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Development of Trypanosoma fallisi in the leech, Desserobdella picta, in toads (Bufo americanus), and in vitro. A light and electron microscopic study.

The development of Trypanosoma fallisi of Bufo americanus from Algonquin Park, Ontario was studied by light and electron microscopy in blood culture, in its leech vector Desserobdella (= Batracobdella) picta, and in its toad host. In culture, bloodstream trypomastigotes transformed within one day to elongate epimastigotes which divided into rosettes. These gave rise to amastigotes, spheromastigotes, stumpy and elongate epimastigotes, and slender metacyclic trypomastigotes over a 4- to 6-day period. Development in the leech crop was similar to that in culture, with fewer amastigotes and no spheromastigotes observed. The stages in the leech were similar in size to their culture counterparts, except for metacyclic trypomastigotes, which were larger in culture. Culture and leech stages possessed a well developed cytostome-cytopharyngeal complex and prominent reservosomes. The kinetoplast of crop stages was small with a rectangular profile, but became larger and basket-like in the proboscis forms. Migration of trypanosomes to the proboscis appeared to depend on the rate of digestion of the bloodmeal. Flagellates in the leech were also characterized by the presence of intracellular microorganisms. Development of culture forms to mature stages in the toad was completed within 8 days postinoculation, with the organisms transforming into the typical "C"-shape with a large square kinetoplast. Natural infection of B. americanus was detected at 3 days postfeeding by D. picta and the resulting bloodstream trypomastigotes developed more slowly than inoculated cultured stages.

Animals↗

Evaluation of antigens for the serodiagnosis of kala-azar and oriental sores by means of the indirect immunofluorescence antibody test (IFAT).

Antigens and corresponding sera were collected from travellers with leishmaniasis returning to Germany from different endemic areas of the old world. The antigenicity of these Leishmania strains, which were maintained in Syrian hamsters, was compared by indirect immunofluorescence (IFAT). Antigenicity was demonstrated by antibody titres in 18 sera from 11 patients. The amastigotic stages of nine strains of Leishmania donovani and four strains of Leishmania tropica were compared with each other and with the culture forms of insect flagellates (Strigomonas oncopelti and Leptomonas ctenocephali). Eighteen sera from 11 patients were available for antibody determination with these antigens. The maximal antibody titres in a single serum varied considerably depending on which antigen was used for the test. High antibody levels could only be maintained when Leishmania donovani was employed as the antigen, but considerable differences also occurred between the different strains of this species. The other antigens were weaker. No differences in antigenicity between amastigotes and promastigotes of the same strain were observed. It is important to select suitable antigens. Low titres may be of doubtful specificity and are a poor baseline for the fall in titre which is an essential index of effective treatment.

Antibody Formation↗

In vitro attachment of trypanosomes to plastic.

Description of an in vitro system for the study of the attachment of trypanosomes to polystyrene flasks by means of hemidesmosomes. This type of attachment, whose significance is so far unknown, reproduces a natural stage in the life cycle of medically important trypanosomatid flagellates in their vector.

Cell Adhesion↗

The antimitotic activities of some benzodiazepines.

Among 9 benzodiazepines, tested on the proliferation of synchronously dividing flagellate cells, only diazepam and medazepam can induce an accumulation of abnormal mitotic figures after 24 h of treatment. It seems that there is not a direct relation between the activity of benzodiazepines on the central nervous system and their ability to inhibit mitosis.

Benzodiazepines↗

Trichomonas species in a subhepatic abscess.

A rare case of Trichomonas tenax/Trichomonas hominis and mixed oral bacterial flora in pus from a subhepatic abscess in a patient with a perforated penetrating ventricular ulcer is reported and the possible pathogenicity of the flagellate is discussed.

Humans↗

Dientamoeba fragilis. An unusual intestinal pathogen.

