The blood parasites of anurans from Costa Rica with reflections on the taxonomy of their trypanosomes

During May 1997, specimens of 7 species of anurans, that included 5 Phrynohyas venulosa Laurenti, 5 Rana forreri Boulenger, 7 Rana vaillanti Brucchi, 6 Eleutherodactylus fitzingeri Schimdt, 4 Smilisca baudinii Duméril and Bibron, 1 Leptodactylus melanonotus, and 3 Bufo marinus Linneaus, from the Guanacaste Conservation Area, Costa Rica were examined for blood parasites. Their hematozoan fauna included intraerythrocytic and intraleukocytic icosahedral viruses, a rickettsia (Aegyptianella sp.), 2 species of Hepatozoon, Lankesterella minima, 2 unknown species of apicomplexans, 9 morphologically distinct types of trypanosomes, and 2 species of microfilariae. Rana vaillanti, the most aquatic species of frog, harbored the most species of parasites. Recent evidence indicates that morphological changes in the highly pleomorphic trypanosomes of anurans from different geographical regions have not kept pace with biochemical (isozyme) and molecular (DNA sequence) changes. Describing new species based solely on bloodstream trypomastigotes is discouraged. Additional criteria described herein should be applied when naming new species of anuran trypanosomes.     

Standard sampling techniques underestimate prevalence of avian hematozoa in willow ptarmigan (Lagopus lagopus)

A total of 68 willow ptarmigan (Lagopus lagopus L.) was collected during September 1995 from two localities in Troms County, northern Norway. Thin blood smears were prepared and examined for blood parasites. Of the 68 willow ptarmigan examined, 94% harbored one or more species of hematozoa. There were four (6%), 44 (65%), 16 (24%), and four (6%) birds infected by zero, one, two, and three species of parasites, respectively. Prevalences at the coastal locality, Kattfjord (n = 43), were Leucocytozoon lovati 86%, Trypanosoma avium (26%), and microfilariae (30%). At the inland locality, Iselvdalen (n = 25), prevalences were L. lovati 96%, T. avium 12%, and microfilariae 0%. We also searched connective tissues for the filaroid nematode Splendidofilaria papillocerca; in Kattfjord this parasite only occurred in adult hosts where prevalence was 94%, but the parasite was not found in Iselvdalen. To estimate the efficiency of parasite detection by standard blood sampling techniques, we sampled peripheral blood from the brachial wing vein and blood from the pulmonary system from willow ptarmigan. Sampling peripheral blood from the brachial vein led to underestimates of the prevalence of microfilariae. There was no significant difference between L. lovati and T. avium prevalence in blood collected from the brachial vein or deep circulation. Age of host had a strong impact on prevalence, especially for S. papillocerca and microfilariae.     

Prevalence of blood parasites in Tyrannidae (flycatchers) in the Eastern plains of Colombia

Blood samples from 159 birds of the New-world family Tyrannidae (the flycatchers) from the eastern plains of Colombia, were examined for haematozoa parasites, in 1999-2000. Haematozoa were detected in six of 20 species. The overall prevalence was 10.1%. The most common parasites detected were microfilariae, followed by Trypanosoma and Plasmodium. The highest prevalence (9.6%) was found in the Ochre-bellied Flycatcher (Mionectes oleaginea). Mixed infections with more than one genus of blood parasite were rare and most infections encountered were of low intensity. The results of this study suggest an important role of ecologically diverse conditions determining composition, transmission, and prevalence of a blood parasite fauna, presumably through host interaction population density. Some new host parasite relationship records are presented.     

