ALBERT

Notizen: Abstract In the Su-Lu ultrahigh-P terrane, eastern China, many coesite-bearing eclogite pods and layers within biotite gneiss occur together with interlayered metasediments now represented by garnet-quartz-jadeite rock and kyanite quartzite. In addition to garnet + omphacite + rutile + coesite, other peak-stage minerals in some eclogites include kyanite, phengite, epidote, zoisite, talc, nyböite and high-Al titanite. The garnet-quartz-jadeite rock and kyanite quartzite contain jadeite + quartz + garnet + rutile ± zoisite ± apatite and quartz + kyanite + garnet + epidote + phengite + rutile ± omphacite assemblages, respectively. Coesite and quartz pseudomorphs after coesite occur as inclusions in garnet, omphacite, jadeite, kyanite and epidote from both eclogites and metasediments. Study of major elements indicates that the protolith of the garnet-quartz jadeite rock and the kyanite quartzite was supracrustal sediments. Most eclogites have basaltic composition; some have experienced variable ‘crustal’contamination or metasomatism, and others may have had a basaltic tuff or pyroclastic rock protolith.The Su-Lu ultrahigh-P rocks have been subjected to multi-stage recrystallization and exhibit a clockwise P-T path. Inclusion assemblages within garnet record a pre-eclogite epidote amphibolite facies metamorphic event. Ultrahigh-P peak metamorphism took place at 700–890° C and P〉28 kbar at c. 210–230 Ma. The symplectitic assemblage plagioclase + hornblende ± epidote ± biotite + titanite implies amphibolite facies retrogressive metamorphism during exhumation at c. 180–200 Ma. Metasedimentary and metamafic lithologies have similar P-T paths. Several lines of evidence indicate that the supracrustal rocks were subducted to mantle depths and experienced in-situ ultrahigh-P metamorphism during the Triassic collision between the Sino-Korean and Yangtze cratons.

Notizen: Abstract Blueschists occurring as layers in calcite marbles of the Meliata unit occur along the so-called Roznava tectonic line situated in the southern part of the Gemericum, Slovakia. Mineral assemblages and compositions from seven blueschists localities and one occurrence of amphibolite facies rocks overprinted by blueschist metamorphism were investigated. The most common minerals in the blueschists are blue amphibole, epidote and albite. Some Fe2+- and Al-rich rocks also contain garnet and chloritoid, respectively. Na-pyroxene with a maximum 50% jadeite component was also found. The blue amphiboles correspond mostly to crossite and also to glaucophane and ferroglaucophane in some samples. Almandine- and spessartine-rich garnet has very low MgO content (〈3 wt%). The Si content in phengite ranges between 3.3 and 3.5 pfu calculated on the basis of 11 oxygens. The zoning patterns of blue amphibole, garnet and chloritoid suggest their formation during a prograde stage of metamorphism. The P-T conditions of metamorphism are estimated to be about 380–460° C and 10–13 kbar. Pressures of 7.5–8.5 kbar and temperatures of 350–370° C were obtained for some actinolite- and aegirine-rich rocks. Apart from chlorite, other mafic minerals formed during retrograde metamorphism are biotite and occasionally also actinolite.

Notizen: Abstract The Port aux Basques gneisses comprise three lithostratigraphic units separated by major fault zones: the Grand Bay Complex; the Port aux Basques Complex; and the Harbor le Cou Group. A similar regionally developed polyphase history of penetrative deformation characterizes each of these units. Thickening during D1 produced rare recumbent folds (F1) and an axial planar schistosity (S1), overprinted by D2 recumbent folds (F2), and transposed during development of a locally penetrative, differentiated crenulation cleavage (S2). In western sectors of the area, D2 was associated with NW-directed reverse shearing. The NE-trending structural grain reflects D3 transpression, partitioned into dextral transcurrent movement along major shear zones and development of upright-to-steeply inclined, periclinal folds (F3) and a variably penetrative schistosity (S3).Amphibolite facies metamorphism increases in grade from west to east across the area. Microstructures, including porphyroblast-matrix foliation relations and internal textural unconformities in garnet, indicate episodic porphyroblast nucleation and growth, which reflect a prograde traverse sequentially across univariant reactions during syntectonic metamorphism. Garnet, kyanite and staurolite porphyroblasts are wrapped by the S2 foliation, but each may contain trails of inclusions that define S1; commonly these trails preserve early stages of S2 crenulation cleavage development. Progressive and sequential reaction out of kyanite, staurolite and muscovite in favour of sillimanite, garnet, biotite and K-feldspar, and the development of an increasing volume of anatectic migmatite in south-eastern sectors of the area record syn- to late-D2 peak metamorphic conditions. Microstructural relationships and petrogenetic grid considerations indicate clockwise trajectories in P-T space for units of the Port aux Basques gneisses. Peak metamorphic conditions are estimated to have been 620–650° C at ≤8kbar in the west and 700–750° C at ≤8 kbar in the east.Titanite from an upper amphibolite facies calc-silicate gneiss yields U-Pb ages of c. 420 Ma, interpreted to date cooling shortly after the thermal peak in these gneisses. Variable D3 strain was associated with some recrystallization of hornblende and micas. 40Ar/39Ar hornblende plateau isotope correlation ages range from c. 419 to c. 393 Ma, from east to west across the area, and are interpreted to record cooling through c. 500° C coeval with or soon after D3 deformation. The range in ages may record the effects of heterogeneous D3 deformation and differential uplift from south-east to north-west associated with displacement on major shear zones. 40Ar/39Ar muscovite plateau ages cluster at c. 390 Ma, and date cooling through c. 375° C during regional exhumation. Cooling rates are moderate to fast and may indicate a component of tectonic exhumation.The Port aux Basques gneisses are a product of Silurian collisional tectonics. The higher grade of metamorphism in comparison with adjacent areas of the Canadian Appalachians is interpreted to reflect greater thickening due to juxtaposition of the St Lawrence promontory (Laurentian margin) with the Cabot promontory (Avalonian margin) during closure of the Iapetus Ocean.

Notizen: Abstract Carbon isotope thermometry has been applied to coexisting calcite and graphite in marbles from throughout the Adirondack Mountains, New York. Eighty-nine calcite-graphite pairs from the amphibolite grade NW Adirondacks change systematically in temperature north-westwards from 680 to 640 to 670° C over a 30-km distance, reflecting transitions from amphibolite facies towards granulite facies to the north-west and to the south-east. Temperature contours based on calcite-graphite thermometry in the NW Adirondacks parallel mineral isograds, with the orthopyroxene isograd falling above 675° C, and indicate that regional metamorphic temperatures were up to 75° C higher than temperatures inferred from isotherms based on cation and solvus thermometry (Bohlen et al. 1985). Fifty-five calcite-graphite pairs from granulite grade marbles of the Central Adirondacks give regional metamorphic temperatures of 670–780° C, in general agreement with cation and solvus thermometry.Data for amphibolite and granulite grade marbles show a 12%oo range in δ13Ccal and δ13Cgr. A strong correlation between carbon isotopic composition and the abundance of graphite (Cgr/Crock) indicates that the large spread in isotopic compositions results largely from exchange between calcite and graphite during closed system metamorphism. The trends seen in δ13C vs. Cgr/Crock and δ13Ccal vs. δ13Cgr could not have been preserved if significant amounts of CO2-rich fluid had pervasively infiltrated the Adirondacks at any time. The close fit between natural data and calculated trends of δ13C vs. Cgr/Crock indicates a biogenic origin for Adirondack graphites, even though low δ13C values are not preserved in marble.Delamination of 17 graphite flakes perpendicular to the c-axis reveals isotopic zonation, with higher δ13C cores. These isotopic gradients are consistent with new graphite growth or recrystallization during a period of decreasing temperature, and could not have been produced by exchange with calcite on cooling due to the sluggish rate of diffusion in graphite. Samples located 〉2km from anorthosite show a decrease of 0.5-0.8%oo in the outer 100 μ of the grains, while samples at distances over 8 km show smaller core-to-rim decreases of c.0.2%oo. Correlation between the degree of zonation and distance to anorthosite suggests that the isotopic profiles reflect partial overprinting of higher temperature contact metamorphism by later granulite facies metamorphism. Core graphite compositions indicate contact metamorphic temperatures were 860–890° C within 1 km of the Marcy anorthosite massif. If samples with a significant contact metamorphic effect (Δ(cal-gr) 〈3.2%oo) are not included, then the remaining 38 granulite facies samples define the relation Δ13C(cal-gr) = 3.56 ± 106T-2 (K).

Notizen: Native grasses may be used for multiple, sometimes conflicting, goals in revegetation projects. Woody plants are frequently desired for moose browse and timber in Alaska, but naturally occurring Calamagrostis canadensis (bluejoint reedgrass) hinders the establishment of these desired species. Seven grass cultivars of Alaskan origin were evaluated for their ability (1) to stabilize the soil, (2) to reduce regeneration of C. canadensis, (3) to allow openings for natural colonization, and (4) to permit establishment of desirable rooted cuttings. Cultivars tested are “Arctared” Festuca rubra (red fescue), “Alyeska”Arctagrostis latifolia (polar-grass), “Nugget”Poa pratensis (bluegrass) “Norcoast”Deschampsia beringensis (Bering hairgrass), “Nortran”Deschampsia caespitosa (tufted hairgrass), “Gruening” Poa alpina (alpine bluegrass), and “Sourdough” Calamagrostis canadensis. These were tested as single species and in multi-species mixtures, with two seeding rates of the multi-species mixture (0.5, 0.25 seeds/cm2). Experimental plots included unfertilized, unfertilized with rooted Salicaceae cuttings, and fertilized (350 kg/ha 20:20:10). A control plot was not seeded. After three growing seasons, Nortran D. caespitosa and Arctared F. rubra were the most successful cultivars. They provided 87% to 98% of the seeded-species cover for soil stabilization and suppressed C. canadensis on the fertilized subplots without reducing species diversity. Gruening Poa alpina was less than 3 cm tall, and it helped stabilize the site without interfering with woody plant establishment. Although cuttings were shorter under some seed treatments compared to the nonseeded control, heights of cuttings were not related to cover of seeded cultivars (r = 0.09, p 〉 0.55) but were positively correlated with total vascular plant cover (r= 0.61, p 〈 0.001).

