A Light and Electron Microscopic Study of Trypanosoma fallisi N. Sp. in Toads (Bufo americanus) from Algonquin Park, Ontario

Trypanosoma fallisi n. sp. is described from Bufo americanus in Ontario. The parasite was observed in 65 of 94 toads examined. The trypanosomes were pleomorphic with respect to the age of infections, being longer and broader in early infections (during spring and summer) and shorter and more slender during late summer and autumn. They ranged in size from 38–76 μm in body length and 3–8 μm in width, with a free flagellum 6–30 μm long. Epizootiological and experimental evidence suggests that this trypanosome is transmitted to the toads by the leech, Batracobdella picta. Trypanosoma fallisi is morphologically similar to T. bufophlebotomi described in Bufo boreas from California, but geographic isolation, host and vector differences as well as slight morphological differences indicate that speciation has occurred. Similar trypanosomes from Bufo americanus (which were identified as T. bufophlebotomi) in Michigan, are probably T. fallisi. This species shares many ultrastructual features with trypanosomes of other lower vertebrates and also of mammals.     

Development of Babesiosoma stableri (Dactylosomatidae; Adeleida; Apicomplexa) in its Leech Vector (Batracobdella picta) and the Relationship of the Dactylosomatids to the Piroplasms of Higher Vertebrates

The sporogonic and merogonic development of Babesiosoma stableri Schmittner & McGhee, 1961 within its definitive host and vector, a leech Batracobdella picta (Verrill, 1872), was studied by light and electron microscopy. Gamonts released from frog erythrocytes in the blood meal of the leech associated in syzygy and fused; the gamonts were isogamous and only 1 microgamete was formed. The ultrastructural appearance of the resulting zygote was similar to that of the gamonts, but it was larger. The zygote had an apical complex (including a polar ring, conoid and 2 pre-conoidal rings and micronemes, but no recognizable rhoptries), triple-membraned pellicle, about 40 subpellicular microtubules and prominent stores of amylopectin. Zygotes penetrated the cells of the intestine and underwent sporogony directly within the cytosplasm of the ieech epithelial cell without the formation of a parasitophorous vacuole. Eight sporozoites budded simultaneously around the periphery of an irregularly shaped oocyst. No oocyst wall was formed. Each sporozoite had a complete apical complex (including rhoptries), abundant amylopectin inclusions and a triple-membraned pellicle with about 32 subpellicular microtubules. The sporozoites initiated merogonic replication primarily within the salivary cells of the leech although other tissues, such as muscle, were infected. Each meront produced 4 merozoites by simultaneous budding, forming a cruciform meront typical of the intraerythrocytic development of this parasite. The meront was located directly within the cytoplasm of the host cell. Merozoites, with abundant amylopectin, had a complete apical complex and triple-membraned pellicle with about 40 subpellicular microtubules. The merozoites either initiated a further cycle of replication, or they moved into the ductules of the leech salivary cells which extend to the tip of the proboscis. Observations on gametogenesis. syngamy and sporogony of B. stableri in its leech host indicate that the family Dactylosomatidae should be placed in the suborder Adeleina (Eucoccidiida: Apicomplexa). Babesiosoma stableri was transmitted to uninfected frogs (Rana spp.) by the bite of infected leeches. Prepatent periods ranged from 26 to 38 days at 25° C. Despite a directed search in laboratory reared tadpoles which had each been injected intraperitoneally with 150,000 merozoites, no pre-erythrocytic developmental stages were observed. Similarities in their biology suggest close phylogenetic affinities of the dactylosomatids, and other adeleid blood parasites, with the piroplasms of higher vertebrates.     

Light and Electron Microscopic Observations on Encystment of Acanthamoeba palestinensis, Reich

SYNOPSIS. The ultrastructural changes occurring during encystment of Acanthamoeba palestinensis have been investigated. The cyst wall consists of endocyst and exocyst, both having the same fine structure. At irregular intervals in the cyst wall ostioles occupied by opercula are present. The nuclear membrane forms bulb-shaped projections and releases vesicles bounded by double membranes into the cytoplasm. Dense nucleolus-like bodies of different sizes and variable numbers are found in the nucleus of every cyst. The importance of the cyst structure as a taxonomic criterion is discussed.     

