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PubMed2003

Phospholipids in parasitic protozoa.

Parasitic protozoa are surrounded by membrane structures that have a different lipid and protein composition relative to membranes of the host. The parasite membranes are essential structurally and also for parasite specific processes, like host cell invasion, nutrient acquisition or protection against the host immune system. Furthermore, intracellular parasites can modulate membranes of their host, and trafficking of membrane components occurs between host membranes and those of the intracellular parasite. Phospholipids are major membrane components and, although many parasites scavenge these phospholipids from their host, most parasites also synthesise phospholipids de novo, or modify a large part of the scavenged phospholipids. It was recently shown that some parasites like Plasmodium have unique phospholipid metabolic pathways. This review will focus on new developments in research on phospholipid metabolism of parasitic protozoa in relation to parasite-specific membrane structures and function, as well as on several targets for interference with the parasite phospholipid metabolism with a view to developing new anti-parasitic drugs.

Animals↗
PubMed2001

Advances and prospects for subunit vaccines against protozoa of veterinary importance.

Protozoa are responsible for considerable morbidity and mortality in domestic and companion animals. Preventing infection may involve deliberate exposure to virulent or attenuated parasites so that immunity to natural infection is established early in life. This is the basis for vaccines against theilerosis and avian coccidiosis. Vaccination may not be effective or practical with diseases, such as cryptosporidiosis, that primarily afflict the immune-compromised or individuals with an incompletely developed immune system. Strategies for combating these diseases often rely on passive immunotherapy using serum or colostrums containing antibodies to parasite surface proteins. Subunit vaccines offer an attractive alternative to virulent or attenuated parasites for several reasons. These include the use of bacteria or lower eukaryotes to produce recombinant proteins in batch culture, the relative stability of recombinant proteins compared to live parasites, and the flexibility to incorporate only those antigens that elicit "protective" immune responses. Although subunit vaccines offer many theoretical advantages, our lack of understanding of immune mechanisms to primary and secondary infection and the capacity of many protozoa to evade host immunity remain obstacles to developing effective vaccines. This review examines the progress made on developing recombinant proteins of Eimeria, Giardia, Cryptosporidium, Toxoplasma, Neospora, Trypanosoma, Babesia, and Theileria and attempts to use these antigens for vaccinating animals against the associated diseases.

Animals↗
PubMed2001

Trends in the use of protozoa in the assessment of wastewater treatment.

Increasing environmental pollution and the continuous development of new chemicals and drugs has led to ever growing concern about the potential effects of these compounds directly or indirectly on human health. As concerns water pollution, protozoa seem to be an excellent tool to assess both toxicity and pollution: they are regarded as biological indicators of pollution when their presence or absence can be related to particular environmental conditions, and they are considered test organisms when a species or population is used to evaluate the toxicity of relevant toxic compounds. Thus, an integrated approach is being developed to assess how toxic compounds affect the different biological levels of organisation--from the community level to the species level--of ciliated protozoa. The present paper reports and discusses the current state of the art of this approach.

Animals↗
PubMed2003

ABC transporters and drug resistance in parasitic protozoa.

Parasitic protozoa are responsible for a wide spectrum of diseases in humans and domestic animals. The main line of defence available against these organisms is chemotherapy. However, the application of chemotherapeutic drugs has resulted in the development of resistance mechanisms, which limit the number of antiprotozoal drugs that are effective in the treatment and control of parasitic diseases. Knowledge about the resistance mechanisms involved may allow the development of new drugs that minimise or circumvent drug resistance or may identify new targets for drug development. This review focuses on the role of protozoal ATP-binding cassette (ABC) transporters in drug resistance. These membrane proteins mediate the ATP-dependent transport of a wide variety of chemotherapeutic drugs away from their targets inside the parasites. The genome sequence of Plasmodium falciparum and Plasmodium yoelii has recently been completed, and the sequencing of other parasitic genomes are now underway. As a result, many new membrane transporters belonging to the ABC superfamily are being discovered. We review the ABC transporters in major parasitic protozoa, including Plasmodium, Leishmania, Trypanosoma and Entamoeba species. Transporters with an established role in drug resistance have been emphasised, but newly discovered transporters with a significant amino acid sequence identity to established ABC drug transporters have also been included.

