Fine Structure and Life Cycle of Lankesteria clarki sp. n. (Sporozoa: Eugregarinida) Parasitic in the Mosquito Aedes sierrensis (Ludlow)

Lankesteria clarki sp. n. from the treehole mosquito, Aedes sierrensis (Ludlow) in California, is described. This species can be separated from the closely related ones, L. culicis (Ross) and L. barretti Vávra, by the totally intracellular nature of the trophozoite, the shape of the gamont, the position of the gamont nucleus and the structure and position of the residual body in the spore. Morphologic and ultrastructural investigations of gametogenesis and sporogony were conducted and the host relationship discussed. Lankesteria clarki was found in mosquitoes collected from all treeholes examined near Novato in Marin County, California and an examination of over 6,000 larvae of A. sierrensis showed an infection rate of 27.5%. The parasite is capable of destroying the midgut epithelial cells during its trophic stage and Malpighian tubule cells during gametogenesis and sporogony.     

The Fine Structure of the Sporozoite of Lankesteria culicis

SYNOPSIS. The sporozoite of Lankesteria culicis was studied by light and electron microscopy, after excystation in the intestine of Aedes aegypti 1st stage larvae. The sporozoite was 9.5–10.0 μ long with a blunt anterior end and a tapered posterior region. The organism was enclosed by a typical pellicle consisting of an outer and an inner membrane with underlying subpellicular microtubules. The anterior end had a conoid with 2 associated rings, a polar ring which served as a termination of the subpellicular microtubules and a flask-shaped structure situated internal and posterior to the conoid. A micropyle consisting of a collar formed from the inner membrane and lacking an invagination of the outer membrane was present near the anterior end of the parasite. The nucleus was centrally located and had a peripheral concentration of chromatin and a central nucleolus. One or more mitochondria were observed in the vicinity of the nucleus.     

Four New Species of Eimeria (Protozoa: Eimeriidae) from the Pika Ochotona princeps from Alberta and O. pallasi from Kazakhstan1

SYNOPSIS. Eimeria worleyi n. sp. and Eimeria barretti n. sp. from the pika Ochotona princeps from Alberta and Eimeria pallasi n. sp. and Eimeria shubini n. sp. from the pika Ochotona pallasi from Central Kazakhstan are described. Oocysts of E. worleyi were 12–16 by 10–15 μ (mean 13.5 by 12.5 μ) and spherical to subspherical. E. barretti oocysts were ellipsoidal to slightly ovoid and 27–36 by 21–27 μ (mean 32.9 by 23.8 μ). Oocysts of E. pallasi were ellipsoidal or ovoid and 19–34 by 17–26 μ (mean 26.3 by 21.3 μ). E. shubini oocysts were spherical and 22 μ in diameter. E. kriygsmanni Yakimoff and Gousseff, 1938 of Svanbaev (1958) in O. pallasi, [non] E. kriigsmanni Yakimoff and Gousseff, 1938 in Mus musculus, is a synonym of E. pallasi. E. musculi Yakimoff and Gousseff, 1938 of Svanbaev (1958) in O. pallasi, [non] E. musculi Yakimoff and Gousseff, 1938 in Mus musculus, is a synonym of E. shubini.     

Coccidia (Protozoa: Eimeriidae) of the Alpaca Lama pacos*

SYNOPSIS. Three new species of coccidia are described from the alpaca Lama pacos from Peru. These are the first species of coccidia to be named from this host. The oocysts of Eimeria lamae n. sp. are ellipsoidal, occasionally ovoid, 30–40 by 21–30 μ (mean 35.6 by 24.5 μ) with elongate ovoid sporocysts 13–16 by 8–10 μ (mean 15.3 by 8.5 μ). The oocysts of Eimeria alpacae n. sp. are ellipsoidal, rarely ovoid, 22–26 by 18–21 μ (mean 24.1 by 19.6 μ), with ovoid sporocysts 10–13 by 7–8 μ (mean 11.0 by 6.8 μ). The oocysts of Eimeria punoensis n. sp. are ellipsoidal, occasionally ovoid, 17–22 by 14–18 μ (mean 19.9 by 16.4 μ), with ovoid sporocysts 8–11 by 5–7 μ (mean 9.2 by 6.1 μ).     

