borne diseases affecting humans, wildlife and livestock have significantly increased their incidence and distribution in the last decades. Because the interaction among vectors-parasite-vertebrate hosts plays a key role driving vector-borne disease transmission, the analyses of the diversity and structure of vector-parasite networks and host-feeding preference may help to assess disease risk. Also, the study of seasonal variations in the structure and composition of vector and parasite communities may elucidate the current patterns of parasite persistence and spread as well as facilitate prediction of how climate variations may impact vector-borne disease transmission. Avian malaria and related haemosporidian parasites constitute an exceptional model to understand the ecology and evolution of vector-borne diseases. However, the characterization of vector-haemosporidian parasite-bird host assemblages is largely unknown in many regions. Here, we analyzed 5859 female mosquitoes captured from May to November in five localities from southwestern Spain to explore the composition and seasonal variation of the vector-parasite-vertebrate host network. We showed a gradual increase in mosquito abundance, peaking in July. A total of 16 different haemosporidian lineages were found infecting 13 mosquito species. Of these assemblages, more than 70% of these vector-parasite associations have not been described in previous studies. Moreover, three Haemoproteus lineages were reported for the first time in this study. The prevalence of avian malaria infections in mosquitoes varied significantly across the months, reaching a maximum in November. Mosquito blood-feeding preference was higher for mammals (62.5%), whereas 37.5% of vectors fed on birds, suggesting opportunistic feeding behavior. These outcomes improve our understanding of disease transmission risk and help tovector control strategies.

Prevalence and genetic diversity of avian haemosporidian parasites in islands within a mega hydroelectric dam in the Brazilian Amazon.

The Brazilian Amazon supports an extremely diverse avifauna and serves as the diversification center for avian malaria parasites in South America. Construction of hydroelectric dams can drive biodiversity loss by creating islands incapable of sustaining the bird communities found in intact forest sites. Besides anthropogenic actions, the presence of parasites can also influence the dynamics and structure of bird communities. Avian malaria (Plasmodium) and related haemosporidian parasites (Haemoproteus and Leucocytozoon) are a globally distributed group of protozoan parasites recovered from all major bird groups. However, no study to date has analyzed the presence of avian haemosporidian parasites in fragmented areas such as land bridge islands formed during artificial flooding following the construction of hydroelectric dams. The aim of this study is to assess the prevalence and molecular diversity of haemosporidians in bird communities inhabiting artificial islands in the area of the Balbina Hydroelectric Dam. The reservoir area covers 443,700 ha with 3546 islands on the left bank of the Uatumã River known to contain more than 400 bird species. We surveyed haemosporidian infections in blood samples collected from 445 understory birds, belonging to 53 species, 24 families, and 8 orders. Passeriformes represented 95.5% of all analyzed samples. We found a low overall Plasmodium prevalence (2.9%), with 13 positive samples (two Plasmodium elongatum and 11 Plasmodium sp.) belonging to eight lineages. Six of these lineages were previously recorded in the Amazon, whereas two of them are new. Hypocnemis cantator, the Guianan Warbling Antbird, represented 38.5% of all infected individuals, even though it represents only 5.6% of the sampled individuals. Since comparison with Plasmodium prevalence data prior to construction of Balbina is not possible, other studies in artificially flooded areas are imperative to test if anthropogenic flooding may disrupt vector-parasite relationships leading to low Plasmodium prevalence.

Avian malaria, haematocrit, and body condition in invasive wetland passerines settled in southwestern Spain

Avian malaria and related haemosporidian parasites can negatively impact fitness in many songbirds. Research on the malaria infection and its physiological costs on their avian hosts is heavily skewed toward native passerines, with exotic species underrepresented. However, introduced species may carry on and spread new pathogens to native species, and play a role on parasite transmission cycle in invaded bird communities as pathogen reservoir. Here, we molecularly assess the prevalence and diversity of haemosporidian parasites in three introduced wetland passerines (the Red Avadavat Amandava amandava, the Yellow-crowned Bishop Euplectes afer, and the Common Waxbill Estrilda astrild) captured during the same season in southwestern Spain. We also explored the relation between parasite infection, body condition, haematocrit, and uropygial gland volume. We detected an overall parasite prevalence of 3.55%, where Common Waxbills showed higher prevalence (6.94%) than Red Avadavats (1.51%). None Yellow-crowned Bishops were infected with haemosporidians. Almost 60% of infections were caused by Leucocytozoon, and about 40% by Plasmodium. We identified four unique lineages of Plasmodium and three of Leucocytozoon. Moreover, 91% of the identified host–parasite interactions represented new host records for these haemosporidian parasites. Parasite infection was not related to body condition, haematocrit, and uropygial gland volume of the wetland passerines. Haematocrit values varied seasonally among bird species. Additionally, haematocrit was positively related to body condition in the Yellow-crowned Bishops, but not in the other species. Red Avadavats had higher haematocrit levels than Yellow-crowned Bishops, whereas Common Waxbills showed the lower haematocrit values. The uropygial gland volume was positively correlated with body condition in all bird species. Common Waxbills showed higher uropygial gland volumes related to their body size than birds from other two species. These outcomes highlight the importance of exotic invasive species in the transmission dynamics of haemosporidian parasites.

