Redescription of Haemoproteus mesnili (Apicomplexa: Plasmodiidae) and its meronts, with description of a second haemosporidian parasite of African cobras

Haemoproteus mesnili (Bouet 1909) Wenyon 1926 is redescribed from the spitting cobra, Naja nigricollis nigricollis, of Tanzania. Mature gametocytes in the acute phase of infection averaged 17.7 X 7.3 jim, with LW 128.1 jim-, and L:W ratio 2.52. Nuclei were visible in both sexes. Both sexes were heavily pigmented, with 31-62 black granules dispersed in macrogametocytes; 20-46 granules were often clumped or concentrated near ends of microgametocytes. The halteridial form was present in 28% of active-phase gametocytes, but in only 8% of those in chronic phase. A few large, possibly first generation, meronts were present in cardiac muscle; uninucleate parasites within parasitophorous vacuoles in splenic cells produced small rounded or ovoid meronts, 12.2 x 9.6 microm, with 12-16 deeply basophilic, square-to-rectangular cytomeres. Meronts with 17-32 cytomeres were 16.9 x 11.9 microm. Meronts, 20 x 16 to 26 x 22 microm, contained 51-57 cytomeres. Mature meronts were ovoid, 13.7 x 11.5 microm, with many rounded merozoites. Haemoproteus balli n. sp, found in an Egyptian cobra, Naja haje haje of Kenya, differs from H. mesnili in average gametocyte dimensions, 10.8 x 7.7 microm; LW, 83.2 microm2; L/W ratio, 1.42; absence of halteridial forms; sparse pigmentation (3-10 granules); and presence of a broad peripheral band, apparently chromatin, along one side of microgametocytes.     

Revision of the Thelastomatoidea,Oxyurida of invertebrate hosts I. Thelastomatidae

Haemoproteus psittaci n. sp., a microsomal halteridial haemoproteid, is described from the African grey parrot Psittacus erithacus. This species differs from Haemoproteus handai, the only other haemoproteid described from the Psittaciformes, in that it is halteridial, not circumnuclear, is much smaller in all dimensions and has only 10–12 pigment granules compared to 20–24 noted in the latter species.     

New avian Haemoproteus species (Haemosporida: Haemoproteidae) from African birds, with a critique of the use of host taxonomic information in hemoproteid classification

Haemoproteus (Parahaemoproteus) micronuclearis n. sp., Haemoproteus (Parahaemoproteus) nucleofascialis n. sp., Haemoproteus (Parahaemoproteus) paranucleophilus n. sp., and Haemoproteus (Parahaemoproteus) homobelopolskyi n. sp. (Haemosporida, Haemoproteidae) are described from African passeriform birds based on the morphology of their blood stages and segments of the mitochondrial cytochrome b gene. Red-billed quelea (Quelea quelea), red-headed malimbe (Malimbus rubricollis), and black-headed weaver (Ploceus melanocephalus) are the type vertebrate hosts of new hemoproteids. It is probable that new species have wide distribution in weavers in sub-Saharan Africa. Both H. micronuclearis and H. nucleofascialis can be readily distinguished from other avian hemoproteids by tiny, compact microgametocyte nuclei that are significantly smaller than macrogametocyte nuclei and are a rare character of hemosporidian parasites. Gametocytes of H. paranucleophilus are closely appressed to the erythrocyte nuclei and do not touch the erythrocyte envelope along their entire margin at all stages of their development, including fully grown gametocytes. A particularly distinctive feature of H. homobelopolskyi development is the presence of circumnuclear dumbbell-shaped macrogametocytes. Illustrations of blood stages of the new species are given, and morphological and phylogenetic analyses identify the DNA lineages that are associated with these parasites. Numerous recent studies show that some lineages of hemoproteids are often present in birds belonging to different families. As a result, the use of the host family as a taxonomic character should be questioned and preferably discouraged in hemoproteid taxonomy, particularly with regard to the parasites of passerine birds. Microscopic identification of avian hemoproteids requires comparison of Haemoproteus species described from birds of different families, as is an established practice with avian Plasmodium spp. Development of bar-coding techniques remains essential in taxonomic and field studies of hemosporidian parasites.     