Ultrastructural study of developmental stages of Mattesia dispora (Neogregarinorida: Lipotrophidae), a parasite of the flour moth Ephestia kuehniella (Lepidoptera)

The ultrastructure of merozoites, gamonts and oocysts of the neogregarine Mattesia dispora and their development in larvae of the flour moth Ephestia kuehniella were studied by electron microscopy. The apical complex of free macronuclear merozoites was very distinct in micrographs of sections, the polar rings being especially prominent. Two gamonts associated in head-to-head syzygy and the apical complexes served as the contact point during pairing. At this stage the rhoptries became reduced and the conoid widened. The gamonts had a foam-like appearance in the light microscope. Paired gamonts formed an envelope and developed into a gametocyst, within which the gamonts were separated by a distinct border. Four gametes and two residual cells developed inside the gametocyst. The gametes were covered with a single membrane. The gametes fused in pairs to form two spherical zygotes, each covered by two membranes and with one large nucleus. The external layer appeared more undulated than the inner one. A single membrane covered each residual cell. Walls were formed around both zygotes to produce two oocysts. Each mature oocyst was lemon-shaped with polar plugs and eight peripheral sporozoites, which had a pellicle similar to that of the merozoites, lay beneath the thick oocyst wall.     

The Oocysts of Eimeria vermiformis sp. n. and E. papillata sp. n. (Protozoa: Eimeriidae) from the Mouse Mus musclus

SYNOPSIS. Eimeria vermiformis sp. n. and E. papillata sp. n. are described from the mouse Mus musculus. The sporulated oocysts of E. vermiformis are 18–26 by 15–21 μ (mean 23.1 by 18.4 μ); its sporocysts are 11–14 by 6–10 μ (mean 12.8 by 7.9 p). The sporulated oocysts of E. papillata are 18–26 by 16–24 μ (mean 22.4 by 19.2 μ); its sporocysts are 10–13 by 6–9 μ (mean 11.2 by 8.0 μ). A substiedal body is present in E. papillata sporocysts. Patent infections were produced in white laboratory mice with both species. Fourteen species of Eimeria have now been described from the genus Mus.     

Eimeria macusaniensis n. sp. (Protozoa: Eimeriidae) of the Alpaca Lama pacos

SYNOPSIS. A new species of coccidium, Eimeria macusaniensis n. sp., is described from the alpaca Lama pacos in Macusani, Peru. The oocysts are ovoid to piriform, 81–107 by 61–80 μ (mean 93.6 by 67.4 μ) with a micropylar cap and elongate ovoid sporocysts 33–40 by 16–20 μ (mean 36.3 by 18.3 μ).     

Gametogenesis and Sporogony of Hepatozoon Mocassini (Apicomplexa: Adeleina: Hepatozoidae) In an Experimental Mosquito Host, Aedes Aegypti

ABSTRACT. The sexual life cycle of the hemogregarine Hepatozoon mocassini was studied in Aedes aegypti , an experimental mosquito host, using transmission electron microscopy. Gamonts were observed leaving the host snake erythrocyte as early as 30 min after mosquitoes ingested infected blood, and some gamonts had penetrated the gut epithelial cells by this time. Six hours post-feeding, gamonts were identified within cells of the abdominal fat body. Twenty-four hours post-feeding, gamonts were often entrapped within the peritrophic membrane, but were no longer observed within the gut wall. Parasites pairing up in syzygy and undergoing sexual differentiatioe were observed within fat cells at this time, and by 48 hours post-feeding, well-developed macro- and microgametocytes as well as microgametes were discernible. Developing zygotes observed 3 days post-feeding were enclosed within a panoitophorous vacuole. By day 6, multinucleate oocysts with crystalloid bodies in the cytoplasm were seen. Sporazoites developing within sporocysts appeared by day 12. Seventeen days post-feeding, mature oocysts with sporocysts containing approximately 16 sporozoites were observed upon dissection of mosquitoes. Large crystalloid bodies no longer bound by rough endopbsmic reticulum were located anterior and posterior to the sporozoite nucleus. Free sporozoites were not observed.     

Eimeria kinsellai sp. n. (Protozoa: Eimeriidae) in a Marsh Rice Rat Oryzomys palustris from Florida*

SYNOPSIS. Eimeria kinsellai sp. n. is described from the marsh rice rat Oryzomys palustris from Florida. Its ellipsoidal oocysts were 23–31 by 15–19 μ (mean 26.7 by 16.9 μ). One to four polar granules were present but an oocyst residuum was absent. The sporocysts were 12–15 by 7–9 μ (mean 13.9 by 8.1 μ), and sporocyst residua were present.     

Development of Caryospora bigenetica n. sp. (Apicomplexa,Eimeriidae) in Rattlesnakes and Laboratory Mice1

During a survey of the coccidian parasites of reptiles from Iowa, three specimens of Crotalus horridus L., the Timber Rattlesnake, and one of Sistrurus catenatus (Rafinesque), the Massasauga Rattlesnake, were found to be passing oocysts of a Caryospora, here described as C. bigenetica n. sp. Since these snakes (family Crotalidae) are known to subsist mainly on small mammals, oocysts from one of the Timber Rattlesnakes were fed to laboratory white mice (Mus musculus L.) to determine if mammals might be involved as alternate hosts in the life cycle. At necropsy, tissues of the tongue and dermis of the mice revealed a sequence of stages which included mature male and female gamonts, fully sporulated sporocysts, “excysted” sporozoites, and “resting” sporozoites that lay individually in solitary, cyst-like host cells termed “caryocysts.” A coccidia-free Massasauga that was fed an infected mouse, at a time when caryocysts in the mouse would have been present, later passed oocysts similar to those of the original inoculum. These results, along with the discovery of endogenous stages (asexual and sexual) in the intestine of the Timber Rattlesnake and the experimentally infected Massasauga, suggest that this parasite has a heteroxenous life cycle pattern, with sexual stages occurring both in the ophidian and the mammalian hosts.     

