Effects of itraconazole on the sporocysts wall of the entomopathogenic fungi Ascosphaera apis as revealed by the scanning electron microscope

The young sporocysts had a wrinkled sporocyst wall, numerous papillae on the wall surface and the wall was granular and porous. In the itraconazole-treated culture, the walls of the young sporocysts were also wrinkled, but the characteristic papillae were replaced by larger lumps which were densely packed and coated the entire surface of the sporocyst wall. The mature sporocysts walls were smooth and possessed numerous papillae. In the itraconazole-treated culture, the walls of mature sporocysts walls were also smooth but possessed densely packed larger lumps instead of papillae. At higher magnification, each of the lumps were found to consist of numerous globules. No pores were observed as they were in the normal sporocyst wall.     

Ultrastructural changes in the spore and mycelia of Ascosphaera apis after treatment with benomyl (Benlate 50 W)

In Ascosphaera apis, after 8 days growth in darkness at 28° C, numerous sporocysts were observed, within which mature spores were seen aggregated into a spore ball. The mature spore of A. apis had a thick spore wall with an electron-opaque outer layer, a spore membrane with many depressions, and sporoplasm containing numerous ribosomes and mitochondria. In the cytoplasm of the mycelium, mitochondria with well-defined cristae and numerous ribosomes were observed. At a concentration of 1 g/ml of culture medium, benomyl appeared to inhibit colony growth of A. apis, but some sporocysts containing deformed spores were found. Deformed spores possessed a thick spore wall with a grainy matrix, and depressions were no longer detected in the spore membrane. Ribosomes were lacking in the sporoplasm and mitochondria appeared degenerate. The mycelium from the treated culture contained mitochondria with an electron-lucid matrix and no well defined cristae, while ribosomes were completely depleted. The significance of these observations in relation to the use of benomyl to control chalkbrood disease in the honey bee is discussed.     

Sarcocystis neurona (Protozoa: Apicomplexa): description of oocysts, sporocysts, sporozoites, excystation, and early development

Equine protozoal myeloencephalitis is a major cause of neurological disease in horses from the Americas. Horses are considered accidental intermediate hosts. The structure of sporocysts of the causative agent, Sarcocystis neurona, has never been described. Sporocysts of S. neurona were obtained from the intestines of a laboratory-raised opossum fed skeletal muscles from a raccoon that had been fed sporocysts. Sporocysts were 11.3 by 8.2 microm and contained 4 sporozoites. The appearance of the sporocyst residuum was variable. The residuum of some sporocysts was composed of many dispersed granules, whereas some had granules mixed with larger globules. Excystation was by collapse of the sporocyst along plates. The sporocysts wall was composed of 3 layers: a thin electron-dense outer layer, a thin electron-lucent middle layer, and a thick electron-dense inner layer. The sporocyst wall was thickened at the junctions of the plates. Sporozoites were weakly motile and contained a centrally or posteriorly located nucleus. No retractile or crystalloid body was present, but lipidlike globules about 1 microm in diameter were usually present in the conoidal end of sporozoites. Sporozoites contained 2-4 electron-dense rhoptries and other organelles typical of coccidian zoites. Sporozoites entered host cells in culture and underwent schizogony within 3 days.     

In vitro activity of 15-azasterol (A25822B) against chalkbrood pathogen Ascosphaera apis in the honey bee

The antifungal agent 15-azasterol A25822B was examined for effects on the growth and development of Ascosphaera apis. The minimum inhibition concentration (MIC) of azasterol against A. apis was 1 m. Growth and development of A. apis was completely controlled at this concentration. At a concentration of 0.01 m growth of A. apis was retarded and although sporocysts were formed developing spores were not be able to reach maturation. A major effect of azasterol at this low concentration was the accumulation of lipid in the hyphae, sporocysts and immature spores. In addition it caused a conformational change in mitochondria and damage to the spore membrane structure. On the basis of these results, further investigations of azasterol for the treatment of chalkbrood disease in the honey bee are warranted.Work was performed during sabbatical leave at the University of California, Davis.     