Haemoproteids of the avian family Dicruridae (the drongos)

Dicrurids are a widespread avian family in Africa and Asia. Earlier surveys of this family in these areas have reported the presence of hematozoa and 1 species of Haemoproteus, i.e., Haemoproteus dicruri (De Mello, 1935). One species of drongo occurs in Madagascar and has not been examined previously. Blood smears collected from wild-caught crested drongos, Dicrurus forficatus, in Madagascar were examined using a compound microscope for the presence of hematozoa. A new species, Haemoproteus khani, is described in this study. This new species has circumnuclear gametocytes, in contrast to the halteridial H. dicruri. In addition, H. dicruri is reported for the first time from the crested drongo and is redescribed. This is the first report of hematozoa in drongos of Madagascar.     

Two new species of Haemocystidium Castellani & Willey (Apicomplexa: Plasmodiidae) from Pakistani lizards, and the support their meronts provide for the validity of the genus

Haemocystidium papernai n. sp., a parasite of Agama nupta fusca in Sind, Pakistan produces large halteridial gametocytes in active infections that do not differ in morphology by sex. In chronic phase macrogametocytes are larger than microgametocytes. In Baluchistan, Agama nupta nupta is parasitised by Haemocystidium quettaensis n. sp. Gametocytes are smaller than those of H. papernai n. sp., round or oval in shape and heavily pigmented. Macrogametocytes are larger than microgametocytes and more heavily pigmented. Merogony of H. papernai n. sp. occurs in endothelium and connective tissue of lungs, heart, liver, spleen and femoral muscles. Meronts form as oval or rounded bodies that apparently are not preceded by the formation of pseudocytomeres that appear in the merogony of Haemoproteus spp. in snakes, turtles and birds. Meronts of Haemocystidium kopki were found in the lungs of the Pakistani gecko Teratoscincus scincus that were similar to those of H. papernai n. sp. The meronts of both species closely resemble phanerozoic meronts of saurian Plasmodium spp., and provide adequate justification for recognition of the genus Haemocystidium Castellani & Willey, 1904. Haemocystidium is distinguished from Plasmodium by the absence of an asexual cycle in circulating blood cells, and from Haemoproteus by meronts that do not form as pseudocytomeres. Sex ratios of H. papernai n. sp. varied from 22–47% microgametocytes, with no evident correlation with parasitemia, infection stage or month of collection, and H. kopki infections were comprised of 61–74% microgametocytes.     

Haemoproteus meleagridis Levine 1961: redescription and developmental morphology of the gametocytes in turkeys

Haemoproteus meleagridis Levine 1961 is redescribed and illustrated from material obtained from wild and domestic turkeys (Meleagris gallopavo) in Florida and Georgia. The mature gametocyte of this haemoproteid surrounds the erythrocytic nucleus, occupies 80 to 90% of the host-cell-parasite complex, and causes atrophy of the host cell's nucleus and hypertrophy of the host cell. The developmental sequence of H. meleagridis was studied critically. Following the entry of merozoites into the erythrocytes, they grew into halteridial and then circumnuclear forms. This was followed by a 10- to 13-day period during which trophozoites were not detectable by blood smear, and after which, the trophozoites returned to the peripheral circulation.     

Fallisia parasites (Haemosporidia: Plasmodiidae) from the flying lizard, Draco maculatus (Agamidae) in Thailand

Plasmodium (Fallisia) siamense n. sp. was described from the flying lizard Draco maculatus (Agamidae) collected near Bangkok, Thailand. Parasitic in thrombocytes, it produces 18-64 merozoites in schizonts that are larger than the usually oval gametocytes. Both gametocyte sexes showed a prominent nucleus, elongate in macrogametocytes and triangular in microgametocytes. This is the first Fallisia parasite described from a host outside the Neotropical Region, although presence of the genus in Australasia was reported 40 years ago, without formal taxonomic designation.     

Molecular phylogenetic analysis of circumnuclear hemoproteids (Haemosporida: Haemoproteidae) of sylviid birds, with a description of Haemoproteus parabelopolskyi sp. nov

