艾美耳球虫对高原鼠兔繁殖的影响

寄生物对宿主繁殖的影响取决于宿主对当前繁殖值和剩余繁殖值的权衡。球虫为微型寄生物,而微型寄生物对宿主当前繁殖值的影响较剩余繁殖值要大。因此,本研究检验了寄生在高原鼠兔肠道内的艾美耳球虫可影响其当前繁殖的假设。在繁殖早、中、晚期,野外共观测高原鼠兔170只。结果表明,不同繁殖期感染率有显著差异。在繁殖中期,未感染雌性的妊娠率显著高于感染雌性。且未妊娠雌性较妊娠雌性有更高的感染强度,但在另外两个繁殖期没有发现此效应。在雄性中,任何繁殖期的感染强度和感染率与睾丸和附睾指数均无显著相关性,且感染和未感染球虫雄性睾丸及附睾指数无显著差异。此外,野外观测实验结果表明,感染雌性的胚胎重较未感染雌性显著降低,与野外感染对胚胎重量影响的实验结果相一致。说明艾美耳球虫感染可影响胚胎的发育。上述结果说明,艾美耳球虫对高原鼠兔繁殖的影响随繁殖期而有不同效应,且存在性别间差异,这种效应可能与不同性别间的繁殖对策有关。     

Ultrastructure of in vivo-produced caryocysts containing the coccidian Caryospora bigenetica (Apicomplexa: Eimeriidae)

A cotton rat was inoculated orally with oocysts of Caryospora bigenetica from the feces of a rattlesnake. Sixteen days later the rat was euthanized, and portions of the scrotum, foot pad and muzzle were processed for histological sections and transmission electron microscopy. Sporozoites within caryocysts had typical coccidian features such as an anterior and posterior refractile body, centrally located nucleus, micronemes, rhoptries, a conoid, a micropore near the anterior refractile body, a posterior pore, amylopectin granules, lipid bodies, a Golgi-like body, a mitochondrion and subpellicular microtubules. The infected host cell was spherical and surrounded by a fibrous wall-like covering, 0.35-1.00 microns thick. This outer covering, when viewed in stained histological sections, was periodic acid-Schiff (PAS)-positive.     

Current views on the life cycle of cyst-forming Coccidia (Eucoccidia, Sporozoa, Apicomplexa)]

The life cycles of cyst-forming coccidia of the genera Sarcocystis and Toxoplasma have been first analysed in terms of generally recognized biological phenomena, such as proliferation, differentiation, dedifferentiation, programmed cell death. The differences, existing between the respective obligatory heteroxenous (Sarcocystis) and facultatively heteroxenous (Toxoplasma) life cycles, have been found to involve the obvious differences in the degree of a zoite's differentiation, occurring in the course of asexual development in the intermediate host. In Sarcocystis spp., the degree of differentiation in merozoites, throughout their development, is much higher than in those of T. gondii. This level of merozoite differentiation in Sarcocystis is thought to determine the irreversible, one-directional way of both pre-cystic and cystic development of the pathogen, starting from the sporozoite stage and terminating in gamont formation within the tissue cyst. Unlike, in T. gondii, the level of merozoite differentiation is not so strong as in Sarcocystis spp., which may account for the reversible merozoite development in the former, which is clearly demonstrated by a ready conversion of pre-cystic tachyzoites into cystic bradyzoites, and the other way round. In the course of endogenous development, the pathogens adversely affect their environment (the infected cells and tissues), which, in its turn, may exert its influence on the particular parasites. Thus, both the parasite and the host represent a unique feed-bach regulatory system.     

Structural and functional characterization of cyst cells in Sarcocystis sp. (Sporozoa, Apicomplexa)

By means of light and electron microscopy, the structural pattern of muscle cysts (sarcocysts) was examined for the four species of the genus Sarcocystis: S. muris (from murine skeletal muscles), Sarcocystis sp. and S. fusiformis (from, respectively, heart and skeletal muscles of buffalo), and S. ovifelis (from ovine tong muscles). The orderly fashion of the interior of the cyst is attained by partitition of its space into numerous compartments with the involvement of the intermediate filaments. These, in their turn, are bound to each other by thin filaments to make eventually a common filamentous net. The net limits separate groups of cells referred to as cyst zoites. The common net of filaments and microtubules (when present) may be regarded not only as the organizer of the cyst interior cytoskeleton, but also as the main mechanism of substance transportation in various directions: from the host cell to the sarcocyst, and within or outside the cyst. The role of dedifferentiation, proliferation and differentiation processes is suggested in the establishment of the fixed sequence of events throughout the unidirectional development of cyst cells and their interaction, from precystic meronts to cyst merozoites (gamonts). Special attention is paid to metrocyte morphogenesis and functioning. In the present work, metrocytes subjected to apoptosis were recognized. It is suggested that phenomenon of programmed cell death in metrocytes may be associated with the control of cell number in mature and ageing sarcocysts.     