Descriptions and phylogenetic systematics of Myxobolus spp. from cyprinids in Algonquin Park, Ontario

Eight species of Myxobolus were collected from four species of cyprinids in Algonquin Park, Ontario. On the basis of spore morphology, five of these species are described as new and two are redescribed. The evolutionary relationships among these eight species were studied using partial small subunit ribosomal DNA (ssu-rDNA) sequence data. The resulting cladograms, which were highly resolved and with strongly supported relationships, allowed for the evaluation of spore morphology, host specificity, and tissue tropism, criteria traditionally used in species identification. These criteria, recently criticized for creating artificial rather than natural taxonomic groupings, were evaluated for their reliability in the systematics of the species examined. The data showed that distantly related species often infect the same host and tissue, and that closely related species often occur in different hosts. Morphologically similar species are more closely related to each other and the taxonomy based on spore morphology is consistent with the relationships depicted in the phylogenies. These results suggest that spore morphology is better than host specificity and tissue tropism as a species character, as well as for determining evolutionary relationships among the species of Myxobolus examined.     

Light and Electron Microscopic Observations on the Heterotrich Ciliate Climacostomum virens

SYNOPSIS. Alveolar membranes and an epiplasm exist under the cell membrane of the noncontractile heterotrich ciliate Climacostomum virens. Postciliary microtubular ribbons join at the right of each somatic kinety to form a Km fiber. Two transverse microtubular fibers occur per kinetosomal pair. A myonemal network interconnects the kinetosomal bases intrakinetally and interkinetally. Ultrastructural comparisons are made between the contractile and noncontractile heterotrichs.
The buccal cortex consists of an adoral zone of membranelles, a peristomal field, a buccal tube, the apical membranelles, and a haplokinety. The kineties of the peristomal field and buccal tube are rows of paired kinetosomes, with a postciliary ribbon of microtubules arising from the posterior kinetosome of each pair, and a transverse ribbon and an oblique ribbon from the anterior kinetosome. No Km fibers exist in this region. The haplokinety is a collar of paired kinetosomes surrounding the cytostome; a postciliary microtubular ribbon descends from each kinetosomal pair into the cytostomal region. Ultrastructural details of the buccal cortex of C. virens and other heterotrichs are compared. The nemadesmata which lie under the membranelles are implicated in the body bending of C. virens.
Algae endosymbiotic in the cytoplasm of C. virens are described.     

Light and Electron Microscopic Observations of Gametogenesis in Hastigerina pelagica (Foraminifera)*

SYNOPSIS. During gametogenesis mother individuals of Hastigerina pelagica (d'Orbigny) undergo significant morphological changes. Thirty h before gamete release, the cytoplasm changes from pale orange to bright red, possibly due to transport of stored lipids from the inner region to more peripheral parts of the cytoplasm. During the next 10 to 15 h the bubble capsule which surounds the calcareous shell is discarded. After all bubbles have disappeared, the individual sheds its spines by resorbing the spine bases close to the shell surface. A single mother nucleus divides into some hundreds of thousands of gamete nuclei within a span of ～ 20 h. A bulge of cytoplasm is extruded from the aperture and increases in size during the next 5 to 10 h. This bulge consists of cytoplasmic strands in which gametes and spherical bodies are embedded. The gametes and spherical bodies mature and are released during the afternoon and early evening. The gametes have 2 unequal acronematic flagella. A previously undescribed structure in foraminiferal reproduction is the spherical body which consists of a large vacuole surrounded by a thin cytoplasmic layer in which several nuclei, various typical cell organelles and multiple flagella are present. The spherical bodies are believed to play a role as receptacles of waste material, possibly including residual digestive enzymes, thereby protecting the gametes from lysis during the reproductive process. Fusion of gametes and further development into the next generation have not been observed.     

