黄牛、水牛体内人肉孢子虫18S rRNA基因研究及虫种鉴定

本文对自然感染的水牛源的人肉孢子虫以及黄牛源人孢子虫ＤＮＡ的１８Ｓ ｒＲＮＡ基因的ＰＣＲ扩增产物进行了测序。对年获的８６３ｂｐ的１８Ｓ ｒＲＮＡ基因分析比较表明,二者有较高的同源性,因此认为二者可能同是一种肉孢子虫－－人肉孢子虫（Ｓａｒｃｏｃｙｓｔｉｓ ｈｏｍｉｎｉｓ Ｒａｉｌｌｉｅｔ ａｎｄ Ｌｕｃｅｔ,１８９１）。由此推断,不仅黄牛可作为人肉孢子虫的中间宿主,水牛也可作为人肉孢子虫的中间宿主。     

Fine Structure of Sarcocystis cruzi Schizonts

SYNOPSIS Schizogony of Sarcocystis cruzi Hasselmann (syn. S. fusiformis Railliet) takes place in vascular endothelial cells 26 to 33 days after cattle ingest sporocysts from dogs. Kidney cortex from a heavily infected, dexamethasone-treated bovine was fixed for electron microscopy to determine the method of schizogonie development. Schizogony takes place by endopolygeny characterized by marked enlargement of the parasite nucleus, formation of nuclear lobes, presence of numerous spindles with adjacent pairs of centrioles along the nucleus, and simultaneous formation of daughter merozoites in the cytoplasm adjacent to the spindle poles. Endopolygeny in S. cruzi differs from that in other Sporozoa in that merozoite anlagen are seen in the cytoplasm before any nuclei divide. The resultant merozoites continue development and, when mature, resemble other sporozoan zoites. Upon release from the host cell into capillaries, they travel to muscle tissue to continue the life cycle by forming sarcocysts.     

Fine structure of the mouthparts of Diostrombus politus and Proutista moesta (Hemiptera: Derbidae)

Insect mouthparts are important sensory and feeding structures, morphological studies of this organ can provide additional data for phylogenetic studies. The planthopper superfamily Fulgoroidea is among the dominant groups of phytophagous insects, however, the mouthparts ultrastructure of this superfamily remains unsatisfactorily studied. This study investigates the ultrastructure of the mouthparts of two species in the family Derbidae (Hemiptera: Fulgoroidea), Diostrombus politus Uhler and Proutista moesta (Westwood), using a scanning electron microscope. The results show that these two derbids are of a typical piercing-sucking type found in Hemiptera. They consist of a cone-shaped labrum, a three-segmented labium and a stylet fascicle with two interlocked maxillary stylets incompletely wrapped by two mandibular stylets. The arrangement of the sensilla on the labial tip differ slightly between the two derbid species, and the subapical labial sensilla are likely different among genera in the family Derbidae.     

Morphology and ultrastructure of definitive males of Arcticotantulus pertzovi and Microdajus tchesunovi (Crustacea: Tantulocarida)

Free-swimming definitive males of two species of Tantulocarida (Crustacea), Arcticotantulus pertzovi (Basipodellidae) and Microdajus tchesunovi (Microdajidae), were reared for the first time. Their morphology and ultrastructure were studied using scanning electron and light microscopy. A detailed analysis of the morphological characters of all currently described species revealed several features typical for most known male tantulocaridans, such as the presence of eight aesthetascs, seven pairs of multifid sensilla on the carapace, paired brush setae on the protopods of the thoracopods, and three furcal setae. The monophyly of the families Microdajidae and Doryphallophoridae is corroborated, while the families Deoterthridae and Basipodellidae are more likely paraphyletic.     

Phylogenetic utility and comparative morphology of the composite scale brushes in male phycitine moths (Lepidoptera, Pyralidae)

Male pyralid moths in the subfamily of Phycitinae are known to possess composite scale brush structures associated with the 8th abdominal sternite, but the histology and the structural morphology of these organs have not been adequately explored. As such, the phylogenetic utility of these structures is unknown. We examine the pre-genitalic abdominal histology of male Dioryctria reniculelloides (Pyralidae: Phycitinae) associated with the composite scale brushes, as well as structural morphology within the genus Dioryctria and two closely related genera. The composite scale brushes are composed of fused scales. The musculature associated with the base of sternum 8 shows considerable modification compared to previously described Lepidoptera. Complex glandular tissue was also found associated with the scale brush structures, suggesting secretory function. Phylogenetic utility of ultrastructure and gross morphology was examined for major Dioryctria species groups. Many characters were homoplasious, but several supported the monophyly of the genus, as well as some internal relationships. In conclusion, the combination of ultrastructural, gross morphological and histological characters can be a rich source of information for elucidating a range of evolutionary relationships within the subfamily.     