Notizen: Throughout tropical moist climates, Dicranopteris linearis fernlands can develop as a result of rain forest clearance followed by frequent burning. In Sri Lanka, D. linearis fern-lands are capable of suppressing the regeneration of rain forest. Field experiments were conducted at Sinharaja Man and Biosphere Reserve, a rain forest where fernlands occupy substantial areas of the reserve boundary. The experiment's objective was to identify methods for initiating forest regeneration in fernlands dominated by D. linearis Three disturbance treatments were used to initiate seedling regeneration: clean weed, root removal, and till. We hypothesized that increasing the severity of the soil disturbance would establish vegetation with higher species richness and diversity, greater above-ground dry biomass, and higher percentage cover and seedling density. Results indicate only partial support for this hypothesis. Dry biomass was greatest in till treatments, the most severe soil disturbance. By comparison, species richness and diversity, seedling density, and percentage cover were greatest in root-removal treatments, though in many instances the differences were not significant. The study clearly demonstrated that any kind of soil disturbance can facilitate the establishment of herbs, shrubs, and trees in a fernland dominated by D. linearis. Results showed that herbs, sedges, grasses, and pioneer shrubs represented greater proportions of seedling recruits than did pioneer trees. Seedlings of primary-forest tree species were nearly nonexistent. In general, results showed that soil disturbance can play an important role in site preparation for the purpose of initiating non-fern vegetation in fernlands dominated by D. linearis.

Notizen: The effect of a pipeline corridor constructed through an ecological reserve in Southern California was investigated by assessing plant species composition and soil chemistry. A homogeneous plant community comprised primarily of exotic annuals was found along the entire length of the corridor. This community has low similarity to the adjacent native plant communities. Soil organic matter was significantly less on the disturbed corridor than in contiguous undisturbed areas. Both available nitrogen and extractable phosphorus values were greater in the disturbed corridor. By contrast, total nitrogen was significantly higher outside the pipeline. The more labile litter of the exotic annuals allows increased mineralization along the corridor than does the more recalcitrant litter of the native perennial shrubs in the undisturbed areas. Once established, the weedy exotic annual litter may completely turn over organic matter and nitrogen, favoring the persistence of the weedy annuals. These exotic annuals appear to be moving into three of the native communities - grassland, coastal sage, and oak woodland - that have less organic matter and a more open plant canopy. Poor restoration efforts can lead to the establishment of such exotics, subsequent invasion into the surrounding undisturbed habitat, and degradation of the reserve.

Notizen: A critical element of the ongoing effort to restore the ecological integrity of Florida's Kissimmee River ecosystem is the reestablishment of pre-channelization habitat structure and function. Restoration of habitat will form the basis for responses by most biological components of the ecosystem and will provide a key indicator of the success of the restoration effort. This paper evaluates the relative importance of a range of abiotic and biotic habitat parameters in the existing and historic Kissimmee River ecosystem and provides a conceptual framework for predicting expected spatial and temporal responses of river and floodplain habitats to the restoration project. Among the ecological factors and process that influenced the development, dynamics, and maintenance of river and floodplain habitat structure, hydrology is expected to be of central importance in eliciting restoration responses in the Kissimmee River Ecosystem. Based on the assumption that the restoration plan will reestablish historic hydrologic characteristics, predictions are made of expected responses by geomorphic and vegetative components of the Kissimmee River's habitat structure. Recommendations are made regarding key habitat parameters requiring long term tracking and analysis and utilization of a geographic information system(GIS). A hierarchical habitat classification scheme is provided as a foundation for all components of the restoration evaluation program.

Notizen: When the Kissimmee River was channelized in the 1960s and 1970s and placed under stage-fluctuation management, the dynamic interactions between the river and the flood-plain were essentially removed. Correspondingly, aquatic invertebrate life in the river and floodplain ecosystem shifted from a riverine to a more lacustrine fauna. A relinkage of the Kissimmee River with the floodplain following restoration will result in numerous changes to such ecologically important factors as streamflow, substrate composition, food quality and quantity, and water quality, all of which will influence invertebrate communities. These factors and their function in the ecosystem as the fauna shifts from predominantly lacustrine back to riverine are presented in a conceptual model. As an integral component of all aquatic ecosystems and a key link between primary producers and higher trophic levels, aquatic invertebrates are a valuable group with which to evaluate the recovery of the Kissimmee River. Utilization of a geographic information system mapping approach linking expected increased habitat heterogeneity and invertebrate richness with restoration efforts is suggested as an economical means of monitoring recovery of the Kissimmee River ecosystem.

Notizen: Despite the critical role insects play in ecosystem functioning, there has been little study of factors affecting their reestablishment in restored ecosystems. The goals of this research were to quantify the nectar resources provided by reclaimed coal surface mines and to examine the role nectar resources play in determining butterfly community composition on these sites. Adult butterfly communities and nectar resources were sampled on 18 reclaimed coal surface-mined sites and five unmined hardwood sites in southwestern Virginia. Recently, reclaimed sites provided an average of 300 times the nectar abundance of the surrounding hardwoods, and nectar abundance and species richness decreased with time since reclamation. Total nectar abundance was highly correlated with total butterfly abundance and species richness for the entire flight season; these variables were also significantly correlated among sites during most of the 12 sampling periods during the flight season. In only a few cases, however, were butterfly and nectar abundance and species richness significantly correlated within individual sites during the flight season. These results suggest that, although adults of many butterfly species move in response to nectar availability, nectar resources are not sufficiently limiting that their life histories have evolved to maximize nectar resources temporally. While planting species in restored areas that provide abundant nectar will likely attract adult butterflies, this is only one of a number of habitat variables that must be considered in efforts to restore butterfly populations. Finally, adult butterflies appear to have limited utility as indicators of revegetation success.

Notizen: The restoration of the northern jarrah (Eucalyptus marginata) forest after bauxite mining is a major objective of Alcoa of Australia Limited. The typically variable and sometimes low emergence of broadcast seed of jarrah-forest plant species may relate to microclimatic changes associated with mining disturbance. This study examined the effect of the presence and absence of a canopy and topographic position in the post-mining landscape on the emergence of four canopy species (E. marginata, E. calophylla, E. patens, and E. diversicolor) and related these patterns to detailed measures of surface soil temperature and moisture. The absence of a canopy in the restoration appeared to result in adverse microclimatic conditions for the successful early establishment of E. marginata and E. calophylla from seed, particularly in the low topographic regions of the restoration. Emergence beneath a canopy compared to that in the open was 17% and 6%, respectively, for E. marginata and 23% and 2%, respectively, for E. calophylla. For both species, emergence was also greater at upland than at lowland open restoration sites (9% and 3%, respectively, for E. marginata; 4% and 0.3%, respectively, for E. calophylla). In contrast, canopy removal and position on the topographic landscape did not reduce the early establishment success of E. patens and E. diversicolor. Field measurements revealed that soils were drier and that diurnal temperature fluctuations were wider in the open restoration sites than beneath a canopy. Furthermore, cold conditions were more frequent at lowland than at upland restoration sites, suggesting the occurrence of cold-air drainage to Jew-lying areas. It is therefore possible that the field emergence patterns reflected the lower tolerance of E. marginata and E. calophylla than both E. diversicolor and E. patens to cold and dry surface-soil conditions. The ecological significance and practical implications of the results are discussed.

Notizen: River and stream restoration projects are increasingly numerous but rarely subjected to systematic post-project evaluation. The few such evaluation studies conducted have indicated a high percentage of failures. Thus, post-project evaluation (and dissemination of results) is essential if the field of river restoration is to advance. Effective evaluation of project success should include: (1) Clear objectives, essential to identity potential incompatibilities among project objectives and to provide a framework for design of project evaluation. (2) Baseline data, needed as an objective basis for evaluating change caused by the project and encompassing as long a pre-project period as possible (including a detailed historical study). (3) Good study design, to demonstrate the effects of restoration projects in the complex riverine environment. (4) Commitment to the long term, to detect effects evident only years following project completion; in general, monitoring should continue for at least a decade, with surveys conducted after each flood above a predetermined threshold. (5) Willingness to acknowledge failures, or rather to recognize that each restoration project constitutes an experiment, so that a failure can be just as valuable to the science as a success, provided we can learn from it (which requires objective, robust post-project evaluation).

Notizen: Festuca idahoensis (Idaho fescue) was a common native perennial bunchgrass in the sagebrush steppe of the western United States until the introductions of domestic livestock and alien plants. Restoration of Idaho fescue to degraded sites will likely involve reseeding, and one of the factors affecting reseeding success is germinability of the seeds employed. We investigated effects of after-ripening and storage temperature on germinability of Idaho fescue seeds collected from a central Oregon site. Six months of after-ripening were required before maximum germination was obtained. Storage of dry seeds at either room temperature (20°C) or at cooler, alternating temperatures (5/15°C) did not alter the rate at which dormancy was lost. Storage at the warmer temperature promoted rapid germination in seeds that had broken dormancy. Seed longevity varied greatly from year to year. Seeds produced in a very dry year had poorer germination and shorter longevity than seeds produced during a year with near normal precipitation. Because seed dispersal occurs in late July and early August for Idaho fescue in central Oregon, a six-month after-ripening requirement ensures that the greatest potential germination coincides with the spring period most likely to provide sufficient moisture for seedling establishment.

Notizen: We conducted an experiment with the dual aims of (1) examining the feasibility of establishing a species-rich grassland using a commercially available grass and wild-flower seed mixture and (2) examining the effects of different defoliation and fertilizer managements on the productivity, species richness, diversity, and composition of a species-rich grassland established on a site reclaimed after opencast coal mining. The use of the seed mixture successfully established a sward of some 18–25 species per square meter. The species composition was enriched to some extent by recruitment of unsown species, principally from the soil seed bank. Hay-type defoliation management produced greater dry matter yield and species richness than grazing defoliation, but grazing defoliation produced greater species diversity. Fertilizer application had no significant effect on dry matter production but reduced species diversity. Ordination analysis revealed that both defoliation and fertilizer management significantly affected species composition. The response obtained by individual species was explicable largely by their comparative biology.