The invasion of Biomphalaria pfeifferi by Schistosoma mansoni miracidia and the development of daughter sporocysts

Laboratory experiments have been carried out to determine the susceptibility of Gezira Biomphalaria pfeifferi snails to S. mansoni miracidia and the relationship between miracidia and daughter sporocyst production at the 10–17 day development stage. The relationship between snail numbers, miracidia numbers and water volume has also been studied. Two non susceptible snails, Bulinus truncatus and Cleopatra bulimoides, both of which occur naturally in Gezira canals, were tested to see if they act as decoys for S. mansoni miracidia.The results showed that the B. pfeifferi are 100% susceptible to S. mansoni invasion, at least to the daughter sporocyst development stage. The more miracidia that penetrated the more daughter sporocysts were produced, however individual variation and overlap were great. When one miracidium was released to find one snail it succeeded in low water volumes (5 m, 50 ml), but failed in 5 litres. When 100 miracidia were released mortality of snails was high suggesting superinfection particularly when only one or five snails were available. Among survivors daughter sporocyst counts were very high. Cleopatra and Bulinus snails do have a decoy effect when present in large numbers. In their presence the number of infected snails was marginally reduced and the number of daughter sporocysts greatly reduced. However, if superinfection is reduced by decoy effect, it is conceivable that Biomphalaria may be protected by decoy snails in circumstances where miracidia counts are high.     

Parthenogenetic generations of Helicometra fasciata Rud., 1819 (Trematoda: Opecoelidae) in the Black Sea molluscs Gibbula adriatica

Morphology of the Helicometra fasciata Rud., 1819 parthenogenetic generation from the Black Sea gastropods Gibbula adriatica (Phil.) was studied for the first time. Data on seasonal dynamics of the hemipopulation of daughter sporocysts are given. Daughter sporocysts of H. fasciata infest 10 +/- 0.2 % of G. adriatica (mainly specimens of larger size and elder age classes). As a rule, local microhemipopulations of daughter sporocysts castrate mollusc hosts. Reproduction of H. fasciata daughter sporocysts is asynchronous: daughter sporocysts born specimens of next sporocyst generation during autumn and winter, and then they begin producing cercaria. In winter development of the cercaria embryo is blocked. Second change of the character of the each sporocyst' posterity is impossible because of the annual life cycle of G. adriatica. Endogenous agglomeration of the H. fasciata daughter sporocysts is extremely little: individuals of next sporocyst generation develop from no more than 2 % of embryonic balls. Energy resources of the mollusc host are used by the H. fasciata daughter sporocysts mainly for producing cercaria; this fact can be interpreted as an adaptation of H. fasciata to using medium-sized, short-living mollusc hosts.     

Ultrastructure of the daughter sporocyst and developing cercaria of Schistosoma japonicum in experimentally infected snails, Oncomelania hupensis hupensis

Ultrastructure of daughter sporocysts and cercariae of Schistosoma japonicum were studied 2 and 4 months after infection of Oncomelania hupensis hupensis. The body walls of daughter sporocysts are similar at all infectious stages. They consist of an external syncytial tegument on a basement membrane, and an internal cellular subtegument surrounding a body cavity containing developing cercariae. The cercariae embryos develop 2 months after infection from germinal balls in the brood chamber of the daughter sporocyst. They are at first enveloped by a primitive epithelium rising from the daughter sporocyst. Four months after infection, the cercariae were almost fully developed and the primitive epithelium had degenerated. The body wall of the cercaria consists of a thin tegument covered by a surface coat of fibrous material and connected to the subtegumental cells by cytoplasmic processes. The matrix of the tegument contains numerous dense bodies, vacuoles, and spines. Two types of sensory structures - uniciliated and multiciliated - are found at the anterior tip of the cercaria. There are five pairs of penetration gland cells of two distinct types differentiated by the morphology of secretory granules. Flame cells are found in both daughter sporocysts and in cercariae. The cilia of the flame cells are characterized by the typical 9 and 2 cilium pattern.     

