Cannibalism and Coprophagy Are Modes of Transmission of Blastocrithidia triatomae (Trypanosomatidae) between Triatomines

Transovarial transmission was not detectable among Blastocrithidia triatomae- infected Triatoma infestans . Rather, B. triatomae was transmitted directiy between triatomines by cannibalism and coprophagy. Cannibalism conditions that excluded coprophagy always resulted in an infection of Dipetalogaster maxima . The efficiency of transmission was not influenced by the blood source—mice or chickens—fed to the infected donor bugs although chicken blood lyses the epimastigotes of the stomach population. Triatoma infestans was infected by coprophagy only if fed, not if unfed. Blastocrithidia triatomae in dry feces was taken up only if the feces were redissolved in fresh feces. Infections also appeared in groups of bugs fed on chickens previously used for feeding infected bugs.     

Thrust reversal by tubular mastigonemes: immunological evidence for a role of mastigonemes in forward motion of zoospores ofPhytophthora cinnamomi

Summary The role of tubular mastigonemes in the reversal of thrust of the anterior flagellum ofPhytophthora cinnamomi was analysed using mastigoneme-specific monoclonal antibodies and immunoflu-orescence and video microscopy. Exposure of live zoospores ofP. cinnamomi to the mastigoneme-specific Zg antibodies caused alterations in the arrangement of mastigonemes on the flagellar surface and at Zg concentrations above 0.3 /ml, mastigonemes became detached from the flagellum. As a consequence of antibody binding to the mastigonemes there were concentration-dependent perturbations in zoospore swimming behaviour and anterior flagellum beat pattern. With increasing antibody concentration zoospores swam more slowly and other parameters of their swimming pattern, such as the wavelength of the swimming helix and the frequency of rotation, were also reduced. The effects of Zg antibodies were specific at two levels: control immunoglobulins or antibodies that bound to other flagellar surface components did not have an effect on motility, and Zg antibodies did not interfere with the motility of zoospores of oomycete species to which they did not bind. The effects of antibody-induced disruption of mastigoneme arrangement strongly support previous hypotheses that tubular mastigonemes are responsible for thrust reversal by the anterior flagellum, enabling it to pull the cell through the surrounding medium.     

Naegleria: A Research Partner For Cell and Developmental Biology1

ABSTRACT. Organisms in the genus Naegleria offer special opportunities for research in contemporary biology. the dramatic cell differentiation from amebae to flagellates is unique among eukaryotes in the rapidity, synchrony, reproducibility, homogeneity, and accessibility of a major phenotypic change. Environmental signals initiate a progressive signal transduction pathway in which genes are turned on, including those for several calcium-binding proteins, and newly synthesized proteins become localized in newly assembled organelles, including the centriole-like basal bodies, with the overall consequence that the cell changes its shape, motility, and behavior. This essay reviews research opportunities for which Naegleria excels, as well as interesting aspects of its biology that provide challenges for future investigations. Because these organisms alternate between two major eukaryotic motility forms, their phylogenetic position is also provocative. Although there are hints that Naegleria is capable of sexual reproduction in nature, mating has not yet been observed in the laboratory. In order to fully exploit the opportunities offered by this wonderful experimental system we are working to develop means to do genetic manipulation, in particular via DNA-mediated transformation.     

Effect of organic matter on growth and cell yield of ammonia-oxidizing bacteria

The effect of various organic compounds on the growth of ammonia-oxidizing bacteria was examined.Nitrosococcus oceanus, a strongly halophilic bacterium, had a very low tolerance to organic matter compared with other organisms tested. Organic compounds scarcely affected the growth of theNitrosomonas strains whereas nitrite formation by bothNitrosococcus mobilis strains was inhibited by nearly all of the substances tested. The growth ofNitrosospira strain Nsp₁ was enhanced more than 30% by acetate and formate, but not growth was detectable in the presence of pyruvate. On the contrary,Nitrosospira strain Nsp₅ was stimulated only by pyruvate. Nitrite formation by the twoNitrosovibrio tenuis strains tested was similar. The growth of both strains was enhanced considerably by formate and glucose; acetate and, to a greater extent, pyruvate inhibited these bacteria.In batch culture, the energy efficiency of autotrophically grown ammonia-oxidizing bacteria varied from strain to strain. The cell yield of mixotrophically grown cultures, per unit of ammonia oxidized, was increased in comparison with autotrophic ones. No heterotrophic growth was detected.     

Combining Molecular Data with Classical Morphology for Uncultured Phagotrophic Euglenids (Excavata): A Single‐Cell Approach

Phagotrophic euglenids are one of the most diverse and important forms of heterotrophic flagellates in sediment systems, and are key to understanding the evolution of photosynthetic euglenids and ‘primary osmotrophs’, yet relatively little is known about their biodiversity and phylogenetic relationships. A wealth of light microscopy‐based information is available, but little progress has been made in associating this with molecular sequence data. We established a protocol to obtain light microscopy data and molecular data from single euglenid cells isolated from environmental samples. Individual cells from freshwater and marine benthic samples were isolated and rinsed by micropipetting, documented using high‐resolution photomicroscopy, then subjected to single‐cell nested PCR using taxon‐specific primers in combination with universal eukaryotic primers, generating > 75% or full‐length SSU rDNA sequences. As a proof‐of‐principle eight individuals were characterised and subjected to phylogenetic analyses. Many of these cells were identified as Anisonema or Dinema, and grouped with existing sequences assigned to these taxa, and with a ‘Peranema sp.’ sequence that we could now clearly demonstrate was misidentified or misannotated. Another cell is Heteronema c.f. exaratum, the first ‘skidding heteronemid’ for which sequence data are available. This is not closely related to Heteronema scaphurum, and intriguingly, branches as the sister group to primary osmotrophs. A cell similar to Ploeotia vitrea (the type of this genus), shows no particular phylogenetic affinity to Ploeotia costata, the best studied Ploeotia species. Our experimental protocol provides a useful starting point for future analyses on euglenid biodiversity (including environmental sequence surveys), and their evolution and systematics.     