Stimuli Followed by Avian Malaria Vectors in Host-Seeking Behaviour.

Simple SummaryVector-borne diseases (VBDs) (e.g., malaria, yellow fever, dengue fever) account for 17% of the estimated global burden of all infectious diseases. They are transmitted to humans and other animals by blood-feeding arthropods. In their pursuit of blood meal, insect vectors use different cues to detect their hosts. The knowledge of these stimuli followed by vectors in this host-seeking behaviour is essential to design strategies to prevent VBD infections. Since its discovery in the late 19th century, avian malaria investigations have allowed significant advances to understand the dynamics and mechanisms of VBD transmission to many organisms, including humans. Here, we review published contributions on the different physical and chemical cues used by mosquitoes and other bird haemosporidian vectors to locate their hosts. This information would be highly valuable for vector surveillance and public health policies.Vector-borne infectious diseases (e.g., malaria, dengue fever, and yellow fever) result from a parasite transmitted to humans and other animals by blood-feeding arthropods. They are major contributors to the global disease burden, as they account for nearly a fifth of all infectious diseases worldwide. The interaction between vectors and their hosts plays a key role driving vector-borne disease transmission. Therefore, identifying factors governing host selection by blood-feeding insects is essential to understand the transmission dynamics of vector-borne diseases. Here, we review published information on the physical and chemical stimuli (acoustic, visual, olfactory, moisture and thermal cues) used by mosquitoes and other haemosporidian vectors to detect their vertebrate hosts. We mainly focus on studies on avian malaria and related haemosporidian parasites since this animal model has historically provided important advances in our understanding on ecological and evolutionary process ruling vector-borne disease dynamics and transmission. We also present relevant studies analysing the capacity of feather and skin symbiotic bacteria in the production of volatile compounds with vector attractant properties. Furthermore, we review the role of uropygial secretions and symbiotic bacteria in bird–insect vector interactions. In addition, we present investigations examining the alterations induced by haemosporidian parasites on their arthropod vector and vertebrate host to enhance parasite transmission. Finally, we propose future lines of research for designing successful vector control strategies and for infectious disease management.

Metacommunity structure reveals that temperature affects the landscape compositional patterns of avian malaria and related haemosporidian parasites across elevations

Metacommunity ecology studies how species compositional patterns and their distributions vary across local and regional scales and provides insights on processes driving the distribution of communities. Avian haemosporidians comprise a diverse and widely distributed parasite taxon; some studies have analyzed their alpha and beta diversity patterns. Yet, metacommunity structures of avian haemosporidians and thus relevant biotic and abiotic variables explaining such structures at the landscape scale (i.e., 10–200 km) have not been assessed. We studied the metacommunity structure of avian haemosporidian mtDNA cyt b lineages and the infected avian host assemblage across four different elevations in Central Veracruz, Mexico. We performed variation-partitioning analyses to evaluate the contribution of host-related traits and climatic variables to the metacommunity. We found a richness of 78 lineages within 38 infected species of birds. At the component community level, we observed that bird species infected with a lower number of parasite lineages (e.g., <3) represented a nested subset of those with a higher number of parasite lineages (e.g., >8) (i.e., nested structure). However, this nested pattern was due to the restricted spatiotemporal co-occurrence of hosts and parasites, given the high degree of turnover across elevations. Host-related traits (functional, transmission-associated, and phylogenetic relationships) only explained a small fraction of the variation (4.4%) in parasite lineage composition across avian hosts. At the habitat level, there was a group turnover by parasite genera across elevation (i.e., quasi-Clementsian structure), which was partly explained by climatic variables (mean annual temperature and annual diurnal range; 27.6%) that may constrain parasite reproduction and vector distribution across the environmental gradient. At the scale of our study, environmental conditions represented a more important driver of avian haemosporidian metacommunity structure than host-related traits, suggesting an important role of environmental filtering structuring parasite assemblages at the landscape level.