Molecular characterization of myxozoan parasites from Lake Sasajewun, Algonquin Park, Ontario, by riboprinting

The small subunit-rRNA genes of 18 myxozoans from Lake Sasajewun, Algonquin Park were amplified and digested with restriction endonucleases for riboprinting analysis. Identical riboprints were not found between the myxosporeans and the actinosporeans. The distinct riboprinting patterns observed among these myxozoans indicate considerable genetic diversity within this group. Identical riboprints were found between Myxobolus pendula and Myxobolus pellicides, and between triactinomyxon 'C' and Triactinomyxon ignotum. Parsimony analysis of the riboprints demonstrated that neither the myxosporeans nor the actinosporeans formed a monophyletic group. Some species of Myxobolus are more closely related to forms of triactinomyxon, echinactinomyxon or raabeia than to other Myxobolus species. These results are consistent with the two-host life cycle hypothesis of myxozoans that myxosporeans and actinosporeans are alternating stages of the same organisms.     

Trichonosema algonquinensis n. sp. (Phylum microsporidia) in Pectinatella magnifica (Bryozoa: phylactolaemata) from Algonquin Park, Ontario, Canada

A new species of microsporidian, Trichonosema algonquinensis, is described from a freshwater bryozoan, Pectinatella magnifica from Ontario, Canada. The parasite develops in epithelial cells and appears as white, spherical masses throughout the tissues. Trichonosema algonquinensis is diplokaryotic, diploblastic and undergoes development in direct contact with the cytoplasm of the host cell. Mature spores are ovoid, tapered at one end, and measure 8.5 +/- 0.3 x 4.4 +/- 0.1 microm. The polar filament is wound in 20 to 23 helical coils. Although the parasite resembles T. pectinatellae described from the same host in Michigan and Ohio, it differs in the length of the spore and number of coils of the polar filament. Analysis of 16S rDNA by maximum likelihood, parsimony and Baysian inference, complements the morphological data in supporting the placement of T. algonquinensis as a sister species of T. pectinatellae.     

A light and electron microscopic study of Trypanosoma fallisi N. Sp. in toads (Bufo americanus) from Algonquin Park, Ontario

Trypanosoma fallisi n. sp. is described from Bufo americanus in Ontario. The parasite was observed in 65 of 94 toads examined. The trypanosomes were pleomorphic with respect to the age of infections, being longer and broader in early infections (during spring and summer) and shorter and more slender during late summer and autumn. They ranged in size from 38-76 microns in body length and 3-8 microns in width, with a free flagellum 6-30 microns long. Epizootiological and experimental evidence suggests that this trypanosome is transmitted to the toads by the leech, Batracobdella picta. Trypanosoma fallisi is morphologically similar to T. bufophlebotomi described in Bufo boreas from California, but geographic isolation, host and vector differences as well as slight morphological differences indicate that speciation has occurred. Similar trypanosomes from Bufo americanus (which were identified as T. bufophlebotomi) in Michigan, are probably T. fallisi. This species shares many ultrastructural features with trypanosomes of other lower vertebrates and also of mammals.     

Distribution of Phoxinus eos,Phoxinus neogaeus,and Their Asexually-Reproducing Hybrids (Pisces: Cyprinidae) in Algonquin Provincial Park,Ontario

Hybrid Phoxinus are one of the few asexually reproducing vertebrates species. The distribution of hybrid Phoxinus among lakes in Algonquin Park, Ontario, was evaluated relative to the distribution of parental species and relative to physiochemical lake characteristics. No association between the distribution of hybrids and the distribution of parental species was found, suggesting that the hybrids can successfully coexist with either parental species. In addition, we found no association between hybrid distribution and the physiochemical characteristics of lakes, suggesting that the hybrids are generalists with respect to the ecological niches available in Algonquin Park. There was a difference between the physiochemical characteristics of lakes with and without the parental species P. neogaeus. The lakes containing P. neogaeus were lower elevation than the lakes containing the other parental species, P. eos. The difference in distribution between the parental species may therefore be due to different dispersal abilities, to later arrival following post-glacial dispersal, or to differences in ecology. These results suggest that asexual reproduction is a successful strategy for hybrid Phoxinus in Algonquin Park because these sperm-dependent asexual hybrids are able to survive and persist regardless of which parental species is present, and regardless of the physiochemical characteristics of their habitat.     