Elucidating the evolution and diversity of Uroglena-like colonial flagellates (Chrysophyceae): polyphyletic origin of the morphotype

ABSTRACT

The Uroglena-like morphotype represents a prototype of a colonial naked chrysophyte, comprising plastid-bearing cells that are arranged as the surface monolayer of the spherical colony. So far, insufficient molecular characterization appears to be the most significant brake on the modern taxonomic revision of this ecologically and morphologically coherent group of organisms. The general aim of this work was to conduct a modern taxonomic revision of Uroglena-like flagellates by using combined molecular, morphological and ultrastructural methodology, complemented by exploring type localities of Uroglena volvox and Uroglenopsis americana in Europe and North America, respectively. On the basis of phylogenetic analysis of concatenated nuclear SSU rDNA and plastid rbcL sequences we show that Uroglena-like colonial flagellates form three genetically and morphologically distinct lineages within the Ochromonadales (Chrysophyceae), distinguished here as Uroglena, Uroglenopsis and Urostipulosphaera gen. nov. The taxonomic status of the other chrysophyte genera with spherical colonies is discussed in light of our findings.     

Changes in the temperature sensitivity of SOM decomposition with grassland succession: implications for soil C sequestration

Understanding the temperature sensitivity (Q₁₀) of soil organic matter (SOM) decomposition is important for predicting soil carbon (C) sequestration in terrestrial ecosystems under warming scenarios. Whether Q₁₀ varies predictably with ecosystem succession and the ways in which the stoichiometry of input SOM influences Q₁₀ remain largely unknown. We investigate these issues using a grassland succession series from free‐grazing to 31‐year grazing‐exclusion grasslands in Inner Mongolia, and an incubation experiment performed at six temperatures (0, 5, 10, 15, 20, and 25°C) and with four substrates: control (CK), glucose (GLU), mixed grass leaf (GRA), and Medicago falcata leaf (MED). The results showed that basal soil respiration (20°C) and microbial biomass C (MBC) logarithmically decreased with grassland succession. Q₁₀ decreased logarithmically from 1.43 in free‐grazing grasslands to 1.22 in 31‐year grazing‐exclusion grasslands. Q₁₀ increased significantly with the addition of substrates, and the Q₁₀ levels increased with increase in N:C ratios of substrate. Moreover, accumulated C mineralization was controlled by the N:C ratio of newly input SOM and by incubation temperature. Changes in Q₁₀ with grassland ecosystem succession are controlled by the stoichiometry of newly input SOM, MBC, and SOM quality, and the combined effects of which could partially explain the mechanisms underlying soil C sequestration in the long‐term grazing‐exclusion grasslands in Inner Mongolia, China. The findings highlight the effect of substrate stoichiometry on Q₁₀ which requires further study.     

Phytate as the sole phosphate source for growth of Tetrahymena

Phytate, the storage form of phosphate in seeds and grains, is a major form of environmental phosphate loading from fertilizer inputs and agricultural runoff. We have investigated the ability of Tetrahymena populations to grow on phytate as their sole phosphate source. Populations grew equally well in chemically defined medium with phosphate and medium in which the phosphate was replaced with phytate in comparable concentrations between 0.5 mM and 6 mM. Intracellular phytate concentrations of cells grown in phytate showed a 4-6-fold increase over those grown in phosphate when measured during the late stage of exponential growth. These results demonstrate that phytate can provide a source of adequate phosphate for sustained growth in phytate-rich environments.     

Structure of the bc1 complex from Seculamonas ecuadoriensis,a jakobid flagellate with an ancestral mitochondrial genome

In eubacteria, the respiratory bc(1) complex (complex III) consists of three or four different subunits, whereas that of mitochondria, which have descended from an alpha-proteobacterial endosymbiont, contains about seven additional subunits. To understand better how mitochondrial protein complexes evolved from their simpler bacterial predecessors, we purified complex III of Seculamonas ecuadoriensis, a member of the jakobid protists, which possess the most bacteria-like mitochondrial genomes known. The S. ecuadoriensis complex III has an apparent molecular mass of 460 kDa and exhibits antimycin-sensitive quinol:cytochrome c oxidoreductase activity. It is composed of at least eight subunits between 6 and 46 kDa in size, including two large "core" subunits and the three "respiratory" subunits. The molecular mass of the S. ecuadoriensis bc(1) complex is slightly lower than that reported for other eukaryotes, but about 2x as large as complex III in bacteria. This indicates that the departure from the small bacteria-like complex III took place at an early stage in mitochondrial evolution, prior to the divergence of jakobids. We posit that the recruitment of additional subunits in mitochondrial respiratory complexes is a consequence of the migration of originally alpha-proteobacterial genes to the nucleus.