日本血吸虫胞蚴期超微结构的初步观察

本文用扫描与透射电镜观察了我国大陆品系37日和45日龄日本血吸虫母胞蚴及其体内未成熟子胞蚴体被的结构。同时观察了取自螺肝组织的62日龄成熟子胞蚴。初次揭示日本血吸虫胞蚴期体被的超微结构,基本上与曼氏血吸虫胞蚴期相似。日本血吸虫母胞蚴和成熟子胞蚴除体被无体棘外,其他很相似。比较未成熟的子胞蚴与未成熟的尾蚴,揭示外质膜由两层结构构成;随后,外层结构溶化消失,而同时出现微绒毛。构成这样母子二代胞蚴及其体内胚胎既相同又有差别,认为与幼虫寄生部位及生殖生理状态有关。     

Caryospora uptoni n. sp. (Apicomplexa: Eimeriidae) from red-tailed hawks (Buteo jamaicensis borealis)

Oocysts of Caryospora uptoni n. sp. were described from the feces of red-tailed hawks, Buteo jamaicensis borealis. Sporulated oocysts were spherical or subspherical and measured 28.1 by 26.4 micron. The oocyst wall was composed of a yellowish outer layer and brownish inner layer and was about 1.5 micron thick. Neither micropyle, polar granules, nor oocyst residuum were present. A single, spherical sporocyst 18.2 by 17.9 micron was present; a Stieda body was absent. A spherical eccentrically located sporocyst residuum was present in many sporocysts, but it degenerated to form a dispersed granular residuum in other sporocysts. Eight randomly arranged sporozoites, 12.6 by 4.2 micron, were present in each sporocyst; they contained a centrally or slightly posteriorly located nucleus.     

New Species of Eimeria (Protozoa: Eimeriidae) from Malaysian Rats*

SYNOPSIS. Four new species of Eimeria were found in a survey of 255 rats of 14 species in Malaysia. E. tikusi n. sp. and E. edwardsi n. sp. are described from Edwards' rat Rattus edwardsi. The ellipsoidal, single-layered oocysts of E. tikusi average 30.3 by 24.4 μ. A micropyle is absent; a polar granule is present. Ovoid sporocysts average 14.2 by 9.8 μ. A sporocyst residuum and Stieda body are present. The ovoid, 2-layered oocysts of E. edwardsi average 29.1 by 21.8 μ. A micropyle is present; a polar granule is absent. Ellipsoidal to ovoid sporocysts average 14.5 by 6.5 μ. A sporocyst residuum is present; Stieda body is small or absent. E. surifer n. sp. is described from the red spiny rat Rattus surifer. Its ellipsoidal 3-layered oocysts average 34.7 by 24.8 μ. A micropyle is absent; a polar granule is present. The ellipsoidal sporocysts average 15.4 by 9.5 μ. A sporocyst residuum, Stieda body and sub-Stieda body are present. E. sabani n. sp. is described from the long-tailed giant rat R. sabanus. Its ellipsoidal 2-layered oocysts average 28.5 by 21.7 μ. A micropyle is absent; a polar granule is present. The ellipsoidal-to-ovoid sporocysts average 11.9 by 8.0 μ. A sporocyst residuum and Stieda body are present.     

Encapsulation response of the marine prosobranch Cerithidea californica to natural infections of Renicola buchanani sporocysts (Trematoda: Renicolidae)

A hyalinocyte-mediated encapsulation reaction is elicited by sporocysts of Renicola buchanani infecting the anterior mantle region of Cerithidea californica. Three phases of capsule formation are recognized: (1) an initial aggregating of hyalinocytes around each sporocyst in which pseudopodia from encapsulating cells loosely interdigitate with the parasite's tegumental microvilli, (2) the infiltrating of numerous hyalinocytes to form a dense matrix of cells which lies in close contact with the sporocyst's tegument, and (3) the horizontal flattening of hyalinocytes against the sporocyst's surface to form a tightly adhering capsule four to eight cell layers deep. Sporocysts are not harmed as a consequence of encapsulation. Capsule formation in response to R. buchanani sporocysts is considered a type of leucocytic encapsulation, specifically designated hyalinocytic encapsulation.     