Notizen: Books reviewed in this article: World Soil Erosion and Conservation. D. Pimentel, editor Environmental Impacts of Mining: Monitoring, Restoration and Control. M. Sengupta

Notizen: We investigate why different states in the United States choose different regulatory plans in their telecommunications industry. We present a simple theoretical model and an empirical analysis of the issue. We find that a state is more likely to replace rate-of-return regulation with incentive regulation when: (1) residential basic local service rates have historically been relatively high; (2) allowed earnings under rate-of-return regulation in the state have been either particularly high or particularly low; (3) the state's leaders tend to come from both major political parties, rather than from a single party; (4) the state's urban population is growing relatively rapidly; and (5) the bypass activity of competitors in the state is less pronounced.

Notizen: After the initial breakthrough in the research phase of R&D, a new product undergoes a process of change, improvement, and adaptation to market conditions. We model the strategic behavior of firms in this development phase. We emphasize that a key dimension to this competition is the innovation that leads to product differentiation and quality improvement. In a duopoly model with a single adoption choice, we derive endogenously the level and diversity of product innovations. We demonstrate the existence of equilibria in which one firm enters early with a low-quality product while the other continues to develop the technology and eventually markets a high-quality good. In such an equilibrium, no monopoly rent is dissipated and the later innovator makes more profits. Incumbent firms may well be the early innovators, contrary to the predictions of the “incumbency inertia” hypothesis.

Notizen: This paper addresses the strategic commitment value of various price and nonprice vertical contracts. Assuming contract offers are sufficiently public to have strategic importance, we focus on commitment as a necessary and separable condition to be met. The analysis has two components. First, considering the relationship between a monopolist manufacturer and his imperfectly competing retailers, enforcement conditions necessary for alternative vertical contracts to have commitment value are identified in the static (reputation-free) context. It is shown that external help (i.e., enforcement by nonsignatories) is necessary for bilateral minimum resale price maintenance (price floors) and exclusive territory contracts to have commitment value, whereas maximum resale price maintenance (price ceilings) do not require such help (i.e., they are internally enforcing). Second, the paper provides an analysis of the U.S. case law and discusses the empirical validity of the “external enforcement” assumption that is shown to underlie recent proposals to make certain vertical restraints illegal.

Notizen: . Experimental evidence has been gathered to show that the life cycle of the myxozoan gallbladder parasite Zschokkella nova Klokacewa, 1914, which infects the fish Carassius carassius, has a complex life cycle with alternation of two hosts (fish and Oligochaeta) and two developmental phases (myxosporean and actinosporean). The gut epithelium of the oligochaete, Tubifex tubifex, exposed experimentally to Z. nova, obtained from C. carassius, became infected with organisms resembling Actinosporea. The spore structure and cube-like network of the interconnected spores is reminiscent of Siedleckiella silesica Janiszewska, 1952, although the spores are very different in size and number of sporoplasm nuclei. The life cycle of Z. nova resembles that of the whirling disease agent Myxosoma cerebralis described by Wolf and Markiw, which also alternates between fish and oligochaete hosts.

Notizen: . Organisms in the phylum Apicomplexa appear to have a large extrachromosomal DNA which is unrelated to the mitochondrial DNA. Based on the apparent gene content of the large (35 kb) extrachromosomal DNA of Plasmodium falciparum, it has been suggested that it is a plastid-like DNA, which may be related to the plastid DNA of rhodophytes. However, phylogenetic analyses have been inconclusive. It has been suggested that this is due to the unusually high A + T content of the Plasmodium falciparum large extrachromosomal DNA. To further investigate the evolution of the apicomplexan large extrachromosomal DNA, the DNA sequence of the organellar ribosomal RNA gene from Toxoplasma gondii, was determined. The Toxoplasma gondii rDNA sequence was most similar to the large extrachromosomal rDNA of Plasmodium falciparum, but was much less A + T rich. Phylogenetic analyses were carried out using the LogDet transformation to minimize the impact of nucleotide bias. These studies support the evolutionary relatedness of the Toxoplasma gondii rDNA with the large extrachromosomal rDNA of Plasmodium falciparum and with the organellar rDNA of another parasite in the phylum Apicomplexa, Babesia bovis. These analyses also suggest that the apicomplexan large extra-chromosomal DNA may be more closely related to the plastid DNA of euglenoids than to those of rhodophytes.

Notizen: . Seabottom sediments from a discontinued Philadelphia-Camden 40-Mile ocean sewage disposal site were cultured for cyst-forming free-living amoebae. Barge delivered wastes were discharged at the site from 1973 until 1980 when the site was closed. One station at the southeast margin of the site was sampled at a depth of approximately 50 m, twice in 1978 and once in 1982, 1983 and 1984. Sediment from the 1978 collection yielded Acanthamoeba polyphaga, Vahlkampfia sp., and an unknown amoeba with stellate endocysts similar to those of A. astronyxis. Trophozoites and cysts of the isolate were typical of those described for the genus Acanthamoeba. Biochemical tests employing enzyme electrophoresis and morphological studies on live and stained specimens showed that the isolate was distinct from other well-described species within the family Acanthamoebidae Sawyer & Griffin, 1975.

Notizen: . Alexandrium tamarense (Lebour) Balech (=Gonyaulax tamarensis Lebour) has been widely distributed and occasionally abundant in coastal waters of Long Island, New York in recent years. However, the distribution on the New Jersey coast has been sparse and this fact cannot be explained by this dinoflagellate's ability to migrate, or by its nutritional and physiological characteristics, or by the region's general suitability for phytoplankton. Therefore, the possibility that New Jersey coastal waters might be chemically exclusionary for A. tamarense seemed worth exploring. In a limited approach, we tested for water quality detrimental to the species in one New Jersey site (Great Bay) with a series of annual assays. Parsonage Creek, Long Island, New York, was assayed for comparison; this creek is assumed to have had at least marginal suitablity for A. tamarense, based on its reported long-term presence. Results provide tentative support for our working hypothesis, i.e. Great Bay chemical water quality is generally unfavorable for A. tamarense. Inhibition of A. tamarense growth, or culture decline, occurred in both assay series, but was substantially greater in Great Bay water. Inimical water quality was the most important factor distinguishing the two sites. Chelation with EDTA had greatest overall benefit in Great Bay assays, suggesting that lower availability of a natural chelator in the bay could be a secondary factor. Assay metal response is problematic, but we believe it permits speculation that essential metals could be partially limiting to A. tamarense in Great Bay, but would not be a critical regulator.

Notizen: . Experimental transmission of two bacterial endosymbionts to symbiont-free isolates of Acanthamoeba spp. was studied to determine specificity of the host-symbiont relationship. Both symbionts originated from amoebic isolates displaying an identical mitochondrial DNA EcoRI fingerprint (group AcUW II). Symbioses were readily established in one amoebic isolate which displayed a homologous mtDNA fingerprint (group AcUW II). Exposure of a heterologous amoebic isolate (group AcUW IV) to the two symbionts resulted in either cell death or encystation without the establishment of symbioses. While symbioses were established with an amoebic isolate from a second heterologous group (AcUW I), a unique membranous sheath appeared and persisted around one of the symbionts which did not exist in the original host. An isolate representing a third heterologous amoebic group (AcUW VI) was variable in its susceptibility with one symbiont unable to infect the host and the other becoming established only after an initial reaction in which trophozoites rounded-up and floated off the substrate. These studies suggest that a specific recognition system exists between particular isolates of Acanthamoeba and their symbionts, and that the appearance of a killer phenotype is related to contact between mismatched, though recognized, pairs.

Notizen: . Microsporidian spores were developed from cells which were grown in vitro from a human liver lesion which was due to larval Echinococcus multilocularis. The microsporidian spores developed in the same fashion as an Encephalitozoon cuniculi. The Encephalitozoon-like spores were completely separated on Percoll gradients. The separated spores contained DNA capable of amplification by two different primer sets designed for the polymerase chain reaction (PCR) of E. multilocularis DNA. However, the cell DNA from which microsporidium developed was thoroughly insensitive to the PCR using the E. multilocularis primer sets. The results strongly suggested that Encephalitozoon should be taken into consideration, when DNA isolated from larval E. multilocularis is analyzed.

Notizen: . We have used the anti-phosphoprotein antibody MPM-2 to examine changes in phosphorytation of cortical proteins during cilia regeneration in Tetrahymena thermophila. Although numerous cortical proteins are phosphorylated in both nondeciliated and deciliated cells, deciliation induces a dramatic increase in the phosphorylation of a 90-kDa cortical protein. The 90-kDa protein remained phosphorylated during cilia regeneration and then gradually became dephosphorylated. The 90-kDa protein was phosphorylated and dephosphorylated normally in Tetrahymena mutants that assemble short cilia, suggesting that achievement of full length is not the signal that triggers dephosphorylation of the 90-kDa protein. When initiation of cilia assembly is blocked, the 90-kDa protein becomes phosphorylated and remains phosphorylated for an extended period of time, suggesting that initiation of cilia elongation triggers eventual dephosphorylation of the 90-kDa protein, regardless of how long the cilia actually become.

Notizen: . Giardia lamblia trophozoites express on their surfaces one of a set of cysteine-rich antigenically variant proteins, called variant-specific surface proteins, which comprise the majority of proteins detected by surface labeling. While these VSP proteins may be immunodominant proteins important in the host immune response to G. lamblia, the ability to switch expression from one VSP to another may provide a means for the trophozoites to avoid the host immune response. The first VSP characterized, VSPA6 (from the A6 clone of the WB isolate, originally termed CRP170), contains 18–23 copies of a 65 amino acid repeat. We have now used the repeat as a probe to isolate from a WBA6 genomic library two genes related to vspA6 (called vspA6-S1, vspA6-S2). Sequence analysis of the vspA6-S1 gene revealed nearly two complete copies of the 195 bp repeat and substantial nucleotide and translated amino acid similarity in the coding regions 5’and 3’to the repeats. The vspA6-S2 gene, while still related, showed greater divergence from vspA6 than vspA6-S1 in the nonrepeat coding region and contained nearly four copies of a 201 bp repeat that was 75% identical to the 195 bp vspA6 repeat. These results suggest that gene duplication followed by divergence has played a key role in the generation of the vsp gene repertoire.