ATP-Binding Cassette Transporters↗
PubMed2001

Targeting polyamines of parasitic protozoa in chemotherapy.

All parasitic protozoa contain polyamines and in recent years they, and their associated enzymes, have attracted attention as drug targets because they might reveal novel antiparasite therapies. How justified is this approach to drug discovery? In this review, Sylke Müller, Graham Coombs and Rolf Walter summarize the current status of research into drugs that exploit polyamine metabolism of trypanosomatid and malaria parasites, and propose priorities for research into such drugs. This review was inspired by an Expert Meeting entitled 'Polyamine Metabolism of Parasitic Protozoa as a Drug Target'.

Animals↗
PubMed2001

Aspartic proteases of Plasmodium falciparum and other parasitic protozoa as drug targets.

All parasitic protozoa contain multiple proteases, some of which are attracting attention as drug targets. Aspartic proteases are already the targets of some clinically useful drugs (e.g. chemotherapy of HIV infection) and a variety of factors make these enzymes appealing to those seeking novel antiparasite therapies. This review provides a critical analysis of the current knowledge on Plasmodium aspartic proteases termed plasmepsins, proposes a definitive nomenclature for this group of enzymes, and compares these enzymes with aspartic proteases of humans and other parasitic protozoa. The present status of attempts to obtain specific inhibitors of the parasite enzymes that will be useful as drugs is outlined and suggestions for future research priorities are proposed.

Amino Acid Sequence↗
PubMed2002

Bacteria and protozoa as integral components of the forest ecosystem--their role in creating a naturally varied soil fertility.

The paper explores interactions between the two first organism groups to appear on earth, the bacteria and protozoa, and their interplay with the rest of the ecosystem focusing upon northern boreal forests. The microbial loop is suggested as a mechanism for local inputs of new N to the ecosystem. The possibility to couple short-term microbial processes with their long-term effects,--as registered in plants, soil and the atmosphere, via the abiotic variables--is explored. The latter are investigated in relation to the environments they create for the micro-organisms, and how this results in varying soil fertility. A chain of events is presented that relate high Ca concentration in the mineral soil and high water availability to increased nitrogen availability for plants via the micro-organisms. An example is given of the influence of these parameters directly upon protozoa along an extreme fertility gradient, and also indirect evidence from a Finnish field study of 30 sites with four fertility levels. Finally, there is a discussion about ways to convert knowledge gained in detailed studies of microbial interactions into forms useful when evaluating the present status of and effects of ameliorative management on ecosystems strongly affected by humans.

Animals↗
PubMed2002

Evasion of innate immunity by parasitic protozoa.

Parasitic protozoa are a major cause of global infectious disease. These eukaryotic pathogens have evolved with the vertebrate immune system and typically produce long-lasting chronic infections. A critical step in their host interaction is the evasion of innate immune defenses. The ability to avoid attack by humoral effector mechanisms, such as complement lysis, is of particular importance to extracellular parasites, whereas intracellular protozoa must resist killing by lysosomal enzymes and toxic metabolites. They do so by remodeling the phagosomal compartments in which they reside and by interfering with signaling pathways that lead to cellular activation. In addition, there is growing evidence that protozoan pathogens modify the antigen-presenting and immunoregulatory functions of dendritic cells, a process that facilitates their evasion of both innate and adaptive immunity.

Animals↗
PubMed2004

Stable isotope probing of rRNA and DNA reveals a dynamic methylotroph community and trophic interactions with fungi and protozoa in oxic rice field soil.

Stable isotope probing (SIP) is a novel technique to characterize structure and in situ function of active microbial populations, which is based on the incorporation of 13C-labelled substrates into nucleic acids. Here, we have traced methylotrophic members of a rice field soil microbial community, which became active upon continuous addition of 13C-methanol (< 22 mM) as studied in microcosms. By combining rRNA- and DNA-based SIP, as well as domain-specific real-time PCR detection of templates in fractions of centrifugation gradients, we were able to detect 13C-labelled bacterial rRNA after 6 days of incubation. Fingerprinting and comparative sequence analysis of 'heavy' bacterial rRNA showed that mostly members of the Methylobacteriaceae and a novel clade within the Methylophilaceae formed part of the indigenous methylotrophic community. Over time, however, the Methylophilaceae were enriched. Unexpectedly, nucleic acids of eukaryotic origin were detected, mostly in intermediately 13C-labelled gradient fractions. These eukaryotes were identified as fungi mostly related to Fusarium and Aspergillus spp., and also Cercozoa, known as predatory soil flagellates. The detection of fungi and protozoa in 13C-enriched nucleic acid fractions suggests a possible involvement in either direct assimilation of label by the fungi, or a food web, i.e. that primary 13C-methanol consuming methylotrophs were decomposed by fungi and grazed by protozoa.