Assessment of Associations between Malaria Parasites and Avian Hosts—A Combination of Classic System and Modern Molecular Approach

Simple SummaryThroughout history, frequent outbreaks of diseases in humans have occurred following transmission from animals. While some diseases can jump between birds and mammals, others are stuck to closely related species. Understanding the mechanisms of host–parasite associations will enable us to predict the outbreaks of diseases and will therefore be important to society and ecological health. For decades, scientists have attempted to reveal how host–parasite associations are formed and persist. The key is to assess the ability of the parasite to infect and reproduce within the host without killing the host. Related studies have faced numerous challenges, but technical advances are providing solutions and are gradually broadening our understanding. In this review, I use bird malaria and related blood parasites as a model system and summarize the important advances in techniques and perspectives and how they provide new approaches for understanding the evolution of host–parasite associations to further predict disease outbreaks.Avian malaria and related haemosporidian parasites are responsible for fitness loss and mortality in susceptible bird species. This group of globally distributed parasites has long been used as a classical system for investigating host–parasite associations. The association between a parasite and its hosts can be assessed by the prevalence in the host population and infection intensity in a host individual, which, respectively, reflect the ability of the parasite to infect the host and reproduce within the host. However, the latter has long been poorly investigated due to numerous challenges, such as lack of general molecular markers and limited sensitivity of traditional methods, especially when analysing naturally infected birds. The recent development of genetic databases, together with novel molecular methodologies, has shed light on this long-standing problem. Real-time quantitative PCR has enabled more accurate quantification of avian haemosporidian parasites, and digital droplet PCR further improved experimental sensitivity and repeatability of quantification. In recent decades, parallel studies have been carried out all over the world, providing great opportunities for exploring the adaptation of haemosporidian parasites to different hosts and the variations across time and space, and further investigating the coevolutionary history between parasites and their hosts. I hereby review the most important milestones in diagnosis techniques of avian haemosporidian parasites and illustrate how they provide new insights for understanding host–parasite associations.

Diversity and host assemblage of avian haemosporidians in different terrestrial ecoregions of Peru.

Characterizing the diversity and structure of host–parasite communities is crucial to understanding their eco-evolutionary dynamics. Malaria and related haemosporidian parasites are responsible for fitness loss and mortality in bird species worldwide. However, despite exhibiting the greatest ornithological biodiversity, avian haemosporidians from Neotropical regions are quite unexplored. Here, we analyze the genetic diversity of bird haemosporidian parasites (Plasmodium and Haemoproteus) in 1,336 individuals belonging to 206 bird species to explore for differences in diversity of parasite lineages and bird species across 5 well-differentiated Peruvian ecoregions. We detected 70 different haemosporidian lineages infecting 74 bird species. We showed that 25 out of the 70 haplotypes had not been previously recorded. Moreover, we also identified 81 new host–parasite interactions representing new host records for these haemosporidian parasites. Our outcomes revealed that the effective diversity (as well as the richness, abundance, and Shannon–Weaver index) for both birds and parasite lineages was higher in Amazon basin ecoregions. Furthermore, we also showed that ecoregions with greater diversity of bird species also had high parasite richness, hence suggesting that host community is crucial in explaining parasite richness. Generalist parasites were found in ecoregions with lower bird diversity, implying that the abundance and richness of hosts may shape the exploitation strategy followed by haemosporidian parasites. These outcomes reveal that Neotropical region is a major reservoir of unidentified haemosporidian lineages. Further studies analyzing host distribution and specificity of these parasites in the tropics will provide important knowledge about phylogenetic relationships, phylogeography, and patterns of evolution and distribution of haemosporidian parasites.