Notizen: . The glycerophosphate oxidase (GPO), the unique terminal oxidase of bloodstream trypanosome (TAO), appears to be functionally similar to the alternative oxidases of some plants and higher fungi. Immunoblotting of mitochondrial proteins of bloodstream trypomastigotes of Trypanosoma brucei brucei with monoclonal or polyclonal antibodies to Sauromatum guttatum (voodoo lily) and Symplocarpus foetidus (skunk cabbage) alternative oxidases respectively revealed two proteins of about 33 kDa (p33) and 68 kDa (p68). These proteins are not present in procyclic trypomastigotes. Electrophoresis under rigorous denaturing conditions indicated p68 to be the dimer of p33. Indirect immunofluorescent studies of bloodstream and procyclic trypomastigotes with monoclonal antibody to plant alternative oxidase also showed the localization of 33 kDa protein in the mitochondria of the bloodstream trypomastigotes. The functional TAO activity could be solubilized efficiently from the mitochondrial membrane of the bloodstream trypomastigotes by 1% NP-40 or 10 mM lauryl maltoside. When fractionated by Superose 12 gel filtration chromatography, p33 was co-purified with the TAO enzymatic activity. The apparent molecular size of the active enzyme complex was found to be 160 kDa. Gradual disappearance of the 33 kDa protein and the TAO enzymatic activity were well correlated during in vitro differentiation of the bloodstream to procyclic trypomastigotes. This study implies that the net biosynthesis of p33, an essential subunit of TAO, is decreased during differentiation from bloodstream to procyclic trypomastigotes.

Notizen: . The microsporidium, Enterocytozoon bieneusi, causes a severe, debilitating, chronic diarrhea in patients with the acquired immunodeficiency syndrome. Specific diagnosis of intestinal microsporidiosis, especially due to Enterocytozoon, is difficult and there is no known therapy that can completely eradicate this parasite. Preliminary studies indicate that a short term (about 6 months) in vitro culture of this parasite yielding low numbers of spores, may be established by inoculating human lung fibroblasts and/or monkey kidney cell cultures with duodenal aspirates and or biopsy from infected patients. The cultures may subsequently be used for the isolation and molecular analysis of parasite DNA.

Notizen: . A new Myxosporea, Ceratomyxa sparusaurati n. sp., was found in the gall bladder and bile of cultured gilthead sea bream (Sparus aurata L.) from different Spanish fish farms. It is clearly different from all the previously reported Ceratomyxa from sparids, and it is distinguished from other members of the genus by the shape and size of the spores. Prevalence of infection was 2.15% in an Atlantic farm, 48.7% in a Mediterranean farm and 28.6% in the facilities of the Instituto de Acuicultura de Torre de la Sal. The cell-in-a-cell pattern was found through all the sporogenesis and the general ultrastructure resembled other Myxosporea. Primary cells with two developing spores, harbored other secondary cells. Disporous sporoblasts contained numerous membrane-bound inclusions, a few lipid droplets and polysaccharides as evidenced by cytochemistry. In mature spores, binucleate sporoplasmic cells contained abundant Thièry-negative sporoplasmosomes.

Notizen: . The immobilization antigens (i-antigens) are a class of highly abundant surface membrane proteins found on a number of holotrich ciliates. In Ichthyophthirius multifiliis (an obligate parasite of fish) these antigens appear to be targets of the host immune response. While the i-antigens of Ichthyophthirius are predominantly membrane-associated proteins, we now find that they are released into the water surrounding the parasite in a highly enriched form. The membrane-associated and water soluble proteins appear indistinguishable by antigenic means, as well as by several biochemical criteria including peptide mapping, mobility in reducing and non-reducing SDS-polyacrylamide gels, and relative glycosylation. Antibodies raised against the membrane-associated antigens react with the water soluble proteins on Western blots. Not surprisingly, immunocytochemical localization studies show binding of these antibodies to surface membranes of the cell. In addition, however, antibody binding is also detectible on the membranes of a secretory organelle (that is, mucocysts) present in the cortical cytoplasm. The significance of these findings with regard to the potential role of the i-antigens in infection and immunity is discussed.

Notizen: . The complete nucleotide sequence has been established for two genes (maeA and maeB) coding for different subunits of the hydrogenosomal malic enzyme [malate dehydrogenase (decarboxylating) EC 1.1.1.39] of Trichomonas vaginalis. Two further genes (maeC and maeD) of this enzyme have been partially sequenced. The complete open reading frames code for polypeptides of 567 amino acids in length. These two open reading frames are similar with less than 12 percent pairwise nucleotide differences and less than 9 percent pairwise amino acid differences. The open reading frames of the two partially sequenced genes correspond to the amino-terminal part of the polypeptides coded and are similar to the corresponding parts of the completely sequenced ones. The deduced translation products of the two complete genes differ in their calculated pI values by 1.5 pH unit. The genes code for polypeptides which contain 12 or 11 amino-terminal amino-acyl residues not present in the proteins isolated from the cell. Other hydrogenosomal enzymes also have similar amino-terminal extensions which probably play a role in organellar targeting and translocation of the newly synthesized polypeptides. A comparison of 19 related enzymes from bacteria and eukaryotes with the maeA product revealed 34–45 percent amino acid identity. Phylogenetic reconstruction based on nonconservative amino acid differences with maximum parsimony (phylogenetic analysis using parsimony, PAUP) and distance based (neighbor-joining, NJ) methods showed that the T. vaginalis enzyme is the most divergent of all eukaryotic malic enzymes, indicating its long independent evolutionary history.

Notizen: Previous investigations have shown that the adhesion of T. cruzi plasma membrane vesicles (PMV) to monolayers of host cell myoblasts and to immobilized heart muscle sarcolemma membranes (PAM) on polyaerylamide beads is mediated by the interaction of T. cruzi attachment sites with the muscarinic cholinergic and β-adrenergic receptors of the host cell membrane. It has also been shown that this interaction is blunted by the specific antagonists of the mammalian receptors atropine and propranol, respectively. In the studies reported here, PAM also rapidly attached to swimming T. cruzi trypomastigotes in a complex, concentration-dependent fashion and binding isotherms showed that the equilibrium between free and bound PAM is rapidly reached within 2 minutes of incubation in physiologically balanced salt solutions. In this time frame, trypomastigote cAMP levels are significantly reduced from steady state values within 30 seconds of the addition of PAM in a buffer system containing a diesterase inhibitor. Maximal attenuation of cAMP levels was measured between 1 and 2 minutes of the addition of PAM to T. cruzi trypomastigotes. The degree of cAMP level attenuation was reduced by blocking PAM attachment with either atropine or propranol. On the basis of these results we propose that a likely pathway for the negative parasite signal generated upon adhesion of host muscle cell membranes to the surface of the flagellates is from the parasite's surface attachment sites directly to a Pertussis toxin sensitive inhibitory protein Gi, thereby blunting adenyl cyclase activity and cAMP formation.

Notizen: Morphology and morphogenesis of Onychodromus grandis (Hypotrichida, Ciliata) were studied in vivo and in protargolimpregnated preparations. The species was isolated from halophyte communities in astatic shallow waters of the Hortobágy National Park, Hungary. The species has two well developed dorsal horns. According to some characteristics (number and situation of ventral cirri) our findings differ from Stein's description in 1859. Individuals of the population studied had 9 ventral cirri in 3 rows. Nine rows of dorsal kineties were found on the back. It was found that the oral primordium develops in the vicinity of the transverse cirri. In O. grandis marginal primordia are formed in the marginal rows with the participation of parental cirri. This is also true for Laurentiella acuminata. Dorsal fragmentation of O. grandis displays similarity to that of L. acuminata, considering especially the development of the numerous small dorso-marginal kineties. There are certain similarities among O. quadricornutus, O. indica, L. acuminata, Stylonychia vorax and O. grandis in the development of the ventral and dorsal pattern and in some morphogenetical events. The remarkable differences in the interphase cells can be related simply to the significantly different numbers of cortical units (number of cirri). and also to the various cell sizes.

Notizen: Isotricha jalaludinii n. sp. found in the rumen of lesser mouse deer, Tragulus javanicus, in Malaysia was described and illustrated. This new species is characterized by the location and direction of the vestibulum, shape of the macronucleus, and absence of a dent at the vestibular opening. The presence of single peculiar isotrichid species in the rumen of mouse deer, which is recognized as one of the most primitive ruminants, suggests that the isotrichid ciliates similar to I. jalaludinii and Isotricha intestinalis were established at a fairly early period during the evolution of ruminants.

Notizen: We have generated a transformation marker for Paramecium using a Paramecium expression vector (pPXV) and the open reading frame (ORF) of the bacterial antibiotic resistance gene aminoglycoside 3'-phosphotransferase-II (APH-3'-II or neor) from the transposon Tn5. The expression vector contained a small multiple cloning site between the 5' and 3' non-coding regions of the calmodulin gene, and Tetrahymena telomere sequences for the stability of the plasmid in Paramecium. After the neor ORF was inserted, the plasmid was referred to as pPXV-NEO. Delivery of approximately 10–20 picoliters of linearized PXV-NEO at 〉 2000 copies/pl into the macronucleus effected 100% transformation. Southern and Northern blot hybridization showed the presence of neor-specific DNA and RNA, respectively, in all of the transformed clones but not in the untransformed clones. The degree of resistance to G-418, and the concentrations of neor-specific DNA and neor-specific RNA in the clones were proportional to the concentration of the vector injected. We have demonstrated that when the linearized plasmid was injected into the macronucleus, the prokaryotic sequence conferred an antibiotic resistance to Paramecium despite codon-usage differences.