Interspecific differences in hematozoan infection in Sonoran desert Aimophila sparrows

Numerous studies have identified factors that control avian hematozoan infections, but the mechanisms that account for host differences in parasitemia remain largely speculative. To address this issue, we compared the prevalence of these parasites in stained blood smears from four conspecific Sonoran desert Aimophila sparrow species sampled during their breeding season: rufous-winged (Aimophila carpalis; RWSP), rufous-crowned (Aimophila ruficeps; RCSP), Cassin's (Aimophila cassinii, CASP), and Botteri's (Aimophila botterii; BOSP) sparrows. Blood smears contained Haemoproteus fringillae (RWSP), Trypanosoma everetti (RWSP, RCSP, BOSP), Trypanosoma avium (CASP), and microfilariae (all species). Most (92.5%) RWSP (n=40) were infected with Haemoproteus, but this parasite was not detected in RCSP (n=20) or BOSP (n=20) and was found only in one (2.5%) CASP (n=40). Trypanosoma spp. and microfilariae were detected in all species, but prevalence differed between these four sparrow species. Species differences in parasite prevalence were not due to difference in sex, age, adult body mass, incubation period, breeding habitat, or plumage colorfulness. However, differences in Haemoproteus sp. prevalence correlated with preferred nesting height, as RWSP generally nest above ground, whereas the other species nest on or close to the ground. Elevated H. fringillae prevalence in breeding-condition RWSP presumably does not result from a seasonal relapse associated with breeding or require new infection because 1) this prevalence did not differ in males sampled during and outside (n=21) the breeding season, and 2) all male RWSP (n=25) that we held in captivity and shielded from new infections and influence of natural photoperiod for 1 yr had viable blood H. fringillae gametocytes. H. fringillae prevalence in fall-sampled hatch-year male RWSP (n=11) was 63.6%, demonstrating that this parasite can be transmitted on the breeding grounds and during the first months of life. T. everetti prevalence in RWSP was lower in winter than in summer and also in long-term captive than in free-ranging adults. Presence of this parasite in the blood of breeding males may depend on recrudescence of existing infections or new infections.     

Chronic wuchereriasis presenting as a vaginoperineal fistula: report of a case with aspiration cytologic diagnosis

BACKGROUND: Filariasis is often an occult disease with myriad presentations. Cytology has an established role in diagnosing clinically unsuspected cases. CASE: A 20-year-old female presented with recurring perineal ulcers, vaginal discharge and inguinal lymphadenopathy. Radiology revealed a vaginoperineal fistula. Fine needle aspiration (FNA) of the ulcer bed and smears prepared from the ulcer discharge showed an adult, gravid, female filarial worm and unsheathed larvae. Similar microfilariae were also seen in FNA smears from inguinal lymph nodes. The atypical clinical presentation and unusual parasite morphology posed initial difficulties in characterization of the microfilarial species. CONCLUSION: This case report highlights the morphologic clues to the dif ferential diagnosis offilarial species on cytologic specimens. Chronic wuchereriasis presenting as a vaginoperineal fistula has not been reported previously.     

Prevalence of avian hematozoa in central Vermont

Peripheral blood smears from 1,547 birds, of 50 species, from 15 families trapped in Northfield, Vermont were examined for hematozoa. Numerous new host-parasite relationships were identified. The prevalence of all species of parasites over the 3 yr of the study was 42.5%. Prevalence of the genera of parasites were: Leucocytozoon--36.5%; Trypanosoma--7.3%; Haemoproteus--6.3%; microfilariae--1.0%; Plasmodium--1.0%. Peak prevalence (78.9%) occurred in the first half of July. Peak intensity was seen in the last half of June. Evidence from immature birds suggested that active transmission of all genera of hematozoans took place in the study area. Leucocytozoon gametocytes, unlike Haemoproteus gametocytes, were detected in smears from birds during all seasons of the year, and showed no period of complete remission. Trypanosoma and microfilariae also were seen throughout the year.     

Premature release of Plasmodium falciparum from swollen erythrocytes induced by some antimalarials

Qinghaosu and chloroquine, but not pyrimethamine, treatment of Plasmodium falciparum cultures resulted in the formation of swollen red blood cells (RBCs) and the expulsion of degenerate trophozoites and schizonts, but not ring-stage parasites, from the infected RBCs. The parasite release resulted in the formation of RBCs with holes, that had otherwise retained their structural integrity. Membranes of swollen RBCs and their ghosts associated with parasites were efficiently visualized by Giemsa staining of thin smears for 18-24 hr but not by standard Giemsa staining for 20 min.     