A phylogeographic survey of brook charr (Salvelinus fontinalis) in Algonquin Park, Ontario based upon mitochondrial DNA variation

Forty-nine populations of brook charr (Salvelinus fontinalis) from Algonquin Park lakes and rivers were analysed for mitochondrial DNA variation. Haplotypic distributions of wild fish in the Algonquin Park region of Ontario, Canada, predominantly reflect postglacial dispersal patterns into the region in spite of substantial hatchery plantings. Two major refugial groupings colonized this region. Northern and eastern watersheds (Amable du Fond, Bonnechere, and northern Petawawa), were colonized primarily by haplotype 1 fish (B1 phylogenetic assemblage), while Oxtongue River, southern Petawawa, and York River populations were colonized predominately by fish from the B2 and A mtDNA phylogenetic assemblages. Fish with haplotypes in the A and B2 phylogenetic assemblages are common in the Lake Huron drainage. All watersheds in the Park drain into the Ottawa River, except the Oxtongue drainage (part of the Lake Huron watershed). This suggests that early glacial outflows south of the Algonquin Park region (Kirkfield-Trent) may have been colonized by fish that initially invaded ‘Ontario island’ (south-western Ontario), while fish which invaded northern Algonquin Park were derived from a different refugial grouping(s) which may have involved colonization both up the Ottawa River drainage, and/or from a more westerly (Mississippian) refugial grouping. A majority of the populations in Algonquin Park have been planted with hatchery reared brook charr since the 1940s. The Hills Lake or ‘Domestic’ strain was used almost exclusively for these plantings. Comparisons of mtDNA haplotypic distributions in hatchery and wild fish suggests that hatchery females had minimal spawning success and/or their progeny had poor survivorship in the wild.     

Taxonomy and molecular phylogeny of Myxobolus bilobus n. sp. (Myxozoa) parasitizing Notemigonus crysoleucas (Cyprinidae) in Algonquin Park, Ontario, Canada

Myxobolus bilobus n. sp. (Myxozoa) is described from golden shiner Notemigonus crysoleucas in Algonquin Park, Ontario, Canada. Plasmodia develop at the distal end of gill filaments, where they form a novel-shaped plasmodium made up of 2 side-by-side hemispheres joined at a central pore. Surrounding gill tissue is vacuolated, necrotic, and hemorrhagic. Spores of M. bilobus n. sp. resemble those of M. aureatus Ward, 1919, M. angustus Kudo, 1934, M. spalli Landsberg & Lom, 1991, and M. pseudokoi Li & Desser, 1985 in parasitizing cyprinids and in their pyriform shape. Spores of M. bilobus n. sp., however, are much larger (20 to 22.1 microm long, 7.5 to 9.3 microm wide, and 6 microm thick) than those of these other species, and the plasmodium is bi-lobed rather than the typical hollow sphere. A phylogenetic analysis of the 18S rDNA (2014 bp) reveals that M. bilobus n. sp. is a member of a clade that includes 11 species of Myxobolus, all of which are parasites of cyprinid fishes in North America or Eurasia. The study concludes that M. bilobus n. sp. is a member of a clade that has undergone radiation within cyprinid fishes of the 2 continents and that this evolution has involved both host and site switching.     

Electron Microscopic Observations of the Bloodstream Form of Cryptobia salmositica Katz, 1951 (Kinetoplastida: Bodonina)1

The ultrastructure of the bloodstream form of Cryptobia salmositica in rainbow trout was examined during the acute phase of experimental infection. The arrangement of the major groupings of cytoplasmic microtubules originating near the basal bodies is similar to that in other bodonids. The cytostome is reinforced both by pellicular microtubules and an electron-dense plaque. Certain microtubules associated with the flagellar pocket serve as nucleating sites for pellicular microtubules. A flagellar rootlet, consisting of two parallel fibers which are bound together intermittently by electron-dense plaques, curves posteriorly from the basal body of the recurrent flagellum towards the kinetoplast. The basal body associated plaque on the kinetoplast membranes is duplicated at the same time as the basal bodies. Cytoplasmic microtubules are found in association with the plaque and the outer kinetoplastic membrane. A pulsatile vacuole, described for the first time in a hemoparasitic cryptobiid, lies adjacent to the post-flagellar pit. Smaller, interconnected vesicles of the spongiome are continuous with the pulsatile vacuole. Since a pulsatile vacuole occurs not only in free-living and ectoparasitic cryptobiids but in the hemoparasitic (=trypanoplasm) forms as well, this is no longer a character by which the genus Trypanoplasma may be separated from the genus Cryptobia. Possession of this osmoregulatory complex may allow the organism to survive outside of a host and fulfill a monoxenous life cycle, in addition to the usual heteroxenous cycle involving a leech as vector.     