Animals↗
PubMed1986

Role of rumen protozoa in nitrogen digestion in sheep given two isonitrogenous diets.

1. The effect of protozoa on digestion in the rumen was studied using either defaunated or faunated sheep. 2. Six wethers, each fitted with rumen and simple duodenal cannulas, were given two isonitrogenous diets containing either lucerne (Medicago sativa) hay (diet L) or sodium hydroxide-treated wheat straw (diet S). The diets were given in eight equal portions per day at 3-h intervals. The mean intake of dry matter, 53 g/kg body-weight0.75 per d, was similar for the two diets and each diet had a similar digestible organic matter content. Diet L promoted a large protozoal population and was rich in nitrogen sources of low rumen-degradability, while diet S supported a smaller protozoal population and was rich in rumen-degradable N. 3. Digesta flow at the duodenum was estimated by means of a dual-marker technique using chromium-mordanted lucerne hay and polyethylene glycol as markers. The microbial flow at the duodenum was estimated using diaminopimelic acid (DAPA), nucleic-acid purine bases (PB) and 35S incorporation simultaneously. The different microbial markers were compared in the defaunated sheep. Protozoal N contribution was estimated in faunated sheep. 4. Defaunated sheep had lower rumen ammonia concentrations and molar proportions of butyric acid than faunated sheep, but they had higher molar proportions of propionic acid. 5. Rumen organic matter digestion was reduced by defaunation, but this decrease was compensated for by increased intestinal digestion. 6. There was a net increase of N flow (approximately 10 g/d) between mouth and duodenum in defaunated sheep. This was explained by increases in both microbial and dietary N flows from the rumen compared with faunated sheep. 7. The influence of protozoa on solid- and liquid-phase retention times in the rumen is discussed, as well as the protozoal contribution to microbial N flow in the duodenum of faunated sheep.

Animals↗
PubMed2004

Effects of dietary sunflower seeds on rumen protozoa and growth of lambs.

Linoleic acid-rich sunflower-seed supplements (SSS) were used in two experiments (experiment 1, high-concentrate diets; experiment 2, high-forage diets) to study effects on rumen protozoa and the growth of lambs. Both experiments consisted of four treatments, two with a low-protein diet (120 g/kg) and two with a high-protein diet (160 g/kg). For both diets, one treatment was without (control) and one with the SSS (140 g/kg dietary DM). The lambs were fed ad libitum for 70 and 140 d in experiments 1 and 2, respectively. Thereafter, the digestibility of organic matter (OM), acid-detergent fibre and neutral-detergent fibre were determined for each diet with four lambs, and then all lambs were slaughtered and rumen fluid samples were collected and analysed. The results showed substantial decreases (P < 0.001) or total elimination of protozoa in the rumen fluid of the SSS-receiving lambs. In the first experiment the SSS also decreased (P < 0.05) feed intake, but an increase in average daily gain (P < 0.06) resulted in an improved (P < 0.05) feed:gain ratio. Also, the SSS increased (P < 0.05) the digestibility of fibre. In the second experiment the SSS decreased (P < 0.05) the OM digestibility, feed intake and growth of lambs. It was concluded that the use of sunflower-seed supplementation in high-concentrate diets of ruminants reduces rumen fauna, resulting in savings on dietary protein supplements and an increased digestion of feed.

Animals↗
PubMed1999

Intestinal protozoa and associated bacteria in captive houbara bustards (Chlamydotis undulata) in the United Arab Emirates.

