Natural Chagas disease in four baboons

Background  Chagas disease is common in Central and South America and the southern United States. The causative agent is Trypanosoma cruzi (order Kinetoplastida, family Trypanosomatidae), a kinetoplastid protozoan parasite of humans and other vertebrates. It is a serious public health issue and the leading cause of heart disease and cardiovascular death in Central and South America. In 1984, a colony baboon was discovered to be infected with T. cruzi .
Methods  As the initial diagnosis was made by microscopic observation of the amastigote forms of T. cruzi in myocardial fibers, T. cruzi amastigotes have been identified in three additional baboons.
Results  The primary findings were similar in all four baboons and were congestive heart failure with edema of dependent areas, hydrothorax, hydropericardium, and multifocal to diffuse lymphoplasmacytic myocarditis.
Conclusions  A baboon animal model of Chagas disease could contribute significantly to the development of therapies for the disease in humans.     

Vertical Micro‐Zonation of Testate Amoebae and Ciliates in Peatland Waters in Relation to Potential Food Resources and Grazing Pressure

The aim of this study was to examine the community structure and vertical micro‐distribution of testate amoebae and ciliates in a raised bog in eastern Poland, as well as to assess the influence of potential food resources (Chl‐a, bacteria, heterotrophic flagellates) and predators (rotifers and copepods) on protozoa communities. Samples were taken from surface, bottom and interstitial waters. At each type of micro‐habitat and each sampling date water was sampled using a plexiglass corer or mini‐piezometers. Additionally, in order to evaluate grazing pressure, field enclosures were used in which metazoan abundance and composition was manipulated by size‐fractionation. Over experiments, metazoan populations shifted from dominance of rotifers to copepods. In the first experiment, with rotifers dominating, metazoa had only a modest predatory impact on the protozoa. In contrast, the second experiment, with copepods prevailing, demonstrated a clear top‐down control of the protozoa communities by metazoan. The density and biomass of protozoa significantly differed between the studied stations, with the lowest numbers in the interstitial water and the highest in the surface water. Surface sampling were dominated by mixotrophic taxa, whereas the deepest sampling level was characterized by increase in the proportion of bacterivore species. These differences between micro‐habitats may be due to differences in environmental conditions (food resources and grazing pressure). Ordination analysis revealed that bacteria can strongly regulate the abundance and taxonomic composition of protozoa in the bottom and interstitial waters. Metazoan predators could be the main regulators of protozoa communities in surface water. (© 2010 WILEY‐VCH Verlag GmbH & Co. KGaA, Weinheim)     

Improved one‐step steam pretreatment of SO2‐Impregnated softwood with time‐dependent temperature profile for ethanol production

In the production of ethanol from lignocellulosic material, pretreatment of the raw material before enzymatic hydrolysis and fermentation is essential to obtain high overall yields of sugar and ethanol. Two‐step steam pretreatment results in higher ethanol yields from softwood than the standard one‐step pretreatment process. However, the difficulty with separation and washing of the material at high pressure between the two pretreatment steps is a major drawback. In this study, a new one‐step pretreatment procedure was investigated, in which the time‐temperature profile was varied during pretreatment. The efficiency of pretreatment was assessed by performing simultaneous saccharification and fermentation on the pretreated slurries. Pretreatment of SO₂‐impregnated softwood performed by varying the temperature (190–226°C), the residence time (5–10 min), and the mode of temperature increase (linear or stepwise), resulted in recovery of about 90% of the mannose and glucose present in the raw material. The highest ethanol yield, 75% of theoretical based on the glucan and mannan content of the raw material, was obtained at pretreatment conditions of 190°C for 12 min. Similar ethanol yields were achieved when running the pretreatment as one‐step (190–200°C), two levels of temperature, at shorter residence time (7 min), which results in lower capital costs for the process. © 2010 American Institute of Chemical Engineers Biotechnol. Prog., 2010     

Cell pairing and methylation in Tetrahymena thermophila are altered by exogenous homocysteine

Homocysteine is causally associated with birth defects such as spina bifida, and with premature vascular disease. We have investigated the effects of homocysteine on a cell-cell interaction in a fundamental eukaryotic system, the free-living ciliate Tetrahymena. Exogenously added homocysteine inhibits cell pairing in a dose-dependent manner. These effects are exacerbated by adenosine, which by itself has little demonstrable influence on pairing. S-adenosylhomocysteine (SAH) is a product of the reaction between adenosine and homocysteine, and is an inhibitor of methyl transferases. We therefore predicted that protein methylation would be significantly inhibited by homocysteine. A direct test of that hypothesis involved a demonstration that incorporation of an isotopically labeled methyl group from methionine into proteins was significantly reduced by homocysteine. The undermethylated proteins are of low molecular weight, and might correspond to known methylatable signaling proteins. We show that vanadate, an inhibitor of protein phosphatase, also inhibits cell pairing, and that the effects of vanadate and homocysteine are additive. This is the first demonstration that methylation and possibly phosphorylation play a regulatory role in cell-cell interactions in ciliates.     

