Genomic and transcriptomic evidence for descent from Plasmodium and loss of blood schizogony in Hepatocystis parasites from naturally infected red colobus monkeys.

Hepatocystis is a genus of single-celled parasites infecting, amongst other hosts, monkeys, bats and squirrels. Although thought to have descended from malaria parasites (Plasmodium spp.), Hepatocystis spp. are thought not to undergo replication in the blood-the part of the Plasmodium life cycle which causes the symptoms of malaria. Furthermore, Hepatocystis is transmitted by biting midges, not mosquitoes. Comparative genomics of Hepatocystis and Plasmodium species therefore presents an opportunity to better understand some of the most important aspects of malaria parasite biology. We were able to generate a draft genome for Hepatocystis sp. using DNA sequencing reads from the blood of a naturally infected red colobus monkey. We provide robust phylogenetic support for Hepatocystis sp. as a sister group to Plasmodium parasites infecting rodents. We show transcriptomic support for a lack of replication in the blood and genomic support for a complete loss of a family of genes involved in red blood cell invasion. Our analyses highlight the rapid evolution of genes involved in parasite vector stages, revealing genes that may be critical for interactions between malaria parasites and mosquitoes.

Haemosporidian parasites from captive Strigiformes in France.

Haemosporidia infections may cause major damage to avian populations and represent a concern for veterinarians working in zoological parks or wildlife rescue centres. Following the fatal infection of 9 Great grey owls (Strix nebulosa) at Mulhouse zoological park, between summer 2013 and 2015, a prospective epidemiological investigation was performed in captive strigiform birds in France in 2016. The purpose was to evaluate the prevalence of haemosporidian parasites in captive Strigiformes and to estimate the infection dynamics around the nesting period. Blood samples were taken from 122 strigiform birds representing 14 species from 15 French zoological parks. Parasites were detected by direct examination of blood smears and by PCR targeting the mitochondrial cytochrome b gene. Haemosporidian parasites were detected in 59 birds from 11 zoos. Three distinct Haemoproteus mitochondrial cytochrome b sequences (haplotypes A and C for H. syrnii and haplotype B for Haemoproteus sp.) as well as two species of Plasmodium were detected. The overall prevalence of Haemoproteus infection was 12.8%. The percentage of birds infected by Haemoproteus varied according to the period of sampling. Nesting season seemed to be at greater risk with an average prevalence of 53.9% compared with winter season with an average prevalence of 14.8%, related to the abundance of the vectors. The prevalence of Plasmodium infection in Strigiformes did not exceed 8% throughout the year. This study confirmed how significant Haemosporidia infection could be in Strigiformes from zoological parks in France. The nesting season was identified as a period of higher risk of infection and consequently the appropriate period to apply prophylactic measures.

[A comparative study of commercial ELISAs for antibody detection in the diagnostic investigation of Neospora caninum-associated abortion in dairy cattle herds in Uruguay]. / Comparación de ELISAs comerciales para la detección de anticuerpos en la investigación diagnóstica del aborto asociado a Neospora caninum en rodeos lecheros de Uruguay.

Bovine abortion causes considerable economic losses to the livestock industry worldwide and is of concern for public health and food safety, given that many abortigenic infectious agents of cattle are zoonotic. Despite its importance, the etiological diagnosis of abortion in cattle is challenging both for veterinary practitioners and laboratory technicians, partly due to the difficulty in recovering aborted fetuses under extensive field conditions for pathological and microbiological diagnostic investigation, and in the early identification of aborted dams. Neospora caninum is a cosmopolitan protozoon identified as one of the main abortigenic agents in cattle worldwide. In this study we propose a comparative seroepidemiological approach for the diagnosis of abortion by N. caninum in dairy cattle. Samples from 12 to 93 cows/heifers with and without recent history of abortion (cases and controls) in four commercial dairy farms were tested. The ratio of controls to cases tested varied from 1:1 to 4.6:1. All samples (n=230) were analyzed by three commercial ELISA kits for the detection of anti-N. caninum antibodies. In all four dairy farms, the proportion of seropositive cows and/or heifers per kit was significantly higher in the cases than in the controls (Odds Ratios=5.13 to 36, p=0.0002 to 0.0485). The agreement among the three kits varied from weak to strong (Cohens kappa coefficients=0.58 to 0.83). We conclude that, despite the imperfect agreement between these kits, all of them allowed to arrive at similar conclusions regarding the statistical association between N. caninum seropositivity and abortion, thus representing a useful tool for the diagnostic approach at the population level under field conditions.