Notizen: . Four methods are commonly used to study cell cycle processes in Paramecium tetraurelia. These include stage frequency analysis in asynchronous cultures, hand selection of synchronous dividing cells, selection of newly divided cells by elutriation centrifugation, and the sister cell method. We have compared the timing and resolution of stages of oral morphogenesis and micronuclear mitosis with each method. The temporal resolution obtainable with the sister cell method was inadequate to position the timing of morphogenesis stages within the cell cycle. Both the asynchronous method and the hand-selected synchronous samples methods are prone to bias. Elutriation centrifuge synchronization provides large samples with resolution comparable to that of hand selected samples. The elutriation method is the least prone to bias when 〈5% of the parent culture of Paramecium is selected.

Notizen: . We compared the expression and localization of the mitochondrial and cytoplasmic hsp70 of the protozoans Trypanosoma cruzi, Trypanosoma brucei and Leishmania major. The mitochondrial protein is encoded by multiple mRNA in all species, while the cytoplasmic protein is encoded by a single mRNA. In all three species, the mitochondrial hsp70 is concentrated in the kinetoplast, a submitochondrial structure that houses the unusual DNA (kDNA) that characterizes this group of organisms, while the cytoplasmic protein is distributed throughout the cell. These results suggest that, in all kinetoplastid species, mt-hsp70 has a specific function in kDNA biology, possibly in the processes of kDNA replication, RNA editing or kinetoplast structure.

Notizen: . Serine is an important amino acid that is utilized in the biosyntheses of proteins and lipids. It is directly incorporated into the head group of phosphatidylserine, which in turn can be converted to other phospholipids. Also, it is required for the formation of long chain bases, precursors of sphingolipids. Uptake and incorporation of radiolabeled serine into both lipids and acid-precipitable material were demonstrated in Pneumocystis carinii carinii organism preparations freshly isolated from infected rat lungs. Radioactivity in proteins was about double that observed in lipids. Liquid scintillation spectrometry of metabolically radiolabeled lipids separated by thin-layer chromatography showed 53% of the total radioactivity were in phosphatidylserine, 12% in phosphatidylethanolamine, 24% in ceramides, and 11% in long chain bases and other compounds. Four long chain bases were detected by thin-layer chromatography in hydrolyzed P. carinii ceramides metabolically labeled with radioactive serine. Phytosphingosine and dihydrosphingosine were tentatively identified by their migrations on thin-layer plates. Radiolabeled ethanolamine was incorporated into P. carinii phosphatidylethanolamine, but relatively low incorporation of radiolabeled choline into phosphatidylcholine occurred. The observations made in this study indicated that P. carinii has the biosynthetic capacity to metabolize phospholipid head groups and to de novo synthesize sphingolipids. L-Cycloserine and β-CI-D-alanine, inhibitors of long chain base synthesis, reduced the incorporation of serine into P. carinii long chain bases and ceramides, which supported the conclusion that the pathogen synthesizes sphingolipids.

Notizen: . The division Chlorarachniophyta comprises amoeboflagellate protists with complex chloroplasts derived from the endosymbiosis of a eukaryotic alga. Analysis of chlorarachniophyte chromosomal DNAs by pulsed-field gel electrophoresis revealed an apparently linear 36-kb chromosome that could not be ascribed to either the host or endosymbiont nuclei. A single eubacterial-like small subunit ribosomal RNA gene is encoded on this chromosome and phylogenetic analyses places this gene within a clade of mitochondrial genes from other eukaryotes. High resolution in situ hybridization demonstrates that transcripts of the small subunit ribosomal RNA gene encoded by the 36-kb chromosome are exclusively located in the mitochondria. The 36-kb chromosome thus likely represents a linear mitochondrial genome. Small amounts of an apparently dimeric (72 kb) form are also detectable in pulsed-field gel electrophoresis.

Notizen: . Developmental processes in multicellular organisms require structural elements, such as adhesion molecules, to stabilize cells at functional positions. In vertebrates, a series of extracellular matrix proteins, e.g. fibronectin and laminin are involved in cell adhesion. These proteins contain Arg-Gly-Asp [RGD] at their binding sites. Here we show that at concentrations above 2 mM the peptide GRGDSPK, comprising the tripeptide RGD (Arg-Gly-Asp), prevents the adhesiveness of cells of the marine amoeba Neopar-amoeba aestuarina. In addition, elevated levels of GRGDSPK cause cells to alter their shapes from those with digitiform subpseudopodia to rounded cells with small lobed pseudopodia. These cells detach from the substratum. These results are specific for the RGD sequence, because incubation in GRGESPK solution at the same concentrations had no effect on cell attachment or structure. From these data we suggest that the structural adhesion molecules identified in vertebrates shows amino acid homologies with those found in unicellular protozoa.

Notizen: . A new diplomonad flagellate, Spironucleus vortens n. sp., is described from the intestinal lumen of the freshwater angelfish, (Pterophyllum scalare), bred in Florida. Live organisms are pyriform, and measure 12.5–20.5 μm long by 5.0–11.2 üm wide. Scanning electron microscopy shows that the trophozoite bears two compound lateral longitudinal ridges, each originating posterior to three emerging anterior flagella, and continuing posteriorly to the emergence of the posterior flagellum. Each ridge comprises a broad central part, surrounded by a peripheral ridge. At the opening of the flagellar pocket, the broader right peripheral ridge crosses to the other side of the body, and then back again. The posterior end of the body bears two papillae. Transmission electron microscopy shows that the compound lateral ridges are supported by microtubules, and bear microfibrillar structures in discrete longitudinal plaques. The serendipitous growth of S. vortens in a culture system with lip tumor tissue, facilitated axenic cultivation in a modified TYM medium (trypticase, yeast extract, maltose). The flagellate is now routinely maintained in an axenic TYI-S-33 medium (trypticase, yeast extract, iron serum), and is stabilized in the cryopreserved state. Spironucleus vortens is an aerotolerant anaerobe that can be cultured at 25° C, 28° C and 30° C.

Notizen: . Microsporidia of the genus Encephalitozoon undergo merogony and sporogony in a parasitophorous vacuole within the host cell. Cultured green monkey kidney cells infected with Encephalitozoon hellem were loaded with the fluorescent dyes fura-2 or BCECF in order to measure intracellular concentrations of calcium and hydrogen ions respectively. Both the parasitophorous vacuole calcium concentration and pH values resembled those of the host cell cytoplasm in infected cells. Calcein entered the parasitophorous vacuole but not other host cell vacuoles or parasite stages within the parasitophorous vacuole. The lack of a pH or calcium concentration gradient across the parasitophorous vacuole membrane and the permeability of this membrane to a large anion such as calcein suggest that the vacuole membrane surrounding E. hellem resembles that surrounding some other intracellular parasites such as Toxoplasma gondii. A potential role is discussed for the parasitophorous vacuole calcium concentration in germination in situ.

Notizen: . The morphology and infraciliature of Siroloxophyllum utriculariae (Penard, 1922) n. g., n. comb. were studied in live cells, with the scanning and transmission electron microscope, as well as in specimens impregnated with protargol and silver carbonate. The new genus, Siroloxophyllum, belongs to the Loxophyllidae and has a specific combination of characters, viz. an oral bulge surrounding almost the entire cell, three perioral kineties, a single brush kinety, and a single right dorsolateral kinety. The ecology and faunistics of S. utriculariae are reviewed. It is a rare and infrequent predator preferring clean freshwaters. The somatic monokinetid of S. utriculariae has typical haptorid ultrastructure, including two transverse microtubular ribbons. The oral bulge is patterned string-like with riffles containing the transverse microtubular ribbons originating from the oral kinetids. Perioral kineties 1 and 2 consist of dikinetids having one basal body each ciliated; the nonciliated basal body is associated with a nematodesmal and a transverse microtubular ribbon. Perioral kinety 3 consists of ciliated monokinetids having a fine structure similar to the somatic kinetids; they form triads with the dikinetids from perioral kinety 2. The classification of pleurostomatid ciliates is reviewed. Two suborders (Amphileptina, Litonotina) and three families (Amphileptidae, Litonotidae, Loxophyllidae n. fam.) are recognized and defined.

Notizen: . Dictyostelium discoideum has a well characterized life cycle where unicellular growth and multicellular development are separated events. Development is dependent upon signal transduction mediated by cell surface, cAMP receptor/G protein linkages. Secreted cAMP acts extracellularly as a primary signal and chemoattractant. There are 4 genes for the distinct cAMP receptor subtypes, CAR1, CAR2, CAR3 and CAR4. These subtypes are expressed with temporally and spatially specific patterns and cells carrying null mutations for each gene have distinct developmental phenotypes. These results indicate an essential role for cAMP signalling throughout Dictyostelium development to regulate such diverse pathways as cell motility, aggregation (multicellularity), cytodifferentiation, pattern formation and cell type-specific gene expression.

Notizen: . Six Leishmania species were studied comparatively, in order to determine the influence of temperature “in vitro” on differentiation, infectivily and protein synthesis. Differentiation ocurred in a heterogeneous manner, even in species that produce similar clinical manifestations. Thus, no association could be found between thermosensitivity and disease. The association between expression of proteins and increasing temperatures was analyzed at 34° C by polyacrylamide gel electrophoresis with sodium dodecyl sulphate (SDS-PAGE), using different incubation times, and employing a technique involving metabolic incorporation of [35S]-methionine. Protein synthesis was very similar in all the New World species apart from L. amazonensis, which expressed a protein of approximately 80 kDa when incubated at 34° C for 2 hours. All the tested species had in common the expression of a 70 kDa protein. Differences, however, were observed in relation to the time interval for protein expression. in L. chagasi, synthesis was detected after 30 minutes of incubation at 34° C, while L. braziliensis required 1 hour at the same temperature. The “in vivo” and “in vitro” infectivity of the differentiated forms was also analyzed, but no significant differences were observed.

Notizen: . Purified preparations of the polypeptide pheromones Er-1 and Er-2 were obtained from type I and II cells of Euplotes raikovi, respectively, radiolabeled, and used to study their ability to compete with each other in binding to the same type I and II cells and to type × cells secreting Er-10, another pheromone homologous to Er-1 and Er-2. It was shown that: 1) These radiolabeled pheromone preparations bind, with similar but not identical affinities, to cells from which they originate as well as to cells of the other types. 2) Every pheromone binding reaction can be completely inhibited by another homologous pheromone added in excess to cells. These results are discussed in relation to phenomena, common in ciliates, of instability of (homotypic) mating pairs formed between genotypically identical cells suspended with a foreign (nonself) pheromone.