Role of fine needle aspiration cytology in detection of microfilariae: report of 2 cases

BACKGROUND: Filariasis is a major public health problem in developing countries, and the diagnosis is conventionally made by demonstrating microfilariae in the peripheral blood smear. However, microfilariae have been incidentally detected in fine needle aspirates of various lesions in clinically unsuspected cases of filariasis with absence of microfilariae in the peripheral blood. CASES: In case 1, a 21-year-old woman presented with multiple left axillary lymphadenopathy of 3 months' duration. In case 2, a 32-year-old woman presented with a thyroid nodule of 7 months' duration. Fine needle aspiration smears from both cases showed sheathed microfilariae of Wuchereria bancrofti. In both cases, microfilariae could not be demonstrated in the peripheral blood smears and the blood eosinophil counts were within normal limits. The histopathologic examination showed neither microfilariae nor adult worm. CONCLUSION: Although microfilariae in cytologic material are considered incidental findings, these cases illustrate the value of routine fine needle aspiration cytology in the detection of asymptomatic and clinically unsuspected cases of bancroftian filariasis. Absence of microfilariae in the peripheral blood does not exdude filarial infection.     

Hematological responses to hematozoa in North American and neotropical songbirds

Hematocrit (Hct), hemoglobin concentration, RBC count, and blood parasite loads (Plasmodium, Leucocytozoon, Haemoproteus, Trypanosoma, microfilariae) were measured in 688 passerine birds captured at four sites in the US and Belize, C.A., to assess physiological consequences of parasitemia. Parasite loads were low overall: 80% of birds had <5 parasites/10000 RBCs; median infection intensity was 1 per 10000 RBCs. Infection prevalence and intensity were higher in Belize than in the US. Analyses across and within taxa revealed no significant linear correlations between hematology values and parasite load, though infected birds were more likely to be anemic. Neotropical migrants reaching the southern US had lower parasite loads than migrants wintering in Belize. Mean Hct was significantly elevated among infected migrants (but not uninfected migrants) reaching the US relative to migrants in Belize. Long-distance migration thus appears to act as a filter for heavily parasitized and anemic birds. Geographic and seasonal variations in hematology indices were similar within and between taxa, suggesting that community-wide factors, rather than phylogenetic attributes or species-specific pathogens, account for the majority of hematological variation. The low parasite loads and weak correlation between parasite load and hematology indices have implications for attempts to test the version of the 'good genes' hypothesis.     

Incidence and Seasonal Variations in Blood and Tissue Parasites of Yellow-Billed Magpies

SYNOPSIS. Blood and tissue smears from 156 yellow-billed magpies showed infection with hematozoa in 154 (99%). Leucocytozoon berestneffi was found in 149 (96%), Haemoproteus picae in 40 (26%), Plasmodium relictum , in 32 (21%), Trypanosoma sp. in 3 (2%), and microfilariae in 42 (27%). Fifty-two birds were infected with two genera of parasites, 27 with 3, and 4 with 4. Leucocytozoon, Plasmodium and microfilariae showed a higher incidence in tissue than in blood smears.     

Dirofilaria repens diagnosed by the presence of microfilariae in fine needle aspirates: a case report

BACKGROUND: Dirofilariasis due to Dirofilaria repens with viable microfilariae outside the worm have not been reported before. CASE: A 40-year-old truck driver from rural Shiraz, Iran, had a firm mass, 2.5 x 2.5 cm, at the dorsolateral aspect of the right forearm. Fine needle aspiration (FNA) was performed on 2 occasions. Several microfilariae with blunt heads, pointed posterior ends and empty caudal spaces resembling microfilariae of Wuchereria bancrofti but longer were seen. Since Iran is a nonendemic area for lymphatic filariae and the patient had a history of contact with a dog, with the impression of dirofilariasis, the mass was excised, and the presence of adult worms in tissue sections confirmed the diagnosis. CONCLUSION: This case ofsubcutaneous dirofilariasis was diagnosed by detecting microfilariae in FNA smears and was confirmed on histopathology.     