Development and Dehiscence of Gametocysts of the Gregarine Pyxinia crystalligera Frenzel

SYNOPSIS. Association of gamonts of Pyxinia crystalligera takes place in the midgut of its beetle host, Dermestes vulpinus. At 25°C. the development of gametocysts to the point of liberation of sporocysts is completed between about 15 hours and 27 hours after the gametocysts are deposited with fecal material. Dehiscence is favored by relative humidities of 0% to 90%, but is not favored by a relative humidity of 100%. During the early development of the gametocysts outside the host, the crystals and paraglycogen granules in the cytoplasm of the associated gamonts become concentrated in large masses. The gametes are formed at the periphery of the gamonts. After fusion of the gametes takes place and the sporoblasts begin to develop, the residual cytoplasm containing the inclusions moves outward to form a continuous layer next to the gametocyst envelope, so that the sporoblasts become crowded into a central core. A few hours before dehiscence is initiated a clear area appears on the upper side of the gametocyst. The contents of the gametocyst begin to shrink away from the envelope except in the region of the clear area. Eventually the sporocysts emerge through the clear area and press against the envelope of the gametocyst, causing formation of a conical papilla in the envelope. With continued pressure from the sporocysts, the papilla ruptures at its tip, and the sporocysts emerge in a continuous thread until dehiscence is completed. The thread of sporocysts may attain a length of about 11 mm.     

Inhibition of Dirofilaria immitis in gregarine-infected Aedes aegypti: preliminary observations.

This study examined some of the effects of Ascocystis culicis infections in Aedes aegypti mosquitoes and on the development of dog heartworm, Dirofilaria immitis, in A. culicis-infected A. aegypti. Infections of 100 or more gregarines had an adverse effect on mosquito larvae and the resulting pupae weighed 1 to 2 mg less than the gregarine-free stock mosquitoes. Intensity of gregarine-induced pathology of adult mosquitoes was best estimated by counting the cells of the Malpighian tubules. Larvae infected with approximately 100 gregarines per larva had at least half the number of cells in their tubules destroyed, and development of D. immitis was greatly reduced. Thus, gregarine infections may be useful in reducing populations of aedine mosquitoes and, in areas endemic for heartworm, may interrupt transmission of D. immitis.     

Uniform Terminology for the Protozoan Subphylum Apicomplexa*

SYNOPSIS. Uniform names for the stages, processes and structures of apicomplexan protozoa are proposed and defined, and names that should be superseded are listed. The same names are used for the same stages of all members of the group. Gregarines are designated as either septate or aseptate rather than as polycystid or monocystid. The gregarine stage often called a sporadin is recognized for what it is, a gamont. The cyst formed around 2 gregarine gamonts in which zygotes are formed is a gametocyst; it contains oocysts which in turn contain sporozoites. The term “spore” is inappropriate for these oocysts. The apical complex includes the polar ring, conoid, rhoptries, micronemes and subpellicular tubules. The gregarine “pseudocyst” is actually a gametocyst residuum. The term micropore is preferred to cytostome for the apicomplexan structure, since it is visible only with the electron microscope.     

Ultrastructure of Infection, Development and Gametocyst Formation of Ascogregarina taiwanensis (Apicomplexa: Lecudinidae) in Its Mosquito Host, Aedes albopictus (Diptera: Culicidae)

ABSTRACT. The life history of the protozoan parasite Ascogregarina taiwanensis in mosquito larvae ( Aedes albopictus , collected in southern Taiwan) was shown to consist of two consecutive stages—intracellular and extracellular. Light microscopy showed that most trophozoites moved into the Malpighian tubules and developed into giant trophozoites during the first day pupa. The locomotion may be associated with bristle-like ridges of the trophozoite. The stage for sexual reproduction, i.e. the gamete, was then formed by segmentation of the giant trophozoite and twisting off the anucleate extremities of the body. Sexual reproduction occurred via fertilization by fusion of two resulting gametes, presumably two opposed sexes. The fused gametes finally generate the formation of the gametocyst, within which oocysts develop by budding from the cytoplasmic mass. This type of sexual reproduction has not been reported previously in any gregarine protozoa. We here proposed it as a new hypothesis for further elucidation of the protozoan reproduction.     