A protozoal survey was carried out in 114 captive houbara bustards (Chlamydotis undulata) in the United Arab Emirates. Trichomonas gallinarum, Chilomastix gallinarum, Giardia spp. and Lophomonas spp. were detected from faecal samples of 49 (43%) captive houbara bustards. Culture and identification studies were carried out to assess associated bacteria. The bacteria isolated from birds negative to protozoa included Enterococcus spp., Aerococcus spp., Micrococci spp., Proteus spp., Enterobacter spp., Escherichia coli and Klebsiella spp., while Enterococcus spp., Micrococci kristinae, Aerococcus spp., Staphylococcus sciuri, Staphylococcus aureus, E. coli, Klebsiella spp. and Alcaligenes faecalis were isolated from protozoa positive birds. Proteus spp., Enterobacter spp., E. coli , Klebsiella spp., Aerococcus spp. and Enterococcus spp. are considered part of the normal intestinal bacterial flora of captive bustards and they were also isolated from the food items used to feed the captive bustards. S. sciuri possibly originated from ingested mice; Alcaligenes faecalis and Micrococcus spp. are contaminants from soil or water.

Journal Article↗
PubMed1976

[The effect of straw in ground and pelleted form on the rumen fermentation and protozoa].

Cows possessing a large ruminal fistula were fed straw meal and pelleted wheat straw to investigate its effect on ruminal fermentation (concentration of NH3 and volatile fatty acids (FFS), pH, molar proportions of FFS, rates of FFS production) and on the protozoa population. The straw-concentrate mixture used in the present trial contained 40% of straw. The feeding of pelleted straw produced a significant rise in FFS concentrations (from 8.8. to 12.3 mMole/100 ml) and a corresponding decline of pH (from 6.7 to 6.1). With high molar proportions of acetate (72 mole%) the influence of the straw diet on molar FFS proportions was low. The NH3 peak observed 1 hr after feeding was higher with the pelleted straw than with the straw meal. The feeding of finely gound straw produced a higher level of FFS production (by 10%) than that of straw pellets. (3.88 and 4.29 mMole per gm DM). The number of protozoa (per ml of ruminal fluid) was 335,000 (straw meal) and 121,833 (pellets). The number of large infusorial cells (Isotricha, Diplodinium, Ophryoscolex) decreased correspondingly from 70,000 (straw meal) to 18,870 per ml (pellets). These results suggest that the feeding of pelleted straw-concentrate mixtures to cows as sole feed will not bring about optimum conditions for ruminal fermentation (FFS formation, protein synthesis) and for the layering of ruminal contents.

Animal Feed↗
PubMed1994

Requirements for glycosylphosphatidylinositol attachment are similar but not identical in mammalian cells and parasitic protozoa.

The general features of the glycosylphosphatidylinositol (GPI) signal have been conserved in evolution. To test whether the requirements for GPI attachment are indeed the same in mammalian cells and parasitic protozoa, we expressed the prototype GPI-linked protein of Trypanosoma brucei, the variant surface glycoprotein (VSG), in COS cells. Although large amounts of VSG were produced, only a small fraction became GPI linked. This impaired processing is not caused by the VSG ectodomain, since replacement of the VSG GPI signal with that of decay accelerating factor (DAF) produced GPI-linked VSG. Furthermore, whereas fusion of the DAF GPI signal to the COOH terminus of human growth hormone (hGH) produces GPI-linked hGH, an analogous hGH fusion using the VSG GPI signal does not, indicating that the VSG GPI signal functions poorly in mammalian cells. By constructing chimeric VSG-DAF GPI signals and fusing them to the COOH terminus of hGH, we show that of the two critical elements that comprise the GPI-signal--the cleavage/attachment site and the COOH terminal hydrophobic domain--the former is responsible for the impaired activity of the VSG GPI signal in COS cells. To confirm this, we show that the VSG GPI signal can be converted to a viable signal for mammalian cells by altering the amino acid configuration at the cleavage/attachment site. We also show that when fused to the COOH terminus of hGH, the putative GPI signal from the malaria circumsporozoite (CS) protein produces low levels of GPI-anchored hGH, suggesting that the CS protein is indeed GPI linked, but that the CS protein GPI signal, like the VSG-signal, functions poorly in COS cells. The finding that the requirements for GPI attachment are similar but not identical in parasitic protozoa and mammalian cells may allow for the development of selective inhibitors of GPI-anchoring that might prove useful as antiparasite therapeutics.

Amino Acid Sequence↗
PubMed1990

Detached ciliary tufts. Comparison with intestinal protozoa and a review of the literature.