Notizen: . Examination of mucus discharged from the blowholes of live odonticete Cetacea and material collected from nasal orifices of dead hosts routinely revealed infestations of a large spindle-shaped ciliate. Kyaroikeus cetarius n. g., n. sp. These ciliates had a prominent posterior podite and were holotrichously ciliated except for a conspicuous bipolar stripe extending along the left margin of the cell. Most specimens were free-swimming and moved slowly through freshly collected mucus, but some individuals were attached to host cells or cellular debris by a transparent thread secreted from the podite. The oral architecture of protargol-impregnated K. cetarius consisted of one preoral and two circumoral kinetics set within a deep oral cavity continuous with an extensive, posteriorly directed cytopharynx. Somatic kineties were composed entirely of monokinetids and formed an expansive right ciliary field covering most of the cell surface, a reduced left ciliary field adjacent to the oral cavity, and a group of four kinetal fragments located mid-ventrally within a shallow pocket. Subkinetal microtubules were associated with the kineties of the right and left fields, and the non-ciliated stripe was underlain by a series of longitudinal fibers. The ciliate's large, heteromeric macronucleus was centrally positioned and clearly evident in living or stained specimens, while the ellipsoid micronucleus was located adjacent to the cytopharynx and often inconspicuous. K. cetarius has several morphological attributes typical of phyllopharyngian ciliates (e.g., adhesive organelle, heteromeric macronucleus, somatic kineties of monokinetids, subkinetal microtubules), and, except for its nearly holotrichous ciliation, most closely resembles dysteriine ciliates.

Notizen: . We developed a method to study the DNA synthetic cycles of Entamoeba histolytica and Entamoeba invadens by flow cytometry (FCM) based on a preparative procedure to reduce both high levels of natural fluorescence and non-specific adsorption of fluorochromes. We modeled G1, S, and G2 phases as a series of overlapping Gaussian curves. Both E. histolytica and E. invadens displayed G1, S, and G2 proportions that are consistent with eukaryotic cell populations in exponential or stationary growth phase. Exponential phase E. histolytica populations contained a hypodiploid subset with a mass of about 20% less than the diploid value which we estimate by FCM to be 24 × 10-14 g DNA/cell. Exponential phase E. invadens populations contained a hypodiploid subset with a mass of about 6% less than the diploid value which we estimate by FCM to be 30 × 10-14 g DNA/cell.

Notizen: . An Entamoeba histolytica gene (hex-A1) that encodes subunit A of the lysosomal enzyme β-hexosaminidase has been cloned and sequenced. The inferred 59 kDa hex-A1 protein has the same molecular weight and 32% amino acid residue identity with the human and mouse proteins and 28% residue identity with the Dictyostelium protein. Northern blot analysis identified a mRNA of approximately 1.6 kb, which is in agreement with the expected size of a mRNA encoding the 522 amino acid hex-A1 protein. Southern blot analysis indicated the presence of at least two β-hexosaminidase A subunit genes.

Notizen: . Epifluorescence microscopy and spectrofluorimetry were investigated as possible non-terminal methods to distinguish live from dead foraminifera. Seven fluorogenic probes (diacetates of fluorescein [FDA], carboxyfluorescein, dichlorofluorescein, and carboxyeosin; AM-esters of biscarboxyethylcarboxyfluorescein [BCECF-AM], calcein, and calcein blue) were tested on Allogromia laticollaris. The probes that consistently produced the brightest fluorescence signals (BCECF-AM and FDA) were judged non-toxic to Allogromia, on the basis of short-term pseudopodial deployment and long-term reproduction assays. Once protocols were established, these two probes were tested on 13 additional benthic foraminiferal species. We found that BCECF-AM is the most suitable probe for direct epifluorescence microscopy of metabolically active foraminifera, especially tectinous and transparent calcareous species. Using spectrofluorimetry, FDA showed promise for opaque species because fluorescence is detected in the incubation media after its release from the cell. However, both approaches could only be used with confidence in light of appropriate controls established for each species examined.

Notizen: . In Euglena gracilis Z, a considerably high activity of mono-ADP-ribosyltransferase occurred and change of it was accompanied by a cell cycle induced by a light-dark cycle. The enzyme activity was strongly inhibited by L-arginine and supported in the presence of poly-L-arginine as a substrate, indicating that ADP-ribosylated amino acid is an arginine residue. Arginine: mono-ADP-ribosyltransferase activity was found in the chloroplasts, mitochondria, microsomes and cytosol as judged from marker enzyme activities and the activity in each organelle fluctuated with the cell cycle.

Notizen: . Tetrahymena thermophila possesses a regulated secretory pathway in which mucin proteins are stored in dense-core granules, called mucocysts. Exocytosis-defective mutants exist that fail to secrete mucin in response to secretagogues. Four of the mutants (SB281, SB283, SB285 and SB715) appear to be blocked at different steps of the regulated secretory pathway. SB281 and SB285 accumulate mucin proteins in heterogeneous cytoplasmic organelles which have not yet been identified; SB283 makes mucocyst-like structures but they contain no immunologically identifiable 80-kDa or 50-kDa mucin proteins; and SB715 has more than normal amounts of immature and undocked mucocysts. The organelles that accumulate in exocytosis-defective mutants could be either normal intermediates in the biosynthetic pathway or aberrant structures that form as a result of the mutations. We have used conjugation rescue to analyze steps in the biogenesis of exocytosis-competent mucocysts and to identify functional intermediates. The cytoplasmic organelles that accumulate in SB281 appear to be unidentified biosynthetic intermediates, and the defect is in a cytosolic protein essential for mucocyst maturation. The organelles which accumulate in the other mutants are likely biosynthetic, but their mutations are in proteins which are labile or not free to diffuse into the mutant conjugant.

Notizen: . We have isolated a cDNA for an α-tubulin mRNA from L. donovani promastigotes and determined its complete nucleotide sequence. Both nucleotide and deduced amino acid sequence analysis of this cDNA showed significant similarity with a previously reported, partial sequence of an L. enriettiiα-tubulin and the complete sequence of human α-tubulin. Further, the in vitro translated L. donovaniaα-tubulin gene product was specifically immunoprecipitated with a monoclonal antibody against human α-tubulin. Northern blot analysis revealed that there was little change in the expression of the L. donovaniα-tubulin RNA during parasite differentiation from promastigote to the in vitro grown “amastigote” form. Southern blot analysis revealed a simple genomic organization for the L. donovaniα-tubulin gene with more than one copy of the α-tubulin gene in the parasite genome. To our knowledge, this is the first complete sequence of an α-tubulin for Leishmania to be reported in the literature.

Notizen: . Entamoeba histolytica causes amebic dysentery (AD) and liver abscess (ALA). Little is known about protective immunity to amebiasis, and studies in this area have been complicated by the paucity of defined ameba antigens. We examined the proliferative responses of peripheral blood mononuclear cells (PBMC) from patients with AD and ALA to a recombinant protein containing a portion of the 170 kDa adhesin of E. histolytica (170CR), and to two synthetic peptides (1 and 2) derived from the 170 kDa sequence that were predicted to contain T cell epitopes. A significant number of patients with AD and ALA had PBMC that proliferated to 170CR molecule, and several individuals with ALA and AD had T cells that recognized one or both peptides. Contrarily, individuals from a non-endemic region for amebiasis did not respond to 170CR protein, or to both peptides. In regard to antibody response, nine of fifteen patients with ALA showed antibodies to 170CR protein. These same patients had antibodies to peptide 2. We identified peptides from 170-kDa adhesin that may contain both T and B cell epitopes recognized by some patients with invasive amebiasis. These peptides may be valuable reagents in studies of the immune response to amebiasis.

Notizen: . The antiproliferative effects and ultrastructural alterations induced in vitro by two antifungal compounds, the azole ketoconazole and the allylamine terbinafine on Leishmania amazonensis are reported. Promastigotes treatment with ketoconazole and terbinafine induced growth arrest and cell lysis in 72 hours. Combination of the two agents produced additive effects on promastigote axenic growth and synergistic effects on intracellular amastigote proliferation. The amastigotes, either axenically grown or infecting murine macrophages, were about 100-fold more sensitive to the drugs. These compounds induced the appearance of large multivesicular bodies, especially after ketoconazole treatment, increased amount of lipid inclusions as well as numerous, polymorphic volutin granules, particularly in terbinafine-treated cells. Multivesicular bodies were observed in close apposition with organelles such as mitochondria, which also showed alterations in the distribution and appearance of cristae, and the formation of paracrystalline arrays within the matrix. Some cells presented large portions of cytoplasm wrapped by endoplasmic reticulum and many parasites also presented myelin-like endoplasmic reticulum profiles. Such alterations together with the strong acid phosphatase activity observed in the multivesicular bodies and volutin granules may indicate the existence of an unusual autophagic process in cells treated with ergosterol biosynthesis inhibitors.

Notizen: . A species of Encephalitozoon has been isolated from the urine of a patient with the acquired immunodeficiency syndrome and maintained in vitro in Madin Darby Canine Kidney cells. When examined by random amplified polymoprhic DNA polymerase chain reaction the new isolate was found to differ from E. hellem and to have amplified products in common with murine and canine E. cuniculi. However, it more closely resembled the canine than the murine isolate. Sodium dodecylsulphate polyacrylamide gel electrophoresis differentiated between all three isolates of E. cuniculi, with a band at 42–45 kDa present in the murine isolate only, bands at 52 kDa present in the canine and human isolates but not the murine, and a single band at 60 kDa (murine) and 65 kDa (canine) replaced by two bands at 55 and 70 kDa in the human isolate. The 55 kDa and 70 kDa antigens were also revealed as characteristic bands of the human isolate by Western blotting. The study has thus revealed that the species Encephalitozoon cuniculi is not a homogeneous entity.