Eimeria bateri Bhatia,Pandey and Pande, 1965 from the Hungarian Quail Coturnix c. coturnix in the United States and Its Attempted Transmission to the Chicken**

SYNOPSIS. Eimeria bateri Bhatia, Pandey, Pande, 1965 is described from the Hungarian quail Coturnix c. coturnix for the first time in the U.S. Its oocysts are either ellipsoidal or ovoid, 14–28 by 12–19 μ with a mean of 20.5 by 15.3 μ. Its sporocysts are elongate ovoid, with a Stieda body, 8–12 by 6–9 μ with a mean of 10.3 by 7.5 μ. Two coccidia-free chicks 8 days old were inoculated with 50,000 oocysts each of E. bateri, but patent infections did not occur. This coccidium is therefore presumably host-specific.     

Susceptibility of Aedes aegypti and Aedes albopictus larvae to Ascogregarina culicis and Ascogregarina taiwanensis (Apicomplexa: Lecudinidae) from Florida

The susceptibility of Aedes aegypti to Ascogregarina culicis and Aedes albopictus to Ascogregarina taiwanensis was examined with mosquito and parasite strains from Tampa, FL. When each host was bioassayed with its natural gregarine, the infection intensity indicated that Ae. aegypti was 59% more susceptible to A. culicis (87 gamonts/larva) than Ae. albopictus to A. taiwanensis (47 gamonts/larva). Infections in single and mixed host populations exposed to 100 oocysts/larva of one and both parasites demonstrated that Ae. aegypti harbors higher A. culicis gamont loads than Ae. albopictus of A. taiwanensis. In dual gregarine exposures of single host populations, the A. culicis infection intensity in Ae. aegypti was reduced by approximately 50%. A. taiwanensis exhibited the same capability of infecting Ae. albopictus in single and dual exposures. In mixed host populations there were no cross infections, but A. taiwanensis in Ae. albopictus produced an infection intensity of approximately 70% lower than that of A. culicis in Ae. aegypti.     

The Life Cycle of Ganymedes oaklandi n. sp., an Acephaline Gregarine of Gammarus fasciatus (Say)*

SYNOPSIS. Ganymedes oaklandi n. sp. (Ganymedidae) from the intestine of Gammarus fasciatus (Say) is described, and its life cycle is given. In the trophic stage the gregarine is acephaline, with an elongated cylindrical body slightly tapered anteriorly, and is up to 298 × 64 μ. The gametocysts are up to 121 × 99 μ, and are covered with a secreted thick sticky gelatinous coat. The spores are released from the gametocyst by rupturing of the cyst wall. The spore body is spherical, approximately 6 μ in diameter, and has 4 episporal rays about 20 μ long.     

Isospora Species of the Cat and Attempted Transmission of I. felis Wenyon, 1923 from the Cat to the Dog*

SYNOPSIS Fecal samples from 130 domestic cats from Illinois were examined for coccidia. Three species of Isospora were found: (1) I. felis Wenyon, 1923, with oocysts 38-51 by 27-39 μ with a mean of 41.6 by 30.5 μ and sporocysts 20-26 by 17-22 μ with a mean of 22.6 by 18.4 μ; it was found in 13% of the cats; (2) I. rivolta (Grassi, 1879) Wenyon, 1923, with oocysts 21-28 by 18-23 μ with a mean of 25.0 by 21.1 μ, and sporocysts 14-16 by 10-13 μ with a mean of 15.2 by 11.6 μ; it was found in 3% of the cats; and (3) I. bigemina (Stiles, 1891) Lühe, 1906, with oocysts 12-15 by 10-13 μ with a mean of 13.2 by 11.8 μ. and sporocysts 8-10 by 6-8 μ with a mean of 8.8 by 6.5 μ it was found in 1.5% of the cats. Four coccidia-free puppies 1.5 months old were inoculated with 100,000 oocysts each of I. felis from the cat, but patent infections did not occur. Partial development of I. felis was not seen in tissue sections of the small intestine of a 5th pup killed 96 hours after inoculation with 150,000 I. felis oocysts. This coccidium is therefore presumably host-specific.     

Fatal Toxoplasmosis and Enteroepithelial Stages of Toxoplasma gondii in a Pallas Cat (Felis manul)1

Toxoplasma gondii was found in tissues of a six-year-old female Pallas cat (Felis manul) from the Milwaukee County Zoo. Toxoplasma gondii meronts (types D and E), gamonts, and oocysts were present in the epithelium of the small intestine. Numerous unsporulated oocysts were present in the intestinal lumen. The cat died of acute, overwhelming toxoplasmosis. Necrotic enteritis, multifocal necrotizing granulomatous hepatitis, and pneumonia were the prominent lesions.