Ultrastructure of Excystment of Toxoplasma gondii Oocysts*

SYNOPSIS. The excystation of sporozoites from intact Toxoplasma gondii oocysts or mechanically released sporocysts was studied by light and electron microscopy. Both intact oocysts and free sporocysts excysted in 5% bovine bile in 0.9% NaCl solution after 30–60 min incubation at 37 C. Sporozoites were first activated in either intact sporocysts or oocysts within 2–12 min of incubation in bile. Sporozoites escaped from sporocysts through 4 plate-like sutures in the sporocyst wall, and from the oocyst as the oocyst wall ruptured at one or more points.     

Exogenous Stages of Isospora serini (Aragão) and Isospora canaria sp. n. in the canary (serinus canarius Linnaeus)*

Exogenous stages of Isospora serini (Aragão) and Isospora canaria sp. n. from the canary (Serinus canarius Linnaeus) are described. Oocysts of I. serini are spheroid and average 19.2 × 20.1 μ m, while those of I. canaria are larger, more ellipsoid, and average 21.8 × 24.6 μ m. No oocyst residuum is present and the oocyst walls of both species are colorless, transparent, and single layered. Sporocysts average 9.4 × 15.2 μ m for I. serini and 11.5 × 18.1 μ m for I. canaria. The I. canaria sporocyst has a substiedal body, but none was found in I. serini sporocysts. Both species have a spherical sporocyst residuum; this was obscured in the I. serini sporocyst by scattered granules. Living sporozoites of I. canaria average 3.6 × 16.9 μ m and have 1 to 3 refractile globules; those of I. serini have 2 globules and average 2.8 × 12.6 μ m. A disseminated infection of the mononuclear phagocytes results from administration of I. serini while I. canaria oocysts give rise to a typical coccidian infection restricted to the intestinal epithelium. Asexual stages of I. serini in macrophages are indistinguishable from parasites previously called avian Toxoplasma, Atoxoplasma, and Lankesterella.     

Lecithochirium furcolabiatum (Jones 1933), Dawes 1947: the miracidium and mother sporocyst.

Experimental infections of the marine topshell Gibbula umbilicalis with Lecithochirium furcolabiatum (Digenea: Hemiuroidea) have allowed the development of a model system which will enable further studies of the molluscan host response. The long-lived intertidal prosobranch host is easily maintained in the laboratory, and experimental infection rates of 98% were consistently achieved. The miracidium and mother sporocyst have been studied at both light and ultrastructural levels, providing the first account of the morphology of these stages in Hemiuridae. The ingested egg hatches within the host intestine, treatment with L-cysteine and alkaline pH stimulating miracidial emergence in vitro. The general body surface of the miracidium is devoid of spines or cilia, the latter being restricted to four plates near the anterior extremity. The miracidium swims actively prior to penetration of the gut wall, the sporocyst being released from the miracidial epidermal coat within the haemocoel. Within 5 weeks of infection, the filamentous mother sporocyst contains 1 to 3 oval germ balls, daughter sporocysts being recorded free within the digestive gland haemocoel 7 weeks later. Twenty three weeks after ingestion of eggs, the daughter sporocyst extends into the host gill filaments, containing cystophorous cercariae ready for emergence.     

Coccidian Parasites (Apicomplexa: Eimeriidae) from Insectivores: New Species from Shrew Moles (Talpidae) in the United States1