Notizen: . The nucleotide sequence of the 16S rRNA gene, part of the 23S rRNA gene and the spacer DNA region was determined for Giardia duodenalis, obtained from humans in The Netherlands (AMC-4) and Washington State (CM). These rDNA sequences differ from other G. duodenalis isolates (Portland-1 and BRIS/83/HEPU/106) both of which have virtually identical rDNA sequences. The most characteristic feature was found close to the 5’end of the 16S rRNA. The Portland-1 - Bris/83/HEPU/106 type has GCG in position 22–24, while AMC-4 and CM have AUC in this position. These two sequences, present in an otherwise conserved region of the 16S rRNA, are “signature” sequences, which divide Giardia isolates into two different groups.

Notizen: . Hoplorhynchus acanthatholius, n. sp. is described from Enallagma civile, the Civil Bluet damselfly. Trophozoites are solitary, lie in the mesenteron between the peritrophic membrane and the epithelium, and attain a maximum length of 850 μm. Epimerite ovoid to broadly ovoid; anterior margin bearing eight equidistant retroarcuate hooks; attached to protomerite by means of a vermicular stalk. Protomerite ovoid; deutomerite narrowly obvoid. Gametocysts spherical; diam 300 μm, sporulating by simple dehiscence in 48–72 h. Oocysts are characteristic of Menosporinae: smooth, biconical, crcscentic, uniform in size and shape. Steganorhynchus dunwoodyi, n. g., n. sp. is described from the damselfly Ischnura verticalis. The genus is characterized by an epimerite comprising an ovoid papilla enclosed in a retractable, globular sheath, borne on a long vermicular stalk. Trophozoites are solitary, lie in the mesenteron between the peritrophic membrane and epithelium, and attain a maximum length of 605 μm. Protomerite very broadly ovoid; deutomerite obvoid. Gametocysts spherical; diam 258 μm, sporulating by simple dehiscence in 48–72 h. Oocysts are characteristic of Menosporinae: smooth, biconical, crescentic, uniform in size and shape. The population dynamics of H. acanthatholius and S. dunwoodyi among damselfly populations in five Nebraska localities are presented.

Notizen: . Twelve monoclonal antibodies were raised that are specific for the membrane skeleton of Tetrahymena. Five were directed against T. pyrifomis and seven were directed against T. thermophila. Some cross-reactivity between species was found. Each monoclonal antibody recognized one of the three major components of epiplasm, i.e. the bands A, B, and C identified in electrophoretic separations of epiplasmic proteins. It was found, using these antibodies, that the epiplasmic proteins A, B, and C have overlapping but independent distributions within the cell.

Notizen: . Spores of Nosema sp. NIS M11, primed with 0.1 N KOH solution, were mixed with either physiological salt solutions or IPL-41 medium, an insect cell culture medium, for germination. In the latter medium, only a few spores germinated, while high percentages of spore germination were obtained in physiological salt solutions, particularly in Rinaldini's solution. By using the salt solutions as inoculation media, KOH-primed NIS M11 spores were inoculated into the Spodoptera frugiperda SF21AEII cell line. The initial infection levels were consistently higher than that obtained by using IPL-41 medium. Among the salt solutions, Rinaldini's solution, containing KCl in place of NaCl, gave the highest percentage of initial cell infection. Increased osmolarity of salt solutions did not improve the efficiency of spore germination and infection of N. sp. NIS M11.

Notizen: . Pancreatic digests of casein are major ingredients of media used in the axenic cultivation of lumen-dwelling parasitic protozoa, especially Entamoeba, Giardia, and trichomonads. The digest used almost exclusively in the development of these media, Medo-Peptone (Trypticase® BBL), has not been available since 1981. Moreover, none of dozens of similar type digests tested since then in our laboratory has proved equal to Medo-Peptone, and in the last two years it has become increasingly difficult to obtain new batches which will support even modest growth of Entamoeba histolytica. In response to this problem we have developed a casein-free medium, YI-S, consisting of a nutrient broth, vitamin mixture and serum. We recommend it as a replacement for the casein-dependent medium TYI-S-33, currently the most widely used for axenic culture of Entamoeba histolytica and other lumen-dwellers.

Notizen: . [35S]methionine incorporation into proteins of either T. cruzi epimastigotes or trypomastigotes was drastically inhibited by low concentrations of crystal violet in a dose-dependent manner. This inhibition was not due to ATP depletion since cellular ATP levels did not change significantly after incubation of epimastigotes with 50 μM crystal violet for similar periods of time, and was unaffected by changes in the extracellular free calcium concentration. Although crystal violet was able to inhibit protein synthesis in a cell-free system from T. cruzi epimastigotes, half maximal inhibition was at 1 mM, a concentration three orders of magnitude higher than those that inhibited protein synthesis in intact cells. On the other hand, crystal violet was able to inhibit total [35S]methionine uptake at similar concentrations to those that inhibited protein synthesis while addition of increasing concentrations of cold methionine to the incubation medium protected the cells against crystal violet inhibition. Crystal violet also inhibited total [3H]proline uptake thus indicating that it has a general inhibitory effect upon the transport of amino acids, and not specifically upon methionine. These results indicate that inhibition of protein synthesis by crystal violet is probably due to inhibition of amino acid uptake.

Notizen: . High resolution sampling of the stratified water column in a fjord-like ecosystem revealed a green-pigmented planktonic ciliate that was found to be a ravenous predator of Euglena proxima. The vertical distributions of both predator and prey were coincident, and maximum populations occurred across the transition from oxic to anoxic water. This ciliate was identified as Perispira ovum (family Spathidiidae; Order Haptorida). P. ovum was observed by transmission electron microscopy to retain not only the chloroplasts, but also the mitochondria and paramylon reserve of its algal prey. A mechanism of sequestration of algal organelles is demonstrated for the first time. This mechanism includes: recognition, capture, and ingestion of prey; rupture and release of algal cell contents; and enrobing of individual organelles and paramylon by the host vacuolar membrane. The structural integrity, peripheral location, and association with host endoplasmic reticulum suggests the sequestered organelles may be functional within P. ovum. The occurrence and high biomass of this aerobic ciliate in an oxygen-limited environment also suggests that the sequestered chloroplasts are photosynthetically active and may provide additional substrates (such as oxygen) and metabolic capabilities that are crucial for its survival.

Notizen: . Some strains of P. caudatum contain macronuclear inclusion bodies that are morphologically distinct from bacteria. They vary in number as well as in size in each macronucleus. The inclusion bodies are basically divided into peripheral and inner areas. The peripheral area consists of fibrillar proteins of 22–24 nm in thickness, which are specifically stained with fast green in 45% acetic acid. On the other hand, chromatin-like granules are within the inner area of large inclusion bodies. The granules within the inner area changed their distribution depending upon the physiological state of their host cells. Transplantation experiments and crossbreeding analyses revealed that genetic factors responsible for the multiplication of the inclusion bodies can ‘infect’ other macronuclei (or cells) via the cytoplasm. These results suggest that the inclusion bodies are a non-bacterial macronuclear endosymbiont, possibly produced by a virus or a virus-like element.

Notizen: . The close contact of Giardia lamblia trophozoites with mucosal surfaces produces surface indentations of epithelial cells, that progress to areas of microvilli depletion. This interaction is mediated by the lateral crest, a specialized contractile structure in the ventral disc of the parasite. We have analyzed the plasma membrane of the ventral disc using freeze fracture electron microscopy to study the distribution of large integral proteins, and freeze fracture cytochemistry with filipin to reveal sterol-containing sites. A previously undetected structural specialization was found at the lateral crest as a horseshoe-shaped membrane domain characterized by a near absence of intramembrane particles, in sharp contrast to the remainder of the ventral disc plasma membrane. In addition, the lateral crest showed a striking paucity of cholesterol-containing complexes in replicas of trophozoites treated with filipin. These observations demonstrate that the plasma membrane of G. lamblia displays a microheterogeneity in the planar distribution of cholesterol and intramembrane particles. Both membrane components are noticeably less abundant at the outer rim of the ventral disc where contraction takes place.

Notizen: . Concanavalin A (≥ 50 μg/ml) inhibits pair formation in both of the two complementary mating types of Euplotes octocarinatus studied in this investigation. This effect can be reversed by methyl-α-d-mannose. Concanavalin A is accessible for methyl-α-d-mannose until pairs are formed. Methyl-α-d-mannose as well as methyl-α-d-glucose and 2-acetamido-2-deoxy-d-glucose alone do not inhibit pair formation unless applied in concentrations ≥ 60 mM. The Concanavalin A-sensitive phase of preconjugant interaction starts 2 h after cells are induced to conjugate. Based on these observations we suggest that Concanavalin A might exhibit its action by binding to carbohydrate moieties of preconjugation-specific adhesion molecules and thereby might allosterically block interactions with their counterparts. To identify preconjugation-specific alterations in number or localization of Concanavalin A-binding glycoconjugates, we probed western blots of total cell proteins or fixed cells, respectively, with digoxigenin-labeled Concanavalin A. On Concanavalin A blots 20 different Concanavalin A-binding glycoconjugates were identified in mating-competent cells. Localization of Concanavalin A-binding sites on mating-competent cells by light microscopy resulted in predominant labeling of a comma-shaped structure near the paroral membranelle. During the preconjugation period no changes in number or localization of Con A-binding glycoconjugates were detected. Possible reasons are discussed.

Notizen: The large, free-living amoebae have been widely used as model cells for studying a variety of biological phenomena, including cell motility, nucleocytoplasmic interactions, membrane function, and symbiosis. Results of studies by our group on amoebae as moving cells, as material for micrurgical manipulations, and as hosts for intracellular symbionts are summarized here. In particular, our recent studies of the amoeba as a microcosm, in which spontaneously infecting foreign microbes have become integrated as necessary cell components, are described in some detail. These processes have involved an initial microbial infection, mutual adaptation by the host and symbionts, and development of obligatory symbiosis. Evidence is presented to show that symbiont-derived macromolecules are involved in the protection of symbionts from digestion, the symbionts have acquired regulatory elements on their chromosomal genes to enhance production of beneficial gene products, and symbionts apparently utilize host-derived macromolecules to their benefit. These studies involved morphological observations both at light and electron microscopic levels, physiological and genetic studies, production and use of poly- and monoclonal antibodies, and molecular-biological approaches including gene cloning and sequencing. It is shown that amoebae are uniquely suited as model cells with which to study these phenomena.