The structural and functional analysis of the surface apparatus in four species of Sarcocystis (Sporozoa, Apicomplexa)

The comparable ultrastructural analysis of the sarcocyst surface apparatus (SSA) was made for four species of Sarcocystis: Sarcocystis muris, S. fusiformis, S. medusiformis, and Sarcocystis sp. from buffalo heart muscles. In all these species, SSA contains a surface membrane, overmembrane complex with glycocalyx, and submembrane complex made of two glycoprotein SSA primembrane layers. SSA makes numerous primary vesicle-like protrusions and pits in between. Some vesicles containing two layers, PM1 and PM2, are pinching off from the totally formed protrusions. Then these vesicles are directed into infected host cell to participate in its degradation. In the SSA pits neither over-, nor submembrane complex is present, the pits being made of the surface membrane only. It is important that fibrillar structures penetrate through the SSA membrane into pits from the host cell. Besides, SSA forms secondary protrusions with different structures in various species of Sarcocystis. They increase the sarcocyst surface and transport different substances along intermediate filaments from the SSA pits membrane to the sarcocyst body. At the same time, deep invaginations are found in the SSA of old sarcocysts. We thought that these structures increased the sarcocyst surface and thus promote to intensify metabolism. This study-defined presence of membranous vesicles in secondary protrusions. According to their structure and localization, the membranous vesicles may be involved in the building of the sarcocyst surface membrane.     

Formation and diversity of parasitophorous vacuoles in parasitic protozoa. The Coccidia (Sporozoa, Apicomplexa)

Data on parasitophorous vacuole (PV) formation in host cells (HC) harbouring different intracellular protozoan parasites have been reviewed and critically analysed, with special reference to the main representatives of the Coccidia. The vacuole membrane (PVM) is the interface between host and parasite, playing a role in nutrient acquisition by the parasite from the HC. The PV phenomenon is regarded as a generalized HC response to the introduction of alien bodies (microorganisms), which eventually reflects the evolutionary established host-parasite relationships at cellular, subcellular and molecular levels. Special attention has been paid to the existing morpho-functional diversity of the PVs within the same genera and species of parasites, and even at different stages of the parasite life cycle. The PVM is generally considered to derive from the HC plasmalemma, whose biochemical composition undergoes significant changes as the intravacuolar parasite grows. The original HC proteins are selectively excluded from the PVM, while those of the parasite are incorporated. As the result, the changed PVM becomes not fusigenic for HC lysosomes. For Toxoplasma gondii and other cyst-forming coccidia (Isospora, Sarcocystis), a definite correlation has been noticed between the extent of rhoptry and dense granule secrets released by a zoite during HC internalization, on the one hand, and the pattern of the PV that forms, on the other one. In T. gondii, tachyzoites, known to discharge abundant secrets, commonly force the development of PVs limited with a single unit membrane and equipped with a tubulovesicular network in the lumen. Unlike, bradyzoites known to be deficient in secretory materials trigger the formation of PVs with a three-membrane lining composed of the changed invaginated plasmalemma in addition to two membranes of endoplasmic reticulum. The two different types of PV harbour, respectively, exoenteric and enteric stages of T. gondii, the latter being confined to the cat intestine only. Unlike, all endogenous stages of the classic intestinal coccidia (Eimeria spp.) develop within PVs limited with a single membrane, with some invaginations extending into the PV lumen. Unusual PV patterns are characteristic of the extracytoplasmic eimerian coccidia (Cryptosporidium, Epieimeria) and adeleid haemogreagarines (Karyolysus). In cyst-forming coccidia, the PVM is actively involved in tissue cyst wall formation, thus protecting the encysted parasites from recognition by the host immune system. All this strongly suggests that the PV is far from being an indifferent membraneous vesicle containing a parasite, but represents a metabolically active compartment in infected cells. Since all the coccidia are obligate intracellular parasites, the mode of their intimate interaction with the HC, largely accomplished via the PV and its membrane, is vital for their survival as biological species.     