Haemoproteus spp., with circumnuclear gametocytes and tentatively belonging to Haemoproteus belopolskyi, are widespread and prevalent in warblers belonging to the Sylviidae, with numerous mitochondrial cytochrome b (cyt b) lineages detected among them.We sampled the hemoproteids from 6 species of warblers adjacent to the Baltic Sea. Parasites were identified to species based on morphology of their gametocytes, and a segment of the parasite's cyt b gene was sequenced. Sixteen mitochondrial cyt b lineages of hemoproteids with circumnuclear gametocytes were recorded. Two clades of lineages (clade A in species of Acrocephalus and Hippolais and clade B in species of Sylvia) with sequence divergence between their lineages >5% are distinguished in the phylogenetic tree. Within the clades A and B, the genetic distance between the lineages is < or = 3.9 and < and = 2.8%, respectively. We compared the morphology of gametocytes of 3 lineages (hHIICT1, hMW1, and hSYAT2) in detail. The lineages hHIICTI and hMW1 (clade A) belong to the morphospecies H. belopolskyi. Parasites of the lineage hSYAT2 (clade B) are described as a new species Haemoproteus parabelopolskyi, which can be readily distinguished from H. belopolskyi by the significantly smaller nuclei of its macrogametocytes. Lineages closely related to H. belopolskyi and H. parabelopolskyi are identified. The sequence divergence between lineages of these 2 morphospecies ranges between 5.3 and 8.1%. It seems probable that avian Haemoproteus spp. with a genetic differentiation of > or =5% in mitochondrial cyt b gene might be morphologically differentiated at the stage of gametocytes. This study establishes the value of both PCR and morphology in identification of avian hemoproteids.     

A Haemocystidium species from the East African gecko Lygodactylus capensis grotei

Haemocystidium lygodactyli n. sp. parasitizes Lygodactylus capensis grotei (Gekkonidae) in Tanzania. Mature gametocytes in acute phase of infection average 16.3 x 5.7 microm (11-20 x 4-9.5 microm), with LW 93.0 (62-140 microm2) and L/W ratio 2.94 (1.2-3.9). Gametocytes usually lateral, lateropolar, or halteridial in position. There was no significant sexual dimorphism in gametocyte dimensions. Nuclei discrete in both sexes at maturity, with a rounded nucleolus usually present in microgametocytes. In chronic infection, gametocytes were 18.1 x 8.7 microm (8-25 x 5-11 microm), with LW 156.8 microm2 (80-250) and L/W 2.16 (1.1-3.6). When gametocytes from the chronic infection were compared with the same sex in acute infection, length did not differ, but differences were present between the same sex in each comparison of width, LW, and L/W. Macrogametocytes and microgametocytes in chronic phase were broader, larger, and less elongate and most commonly halteridial. Meronts were found only in endothelium and connective tissue of lung. Elongate to oval in shape, the larger meronts filled with nuclei were 12.2 x 6.9 microm (10.0 x 5.0-16.0 x 9.0), with LW 50-144 microm2 (85.1). In 1 initial infection followed for 49 days, apparently mature gametocytes appeared by day 28 postcapture. Binucleate parasites were present from day 14 throughout the course of infection, with their frequency increasing from 5% of immature parasites to 34% of mature gametocytes. Binucleate mature gametocytes were found in 1 other infection, where 14% had 2 nuclei. Sex ratio varied from 51 to 63% in favor of macrogametocytes.     

Electron Microscope Studies on Eimeria perforans (Sporozoa)

SYNOPSIS. By means of electron microscopy, a study has been made of the fine structure of the macrogametocytes, microgametocytes and oocysts of Eimeria perforans from the intestine of the wild rabbit (Oryctolagus cuniculus). The parasites lie in a vacuole within the host cell. The surface of the gametocytes is not plain, but displays irregular protrusions. A large intranuclear body can be detected within the macrogametocytes. Similar structures are also found within the cytoplasm. Within the latter there exists a large spread out reticulum, the channels and vesicles of which concentrate especially close to the nuclear membrane. Tubuli are seen in the numerous mitochondria, which often have a dumb-bell shape.
In most of the gametocytes irregular, strongly osmiophilic lipid inclusions are observed, which always are surrounded by the endoplasmic reticulum. Strange folded ovoid bodies are found within the cytoplasm of the oocysts. Nothing can be told with certainty of their nature and function. Probably they represent specific storage bodies.     