Divergence between the Anoas of Sulawesi and the Asiatic Water buffaloes, inferred from their complete amino acid sequences of hemoglobin ß chains

Four complete amino acid sequences of hemoglobin β chains were determined for the swamp and the river types of the Asiatic water buffalo (Bubalus bubalis) and two species of the subgenus Anoa in Bubalus; B. (A.) depressicornis (H. Smith, 1827), the lowland anoa, and B. (A.) quarlesi (Ouwens, 1910), the mountain anoa. The two types of the bubalis were identical in the 145 amino acid residues of the β chains and, compared to this sequence, the two residues were substituted in the depressicornis (β49Thr → Ser and 134Ala → Thr) and the five were in the quarlesi (β53Val → Ile, 74Met → Ile, 111Val → Ile, 115Arg → His and 134Ala → Thr). While both Anoa species diverged from the bubalis by the β134Ala → Thr, they differed from each other by the five substitutions. The Anoa species are endemic to Sulawesi of Indonesia. Their speciation and the present coexistence were discussed with reference to probable immigrations of two ancestral Anoa species to Sulawesi at so long interval that had caused a reproductive isolation between the two wild animals. The earlier immigrants were postulated to be ancestral to the quarlesi and the later ones to the depressicornis.     

Two new species of the genus Stenamoeba (Discosea,Longamoebia): Cytoplasmic MTOC is present in one more amoebae lineage

Two new species of the recently described genus Stenamoeba, named S. berchidia and S. sardiniensis were isolated from a single soil sample on Sardinia, Italy. Both share morphological features characteristic to Stenamoeba and form in phylogenetic analyses together with other Stenamoeba spp. a highly supported clade within the family Thecamoebidae. The ultrastructural investigation of Stenamoeba sardiniensis revealed the presence of cytoplasmic microtubule-organizing centers (MTOCs), located close to one of several dictyosomes found inside the cell. This is the first report of cytoplasmic MTOCs among Thecamoebidae. The presence of MTOCs is now shown in five of nine orders comprising the class Discosea and potentially could be a phylogenetic marker in this group. We re-isolated Stenamoeba limacina from German soils. This strain shows a similar morphology and an almost complete SSU rDNA sequence identity with the type strain of S. limacina originating from gills of fishes, collected in Czech Republic.     

Ultrastructural studies of Sarcocystis cruzi (Hasselmann, 1926) Wenyon, 1926 infection in cattle (Bos taurus): Philippine cases

This paper documents the first report of Sarcocysti s cruzi infection in domesticated cattle (Bos taurus) in the Philippines. Fusiform-shaped microscopic sarcocysts (183-578 microns long and 20-98 microns wide) with distinct septae were found in the skeletal, striated and heart muscle. The sarcocyst wall or parasitophorous vacuolar membrane, 1.37-2.75 microns thick consisted of closely-packed villar protrusions 80-400 nm in dm. Middle and distal segments of VP were bent approximately 90 degrees parallel to the cyst wall surface. The villar core lacked microtubules, and at some points, the distal ends of the VP collectively formed conical tufts. Primary cyst wall had numerous 70-100 nm bubble-like undulations, and the ground substance was 0.25-0.5 micron in thickness. The ultrastructure of S. cruzi cyst wall typifies the Type 7 sarcocyst wall, and bears close similarities with the Philippine and the Vietnam strain of bubaline Sarcocystis levinei.     

Sarcocystis neurona infections in sea otter (Enhydra lutris): evidence for natural infections with sarcocysts and transmission of infection to opossums (Didelphis virginiana).

Although Sarcocystis neurona has been identified in an array of terrestrial vertebrates, recent recognition of its capacity to infect marine mammals was unexpected. Here, sarcocysts from 2 naturally infected sea otters (Enhydra lutris) were characterized biologically, ultrastructurally, and genetically. DNA was extracted from frozen muscle of the first of these sea otters and was characterized as S. neurona by polymerase chain reation (PCR) amplification followed by restriction fragment length polymorphism analysis and sequencing. Sarcocysts from sea otter no. 1 were up to 350 microm long, and the villar protrusions on the sarcocyst wall were up to 1.3 microm long and up to 0.25 microm wide. The villar protrusions were tapered towards the villar tip. Ultrastructurally, sarcocysts were similar to S. neurona sarcocysts from the muscles of cats experimentally infected with S. neurona sporocysts. Skeletal muscles from a second sea otter failed to support PCR amplification of markers considered diagnostic for S. neurona but did induce the shedding of sporocysts when fed to a laboratory-raised opossum (Didelphis virginiana). Such sporocysts were subsequently fed to knockout mice for the interferon-gamma gene, resulting in infections with an agent identified as S. neurona on the basis of immunohistochemistry, serum antibodies, and diagnostic sequence detection. Thus, sea otters exposed to S. neurona may support the development of mature sarcocysts that are infectious to competent definitive hosts.     