Penguins are competent hosts of Haemoproteus parasites: the first detection of gametocytes, with molecular characterization of Haemoproteus larae

BackgroundThe majority of penguins (Sphenisciformes) have evolved in areas with weak or absent transmission of haemosporidian parasites and are usually naïve to avian haemosporidian infections. Plasmodium parasites are transmitted by mosquitoes, and lethal avian malaria has been often reported in captive penguins in many countries. The related haemosporidian parasites belonging to Haemoproteus and Leucocytozoon have also been detected in penguins but less often than Plasmodium infections. The majority of Haemoproteus infection reports in penguins are based solely on PCR-based diagnostics. It remains unclear if haemoproteids can complete their life-cycle and produce infective stages (gametocytes) in penguins or whether these infections are abortive in penguins, and thus dead ends for transmission. In other words, it remains unknown if penguins are competent hosts for Haemoproteus parasites, which cause disease in non-adapted birds.MethodsTwo captive African penguins (Spheniscus demersus) and two Magellanic penguins (S. magellanicus) were found to be positive for Haemoproteus infection in two open-air aquariums in Japan, and the parasites were investigated using both PCR-based testing and microscopical examination of blood films. Samples from a black-tailed gull (Larus crassirostris) and previously tested gulls were used for comparison.ResultsThe lineage hSPMAG12 was detected, and gametocytes of Haemoproteus sp. were seen in the examined penguins and gull. Observed gametocytes were indistinguishable from those of Haemoproteus larae, which naturally parasitize birds of the genus Larus (Laridae). The detected sequence information and Bayesian phylogenetic analysis supported this conclusion. Additionally, morphologically similar gametocytes and closely related DNA sequences were also found in other gull species in Japan. Phylogenetic analysis based on partial cytb sequences placed the lineage hSPMAG12 of H. larae within the clade of avian haemoproteids which belong to the subgenus Parahaemoproteus, indicating that Culicoides biting midges likely transmit the parasites between penguins and gulls.ConclusionsThis study shows that some species of Haemoproteus parasites complete their development and produce gametocytes in penguins, which may be source of infection for biting midges transmitting haemoproteosis. To prevent haemosporidiosis in zoos, we call for control not only of mosquitoes, but also biting midges.

Avian haemosporidian prevalence and its relationship to host traits in Western Tennessee

Avian malaria and related haemosporidian parasites (genera Plasmodium and Parahaemoproteus) are the most common parasites in many bird populations and are known to affect survival and reproduction. We analyze how species-level and individual-level traits influence parasite prevalence among species and infection status among individuals. We collected blood samples of 625 individuals from 35 host species and used PCR to screen for infection status. We found that 44% of the individuals were infected, and 38 unique lineages of haemosporidian parasites were obtained. Total prevalence and prevalence of Plasmodium and Parahaemoproteus separately were significantly heterogeneous across species, ranging from 0 to 100%. Total and Plasmodium prevalence increased significantly with host species abundance, but Parahaemoproteus prevalence did not. Parasite prevalence did not vary with other species-level traits, including species mass, annual survival, nest type, nesting or foraging height, and degree of sexual dimorphism. Individual-level traits, such as age and sex, did not predict infection status of individuals. Our research documents a high diversity of haemosporidian parasites and substantial variation in parasite prevalence across host species. However, contrary to expectations, haemosporidian prevalence is not strongly related to many host life history traits. Future studies that examine vector abundance and parasite prevalence across habitat types might be especially productive.

The nuclear 18S ribosomal DNAs of avian haemosporidian parasites

BackgroundPlasmodium species feature only four to eight nuclear ribosomal units on different chromosomes, which are assumed to evolve independently according to a birth-and-death model, in which new variants originate by duplication and others are deleted throughout time. Moreover, distinct ribosomal units were shown to be expressed during different developmental stages in the vertebrate and mosquito hosts. Here, the 18S rDNA sequences of 32 species of avian haemosporidian parasites are reported and compared to those of simian and rodent Plasmodium species.MethodsAlmost the entire 18S rDNAs of avian haemosporidians belonging to the genera Plasmodium (7), Haemoproteus (9), and Leucocytozoon (16) were obtained by PCR, molecular cloning, and sequencing ten clones each. Phylogenetic trees were calculated and sequence patterns were analysed and compared to those of simian and rodent malaria species. A section of the mitochondrial CytB was also sequenced.ResultsSequence patterns in most avian Plasmodium species were similar to those in the mammalian parasites with most species featuring two distinct 18S rDNA sequence clusters. Distinct 18S variants were also found in Haemoproteus tartakovskyi and the three Leucocytozoon species, whereas the other species featured sets of similar haplotypes. The 18S rDNA GC-contents of the Leucocytozoon toddi complex and the subgenus Parahaemoproteus were extremely high with 49.3% and 44.9%, respectively. The 18S sequences of several species from all three genera showed chimeric features, thus indicating recombination.ConclusionGene duplication events leading to two diverged main sequence clusters happened independently in at least six out of seven avian Plasmodium species, thus supporting evolution according to a birth-and-death model like proposed for the ribosomal units of simian and rodent Plasmodium species. Patterns were similar in the 18S rDNAs of the Leucocytozoon toddi complex and Haemoproteus tartakovskyi. However, the 18S rDNAs of the other species seem to evolve in concerted fashion like in most eukaryotes, but the presence of chimeric variants indicates that the ribosomal units rather evolve in a semi-concerted manner. The new data may provide a basis for studies testing whether differential expression of distinct 18S rDNA also occurs in avian Plasmodium species and related haemosporidian parasites.