Some Eugregarines (Sporozoa, Apicomplexa) from the Stored Fruit Beetle, Oryzaephilus mercator Fauv. (Coleoptera) from India

Morphology and life history of 3 cephaline gregarines found in the gut of the pest of stored fruit, Oryzaephilus mercator. are described. Of these 3, 2 are new species. The 3 species are (1) Hirmocystis minuta (Ishii, 1914) (LP TL = 1:7 – 1:22, WP/WD = 1:1, 6–1:7); (2) Amsotobus indicus n. sp. (LP/TL = 1:3–1:6, WP/WD = 1:1 – 1:1.3); (3) Leidyana oryzaephili n. sp. (LP/TL = 1:2 – 1:12; WP/WD = 1:1–1:1.6).     

DNA barcoding identifies Eimeria species and contributes to the phylogenetics of coccidian parasites (Eimeriorina, Apicomplexa, Alveolata)

Partial (～ 780 bp) mitochondrial cytochrome c oxidase subunit I (COI) and near complete nuclear 18S rDNA (～ 1,780 bp) sequences were directly compared to assess their relative usefulness as markers for species identification and phylogenetic analysis of coccidian parasites (phylum Apicomplexa). Fifteen new COI partial sequences were obtained using two pairs of new primers from rigorously characterised (sensu Reid and Long, 1979) laboratory strains of seven Eimeria spp. infecting chickens as well as three additional sequences from cloned laboratory strains of Toxoplasma gondii (ME49 and GT1) and Neospora caninum (NC1) that were used as outgroup taxa for phylogenetic analyses. Phylogenetic analyses based on COI sequences yielded robust support for the monophyly of individual Eimeria spp. infecting poultry except for the Eimeria mitis/mivati clade; however, the lack of a phenotypically characterised strain of E. mivati precludes drawing any firm conclusions regarding this observation. Unlike in the 18S rDNA-based phylogenetic reconstructions, Eimerianecatrix and Eimeria tenella formed monophyletic clades based on partial COI sequences. A species delimitation test was performed to determine the probability of making a correct identification of an unknown specimen (sequence) based on either complete 18S rDNA or partial COI sequences; in almost all cases, the partial COI sequences were more reliable as species-specific markers than complete 18S rDNA sequences. These observations demonstrate that partial COI sequences provide more synapomorphic characters at the species level than complete 18S rDNA sequences from the same taxa. We conclude that COI performs well as a marker for the identification of coccidian taxa (Eimeriorina) and will make an excellent DNA 'barcode' target for coccidia. The COI locus, in combination with an 18S rDNA sequence as an 'anchor', has sufficient phylogenetic signal to assist in the resolution of apparent paraphylies within the coccidia and likely more broadly within the Apicomplexa.     

A new species of coccidian (Apicomplexa: Eimeriidae) from the prairie racerunner, Cnemidophorus sexlineatus viridis (Sauria: Teiidae), in Arkansas.

Feces from 26 prairie racerunners, Cnemidophorus sexlineatus viridis Lowe, 1966, from Arkansas, were examined for coccidian parasites. One of these was found to be infected with oocysts of an undescribed eimerian, which is described herein as new. Sporulated oocysts of Eimeria sexlineatus n. sp. were cylindrical, 30.4 x 17.1 (28-32 x 16-19) microns, with a shape index (length/width) of 1.8 (1.6-2.0). A micropyle and oocyst residuum were absent but 1 (to several) polar granule(s) was present. Sporocysts were ellipsoidal, 10.7 x 8.5 (9.6-11.2 x 8.0-8.8) microns, with a shape index of 1.3 (1.2-1.4). A sporocyst residuum was present but Stieda, substieda, and parastieda bodies were absent. Sporozoites were elongate, 13.2 x 2.7 microns (12.0-14.4 x 2.4-3.2) in situ, containing a single, spherical posterior refractile body. Oocysts and endogenous developmental stages were found within the gall bladder epithelium of the infected lizard. This represents the first time a coccidian has been reported from a North American whiptail lizard.     

Isospora mcquistioni and Isospora bioccai (Apicomplexa, Eimeriidae): two new coccidian parasites from Carduelis sinica (Passeriformes, Fringillidae).