Innate immunity and environmental correlates of Haemoproteus prevalence and intensity in an opportunistic breeder.

While parasite infection can have substantial fitness consequences in organisms, the predictors of parasite prevalence and intensity are often complex and vary depending on the host species. Here, we examined correlates of Haemoproteus (a common malaria parasite) prevalence and intensity in an opportunistically breeding songbird, the red crossbill (Loxia curvirostra). Specifically, we quantified Haemoproteus prevalence and intensity in crossbills caught in the Grand Teton National Park from 2010 to 2013. We found that parasite prevalence varies seasonally and across years, with the highest number of infected individuals occurring in the summer, although there was variation across summers sampled, and that prevalence was positively related to annual mean cone crop sizes (a measure of crossbill food abundance) and daily ambient temperature (a correlate of vector abundance). Parasite intensity was significantly and positively related to one measure of innate immunity, leucocyte counts per blood volume. Finally, neither crossbill age, ecomorph, nor sex had significant effects on parasite infection intensity; however, parasite prevalence did significantly vary among ecomorph and age classes. These results support the interpretation that a combination of physiological (specifically immune activity) and environmental factors affects parasite prevalence and infection intensity in this opportunistically breeding avian species.

Adaptation and evaluation of an ELISA for Trypanosoma evansi infection (surra) in elephants and its application to a serological survey in Thailand.

Trypanosoma evansi, the causative agent of surra, is widespread in domestic livestock and wildlife in South East Asia. Surra can affect cattle, buffaloes, horses and also Asian elephants (Elephas maximus). Despite the 'threatened to extinction' CITES status of elephant, surra's impact has not been thoroughly assessed yet in this species. This work offers to adapt an antibody enzyme-linked immunosorbent assay (ELISA) protocol, to detect Trypanosoma evansi antibodies in elephant serum. The test was validated with 365 negative-reference samples, which allowed the determination of a 16% positive threshold. The test was applied to a serological survey including 375 individuals. The estimated global seroprevalence was 2·1% (95% CI 1·1-4·2%). Therefore, surra does not appear to be endemic in Thai domestic elephants, but occasional outbreaks were reported to our laboratory during the survey period. These outbreaks seemed to be linked to close proximity to cattle or buffaloes, and led to severe clinical signs in elephants. Frequent relapses were observed after treatment with diminazene aceturate, the only trypanocide drug currently available in Thailand. Therefore, care should be taken to keep elephants away from bovine reservoirs, and to monitor the disease in this endangered species. ELISA proved to be reliable for screening purposes as well as for post-treatment monitoring.

Blood transcriptomes reveal novel parasitic zoonoses circulating in Madagascar's lemurs.

Zoonotic diseases are a looming threat to global populations, and nearly 75% of emerging infectious diseases can spread among wildlife, domestic animals and humans. A 'One World, One Health' perspective offers us an ideal framework for understanding and potentially mitigating the spread of zoonoses, and the island of Madagascar serves as a natural laboratory for conducting these studies. Rapid habitat degradation and climate change on the island are contributing to more frequent contact among humans, livestock and wildlife, increasing the potential for pathogen spillover events. Given Madagascar's long geographical isolation, coupled with recent and repeated introduction of agricultural and invasive species, it is likely that a number of circulating pathogens remain uncharacterized in lemur populations. Thus, it is imperative that new approaches be implemented for de novo pathogen discovery. To this end, we used non-targeted deep sequencing of blood transcriptomes from two species of critically endangered wild lemurs (Indri indri and Propithecus diadema) to characterize blood-borne pathogens. Our results show several undescribed vector-borne parasites circulating within lemurs, some of which may cause disease in wildlife, livestock and humans. We anticipate that advanced methods for de novo identification of unknown pathogens will have broad utility for characterizing other complex disease transmission systems.

Prevalence of Haemoproteus spp. (Apicomplexa: Haemoproteidae) in tortoises in Brazil and its molecular phylogeny.

Captive terrestrial tortoises of the species Chelonoidis carbonaria (n = 17) and Chelonoidis denticulata (n = 37) in the state of Minas Gerais, southeastern Brazil, were examined for hematozoans by using a combination of microscopic and molecular methods. Microscopic examination revealed young intra-erythrocytic forms in blood smears from both species of tortoises. The results of PCR, sequencing, and phylogenetic analysis indicated that these parasites belonged to the Haemoproteus spp., whose observed prevalence was 17.6 % in C. carbonaria and 13.5 % in C. denticulata. Phylogenetic analysis indicated that these sequences formed a clade that was grouped with other sequences of Haemoproteus spp. parasites in birds, separate from the clade formed by Haemoproteus spp. of reptiles. This study expands the information regarding the occurrence and distribution of hemosporidia in turtles and is the first study of blood parasites in C. carbonaria.