All of 18 shrew moles, Neurotrichus gibbsii, collected in Oregon and Washington were infected with one or more species of coccidia. Three eimerians and one isosporan were identified and described as new species. Sporulated oocysts of Eimeria heterocapita n. sp. were subspheroid to ellipsoid, 25.5 times 21.4 (23–27 times 18–23) μm. A membranous, cap-like structure was present at one pole of the oocyst, but a micropyle, oocyst residuum, and polar body were absent. Ovoid sporocysts were 13.6 times 10.0 (12–15 times 9–11) μm; a compact sporocyst residuum was present, but Stieda, sub-, and parastieda bodies were absent. This species was found in 2 of 18 (11%) hosts. Sporulated oocysts of Eimeria neurotrichi n. sp. were avoid, 17.6 times 13.6 (16–20 times 11–16) μm; micropyle and oocyst residuum were absent, but a polar body was present. Ovoid sporocysts were 10.7 times 5.5 (9–12 times 5–6) μm; Stieda body and sporocyst residuum were present, but sub- and parastieda bodies were absent. This species was found in 2 of 18 (11%) hosts. Sporulated oocysts of Eimeria parastiedica n. sp. were subspheroid, 27.4 times 25.5 (25–30 times 22–28) μm; micropyle, oocyst residuum, and polar body were absent. Ovoid sporocysts, pointed at both ends, were 18.3 times 10.4 (16–20 times 9–11) μm; Stieda, sub-, and parastieda bodies were present as was a sporocyst residuum. This species was found in 2 of 18 (11%) hosts. Sporulated oocysts of Isospora neurotrichi n. sp. were subspheroid, 13.9 times 12.0 (11–16 times 10–15) μm; micropyle and oocyst residuum were absent, but 1–3 polar bodies were present. Ellipsoid sporocysts were 9.2 times 6.1 (8–11 times 5–8) μm; sub- and parastieda bodies were absent, but a Stieda body and sporocyst residuum were present. This species was found in 17 of 18 (94%) hosts. Ten of 18 (56%) hosts were seen to be naturally infected with only one coccidium.     

Eimeria (Protozoa: Eimeriidae) of the Cottontail Rabbit Sylvilagus audubonii in Northeastern Colorado, with Descriptions of Three New Species

SYNOPSIS. All of 100 cottontail rabbits Sylvilagus audubonii were found to be infected with 1-6 species of Eimeria. Three new species, E. audubonii, E. neoirresidua and E. poudrei are described from this host. Sub-spherical oocysts of E. audubonii average 21.2 by 17.1 μ; polar body, micropyle, oocyst residuum and sporocyst residuum are all absent; ellipsoidal sporocysts average 12.9 by 5.8 μ. Ovoid to ellipsoidal oocysts of E. neoirresidua average 25.7 by 17.9 μ; polar body and oocyst residuum are absent; micropyle and sporocyst residuum are present; ellipsoidal sporocysts average 14.5 by 6.4 μ. Ovoid to ellipsoidal oocysts of E. poudrei average 26.0 by 18.1 μ; polar body is lacking; micropyle, oocyst residuum and sporocyst residuum are present; ellipsoidal sporocysts average 14.4 by 6.4 μ. Three species of Eimeria previously described in the literature, E. maior Honess, 1939, E. media form honessi Carvalho, 1943 and E. environ Honess, 1939 are redescribed. A detailed structural and statistical analysis of each species is presented with at least 200 sporulated oocysts measured in each instance. A host list and a key to the Eimeria of cottontails is given. The use of detailed studies of oocyst size and structure as a tool for specific diagnosis of the Eimeria of cottontails is discussed.     

Parthenogenetic generations of Sanguinicola armata (Trematoda: sanguinicolidae)

Daughter sporocysts of Sanguinicola armata are represented by several generations, changes of which goes synchronously with the changes of year seasons. Young individuals beginning the reproductions form exclusively cercariae. The old sporocysts begin to produce sporocysts only. These young sporocysts do not quite the organism of the old sporocyst. Therefore, series of sporocysts inside other sporocysts are often observed in hystological cross-sections. Germinal masses of daughter sporocysts of S. armata have some specific characters, which are not observed in analogous organs in daughter sporocysts of other trematode species.     

Eimeria cicaki sp. n. and Isospora thavari sp. n. from the House Lizard Gehyra mutilata Boulenger in Malaysia*

SYNOPSIS. Oocysts and endogenous stages of new species of Eimeria and Isospora from the house lizard, Gehyra mutilata, are described. The ellipsoid to subspherical 2-layered oocysts of E. cicaki averaged 24.0 × 21.0 μm. Polar granules are present. Micropyle and oocyst residuum are absent. Ellipsoid sporocysts average 12.2 × 9.0 μm. A sporocyst residuum is present, but the Stieda body is absent. Endogenous stages are in epithelial cells of the small intestine. The subspherical single-layered oocysts of I. thavari average 23.8 × 22.8 μm. The polar granule is present; micropyle and oocyst residuum are absent. Ellipsoid sporocysts average 12.8 × 9.4 μm. Stieda body and sporocyst residuum are present. There are endogenous stages in epithelial cells of the small intestine.     