This is a case report of a gastrointestinal infection caused by Dientamoeba fragilis. It is a flagellate protozoan that is an uncommon etiology of gastrointestinal disease. Primarily characterized by diarrhea and abdominal pain, other symptoms such as flatulence, nausea, vomiting, fatigue, malaise, and weight loss occur. Diagnosis is made using multiple fresh stool samples that are preserved and permanently stained looking for the typical binucleate trophozoite. Since there is a distinct association with Enterobius vermicularis (possibly the mode of protozoan transmission), the human pinworm is also sought. Treatment of choice consists of diiodohydroxyquin in adults and metronidazole in children.

Abdominal Pain↗

Eukaryotic origins: string analysis of 5S ribosomal RNA sequences from some relevant organisms.

Using the PHYLOGEN tree-forming programs, we evaluate the published 5S rRNA sequences in certain of the files in the Berlin DataBank in an attempt to identify the connection between archaebacteria and the eukaryotic protists. These programs are based on methods of string analysis developed by Sankoff and others. Their discriminatory power is derived from their continuous realignment of sequences through repeated assessment of insertions and deletions as well as substitutions. The programs demonstrate that even these small molecules (ca. 120 bases) retain substantial records of evolutionary events that occurred over a billion years ago. The eukaryotes seem to have been derived from ancestors near the common origins of the halobacterial and Methanococcales groups. Identifying what might have been a primordial eukaryote is more difficult because several of the species considered as early derivatives from the common root are isolated species with large genetic differences from each other and from all other extant forms that have been sequenced. The ameboid, flagellated, and ciliated protists seem to have emerged nearly simultaneously from an ancient cluster, but the sarcodinid protozoa have preference as the group of most ancient origin. The euglenozoa and the ciliates are of later derivation. Our ability to tease plausible trees from such small molecules suggests that the mode of analysis rather than the size of the molecule is often a major limitation in the reconstruction of acceptable ancient phylogenics.(ABSTRACT TRUNCATED AT 250 WORDS)

Animals↗

Molecular phylogenetic analysis of actin genic regions from Achlya bisexualis (Oomycota) and Costaria costata (Chromophyta).

Actin genic regions were isolated and characterized from the heterokont-flagellated protists, Achlya bisexualis (Oomycota) and Costaria costata (Chromophyta). Restriction enzyme and cloning experiments suggested that the genes are present in a single copy and sequence determinations revealed the existence of two introns in the C. costata actin genic region. Phylogenetic analyses of actin genic regions using distance matrix and maximum parsimony methods confirmed the close evolutionary relationship of A. bisexualis and C. costata suggested by ribosomal DNA (rDNA) sequence comparisons and reproductive cell ultrastructure. The higher fungi, green plants, and animals were seen as monophyletic groups; however, a precise order of branching for these assemblages could not be determined. Phylogentic frameworks inferred from comparisons of rRNAs were used to assess rates of evolution in actin genic regions of diverse eukaryotes. Actin genic regions had nonuniform rates of nucleotide substitution in different lineages. Comparison of rates of actin and rDNA sequence divergence indicated that actin genic regions evolve 2.0 and 5.3 times faster in higher fungi and flowering plants, respectively, than their rDNA sequences. Conversely, animal actins evolve at approximately one-fifth the rate of their rDNA sequences.

Actins↗

Primary and secondary structure of the nuclear small subunit ribosomal RNA of the cryptomonad Pyrenomonas salina as inferred from the gene sequence: evolutionary implications.

The cryptomonad Pyrenomonas salina presumably has arisen from a symbiotic event involving a flagellated phagotrophic host cell and a photosynthetic eukaryote as the symbiont. Correspondingly, in this unicellular alga there are four different genomes, e.g., the nuclear and the mitochondrial genomes of the host cell as well as the plastid genome and the genome contained in the vestigial nucleus of the endocytobiont (nucleomorph). To analyze the origin of one of the symbiotic partners the small subunit rRNA gene sequence of the host cell nucleus was determined, and a secondary structure model has been constructed. This sequence is compared to those of 40 other eukaryotes. A phylogenetic tree constructed using the neighborliness method revealed a close relationship between the host cell of P. salina and the chlorophytes, whereas the rhodophytes diverge more deeply in the tree.