In vitro development of Haemoproteus columbae (Haemosporida: Haemoproteidae), with perspectives for genomic studies of avian haemosporidian parasites.

The evolutionary origin of wildlife and human malaria parasites (Plasmodium spp.) has been discussed for several decades. The lack of genomic data about species of wildlife haemosporidian parasites related to Plasmodium limits the number of taxa available for phylogenetic analysis. Genomic data about avian parasites of the genus Haemoproteus parasites, the sister genus to Plasmodium are still not available, mainly due to difficulties in obtaining pure DNA of parasites inhabiting nucleated avian host cells. Recent studies show that microgametes of Haemoproteus (Parahaemoproteus) spp. develop in vitro and can be isolated by simple centrifugation, allowing the isolation of pure parasite DNA for genomic studies. However, in vitro development of Haemoproteus (Haemoproteus) spp. has not been investigated, and it is unclear if microgametes of these parasites also can be obtained under in vitro conditions. Here, we provide the first data about the in vitro development of Haemoproteus (Haemoproteus) columbae, a widespread avian haemosporidian parasite, which is specific to pigeons and doves (Columbiformes) and is transmitted by hippoboscid flies (Diptera, Hippoboscidae). In vitro gametogenesis and ookinete development of H. columbae were studied using a strain isolated from a feral Rock Pigeon (Columba livia) in Bogotá-Colombia. The morphological events leading to exflagellation, fertilization and ookinete formation, as well as the rate of development of these stages were followed in vitro at 40 °C, 19 °C and 15 °C for 48 h. Macrogametes, microgametes, zygotes and initial stages of ookinete development were observed in all temperatures, but mature ookinetes were seen only at 40 °C. The largest diversity of sporogonic stages of H. columbae were present at 40 °C however, exflagellation, fertilization of macrogametes and development of immature ookinetes were also observed at 15 °C and 19 °C. Morphological and morphometric features of these stages in vitro were described and illustrated. This study demonstrates a requirement of high temperature for the successful development of mature ookinetes of H. columbae, but not gametes. We show that 1) parasites of the H. (Haemoproteus) subgenus exflagellate in vitro at 15-19 °C, as is the case in H. (Parahaemoproteus) spp. and 2) in vitro exflagellation can be used to obtain pure DNA for genomic studies.

Molecular characterization of five widespread avian haemosporidian parasites (Haemosporida), with perspectives on the PCR-based detection of haemosporidians in wildlife.

Haemosporidians (Haemosporida) are cosmopolitan in birds. Over 250 species of these blood parasites have been described and named; however, molecular markers remain unidentified for the great majority of them. This is unfortunate because linkage between DNA sequences and identifications based on morphological species can provide important information about patterns of transmission, virulence, and evolutionary biology of these organisms. There is an urgent need to remedy this because few experts possess the knowledge to identify haemosporidian species and few laboratories are involved in training these taxonomic skills. Here, we describe new mitochondrial cytochrome b markers for the polymerase chain reaction (PCR)-based detection of four widespread species of avian Haemoproteus (Haemoproteus hirundinis, Haemoproteus parabelopolskyi, Haemoproteus pastoris, Haemoproteus syrnii) and 1 species of Plasmodium (Plasmodium circumflexum). Illustrations of blood stages of the reported species are given, and morphological and phylogenetic analyses identify the DNA lineages that are associated with these parasites. This study indicates that morphological characters, which have been traditionally used in taxonomy of avian haemosporidian parasites, have a phylogenetic value. Perspectives on haemosporidian diagnostics using microscopic and PCR-based methods are discussed, particularly the difficulties in detection of light parasitemia, coinfections, and abortive parasite development. We emphasize that sensitive PCR amplifies more infections than can be transmitted; it should be used carefully in epidemiology studies, particularly in wildlife parasitology research. Because molecular studies are describing remarkably more parasite diversity than previously expected, the need for traditional taxonomy and traditional biological knowledge is becoming all the more crucial. The linkage of molecular and morphological approaches is worth more of the attention of researchers because this approach provides new knowledge for better understanding insufficiently investigated lethal diseases caused by haemosporidian infections, particularly on the exoerythrocytic (tissue) and vector stages. That requires close collaboration between researchers from different fields with a common interest.