Ultrastructural Features of Cystic and Merogonic Stages of Hepatozoon sipedon (Apicomplexa: Adeleorina) in Northern Leopard Frogs (Rana pipiens) and Northern Water Snakes (Nerodia sipedon) from Ontario, Canada

The cystic and merogonic stages of the haemogregarine Hepatozoon sipedon , infecting Northern water snakes ( Nerodia sipedon sipedon ) and Northern leopard frogs ( Rana pipiens ), respectively, in Ontario, Canada, were investigated by transmission electron microscopy. Cysts, which were observed in the liver of Northern leopard frogs ( Rana pipiens ) after these anurans ingested mosquitoes ( Culex pipiens ) containing oocysts of the parasite, harboured two cystozoites, each of which contained a large crystalloid inclusion anterior to the nucleus. Two types of meronts were observed in snakes that were fed the liver of infected frogs. Macromeronts, which matured in endothelial cells of the liver approximately 16 d after snakes ingested infected frogs, contained about 50 large macromerozoites. Macromerozoites emerged from macromeronts, entered the bloodstream of the snake, and reinfected endothelial cells. Micromeronts, which matured about 34 d post-inoculation, contained about 150 micromerozoites that infected erythrocytes and transformed into gamonts. The ultrastructural features of micromeronts and macromeronts differed only slightly: immature macromeronts and macromerozoites contained numerous amylopectin and lipid inclusions, whereas immature micromeronts and micromerozoites did not contain amylopectin inclusions and featured fewer, smaller lipid inclusions. A comparison of cystic stages among Hepatozoon species in different groups of vertebrates is presented with respect to their structure and evolutionary significance.     

The longevity of actinosporean spores from oligochaetes of Lake Sasajewun, Algonquin Park, Ontario, and their reaction to fish mucus

The longevity of 7 forms of actinosporean spores and the reaction of 6 forms of actinosporeans to fish mucus were investigated. The maximum longevity of actinosporean spores kept at ambient laboratory temperatures was 14 days. Spore longevity ranged from 11 to 14 days among actinosporeans. The reaction of spores to fish mucus varied among the actinosporeans. Triactinomyxon F of Xiao and Desser, 1998 reacted only to the mucus of the common shiner Luxilus cornutus, and golden shiner Notemigonus crysoleucas, whereas the aurantiactinomyxon form of Xiao and Desser, 1998, and raabeia B of Xiao and Desser, 1998 reacted readily to mucus of all fish species tested. The differences in reaction to fish mucus among actinosporeans may indicate their different host range. These results indicate that actinosporean spores are short-lived and that actinosporeans respond to their hosts by chemodetection.     

斑马鱼鳃的光镜和透射电镜观察

应用光学显微镜和透射电镜对斑马鱼(Danio rerio)鳃的组织结构及鳃丝、鳃小片超微结构进行了观察。结果表明,斑马鱼有4对全鳃,鳃耙呈长锥状,鳃丝呈梳状排列在鳃弓上,鳃小片均匀排列在鳃丝两侧。鳃小片由上皮细胞、柱细胞、内皮细胞和毛细血管网组成,鳃小片基部和血管周围分布有泌氯细胞,胞内有丰富的线粒体和排泄小泡,根据线粒体形态特征和细胞质电子密度可将其分为两个亚型。黏液细胞通常与泌氯细胞对生存在,并且有通外的开口。斑马鱼鳃组织结构与其他硬骨鱼鳃结构相似,其结构和功能有密切的关系。