Novel Haemoproteus species (Haemosporida: Haemoproteidae) from the swallow-tailed gull (Lariidae), with remarks on the host range of hippoboscid-transmitted avian hemoproteids

Haemoproteus (Haemoproteus) jenniae n. sp. (Haemosporida: Haemoproteidae) is described from a Galapagos bird, the swallow-tailed gull Creagrus furcatus (Charadriiformes, Laridae), based on the morphology of its blood stages and segments of the mitochondrial cytochrome b (cyt b) gene. The most distinctive features of H. jenniae development are the circumnuclear gametocytes occupying all cytoplasmic space in infected erythrocytes and the presence of advanced, growing gametocytes in which the pellicle is closely appressed to the erythrocyte envelope but does not extend to the erythrocyte nucleus. This parasite is distinguishable from Haemoproteus larae, which produces similar gametocytes and parasitizes closely related species of Laridae. Haemoproteus jenniae can be distinguished from H. larae primarily due to (1) the predominantly amoeboid outline of young gametocytes, (2) diffuse macrogametocyte nuclei which do not possess distinguishable nucleoli, (3) the consistent size and shape of pigment granules, and (4) the absence of rod-like pigment granules from gametocytes. Additionally, fully-grown gametocytes of H. jenniae cause both the marked hypertrophy of infected erythrocytes in width and the rounding up of the host cells, which is not the case in H. larae. Phylogenetic analyses identified the DNA lineages that are associated with H. jenniae and showed that this parasite is more closely related to the hippoboscid-transmitted (Hippoboscidae) species than to the Culicoides spp.-transmitted (Ceratopogonidae) species of avian hemoproteids. Genetic divergence between morphologically well-differentiated H. jenniae and the hippoboscid-transmitted Haemoproteus iwa, the closely related parasite of frigatebirds (Fregatidae, Pelecaniformes), is only 0.6%; cyt b sequences of these parasites differ only by 1 base pair. This is the first example of such a small genetic difference in the cyt b gene between species of the subgenus Haemoproteus. In a segment of caseinolytic protease C gene (ClpC), genetic divergence is 4% between H. jenniae and H. iwa. This study corroborates the conclusion that hippoboscid-transmitted Haemoproteus parasites infect not only Columbiformes birds but also infect marine birds belonging to Pelecaniformes and Charadriiformes. We conclude that the vertebrate host range should be used cautiously in identification of subgenera of avian Haemoproteus species and that the phylogenies based on the cyt b gene provide evidence for determining the subgeneric position of avian hemoproteids.     

Hematozoa and a new haemoproteid species from Cathartidae (New World vulture) in South Carolina

A survey of turkey vultures (Cathartes aura) in South Carolina revealed the presence of a novel haemoproteid. Haemoproteus catharti n. sp. is described from the slides developed from the blood of this vulture. The new species is a thick, halteridial form with complete margins, and is considered distinct from the other recognized haemoproteids from the diurnal raptors in shape, pigment number, parasite outline, and host-family specificity. While reviewing blood films, a series of unusual immature schizonts of Plasmodium sp. were also observed, and these are illustrated, along with another distinct haemoproteid from old slides of poor quality produced from turkey vulture blood.     

Malaria Parasites of the “Borriguerro'’Lizard,Ameiva ameiva (Sauria: Teiidae) in Panama*

SYNOPSIS. The teiid lizard Ameiva ameiva praesignis in Panama is parasitized by 2 species of Plasmodium: P. cnemidophori Carini and P. diminutivum sp. n. The Panamanian strain of P. cnemidophori has thick macrogametocytes which are larger than the oval microgametocytes, and schizonts which contain 42–119 nuclei. P. diminutivum is characterized by round to oval gametocytes which are usually smaller than the host cell nucleus; fan-shaped schizonts with 4–6 nuclei; and prominently vacuolated young stages, with a thick band of chromatin along the periphery of the vacuole.     

Two new species of Haemoproteus Kruse, 1890 (Haemosporida,Haemoproteidae) from European birds,with emphasis on DNA barcoding for detection of haemosporidians in wildlife

Two new species of Haemoproteus Kruse, 1890 (Haemosporida, Haemoproteidae) are described: Haemoproteus (Parahaemoproteus) homovelans n. sp. from Grey-faced Woodpecker, Picus canus Gmelin, and Haemoproteus (Parahaemoproteus) concavocentralis n. sp. recorded in Hawfinch, Coccothraustes coccothraustes (Linnaeus), both sampled in Bulgaria. The morphology of the gametocytes and their host-cells are described and mitochondrial cytochrome b (cyt b) gene sequences are generated. Haemoproteus homovelans possesses circumnuclear gametocytes lacking volutin granules. This parasite is particularly similar to Haemoproteus velans Coatney & Roudabush, 1937 also possessing circumnuclear gametocytes that are, however, overfilled with volutin. Haemoproteus concavocentralis can be readily distinguished from all described avian haemoproteids due to the presence of an unfilled concave space between the central part of advanced gametocytes and erythrocyte nucleus. Bayesian phylogenetic analyses of 40 haemosporidian cyt b lineages showed close relationships of H. concavocentralis (hHAWF2) with a group of Haemoproteus spp. possessing gametocytes that are pale-stained with Giemsa. The lineage hPICAN02 of H. homovelans clustered with parasites infecting non-passerine birds. Phylogenetic analyses support the current subgeneric classification of the avian haemoproteids and suggest that cyt b lineage hPIPUB01 (GenBank EU254552) has been incorrectly assigned to Haemoproteus picae Coatney & Roudabush, 1937, a common parasite of corvid birds (Passeriformes). This study emphasises the importance of combining molecular techniques and light microscopy in the identification and field studies of avian haemosporidian parasites. Future development of barcodes for molecular identification of haemoproteids will allow better diagnostics of these infections, particularly in veterinary studies addressing insufficiently investigated tissue pathology caused by these parasites.     