Avian blood parasite infection during the non-breeding season: an overlooked issue in declining populations?

BACKGROUND: Pathogens and parasites can have major impacts on host population dynamics, both through direct mortality and via indirect effects. Both types of effect may be stronger in species whose populations are already under pressure. We investigated the potential for blood parasites to impact upon their hosts at the immunological, physiological and population level during the non-breeding season using a declining population of yellowhammers Emberiza citrinella as a model. RESULTS: Yellowhammers infected by Haemoproteus spp. showed both a reduced heterophil to lymphocyte (H:L) ratio, and an elevated standardised white blood cell (WBC) count compared to uninfected birds, indicating an immunological response to infection. Infected birds had shorter wings during the first winter of sampling but not during the second, colder, winter; survival analysis of 321 birds sampled across four winters indicated that increased wing length conferred a survival advantage. CONCLUSIONS: We suggest that the potential impacts of blood parasite infections on over-wintering birds may have been underestimated. Further research should consider the potential impacts of sub-clinical parasite infections on the dynamics of vulnerable populations, and we suggest using declining populations as model systems within which to investigate these relationships as well as examining interactions between sub-clinical disease and other environmental stressors. JEL CODE: Q5.

A new method for isolation of purified genomic DNA from haemosporidian parasites inhabiting nucleated red blood cells.

During the last 10 years, whole genomes have been sequenced from an increasing number of organisms. However, there is still no data on complete genomes of avian and lizard Plasmodium spp. or other haemosporidian parasites. In contrast to mammals, bird and reptile red blood cells have nuclei and thus blood of these vertebrates contains high amount of host DNA; that complicates preparation of purified template DNA from haemosporidian parasites, which has been the main obstacle for genomic studies of these parasites. In the present study we describe a method that generates large amount of purified avian haemosporidian DNA. The method is based on a unique biological feature of haemosporidian parasites, namely that mature gametocytes in blood can be induced to exflagellate in vitro. This results in the development of numerous microgametes, which can be separated from host blood cells by simple centrifugation. Our results reveal that this straight forward method provides opportunities to collect pure parasite DNA material, which can be used as a template for various genetic analyses including whole genome sequencing of haemosporidians infecting birds and lizards.

Proportion and seasonality of blood parasites in animals in Mosul using the Veterinary Teaching Hospital Lab data.

Several local studies have examined evidence of blood parasites in different animals in Mosul; however, information about the most prevalent parasite and the seasonality of the infection remains limited. The objective of the study conducted here was to investigate the proportion and seasonality of blood parasites in animals in Mosul using the Veterinary Teaching Hospital Lab data. Laboratory records for a period of 25 months were used for data retrieval. In all included animals, Giemsa-stained blood smears were examined by an attending clinical pathologist for the presence of parasites. Seasons were assigned on a basis of examination date, and the seasonality was quantified by estimating season-to-season ratio. The results indicated that 61.77% of examined animals were tested positive for blood parasites. The most evident parasites were Trypanosoma spp., Theileria spp., Babesia spp., and then Anaplasma spp., with evidence of mixed infection. The odds of the infection did not significantly vary in different age groups. There was a marked linear pattern in the seasonality of the infection with Trypanosoma spp. and Anaplasma spp. An increase of the infection during spring and autumn with Theileria spp. and Babesia spp. was also evident. In conclusion, infection with blood parasites in different animals in Mosul is common with substantial burden, the effect of age-related infection is negligible, and the seasonality of the infection is evident.

Spatially explicit predictions of blood parasites in a widely distributed African rainforest bird.

Critical to the mitigation of parasitic vector-borne diseases is the development of accurate spatial predictions that integrate environmental conditions conducive to pathogen proliferation. Species of Plasmodium and Trypanosoma readily infect humans, and are also common in birds. Here, we develop predictive spatial models for the prevalence of these blood parasites in the olive sunbird (Cyanomitra olivacea). Since this species exhibits high natural parasite prevalence and occupies diverse habitats in tropical Africa, it represents a distinctive ecological model system for studying vector-borne pathogens. We used PCR and microscopy to screen for haematozoa from 28 sites in Central and West Africa. Species distribution models were constructed to associate ground-based and remotely sensed environmental variables with parasite presence. We then used machine-learning algorithm models to identify relationships between parasite prevalence and environmental predictors. Finally, predictive maps were generated by projecting model outputs to geographically unsampled areas. Results indicate that for Plasmodium spp., the maximum temperature of the warmest month was most important in predicting prevalence. For Trypanosoma spp., seasonal canopy moisture variability was the most important predictor. The models presented here visualize gradients of disease prevalence, identify pathogen hotspots and will be instrumental in studying the effects of ecological change on these and other pathogens.