Different paths – the same virulence: experimental study on avian single and co-infections with Plasmodium relictum and Plasmodium elongatum

Co-infections are prevalent worldwide, however, we are still struggling to understand interactions between different parasites and their impacts on host fitness. In the present experimental study we analysed the infection dynamics of two avian malarial parasites Plasmodium elongatum (genetic lineage pERIRUB01) and Plasmodium relictum (genetic lineage pSGS1) and their impacts on host health during single and co-infections. We reveal that P. elongatum intensity of parasitemia is enhanced by the presence of P. relictum during co-infection, while the parasitemia of P. relictum stays the same. This illustrates how development of a parasite (P. elongatum) which infects both mature and young (polychromatic) red blood cells (RBCs) is facilitated during co-infection with a parasite which specialises in adult RBCs only (P. relictum). The virulence of co-infections was similar to that of the more virulent parasite (P. elongatum). However, the profile of infection and the mechanisms that caused mortality were different. Birds infected with P. elongatum only start to die due to non-regenerative anaemia, when intensity of parasitemia is light and the number of polychromatic RBCs decrease dramatically. Meanwhile, co-infected birds start to die when the mean intensity of parasitemia reaches 10% and the number of polychromatic RBCs increases abnormally, reflecting regenerative anaemia. Our findings reveal that typically measured parameters of virulence (e.g., mortality rate, level of hematocrit) can be the same during single and co-infections, but the mechanisms responsible for the observed virulence can be different. This information serves a better understanding of the processes underpinning the interactions of co-infected parasite species.     

Cytomorphology of filariasis revisited. Expansion of the morphologic spectrum and coexistence with other lesions

OBJECTIVE: To review the cytomorphologic spectrum of the filarial worm and associated tissue response in 33 cases. STUDY DESIGN: Retrospective analysis was carried out in clinically unsuspected cases of filariasis diagnosed on cytology over a period of 10 years. Twenty-nine aspirate smears from 28 patients were air dried and stained with May-Grünwald-Giemsa stain. Four routine cervical smears and one centrifuged smear of urine were stained with Papanicolaou stain. RESULTS: Microfilariae alone and along with adult gravid females were present in 25 and 4 cases, respectively. In one case both adult male and female worms with microfilariae and eggs were seen. The diagnosis was based on the presence of eggs alone in one case and fragments of female worms in two. Four of these cases were neoplastic lesions, and microfilariae were found incidentally. In one case of splenomegaly microfilariae were seen along with Leishman-Donovan bodies. CONCLUSION: Filariasis can be diagnosed on cytology by demonstrating microfilariae, a male or female worm, or eggs alone. It can be seen in association with neoplastic lesions and rarely with other parasitic infections.     

A new filarial nematode (Onchocercidae) from warthogs (Phacochoerus aethiopicus) of the Kruger National Park

Fifty-five warthogs [Phacochoerus aethiopicus (Suidae: Artiodactyla)] from the Kruger National Park, Republic of South Africa, were examined for parasites. Adult filarial nematodes were found in lymphatic vessels adjacent to peripheral and visceral lymph nodes, and microfilariae were found in lymph nodes and circulating blood. Both the adult parasite and the microfilaria are described. Specific identification is pending confirmation and recovery of intact adult specimens and microfilariae identical to those described herein.     

Bloodmeal microfilariae density and the uptake and establishment of Wuchereria bancrofti infections in Culex quinquefasciatus and Aedes aegypti.