Eimeria clethrionomyis sp. n., Eimeria gallatii sp. n., Eimeria pileata sp. n. and Eimeria marconii sp. n. from the Red-Backed Vole Clethrionomys gapperi Vigors,from Pennsylvania§

SYNOPSIS Four new eimerian species are described from red-backed voles. Clethrionomys gapperi in Pennsylvania. Sporulated oocysts of Eimeria clethrionomyis sp. n. are ellipsoidal, 18.8 (16.5–21.5) × 14.9 (14.0–16.5) with elongate, ovoid sporocysts, 10.6 (9.5–12.0) × 6.1 (5.5–7.0). The oocyst wall is smooth, with 2 layers, and thins, with terminal cap at one or both ends. Polar granules, dark Stieda body and sporocyst residuum are present. The occyst residuum is absent. Sporulated oocysts of Eimeria gallatii sp. n. are ellipsoidal, 27.7 (21–32) × 19.3 (17–24) with ovoid sporocysts, 13.5 (12–15) × 8.8 (8–10). The oocyst wall is smooth, 2-layered, with a micropyle and thin wall at the end opposite the micropyle. Polar granules. Stieda body and sporocyst residuum are present. The oocyst residuum is atypical, of cobwebby material. Sporulated oocysts of Eimeria pileata sp. n. are subspherical to spherical, 25.2 (20.5–29.5) × 22.5(19.5–25.5) with ellipsoidal sporocysts, 13.4(10.5–15.0) × 8.4 (7.5–9.5). The oocyst wall is rough, pitted, striated, 2-layered, with no micropyle. Polar granules, oocyst and sporocyst residuum. Stieda body and stiedal cap are present. Sporulated oocysts of Eimeria marconii sp. n. are ellipsoidal, 13.0 (10.5–15.0) × 10.6 (9.5–12.0) with elongate, ovoid sporocysts, 7.7 (7.0–8.5) × 4.2 (3.0–4.5). The oocyst wall is smooth, single-layered, with no micropyle. Polar granules, dark Stieda body and sporocyst residuum are present. There is no oocyst residuum.     

Eimeria procyonis sp. n., an Isospora sp., and a Redescription of E. nuttalli Yakimoff and Matikaschwili, 1932 (Protozoa: Eimeriidae) from the American Raccoon (Procyon lotor)

SYNOPSIS. Thirty-two of 48 raccoons examined were infected with a previously undescribed species of Eimeria which is herein named E. procyonis. Of the 32 infected animals, 10 also harbored E. nuttalli and 1 had Isospora sp. oocysts.
The ellipsoid to ovoid oocysts of E. procyonis measured 23.4 × 18.0 (16–29 × 13–24) μm; its sporocysts measured 12.1 × 9.3 (11.5–15 × 7–10) μm, each containing a slightly flattened substiedal body. The sporocyst residuum consisted of numerous scattered granules each ∼1 μm in diameter. The oocyst wall was double-layered. The outer layer appeared rough and pitted, measuring 1.5 μm, except at the micropyle where it was 1 μm thick.
The oocysts of the Isospora sp. measured 16.8 × 13.7 (16–18.5 × 12.5–15.5) μm. The wall consisted of a single layer ∼0.5 μm thick. The sporocysts measured 11.2 × 9.1 (9.5–11.5 × 8–10) μm, and each contained 4 elongate sporozoites. The oocysts of E. nuttalli measured 17.5 × 13.6 (12-21 × 11-15) μm, with a smooth single-layered wall approximately 0.7 μm thick. The sporocysts measured 12.2 × 7.1 (9-13 × 5.5–11) μm. Each sporocyst had a thin, dark, Stieda body and the sporocyst residuum consisted of many fine granules.