The following two species are described from Carduelis sinica (Greenfinch) from Italy. The oocysts of Isospora mcquistioni n. sp. were 26.0 x 22.6 (24.0-28.5 x 20.0-24.2) microns and ovoid with a smooth bilayered wall. Neither micropyle nor oocyst residuum were observed. One polar granule was found. Sporocysts were oval, 18.1 x 11.4 (16.0-19.8 x 11.0-12.0) microns, and with a symmetrical Stieda complex. The residuum was compact and spherical. Isospora bioccai n. sp. oocysts were spherical to subspherical and 24.0 x 23.6 (22.0-26.0 x 21.0-25.8) microns. The oocyst wall was smooth and bilayered. A micropyle and oocyst residuum were absent; 4 to 10 elongate polar granules were present. Sporocysts were 19.5 x 11.6 (18.0-20.0 x 10.0-12.4) microns, ellipsoidal, and with a symmetrical Stieda complex. The sporocyst residuum was diffuse and composed of a few granules.     

Studies of coccidian parasites of fish from the coast of Senegal (West Africa): new species of the genus Goussia (Apicomplexa, Eucoccidiida, Calyptosporidae)

Six new species of coccidian parasites of fish are described from the coast of Senegal: Goussia dakarensis sp.n. from Brachydeuterus auritus, Pomadasys peroteti and Galeoides decadactylus; Goussia decapteri sp.n. from Decapterus rhonchus; Goussia exoceti sp.n. from Hirundichthys affinis; Goussia emissolei sp.n. and Goussia soumbediounensis sp.n. from Leptocharias smithii; and Goussia trachinoti sp.n. from Trachinotus ovatus. Goussia clupearum (Thélohan, 1894) was found in eight new host species and G. senegalensis Faye, 1988 in one new host species.     

Current conception of the Sarcosporidia (Sarcocystis, Eimeriidae, Sporozoa, Apicomplexa): their morphofunctional organization, life cycle and practical importance

Advances in sarcosporidian research within the latest decade are critically reviewed and analysed in respect to some recent cytological findings of the author and her colleagues on the subject. Three rather than two morpho-functional cell types (metrocytes and merozoites) are distinguished within the sarcocyst, the third one being the intermediate cell. Division by endodyogeny in the cyst of Sarcocystis is very likely confined to the latter cell type. The pattern of nuclear chromatin and the constancy in DNA value per nucleus in the cystic merozoites, revealed by flow cytometry, is rather indicative of their incapability of dividing within the cyst, i.e. in the intermediate host. This enabled us to consider these merozoites as homologs of coccidian gamonts. The obvious differences between cysts and cystic stages in Sarcocystis and Toxoplasma may account in part for the rare, if any, reported cases of congenital sarcocystosis in the intermediate host.     

The phylogeny of the Gregarinia (Sporozoa)

The Gregarinia, all parasitic in invertebrates have appeared in marine hosts (Polychaeta). The most primitive are the Archigregarinida (Selenidium) which are intestinal parasites. The Eugreganida (intestinal or coelomic) are found in various marine invertebrates as well as in terrestrial or fresh water arthropods. The Neogregarinida which parasitize only insects have a very reduced number of spores compensated by a schizogony (merogony). The Gregarinia constitute a perfect example of a concomitant evolution between a group of parasites and their hosts.     

Isospora geospizae,a new coccidian parasite (Apicomplexa: Eimeriidae) from the small ground finch (Geospiza fuliginosa) and the medium ground finch (Geospiza fortis) from the Galapagos Islands

Isospora geospizae n. sp. is described from the faeces of the small ground finch Geospiza fuliginosa and the medium ground finch Geospiza fortis from Santa Cruz Island in the Galapagos archipelago. I. geospizae oocysts are spherical to subspherical, with a one-layered, smooth, colourless wall, 15.5×14.5 (13–17×12–17) m with a large polar body and no micropyle or residuum. Sporocysts are ovoid, 10×7.5 (10–12×6–9) m with small Stieda and subtieda bodies and a smoothly contoured, but irregular-shaped residuum containing coarse, non-uniform granules. Sporozoites are sausage-shaped with large refractile bodies at each end and randomly arranged in the sporocysts. Three of 34 (8.8%) G. fuliginosa and one of nine (11.1%) G. fortis were infected with I. geospizae.     

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.     

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.