Haemosporidian parasites and leukocyte profiles of pre-migratory rosy starlings (Pastor roseus) brought into captivity

Abstract Immune response to pathogens such as avian malaria (Plasmodium spp.) and related haemosporidian parasites (Haemoproteus spp. and Leucocytozoon spp.) can lead to increases in energy demands of birds. Migrations are energetically costly for birds and require resources that could be otherwise invested in immune defence against pathogens. Here, we examined the prevalence and intensity of haemosporidian parasites (Apicomplexa, Haemosporida) in rosy starlings (Pastor roseus), an irruptive migrant on the Balkans, during their pre-migratory period. We further evaluated if these infections affected the stress levels of birds brought into overnight captivity, using leukocyte profiles (ratios of heterophils to lymphocytes, or H/L ratios). Haemosporidian parasites were detected based on microscopic examination, PCR and sequencing. We found high prevalence of haemosporidian parasites (83.9%) in rosy starlings (n=31), with predominant infections of Haemoproteus pastoris (cytochrome b lineage LAMPUR01) (80.1%). All birds had high heterophil/lymphocyte ratios (mean = 3.86), indicating the transition to captivity was stressful. However, there was no added stress from blood parasite infection during captivity, since infected birds did not have significantly higher heterophil/lymphocyte ratios than uninfected birds, despite all birds facing the same stress conditions. There was a negative correlation between intensity of H. pastoris and monocyte proportions, which may indicate an association with infection, or an artefact of the stress-induced shift in leukocyte profile following captivity. Our findings suggest that blood parasites have minimal additional impact on rosy starlings during an acute stress scenario (overnight captivity), arguing that they carry little cost to the energetically challenging migratory flight.

Recognition of the 4th International Conference on Malaria and Related Haemosporidian Parasites of Wildlife, held Nov. 1-5, 2018 in Beijing China.

Over the past 10 years, much research has been done and remarkable discoveries have been made in various fields of research regarding wildlife malaria parasites (Plasmodium, Plasmodiidae) and related haemosporidians, the agents of infections, which affect all major groups of terrestrial vertebrates. Recent molecular studies show that these blood parasites cause more harm to wildlife than formerly believed, calling for new fundamental and applied research on this subject. The 4th International Conference on Malaria and Related Haemosporidian Parasites of Wildlife highlighted the recent accomplishments of many research groups throughout the world at the Beijing Normal University, with additional hosts being the Beijing Zoo and the China Ornithological Society. Please see: http://malariaconference2018.com/. We are grateful to the organizers of the meeting and Drs. Gediminas Valkiūnas, Staffan Bensch, and Lu Dong for their contributions to this review.

Malaria parasites and related haemosporidians cause mortality in cranes: a study on the parasites diversity, prevalence and distribution in Beijing Zoo