Experimental infections with African trypanosomes: IV. Immunization of cattle with gamma-irradiated Trypanosoma rhodesiense

Cattle immunized with gamma-irradiated Trypanosoma rhodesiense were protected against a challenge inoculation of unirradiated trypanosomes given 1 wk later. These animals were still resistant when rechallenged after 8 mo and some residual immunity persisted for 14 mo after immunization. Animals which had undergone self-cure after infection were resistant when challenged 14 mo later. No resistance was observed in animals challenged with a heterologous strain of T. rhodesiense. The infections observed in unprotected bovines were of a mild nature. Leukopenia and fever were common early in the course of infection, but no anemia was evident at any time.     

Comparative ultrastructural study of the cuticle and spermatozoa in Propappus volki Michaelsen, 1916 (Annelida: Clitellata)

The ultrastructure of the cuticle and mature spermatozoa of the oligochaete Propappus volki Michaelsen, 1916 is described with the aim of providing additional data for clarifying the systematic position of the taxon. P. volki is a fresh-water species living in streams, and is easily recognized by its proboscis on the pre-segmental prostomium and, in mature specimens, by a clitellum covering the segments XII–XIV. The cuticle is composed of a proximal fibre zone and a distal layered epicuticle covered with membrane-bound epicuticular projections. The fibre zone consists of collagenous fibres in a matrix, arranged in either densely packed parallel layers with the fibres oriented in the same direction, or with more loosely distributed fibres, although with the same main orientation. The epicuticular projections are pyramidal with the base leaning on the outer surface of the epicuticle. The cuticle covering the proboscis differs in morphology from that of the rest of the worm; the fibre zone is composed of thin and short fibrils running in all directions, and the epicuticular projections are longer and more narrow than the projections in other regions of the worm.

The spermatozoa are filiform cells formed, in sequence, by an acrosome, an elongated nucleus, a long midpiece, and a flagellum. The acrosomal tube is short and straight with a completely external acrosomal vesicle. Following the acrosome is a apically corkscrew-shaped and basally straight nucleus. The midpiece is twisted and formed by five mitochondria. The flagellum shows a prominent central sheath arrangement.

A comparison with ultrastructurally described cuticles and spermatozoa from other clitellate species reveals most similarities with enchytraeids.     

Ultrastructural cytochemistry of the sex pheromone glands of Lutzomyia cruzi male sand flies (Diptera: Psychodidae: Phlebotominae)

The sex pheromone glands of Lutzomyia cruzi male sand flies (Diptera: Psychodidae) were analyzed by cytochemical techniques. In adult males, the epithelium at the fourth abdominal tergite is modified into a glandular epithelium, with large columnar gland cells located side by side. The gland cell cytoplasm contains a large number of mitochondria and peroxisomes, the latter with positive (electron-dense) reaction for catalase, a typical peroxisomal enzyme marker. The gland cell cytoplasm also contains a central vacuolated area, with a large number of electron-lucent vacuoles, not limited by a unit membrane. In well-preserved preparations such vacuoles present a homogenous and slightly electron-dense content, typical of lipid droplets. Indeed, incubation of the tergites with imidazole-buffered osmium tetroxide (to detect lipids) resulted in positive reaction in these vacuoles, as well as in between the microvilli of the gland cells. Use of the osmium–potassium iodide (Os–KI) technique allowed to demonstrate the presence of several endoplasmic reticulum (ER) profiles, as expected in secretory cells. Our data suggest that ER, lipid droplets and peroxisomes are involved in the sand fly pheromone biosynthesis.     

The sperm ultrastructure of Caurinus dectes Russell (Mecoptera: Boreidae) and its phylogenetic implications

The sperm structure of the enigmatic mecopteran species Caurinus dectes (Boreidae) is described for the first time. Diagnostic features are the bi-layered acrosome, a cylindric nucleus provided with two longitudinal opposite grooves, and a simple 9 + 2 axoneme which degenerates in the posterior tail end. The results are conform with the monophyly of Mecoptera including Boreidae. A possible autapomorphy of the order is the presence of the two longitudinal opposite grooves along the nucleus, and the presence of two electron-dense fibres beneath the axoneme. Some apparently plesiomorphic features are preserved in the sperm of Caurinus. Features characterizing the distal part of the flagellum, including the presence of an axial cylindrical structure and the distinctive type of axoneme degeneration, are potential synapomorphies of Caurinus and Boreus, i.e. autapomorphic traits of Boreidae.     