Pelagostrobilidium liui n. sp. (Ciliophora,Choreotrichida) from the Coastal Waters of Northeastern Taiwan and an Improved Description of Pelagostrobilidium minutum Liu et al., 2012

The number of somatic kineties in Pelagostrobilidium ranges from 4 to 6 according to the present state of knowledge. This study investigates Pelagostrobilidium liui n. sp. using live observation, protargol stain, and small subunit rDNA data sequencing. Pelagostrobilidium liui n. sp. is characterized by having a spherical‐shaped body, four somatic kineties, with kinety 2 spiraled around the left side of body, about six elongated external membranelles, and invariably no buccal membranelle. It differs from its most similar congener, Pelagostrobilidium minutum Liu et al., 2012 , in (i) cell shape; (ii) macronucleus width; (iii) oral apparatus; (iv) anterior orientation of kinety 2; (v) location where kinety 2 commences; (vi) arrangement of kinety 1; (vii) distance between the anterior cell end and the locations where kineties commence; and (viii) the presence of 12 different bases (including two deletions) in the small subunit rDNA sequences. The diagnosis of P. minutum Liu et al., 2012 is also improved to include the following new characteristics: invariably four somatic kineties; kineties 2 and 4 alone commence at the same level; kinety 2 originates from right anterior cell half on ventral side, extends sinistrally posteriorly, over kinety 1, around left posterior region, terminates near posterior cell end on dorsal side; kinety 1 commences below anterior third of kinety 2.     

Influence of rumen protozoa on methane emission in ruminants: a meta-analysis approach

A meta-analysis was conducted to evaluate the effects of protozoa concentration on methane emission from ruminants. A database was built from 59 publications reporting data from 76 in vivo experiments. The experiments included in the database recorded methane production and rumen protozoa concentration measured on the same groups of animals. Quantitative data such as diet chemical composition, rumen fermentation and microbial parameters, and qualitative information such as methane mitigation strategies were also collected. In the database, 31% of the experiments reported a concomitant reduction of both protozoa concentration and methane emission (g/kg dry matter intake). Nearly all of these experiments tested lipids as methane mitigation strategies. By contrast, 21% of the experiments reported a variation in methane emission without changes in protozoa numbers, indicating that methanogenesis is also regulated by other mechanisms not involving protozoa. Experiments that used chemical compounds as an antimethanogenic treatment belonged to this group. The relationship between methane emission and protozoa concentration was studied with a variance−covariance model, with experiment as a fixed effect. The experiments included in the analysis had a within-experiment variation of protozoa concentration higher than 5.3 log₁₀ cells/ml corresponding to the average s.e.m. of the database for this variable. To detect potential interfering factors for the relationship, the influence of several qualitative and quantitative secondary factors was tested. This meta-analysis showed a significant linear relationship between methane emission and protozoa concentration: methane (g/kg dry matter intake)=−30.7+8.14×protozoa (log₁₀ cells/ml) with 28 experiments (91 treatments), residual mean square error=1.94 and adjusted R²=0.90. The proportion of butyrate in the rumen positively influenced the least square means of this relationship.     

Morphological and Molecular Characterization of a New Species of Stephanopogon,Stephanopogon pattersoni n. sp.

Stephanopogon is a taxon of multiciliated protists that is now known to belong to Heterolobosea. Small subunit ribosomal DNA (SSU rDNA) phylogenies indicate that Stephanopogon is closely related to or descended from Percolomonas, a small tetraflagellate with a different feeding structure, thus these morphologically dissimilar taxa are of ongoing evolutionary interest. A new strain of Stephanopogon, KM041, was cultured, then characterized by light microscopy, electron microscopy, and SSU rDNA sequencing. KM041 is 18–35 μm (mean 26.8 μm) long, with six main ventral ciliary rows, one ventro‐lateral ciliary row, and three anterior barbs. It closely resembles Stephanopogon minuta Lei et al. 1999 in morphology, and is very closely related to an extinct culture “S. aff. minuta”, yet is markedly dissimilar in SSU rDNA sequence from a different isolate identified as S. minuta. This confirms that there are at least two distinct lineages of S. minuta‐like cells, and we describe KM041 as a new species, Stephanopogon pattersoni n. sp. The ultrastructure of KM041 resembles that of previously studied Stephanopogon species, though it has a novel paraxonemal structure in a few cilia. We note that a sub‐basal‐body pad and bulbous axosome are unlikely to be apomorphies for the Stephanopogon–Percolomonas clade.     

Morphological and Molecular Characterisation of Notosolenus urceolatus Larsen and Patterson 1990, a Member of an Understudied Deep‐branching Euglenid Group (Petalomonads)

Petalomonads are particularly important for understanding the early evolution of euglenids, but are arguably the least studied major group within this taxon. We have established a culture of the biflagellate petalomonad Notosolenus urceolatus, and conducted electron microscopy observations and molecular phylogenetic analysis. Notosolenus urceolatus has eight pellicular strips bordered by grooves and underlain by close‐set microtubules. There are ventral and dorsal Golgi bodies. Mitochondria apparently contain fibrous inclusions, as in Petalomonas cantuscygni. A previously undocumented type of large, globular extrusome is present instead of the tubular extrusomes characteristic of Euglenozoa. The feeding apparatus lacks rods and vanes, and is partly supported by an “MTR”. The flagella have complex transition zones that are extremely elongated but unswollen. Only the emergent portion of the anterior flagellum has an organised paraxonemal rod, and also has very fine mastigonemes. The basal bodies are offset and lack connecting fibres. 18S rRNA gene phylogenies show that N. urceolatus is closely related to Petalomonas sphagnophila and P. cantuscygni, not Notosolenus ostium, confirming that current generic assignments based on the number of emergent flagella are phylogenetically unreliable, and making it difficult to infer whether features shared by N. urceolatus and P. cantuscygni (for example) are general for petalomonads.