BackgroundMalaria parasites and related haemosporidian parasites are widespread and may cause severe diseases in birds. These pathogens should be considered in projects aiming breeding of birds for purposes of sustained ex situ conservation. Cranes are the ‘flagship species’ for health assessment of wetland ecosystems, and the majority of species are endangered. Malaria parasites and other haemosporidians have been reported in cranes, but the host-parasite relationships remain insufficiently understood. Morbidity of cranes due to malaria has been reported in Beijing Zoo. This study report prevalence, diversity and distribution of malaria parasites and related haemosporidians in cranes in Beijing Zoo and suggest simple measures to protect vulnerable individuals.MethodsIn all, 123 cranes (62 adults and 61 juveniles) belonging to 10 species were examined using PCR-based testing and microscopic examination of blood samples collected in 2007–2014. All birds were maintained in open-air aviaries, except for 19 chicks that were raised in a greenhouse with the aim to protect them from bites of blood-sucking insects. Bayesian phylogenetic analysis was used to identify the closely related avian haemosporidian parasites.ResultsSpecies of Plasmodium (5 lineages), Haemoproteus (1) and Leucocytozoon (2) were reported. Malaria parasites predominated (83% of all reported infections). The overall prevalence of haemosporidians in juveniles was approximately seven-fold higher than in adults, indicating high susceptibility of chicks and local transmission. Juvenile and adult birds hosted different lineages of Plasmodium, indicating that chicks got infection from non-parent birds. Plasmodium relictum (pSGS1) was the most prevalent malaria parasite. Mortality was not reported in adults, but 53% of infected chicks died, with reports of co-infection with Plasmodium and Leucocytozoon species. All chicks maintained in the greenhouse were non-infected and survived. Species of Leucocytozoon were undetectable by commonly used PCR protocol, but readily visible in blood films.ConclusionCrane chicks often die due to malaria and Leucocytozoon infections, which they likely gain from wild free-living birds in Beijing Zoo. Molecular diagnostics of crane Leucocytozoon parasites needs improvement. Because the reported infections are mainly chick diseases, the authors recommend maintaining of juvenile birds in vector-free facilities until the age of approximately 6 months before they are placed in open-air aviaries.

Diversity, abundance, and host relationships of avian malaria and related haemosporidians in New Mexico pine forests.

Avian malaria and related haemosporidian parasites (genera Haemoproteus, Plasmodium, and Leucocytozoon) affect bird demography, species range limits, and community structure, yet they remain unsurveyed in most bird communities and populations. We conducted a community-level survey of these vector-transmitted parasites in New Mexico, USA, to describe their diversity, abundance, and host associations. We focused on the breeding-bird community in the transition zone between piñon-juniper woodland and ponderosa pine forests (elevational range: 2,150–2,460 m). We screened 186 birds representing 49 species using both standard PCR and microscopy techniques to detect infections of all three avian haemosporidian genera. We detected infections in 68 out of 186 birds (36.6%), the highest proportion of which were infected with Haemoproteus (20.9%), followed by Leucocytozoon (13.4%), then Plasmodium (8.0%). We sequenced mtDNA for 77 infections representing 43 haplotypes (25 Haemoproteus, 12 Leucocytozoon, 6 Plasmodium). When compared to all previously known haplotypes in the MalAvi and GenBank databases, 63% (27) of the haplotypes we recovered were novel. We found evidence for host specificity at the avian clade and species level, but this specificity was variable among parasite genera, in that Haemoproteus and Leucocytozoon were each restricted to three avian groups (out of six), while Plasmodium occurred in all groups except non-passerines. We found striking variation in infection rate among host species, with nearly universal infection among vireos and no infection among nuthatches. Using rarefaction and extrapolation, we estimated the total avian haemosporidian diversity to be 70 haplotypes (95% CI [43–98]); thus, we may have already sampled ∼60% of the diversity of avian haemosporidians in New Mexico pine forests. It is possible that future studies will find higher diversity in microhabitats or host species that are under-sampled or unsampled in the present study. Fortunately, this study is fully extendable via voucher specimens, frozen tissues, blood smears, parasite images, and documentation provided in open-access databases (MalAvi, GenBank, and ARCTOS).

Diversity and distribution of avian malaria and related haemosporidian parasites in captive birds from a Brazilian\xa0megalopolis