The natural heterogeneity of trypanosoma cruzi: Biological and medical implications

Trypanosoma cruzi is a heterogeneous group of parasites. The imposition of natural or artificial pressures can result in the selection of subsets of the population with concomitant changes in characteristics used to evaluate the group. In order to ascertain the extent of heterogeneity, stocks of single-cell clones were prepared from various sources. Selected cell biological, biochemical, immunochemical, parasitological, and histopathological parameters of these clones have been studied. A ten-fold difference in the rate of growth of the epimastigote stage of T cruzi clones has been observed. The extracellular growth rates of the clones correlate with the rate of growth of the obligate intracellular amastigote stage and consequently, the length of intracellular cycle of the parasite. A 40% difference in the amount of total DNA/parasite has been found between clones. Although the amount of DNA/kinetoplast and nucleus varies between clones, the major contribution to the differences in total DNA/parasite appears to be the nucleus. From 16 to 35 antigens have been demonstrated in the T cruzi clones assayed to date. Five to seven of these antigens are common to all of the stocks assayed. However, both isolate- and clone-specific antigens have also been demonstrated. The susceptibility of inbred strains of mice to T cruzi clones varies with the clone of the parasite. These data imply that the genetics of the parasite as well as the host modulate both the course and outcome of a T cruzi infection. The influence of monosaccharides on the receptor-mediated infection of vertebrate cells by trypomastigotes of T cruzi also varies between clones. The implications of these findings upon our concept and understanding of present and future problems in Chagas disease are discussed.     

Giant females and dwarf males: a comparative study of nuptial organs in female Chondracanthidae (Crustacea: Copepoda)

Male chondracanthids attach to specific structures located near the genital apertures of the female and the term nuptial organ is proposed for these structures. The morphology of the nuptial organ in Acanthochondria cornuta, A. limandae, Acanthochondrites annulatus, Blias prionoti, Chondracanthodes deflexus, Chondracanthus lophii, Lernentoma asellina and Pseudoblias lyrifera is described. Nuptial organs in C. lophii contain glandular tissue and appear capable of producing a secretion. In addition to providing a site for male attachment these organs may have other possible functions, which are discussed.     

Morphological studies of possible neuroendocrine structures in Helisoma tenue (Gastropoda: Pulmonata)

Summary Studies of possible neuroendocrine structures in the pulmonate gastropod Helisoma tenue show that cerebral fuchsinophilic neurons with electron-dense granules (mean diameter 1,500 Å) apparently release their secretory content in an intercerebral commissure neurohemal area near the mediodorsal bodies (MDB) or in the median labial nerve neurohemal area.The MDB cells have axon-like processes which branch and end at the cerebral surface, separated by a thin capsule from the cerebral fuchsinophilic cells. The presence of granules (mean diameter 800 Å) in the terminals of the MDB cells suggests cell body origin, distal transport and release of the granular secretory material. The secretory product may have an influence on cerebral fuchsinophilic neurons.Electron microscopy reveals the presence of granules of different sizes and densities in expanded neurites at the periphery of the intestinal nerve of the visceral ganglion which may indicate the presence of a neurohemal area. However, the granules in the intestinal nerve neurites and in the visceral ganglion fuchsinophilic cells are similar to granules found in the heart which also suggests that the granules may carry a neurotransmitter.This study was supported by NTH traineship USPHS T1 CA 5045 and NSF grant GB-6424X. The author would like to thank Professor Howard A. Bern for guidance and support during the course of this work, Mr. John Underhill and Mr. Albert Blaker for photographic assistance, and Mrs. Emily Reid for preparation of the diagram in Fig. 16.     

Herbivorous insects: model systems for the comparative study of speciation ecology

Does ecological divergence drive species-level evolutionary diversification? How so and to what degree? These questions were central to the thinking of the evolutionary synthesis. Only recently, however, has the ecology of speciation become an important focus of empirical study. Here, we argue that ecologically specialized, phylogenetically diverse, and experimentally tractable herbivorous insect taxa offer great opportunities to study the myriad mechanisms by which ecology may cause reproductive isolation and promote speciation. We call for the development and integrated experimental study of a taxonomic diversity of herbivore model systems and discuss the availability and recent evaluation of suitable taxa. Most importantly, we describe a general comparative framework that can be used to rigorously test a variety of hypotheses about the relative contributions and the macroevolutionary generality of particular mechanisms. Finally, we illustrate important issues for the experimental analysis of speciation ecology by demonstrating the consequences of specialized host associations for ecological divergence and premating isolation in Neochlamisus bebbianae leaf beetles.