Detached ciliary tufts are structures of doubtful pathologic significance that are normally present in many body fluids. However, their existence is not widely appreciated by pathologists practicing routine laboratory medicine. If they are observed, their practical clinicopathologic importance is that, because of their vigorous ciliary motion in wet mounts, they can imitate ciliated protozoa and pose a diagnostic dilemma. The authors present a brief review of the literature on detached ciliary tufts and a morphologic comparison with the more commonly encountered intestinal protozoa.

Adolescent↗
PubMed1993

N-glycosylation in trypanosomatid protozoa.

Trypanosomatid protozoa are parasites of considerable medical and economic importance in developing countries. The pathway leading to N-glycosylation in these microorganisms is characterized by the following features: (i) dolichols are composed of only 10-13 isoprene units; (ii) oligosaccharides transferred in N-glycosylation have the compositions Man(6,7,9)GlcNAc2, depending on the species; (iii) trypanosomatids are unable to synthesize dolichol-P-Glc and, in addition, some species lack certain dolichol-P-Man-dependent mannosyltransferases; (iv) the oligosaccharyltransferase does not require the presence of glucose units in the oligosaccharide in order to catalyse an efficient transfer reaction; (v) trypanosomatids have a glucosidase II-like enzyme, but lack glucosidase I; (vi) glucosidase II is required for deglucosylation of oligosaccharides glucosylated by the UDP-Glc:glycoprotein glucosyltransferase, an activity first detected in those parasites; (vii) the structures of polymannose-type compounds in these protozoa have no significant differences with those of their mammalian counterparts except for the presence, in certain species, of oligosaccharides having galactofuranose units linked to external mannose residues; (viii) biantennary complex-type oligosaccharides having in some cases terminal alpha-linked galactose units or poly-N-acetylactosamine extensions, but lacking sialic acid units, have been described in Trypanosoma brucei; (ix) complex-type oligosaccharides having alpha-linked galactose, fucose and sialic acid residues have been described in Trypanosoma cruzi. In this parasite, addition of sialic acid units to glycoproteins and glycolipids is mediated by a trans-sialidase located on the external surface of the parasite and not by an intracellular CMP-sialic acid-dependent sialyltransferase.

Animals↗
PubMed2006

Niche metabolism in parasitic protozoa.

Complete or partial genome sequences have recently become available for several medically and evolutionarily important parasitic protozoa. Through the application of bioinformatics complete metabolic repertoires for these parasites can be predicted. For experimentally intractable parasites insight provided by metabolic maps generated in silico has been startling. At its more extreme end, such bioinformatics reckoning facilitated the discovery in some parasites of mitochondria remodelled beyond previous recognition, and the identification of a non-photosynthetic chloroplast relic in malarial parasites. However, for experimentally tractable parasites, mapping of the general metabolic terrain is only a first step in understanding how the parasite modulates its streamlined, yet still often puzzlingly complex, metabolism in order to complete life cycles within host, vector, or environment. This review provides a comparative overview and discussion of metabolic strategies used by several different parasitic protozoa in order to subvert and survive host defences, and illustrates how genomic data contribute to the elucidation of parasite metabolism.

Animals↗
PubMed1978

Some factors controlling the attachment of the rumen holotrich protozoa Isotricha intestinalis and I. prostoma to plant particles in vitro.

The rumen holotrich protozoa Isotricha intestinalis and I. prostoma showed chemotaxis to sucrose, glucose and fructose. They attached themselves, by means of an organelle on the anterior cell surface, to particulate sources of these carbohydrates provided soluble protein was present in the medium. The concentration of protein eliciting attachment varied with the species and the state of nutrition of the cell, but was between 20 and 150 microgram ml-1. Attachment occurred only if the concentration of carbohydrate, at its source, exceeded the chemotaxis threshold concentration (50 micrometer for sucrose) and if it was less than 1 mM. At concentrations exceeding 1 mM, indiscriminate attachment to gas-liquid and solid-liquid interfaces occurred, provided the protein concentration was high enough to elicit attachment. In the rumen, soluble carbohydrates diffusing from food particles may attract the protozoa which attach themselves to the particles in the presence of soluble plant protein at less than 20 microgram ml-1; these conditions exist in the host animal soon after feeding when fed infrequently. The attachment mechanism may confer an ecological advantage on the Isotricha spp. over other rumen organisms dependent on soluble carbohydrates as energy and carbon sources.

Animals↗