BackgroundThe role of zoos in conservation programmes has increased significantly in last decades, and the health of captive animals is essential to guarantee success of such programmes. However, zoo birds suffer from parasitic infections, which often are caused by malaria parasites and related haemosporidians. Studies determining the occurrence and diversity of these parasites, aiming better understanding infection influence on fitness of captive birds, are limited.MethodsIn 2011–2015, the prevalence and diversity of Plasmodium spp. and Haemoproteus spp. was examined in blood samples of 677 captive birds from the São Paulo Zoo, the largest zoo in Latin America. Molecular and microscopic diagnostic methods were used in parallel to detect and identify these infections.ResultsThe overall prevalence of haemosporidians was 12.6%. Parasites were mostly detected by the molecular diagnosis, indicating that many birds harbour subclinical or abortive infections. In this project, birds of 17 orders (almost half of all the orders currently accepted in taxonomy of birds), 29 families, and 122 species, were tested, detecting positive individuals in 27% of bird species. Birds from the Anatidae were the most prevalently infected (64.7% of all infected animals). In all, infections with parasites of the genus Plasmodium (overall prevalence 97.6%) predominated when compared to those of the genus Haemoproteus (2.4%). In total, 14 cytochrome b (cytb) lineages of Plasmodium spp. and 2 cytb lineages of Haemoproteus spp. were recorded. Eight lineages were new. One of the reported lineages was broad generalist while others were reported in single or a few species of birds. Molecular characterization of Haemoproteus ortalidum was developed.ConclusionThis study shows that many species of birds are at risk in captivity. It is difficult to stop haemosporidian parasite transmission in zoos, but is possible to reduce the infection rate by treating the infected animals or/and while keeping them in facilities free from mosquitoes. Protocols of quarantine should be implemented whenever an animal is transferred between bird maintaining institutions. This is the first survey of haemosporidians in captive birds from different orders maintained in zoos. It is worth emphasizing the necessity of applying practices to control these parasites in management and husbandry of animals in captivity.

Nycteria parasites of Afrotropical insectivorous bats

Parasitic protozoan parasites have evolved many co-evolutionary paths towards stable transmission to their host population. Plasmodium spp., the causative agents of malaria, and related haemosporidian parasites are dipteran-borne eukaryotic pathogens that actively invade and use vertebrate erythrocytes for gametogenesis and asexual development, often resulting in substantial morbidity and mortality of the infected hosts. Here, we present results of a survey of insectivorous bats from tropical Africa, including new isolates of species of the haemosporidian genus Nycteria. A hallmark of these parasites is their capacity to infect bat species of distinct families of the two evolutionary distant chiropteran suborders. We did detect Nycteria parasites in both rhinolophid and nycterid bat hosts in geographically separate areas of Sub-Saharan Africa, however our molecular phylogenetic analyses support the separation of the parasites into two distinct clades corresponding to their host genera, suggestive of ancient co-divergence and low levels of host switching. For one clade of these parasites, cytochrome b genes could not be amplified and cytochrome oxidase I sequences showed unusually high rates of evolution, suggesting that the mitochondrial genome of these parasites may have either been lost or substantially altered. This haemosporidian parasite-mammalian host system also highlights that sequential population expansion in the liver and gametocyte formation is a successful alternative to intermediate erythrocytic replication cycles.

Invasive avian malaria as an emerging parasitic disease in native birds of Peru

Some species of avian malaria parasites are invaders and responsible for diversity losses worldwide. Here we analyze the prevalence and genetic characterization of avian malaria and related haemosporidian parasites in Neotropical birds from two different regions of Peru. We detected an overall prevalence of 32.4 % comprising 12 infected bird species. The pathogen Plasmodium relictum SGS1 was widespread and the most prevalent parasite found in our study (39 % of the total infections), infecting 8 host species in both localities. To the best of our knowledge, this is the first report of this invasive pathogen in the mainland Americas, thus representing a possible menace to over one-third of all bird species in the world.

Co‐infections by malaria parasites decrease feather growth but not feather quality in house martin

During moult, stressors such as malaria and related haemosporidian parasites (e.g.PlasmodiumandHaemoproteus) could affect the growth rate and quality of feathers, which in turn may compromise future reproduction and survival. Recent advances in molecular methods to study parasites have revealed that co‐infections with multiple parasites are frequent in bird–malaria parasite systems. However, there is no study of the consequences of co‐infections on the moult of birds. In house martinsDelichon urbicacaptured and studied at a breeding site in Europe during 11 yr, we measured the quality and the growth rate of tail feathers moulted in the African winter quarters in parallel with the infection status of blood parasites that are also transmitted on the wintering ground. Here we tested if the infection with two haemosporidian parasite lineages has more negative effects than a single lineage infection. We found that birds with haemosporidian infection had lower body condition. We also found that birds co‐infected with two haemosporidian lineages had the lowest inferred growth rate of their tail feathers as compared with uninfected and single infected individuals, but co‐infections had no effect on feather quality. In addition, feather quality was negatively correlated with feather growth rate, suggesting that these two traits are traded‐off against each other. We encourage the study of haemosporidian parasite infection as potential mechanism driving this trade‐off in wild populations of birds.