Nest ectoparasites increase physiological stress in breeding birds: an experiment.

Parasites are undoubtedly a biotic factor that produces stress. Heat shock proteins (HSPs) are important molecules buffering cellular damage under adverse conditions. During the breeding season, blue tit Cyanistes caeruleus (L.) adults are affected by blood parasites, nest-dwelling parasites and biting flies, potentially affecting their HSP-mediated responses. Here, we treated females with primaquine to reduce blood parasites and fumigated nests with permethrin to reduce nest-dwelling parasites to test whether these treatments affect HSP60 level during the breeding season. Medicated females, but not controls, had a significant reduction of the intensity of infection by Haemoproteus spp. blood parasites. However, final intensity of infection did not differ significantly between groups, and we did not find an effect of medication on change in HSP60 level. Fumigation reduced the abundance of nest-dwelling parasites (mites, fleas and blowfly larvae) and engorged biting midges in nests. Females breeding in non-fumigated nests increased HSP60 levels during the season more than those breeding in fumigated nests. Furthermore, the change in HSP60 level was positively correlated with the abundance of biting midges. These results show how infections by nest ectoparasites during the breeding period can increase the level of HSPs and suggest that biting midges impose physiological costs on breeding female blue tits. Although plausible, the alternative that biting midges prefer to feed on more stressed birds is poorly supported by previous studies.

High seroprevalence of Histomonas meleagridis in Dutch layer chickens.

Histomona meleagridis is a protozoan parasite that may cause outbreaks of histomonosis with high mortality, especially in turkey flocks. Chickens are less susceptible to the disease than are turkeys, but are considered to act as an important reservoir. To determine the seroprevalence of H. meleagridis in Dutch layer chicken flocks, a large scale seroepidemiologic study (3376 samples) was performed by sampling 12 organic flocks, 24 free-ranging flocks, 40 flocks with floor housing, and 40 flocks with cage housing. At the end of the laying period, approximately 30 blood samples per flock were collected for serology. The seroprevalence found was high. In every flock, at least one of the samples tested positive while in 87% of the flocks, at least one of the samples was strongly positive. There were no significant statistical differences in seropositivity between the housing types. To confirm the enzyme-linked immunosorbent assay (ELISA) results, a small-scale seroepidemiologic study (576 samples) was performed in 29 additional layer chicken flocks kept in different housing systems. Subsequently, a subset of five seropositive flocks was selected. Five birds were obtained from each of these flocks in order to detect the parasite using culture and PCR. In all five flocks, H. meleagridis was either isolated from (culture), detected in (PCR), or both, the birds sampled. Together with the previously performed validation studies, the latter results confirm that the positive ELISA serology found is genuine. We conclude that the seroprevalence of H. meleagridis in layers is, as anticipated, high.

Chronic exposure and decontamination of a marine sculpin (Myoxocephalus scorpius) to polychlorinated biphenyls using selected body indices, blood values, histopathology, and parasites as bioindicators.

This study was conducted to assess the health of a subtidal marine sculpin, Myoxocephalus scorpius, naturally exposed in a shipping terminal where polychlorinated biphenyls (PCBs) had been spilled, as well as that of others sculpins after decontamination in the laboratory for 16 weeks. Reference samples were captured at a pristine site upcurrent from the contaminated terminal. Examination of field samples caught by scuba divers showed external and toxicopathic lesions in several tissues; decreased body condition and organ indices; and decreased hemoglobin and lymphocyte levels. No difference in the diversity of parasites was observed between reference and contaminated samples, but abundance was generally greater in fish from the latter site. Although the total number of species of parasites decreased in both groups of depurated sculpins, the abundance of infections on gill and gall bladder was significantly greater in fish from the polluted site. These results suggest that M. scorpius was affected adversely by PCBs and that its responses are ecologically significant as a bioindicator species in degraded habitats. Moreover, remediation might not mitigate the observed anomalies in fish inhabiting the impacted site.

Response of Nkedi Zebu and Ankole cattle to tick infestation and natural tick-borne, helminth and trypanosome infections in Uganda.