The relationship between ingestion of microfilariae (mf), production of infective larvae (L3) and mf density in human blood has been suggested as an important determinant in the transmission dynamics of lymphatic filariasis. Here we assess the role of these factors in determining the competence of a natural vector Culex quinquefasciatus and a non vector Aedes aegypti to transmit Wuchereria bancrofti. Mosquitoes were infected via a membrane feeding procedure. Both mosquito species ingested more than the expected number of microfilariae (concentrating factor was 1.28 and 1.81 for Cx. quinquefasciatus and Ae. aegypti, respectively) but Cx. quinquefasciatus ingested around twice as many mf as Ae. aegypti because its larger blood meal size. Ae. aegypti showed a faster mf migration capacity compared to Cx. quinquefasciatus but did not allow parasite maturation under our experimental conditions. Similar proportions of melanized parasites were observed in Ae. aegypti (2. 4%) and Cx. quinquefasciatus (2.1%). However, no relationship between rate of infection and melanization was observed. We conclude that in these conditions physiological factors governing parasite development in the thorax may be more important in limiting vectorial competence than the density of mf ingested.     

Interspecific variability of prevalence in blood parasites of adult passerine birds during the breeding season in Alaska

Blood parasite prevalence based on microscopic examination of stained blood smears was determined in adults of 11 passerine bird species sampled during their breeding season (May and June 1997-98) in interior Alaska (USA). These species included primarily Nearctic migratory species such as the dark-eyed junco (Junco hyemalis) and neotropical migratory species such as the blackpoll warbler (Dendroica striata), alder flycatcher (Empidonax alnorum), Swainson's thrush (Catharus ustulatus), northern waterthrush (Seiurus noveboracensis), and bank swallow (Riparia riparia) as well as one long-distance palearctic migrant, the arctic warbler (Phylloscopus borealis). The more prevalent parasites were Leucocytozoon dubreuili (73% of the sampled turdinids), L. fringillinarum (42% of the sampled fringillids and parulids), and Trypanosoma avium (39% of the sampled hosts). Other parasites (H. fallisi: 18% of the sampled turdinids; Haemoproteus paruli: 14% of the sampled parulids; H. fringillae: 5% of the sampled fringillids; microfilariae: 4% of the sampled hosts) were observed less frequently. Plasmodium vaughani was found only in two yellow warblers (Dendroica petechia). Overall parasite prevalence varied between 0% in the alder flycatcher to >80% in Swainson's thrush, arctic warbler, and Townsend's warbler (Dendroica townsendi). Prevalence of various hematozoa also was bird species-dependent. No relationship was observed between prevalence and either foraging (aerial versus trees/shrubs) or nesting habits (ground versus arboreal) or general location of the wintering area of the different species examined. Prevalence also was unrelated to average dates of arrival on breeding grounds and, therefore, to potential duration of exposure to local insect vectors before capture. Differences in blood parasite prevalence among species breeding in a same region and in the same type of habitat may result from differences in host specificity such as immunological resistance to infection or blood meal preference by potential vectors and/or in behavioral adjustments/physiological traits that alter exposure to vectors.     

Molecular and morphological description of a Hepatozoon species in reptiles and their ticks in the Northern Territory, Australia

Ticks, representing 3 species of Amblyomma, were collected from the water python (Liasis fuscus) and 3 additional reptile species in the Northern Territory, Australia, and tested for the presence of Hepatozoon sp., the most common blood parasites of snakes. In addition, blood smears were collected from 5 reptiles, including the water python, and examined for the presence of the parasite. Hepatozoon sp. DNA was detected in all tick and reptile species, with 57.7% of tick samples (n = 187) and 35.6% of blood smears (n=35) showing evidence of infection. Phylogenetic analysis of the 18S rRNA gene demonstrated that half of the sequences obtained from positive tick samples matched closest with a Hepatozoon species previously identified in the water python population. The remaining sequences were found to be more closely related to mammalian and amphibian Hepatozoon species. This study confirms that species of Amblyomma harbor DNA of the same Hepatozoon species detected in the water pythons. The detection of an additional genotype suggests the ticks may be exposed to 2 Hepatozoon species, providing further opportunity to study multiple host-vector-parasite relationships.