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PubMed2004

Phylum-specific environmental DNA analysis reveals remarkably high global biodiversity of Cercozoa (Protozoa).

This study presents the first 18S rRNA multi-library environmental PCR survey of a single protozoan phylum, Cercozoa Cavalier-Smith 1998, from a range of different habitats. Phylogenetic analysis reveals at least nine novel clades within the phylum, several possibly at the level of order or above. Further experiments are described to ascertain the true ecological and geographical distributions of some clades that might be inferred from the tree to be restricted in either or both ways. These results suggest that the diversity of cercozoan taxa may run into thousands of lineages, making it comparable in diversity to the largest better-characterized protozoan phyla, e.g. Ciliophora (ciliates and suctorians) and Foraminifera. New sequences of cultured Spongomonas, Metromonas and Metopion are also presented. In the light of these additions, and the increased taxon sampling from the environmental libraries, some revisions of cercozoan classification are made: the transfer of Spongomonadea from Reticulofilosa to Monadofilosa; the removal of Metopiida from Sarcomonadea; and the creation of the new order Metromonadida, currently containing the single genus Metromonas. Although Metromonas groups with weak to moderate support with Chlorarachnea, it is here placed in superclass Monadofilosa, to which it is morphologically more similar.

Animals↗
PubMed2004

Evolution of Hsp90 expression in Tetrahymena thermophila (Protozoa, Ciliata) populations exposed to thermally variable environments.

Evolutionary consequences of thermally varying environments were studied in the ciliated protozoan Tetrahymena thermophila. Replicated lines were propagated for 60 days, a maximum of 500 generations, in stable, slowly fluctuating (red spectrum), and rapidly fluctuating (blue spectrum) temperatures. The red and blue fluctuations had a dominant period length of 15 days and two hours, respectively. The mean temperature of all time series was 25 degrees C and the fluctuating temperatures had the same minimum (10 degrees C), maximum (40 degrees C), and variance. During the experiment, population sizes and biomasses were monitored at three-day intervals. After the experiment, carrying capacity and maximum growth rate were measured at low (15 degrees C), intermediate (25 degrees C), and high (35 degrees C) temperatures for each experimental line. Physiological changes in the lines were assessed by measuring the expression of stress-induced heat shock protein Hsp90 at 25 degrees C, 35 degrees C, and 39 degrees C. Population sizes and biomasses showed no differences between stable, blue, or red temperature treatments during the experiment. Also, after the experiment, mean carrying capacities and maximum growth rates were comparable in the stable, blue, and red temperature treatments. The expression of Hsp90 was higher in lines from the blue environment than in lines from the stable environment. Lines from the red environment had an intermediate level of Hsp90 expression. This supports the hypothesis that inducible thermotolerance and expression of canalizing genes can evolve in response to rapidly varying environments. Furthermore, we found correlative evidence of benefits and disadvantages of high Hsp90 expression. Lines with high expression of Hsp90 had an increased growth rate at the highest temperature when food resources were not limiting growth. At low and intermediate temperatures the same lines had the lowest carrying capacities.

Analysis of Variance↗
PubMed1982

The relative efficiency of various fluids in the rapid freezing of protozoa.

The cooling efficiencies of various fluids at low temperature were compared by measuring the temperature decay in 3 microliter water samples plunged into them. A simple model of cooling was used in order to discuss the results. Liquid ethane was found to produce a cooling rate of 660 KS-1, about twice that of liquid propane, while ethanol was almost as effective as ethane between 273 to 223 K. The heat-transfer coefficient of liquid ethane was estimated to be between 1500 and 5000 W m-2 K-1, depending on the physical state assumed for the water sample. Samples of flagellated organisms, after being frozen rapidly in the above way, were freeze-substituted by the method of Barlow & Sleigh (1979). Although this fixation did not give good definition of the microtubules of the flagellar axoneme, it exhibited reasonable tissue preservation in thin sections of the cell body. The fixation method resulted in preserved flagellar wave shapes, which were observed under the light microscope and in critical-point dried cells examined by scanning and conventional electron microscopy. It was concluded (a) that methods for preserving the wave shape of the flagellum and for preserving its internal structure may not be compatible, and (b) that although the present cooling method (with ethane) approaches the speed required to arrest a flagellar wave, further improvements in the speed of the method are desirable.

Animals↗
PubMed2004

Rafts and sphingolipid biosynthesis in the kinetoplastid parasitic protozoa.

Summary relatively large rafts are a feature of activated mammalian cells. These studies allow us to consider the functional role of lipid rafts in kinetoplastid parasites, which are particularly rich in lipid-anchored surface molecules. Morphological, biochemical and genetic studies indicate that lipid rafts (and sphingolipid biosythesis) are important in the differentiation of extracellular Leishmania to mammalian-infective metacyclic promastigotes, perhaps orchestrating the clearly observable reorganization of the plasma membrane during this process that leads to an activated metacyclic primed for invasion. However, the first step in the sphingolipid biosynthetic pathway (mediated by serine palmitoyltransferase), and at least regulated, de novo sphingoid base and ceramide synthesis, are not essential for the pathogenesis of intramacrophage Leishmania amastigotes.

Animals↗
PubMed1998

Inactivation of viruses, bacteria, protozoa, and leukocytes in platelet concentrates.

Despite the increased safety of blood achieved through continued improvements in donor testing, concern remains about the safety of blood components. Transfusion of cellular components has been implicated in transmission of viral, bacterial, and protozoan diseases. While it is commonly recognized that hepatitis B virus (HBV), hepatitis C virus (HCV), cytomegalovirus (CMV), and the retroviruses, such as human immunodeficiency virus (HIV) and the human lymphotrophic viruses (HTLV) can be transmitted through cellular components, other pathogens are emerging as potentially significant transfusion-associated infectious agents. For example, transmission of protozoan infections due to trypanosomes and babesia have been reported. In addition to viral and protozoal infectious agents, bacterial contamination of platelet concentrates continues to be reported; and may be an under reported transfusion complication. More importantly, new infectious agents may periodically enter the donor population before they can be definitively identified and tested for to maintain consistent safety of the blood supply. The paradigm for this possibility is the HIV pandemic which erupted in 1979. During the past decade a number of methods to inactivate infectious pathogens in blood components, including platelets, have been developed. This technology is now entering the clinical trial phase.

Animals↗