A cross-sectional study was conducted in Soroti district of Uganda to establish important traits of Nkedi Zebu and Ankole cattle regarding their production performance responses to natural infections of trypanosomes, gastrointestinal nematodes, Theileria parva, Babesia bigemina, Anaplasma marginale and tick infestations. Over four visits between October 2006 to August 2007, tick counts were performed and blood, faecal samples and sera were collected from the Nkedi Zebu (295) and Ankole (165) cattle from 86 herds in six locations per visit. Low parasitological prevalence of trypanosome infection (<6%) and high prevalence of gastrointestinal nematode infections (>30%) with low faecal egg counts (110-300 eggs per gramme (EPG)) were observed in the Nkedi Zebu and Ankole cattle. Both breeds had high, moderate and low mean counts of Rhipicephalus appendiculatus (18.0-24.0), Rhipecephalus (Boophilus) decoloratus (3.6-10.3) and Amblyomma variegatum ticks (1.7-4.3), respectively. In addition, both breeds had similar mean packed cell volumes (26.4-31.2) and a similar percentage of animals were anaemic (14.5-36.6%). The Nkedi Zebu cattle further had higher mean optical density (OD) values for antibodies against T. parva (1.093-1.445) and A. marginale infections (0.573-0.583), and significantly (P < 0.001) higher mean OD values of antibodies against B. bigemina infections (1.07-2.175) than the Ankole cattle: T. parva (1.030-1.302); A. marginale (0.442-0.603) and B. bigemina infections (0.863-2.154). The Ankole cows produced significantly more (P < 0.001) milk per day (2.68 L) than the Nkedi Zebu cows (1.98 L), and the Ankole oxen had significantly higher (P < 0.05) draught power output (2.57 days/acre) than the Nkedi Zebu oxen (2.93 days/acre). Liveweights of calves aged 0-12 months of both breeds were comparable, suggesting that the Nkedi Zebu and Ankole cattle under similar disease challenge exhibited similar growth rates. In conclusion, the Nkedi Zebu cattle seem to possess a higher degree of disease resistance against endemic parasitic diseases, while the Ankole cattle seem to possess a moderate degree of disease resistance coupled with a moderate production potential.

A preliminary disease survey in the wild Nile crocodile (Crocodylus niloticus) population in the Okavango Delta, Botswana.

The objective of this study was to conduct a preliminary survey of diseases that might be present in the wild Nile crocodile population in the Okavango Delta, Botswana. Blood samples were collected from crocodiles ranging in size from 34.0 cm to 463.0 cm total length. Samples were examined for blood parasites and underwent a haematological analysis. Before release the crocodiles were examined for various clinical abnormalities. Of the 144 crocodiles examined, none were visibly sick or displayed any signs of disease. No antibodies to Mycoplasma crocodyli were detected. Hepatozoon pettiti was present in 55.3% of blood smears examined, but there was no significant difference in any of the haematological values between the infected and uninfected crocodiles, and a high prevalence of Hepatozoon infection is not uncommon in other species. Only 7.6% of the examined crocodiles were infested with leeches. Further research is required for several of the crocodilian diseases, in particular to elucidate the role of wild crocodilians as reservoirs of infection.

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.

LOW PREVALENCE OF HAEMOSPORIDIANS IN BLOOD AND TISSUE SAMPLES FROM HUMMINGBIRDS.

Hummingbirds are vital members of terrestrial ecosystems, and because of their high metabolic requirements, they serve as indicators of ecosystem health. Monitoring the parasitic infections of hummingbirds is thus especially important. Haemosporidians, a widespread group of avian blood parasites, are known to infect hummingbirds, but little is known about the prevalence and diversity of these parasites in hummingbirds. The prevalence of haemosporidians in several hummingbird species was examined and we compared 4 different tissue types in detecting parasites by polymerase chain reaction (PCR). Blood samples from 339 individuals of 3 different hummingbird species were tested, and 4 individuals were found positive for haemosporidian infection, a prevalence of 1.2%. Hummingbird carcasses (n = 70) from 5 different hummingbird species were also sampled to assess differences in detection success of haemosporidians in heart, kidney, liver, and pectoral muscle tissue samples. Detection success was similar among tissue types, with haemosporidian prevalence of 9.96% in heart tissue, 9.52% in kidney tissue, 10.76% in liver tissue, and 11.76% in pectoral muscle tissue. All tissue samples positive for haemosporidian infection were from the Black-chinned Hummingbird (Archilochus alexandri). Possible reasons for low prevalence of these blood parasites could include low susceptibility to insect vectors or parasite incompatibility in these hummingbirds.