Notizen: Two distinct sequevars, denoted Pc1 and Pc2, of the opportunistic pathogen Pneumocystis carinii have been previously identified based on the sequence of their 26S rRNA genes, the location of group I self-splicing introns and pulsed field electrophoretic patterns of chromosomal DNA. This study shows that the sequences of 16S and 5.8S rRNA genes also vary between these sequevars, and that greater variation was seen in the internal transcribed spacer regions. Polymerase chain reaction and restriction analysis can distinguish between these sequevars.

Notizen: Trifluralin, a herbicide which is known to bind to plant and algal tubulin, induced ultrastructural changes in the microtubules of the mature Plasmodium falciparum gametocytes in vitro. Trifluralin treatment led to disassembly of the well ordered subpellicular microtubules, whereas it had no effect on microtubules of human platelets or of rat neuronal cells in vitro. The disassembled microtubules showed fragmented large tubular structures, which frequently were associated with the pellicular membranes. Electron microscopic autoradiography showed radioactive trifluralin associated with the microtubule fragments. These results provide evidence that trifluralin selectively binds to microtubules in malaria parasites and causes disruption of their structure.

Notizen: Although Paramecium has been widely used as a model sensory cell to study the cellular responses to thermal, mechanical and chemoattractant stimuli, little is known about their responses to chemorepellents. We have used a convenient capillary tube repellent bioassay to describe 4 different compounds that are chemorepellents for Paramecium and compared their response with those of Tetrahymena. The classical Paramecium t-maze chemokinesis test was also used to verify that this is a reliable chemorepellent assay. The first two compounds, GTP and the oxidant NBT, are known to be depolarizing chemorepellents in Paramecium but this is the first report of them as repellents in Tetrahymena. The second two compounds, the secretagogue alcian blue and the dye cibacron blue, have not previously been described as chemorepellents in either of these ciliates. Two other compounds, the secretagogue AED and the oxidant cytochrome c, were found to be repellents to Paramecium but not to Tetrahymena. The repellent nature of each of these compounds is not related to toxicity because cells are completely viable in all of them. More importantly, all of these repellents are effective at micromolar to nanomolar concentrations, providing an opportunity to use them as excitatory ligands in future works concerning their membrane receptors and possible receptor operated ion channels.

Notizen: . The DNA sequences of a portion of the 5-enolpyruvyl shikimate phosphate synthase domain of the arom gene, encoding the pentafunctional AROM protein, were determined from isolates of Pneumocystis carinii from five mammalian host species (rat, human, ferret, rabbit and mouse). High levels of genetic divergence were found among P. carinii derived from different hosts species, 7–22% at the DNA sequence level, and 7–26% at the derived amino acid sequence level. Two separate and distinct sequences were isolated from infected ferret lungs. Low levels of divergence were seen in human-derived organisms.

Notizen: . In hypotrichous ciliates such as Euplotes crassus genes in the transcriptionally active macronucleus are present on individual minichromosomes which occur in gene-specific copy numbers. This different degree of gene amplification can be understood as a means to preset the expression potential of the respective genetic information. In addition, the actual steady state transcript amounts are governed by the transcription rates and transcript stabilities. To establish the relative effects of these three parameters the copy numbers of genes transcribed by the three different polymerases were determined. The transcript levels of growing or starving vegetative cells were then determined, and nuclear run-on assays were performed to determine the transcription rates of the genes in the different nutritional states. A weak correlation between the gene copy numbers and transcription rates was found. The transcripts of genes synthesized by RNA polymerase II exhibited different stabilities upon starvation of the cells, compared to the supposedly stable ribosomal 5S and 26S RNA. Refeeding of the cells after starvation also resulted in a differential response with respect to the accumulation of the transcripts of different genes transcribed by RNA polymerase II, which can be interpreted in the context of the gene functions.

Notizen: . Tritrichomonas foetus is a flagellated protozoon found in urogenital tract of cattle. Its free movement in liquid medium is powered by the coordinated movement of three flagella projecting towards the anterior region of the cell, and one recurrent flagellum that forms a junction with the cell body and ends as a free projection in the posterior region of the cell. We have used video microscopy and digital image processing to analyze the relationships between the movements of these flagella. The anterior flagella beat in a ciliary type pattern displaying effective and recovery strokes, while the recurrent flagellum beats in a typical flagellar wave form. One of the three anterior flagella has a distinctive pattern of beating. It beats straight in its forward direction as opposed to the ample beats performed by the others. Frequency measurements obtained from cells swimming in a viscous medium shows that the beating frequency of the recurrent flagelium is approximate twice the frequency for the three anterior flagella. We also observed that the costa and the axostyle do not show any active motion. On the contrary, they form a cytoskeletal base for the anchoring and orientation of the flagella.

Notizen: Book reviewed in this article: Aescht, E. (ed.) 1994. Die Urtiere: eine verborgene Welt. Foissner, W., Berger, H., Blatterer, H. & Kohmann, F. 1995. Taxonomische und okologische Revision der Ciliaten des Sap-robiensystems. Band IV: Gymnostomatea, Loxodes, Suctoria.

Notizen: . The order Euplotida represents a monophyletic order of five families of hypotrich ciliates united by morphology, stomatogenesis, ultrastructure, cyst structure, and behavior. A review of variability of ciliation and nuclei among the 14 genera suggests that lines of evolution may have involved both the loss of cirri and nuclear simplification. We present a binary key to genera in the families Aspidiscidae (Aspidisca and Euplotaspis), Certesiidae n. fam. (Certesia), Gastrocirrhidae (Cytharoides, Euplotidium, and Gas-trocirrhus), Uronychiidae (Diophryopsis, Diophrys, Paradiophrys, and Uronychia), and Euplotidae. The latter family contains species formerly in the genus Euplotes. Based primarily on cortical structure, endosymbionts, data from morphometric analysis, and ecology, we recognize four different groups. The first group of species remains in Euplotes with Euplotes charon as type. We place a second group of species into the genus Moneuplotes Jankowski 1979 with Moneuplotes vannus (Müller, 1786) as type. We erect two new genera: Euplotoides n. g. and Euplotopsis n. g. with Euplotoides patella (Müller, 1773) n. comb. and Euplotopsis affinis (Dujardin, 1841) n. comb. as type species respectively. We discuss possible phylogenetic relationships within the order.

Notizen: . The ultrastructure of two species of Holomastigotoides is presented. The basic unit of organization of these large cells is the flagellar band. Each flagellar band consists of a row of flagellar basal bodies linked by three fiber systems. The number of flagellar bands is species dependent. The flagellar bands originate at the cell apex and are arranged in parallel spirals of increasing gyre, thus defining the conical shape of the cell. In the cell apex a striated root called a parabasal fiber is juxtaposed with the basal bodies of each flagellar band. Linear extensions of two parabasal fibers function as the spindle poles for the persistent extra-nuclear spindle. The nucleus is in close contact with the spindle poles and spindle microtubules. Parallel sheets of microtubules which constitute axostyles are nucleated along the underside of the parabasal fibers. The axostyles extend away from the cell apex, with many reaching the basal region of the cell. Some of the axostyles follow the spiral pattern of the flagellar bands. Numerous Golgi bodies are spaced regularly along the flagellar bands. Together the parabasal fiber, axostyles and Golgi bodies associated with a flagellar band are termed a parabasal complex.

Notizen: . A new species of myxosporean from the gill filaments of the freshwater teleost fish, Acestrorhynchus falcatus collected in the Amazon river is described from light and transmission electron microscope observations. The mature spores (total length 32.3 [30.7–35.1] μ) and all developmental stages were found in the same sporogonic plasmodium. The ellipsoidal spore body consists of 2 unequal shell valves adhering together along the suture lines. Each valve, tapering as a caudal projection, forms a long tail (length 20.5 [18.0–21.7] μm). The tail was surrounded by a homogeneous sheath on its length. The polar capsules measuring 3.1 × 1.2 μm contain 3–4 coils of the polar filament. All surfaces of the immature and mature spores were surrounded by a closely adherent homogenous structural sheath, mainly thicker around the tails. The taxonomic affinities of this parasite to other species are discussed.

Notizen: . The mechanisms that signal gregarine excystation are unknown. Previous authors have demonstrated that gregarine sporozoites excyst from their surrounding oocyst in response to stimuli contained in host digestive fluids, but the role of host intestinal pH in this signaling system has not been investigated. In this study, an in vitro assay is used to quantify the effects of 3 pH levels (6.1, 7.0, 8.0) on the excystation of two gregarine species, Gregarina cuneata and Gregarina polymorpha. Both gregarine species excyst at all three pH levels, but there are significant within and among species differences in excystation rate and cumulative excystation over time. Gregarina cuneata excysts more rapidly at pH 6.0 and G. polymorpha excysts more rapidly at pH 8.0. Cumulative excystation is maximized at pH 6.0 for G. cuneata and at pH 7.0 for G. polymorpha. Hydrogen ion mediated excystation may lead to the formation of foci for subsequent establishment or migration and may play a role in parasite site specificity.

Notizen: . Giardia lamblia trophozoites undergo antigenic variation by modulating the expression of variant-specific surface proteins (VSP), which are encoded by a number of small multigene families. We characterized the genomic copy of the VSP gene expressed by the cloned trophozoite line H7, derived from the isolate GS/M, in addition to a related, but nonexpressed, family member. The coding regions of the two genes encode closely related polypeptides (86% identity). However, differences in the coding region of these genes reside solely in an 873 bp segment. Only four differences were found between the 5’flanking sequences (465 bp). The proximal 205 base pairs downstream from the coding regions were identical, but thereafter the sequences diverged (37% identity over the next 391 bp). Mapping studies indicated that no other VSP gene was located within 4 kb pairs of the expressed H7 VSP gene, and transcripts from the nonexpressed gene were detected in neither GS/H7 nor heterogeneous trophozoites populations derived from this cloned line. Any mechanisms responsible for the differential expression of VSP genes must reconcile the near identity of DNA sequences that flank the coding regions of expressed and nonexpressed VSP genes.