Cytochemical study of the different stages in the life cycle of Toxoplasma gondii. IX. The polysaccharides and lipids in the parasites at developmental stages from the cat intestine]

Amylopectin was detected in all the stages examined. In the oval stages the minute granules of PAS-positive material were seen in the cytoplasm when examined on fresh-frozen sections. In merozoites, amylopectin was more conspicuous with maturation. The residual body of microgametocytes contain large amounts of amylopectin; no polysaccharide was visualized in microgamete bodies. Amylopectin was most abundant in macrogametocytes and zygotes. However, no peripheral position of PAS-positive "plastic granules" (wall-forming bodies), so characteristic of other coccidia and revealed by the electron microscopy for T. gondii macrogametocytes, was seen. Acid mucopolysaccharides in the macrogametocyte were detected in the central zone, leaving the periphery of the cell unstained. Very small, if any, amounts of lipids were detected in asexual stages of T. gondii. Unlike, large accumulation of lipid droplets were seen in growing macrogametocytes suggesting the involvement of lipids along with amylopectin in the metabolism of oocysts later discharged from the host body.     

Haemoproteus gabaldoni n. sp. (Apicomplexa: Haemoproteidae) from the Muscovy duck Cairina moschata (Aves: Anatidae)

Haemoproteus gabaldoni n. sp. is described from the Muscovy duck Cairina moschata from Caracas, Venezuela and is compared to H. greineri and H. nettionis which have been described previously from the Anatidae. The highly amoeboid outline, volutin granules and small number of pigment granules of H. gabaldoni serve to readily separate this species from the other two.     

Myopathy associated with megaloschizonts of Haemoproteus meleagridis in a wild turkey from Florida

Necropsy of an emaciated adult wild turkey (Meleagris gallopavo osceola) that died in captivity soon after capture revealed numerous macroscopic 1-2 mm white cysts in the pectoral muscles. Microscopic, aseptate protozoan megaloschizonts, 50-150 microns in diameter, corresponded to the cysts in histological sections. The megaloschizonts were surrounded by a thick, hyaline wall and packed with spherical merozoites less than 1 micron in diameter. Muscle fibers surrounding most of the megaloschizonts exhibited early signs of dystrophic calcification. The fibers were swollen, pale and hyaline and contained scattered basophilic granules. The megaloschizonts were morphologically distinct from sarcocysts of Sarcocystis sp. and Besnoitia sp. and the thin-walled tissue cysts of Toxoplasma gondii. They were identical in structure and host reaction to the second-generation megaloschizonts of Haemoproteus meleagridis, reported previously from experimentally infected domestic turkeys. While the precise cause of death of the wild turkey could not be determined, the most prominent lesions were associated with the numerous intramuscular megaloschizonts.     

A survey of coccidian infection of freshwater fishes in South Africa,with the description of Goussia anopli n. sp. (Apicomplexa: Eimeriidae)

Seventy-seven specimens of seven freshwater fish species harvested in four sites from rivers and ponds of the Gauteng, North West and Limpopo Provinces (South Africa) were surveyed for coccidian infections. Two fish species were infected with apicomplexans belonging to Goussia Labbé, 1896. In banded tilapia Tilapia sparrmanii Smith unsporulated oöcysts of G. vanasi (Landsberg &; Paperna, 1987) were found which became sporulated in tap-water within 24 hours. Another species in the gut of chubbyhead barb Barbus anoplus Weber harboured sporulated oöcysts in the faeces and in the intestinal epithelium. The latter species has been described as G. anopli n. sp.