Archaea↗

Structure and function of the spermathecal complex in the phlebotomine sandfly Phlebotomus papatasi Scopoli (Diptera: Psychodidae): II. post-copulatory histophysiological changes during the gonotrophic cycle.

The spermathecal complex of Phlebotomus papatasi Scopoli (Diptera: Psychodidae) undergoes histological and physiological changes during its gonotropic cycle. The present histochemical study revealed a mucopolysaccharide secretory mass in the spermathecae of the newly emerged sandfly. Sperm competition occurs when two or more males compete to fertilize an ovum in the female reproductive tract. In this study, spermatophores of two or more competing males were deposited at the base of the spermathecal ducts, which originate from the female bursa copulatrix. This suggests that females play a role in sperm displacement, which is defined as any situation in which the last male to mate with a female fertilizes maximum number her eggs. A blood meal ingested by the female for ovary development and egg laying stimulates the release of sperm from the spermatophore. The spermatozoa then migrate to the lumen of the spermatheca. The ultrastructure of spermatozoa comprises a head with double-layered acrosomal perforatorium, an elongate nucleus, and the axoneme with a 9 + 9 + 0 flagellar pattern. This axomene differs from the flagellate axoneme of other Psychodinae. Morphological changes, such as the casting off of the acrosomal membrane, and histological changes in the spermatophore are also described. Mating plugs that have been described previously in sandflies appear to be artefacts. Females of P. papatasi may be inseminated more than once during each gonotrophic cycle, and additional inseminations may be necessary for each cycle. The relationships between the volumes of the sperm and the spermatheca were calculated to determine sperm utilization and fecundity of P. papatasi. As the females of P. papatasi mate polyandrously, the anatomical and physiological complexity of the spermathecal complex may be related to post-copulatory sexual selection.

Animal Structures↗

Regulation of enzymatic activity: one possible role of dietary boron in higher animals and humans.

It is well established that vascular plants, diatoms, and some species of marine algal flagellates have acquired an absolute requirement for boron (B), although the primary role remains unknown. Discovery of naturally occurring organoboron compounds, all ionophoric macrodiolide antibiotics with a single B atom critical for activity, established at least one biochemical role of B. The unusual nature of B chemistry suggests the possibility of a variety of biological roles for B. At physiological concentrations and pH, B may react with one N group or one to four hydroxyl groups on specific biological ligands with suitable configuration and charge to form dissociable organoboron compounds or complexes. Suitable ligands include pyridine (e.g., NAD+ or NADP) or flavin (e.g., FAD) nucleotides and serine proteases (SP). B reacts with the cis adjacent hydroxyls on the ribosyl moiety of the nucleotides or, in the serine proteases, the N on the imidazole group of histidine or the hydroxyl group on the serine moiety. Reversible inhibition by B of activity of SP or oxidoreductases that require pyridine or flavin nucleotides is well known. Therefore, a proposed essential role for B is as a regulator of relevant pathways, including respiratory burst, that utilize these enzymes.

Animals↗

Antimutagenicity of milk fermented by Enterococcus faecium.

The diethyl ether extracts isolated from unfermented milk and milk fermented by Enterococcus faecium exhibited dose-dependent inhibition of mutagenesis induced by N-methyl-N'-nitro-N-nitrosoguanidine (MNNG), nitrovin (NIT), 5-nitro-2-furylacrylic acid (NFA) and UV-irradiation on the Ames bacterial test (Salmonella typhimurium strains TA97 and TA100) and the unicellular flagellate Euglena gracilis. Overall, the fermented milk extract was the most active against UV-irradiation, less active against NIT and MNNG, and the least active against NFA on bacteria. The highest antibleaching effects were observed against MNNG. The differences between antimutagenic effects from fermented and unfermented milk extracts were determined to be statistically significant at the 0.95 CI level.

Acrylates↗

Effect of elevated temperature on genotoxicity of chemotherapeuticals toward Euglena gracilis.

A collection of 20 compounds was tested for their ability to induce a permanent loss of chloroplasts from Euglena gracilis cells under conditions increasing the sensitivity of the flagellate to genotoxic compounds, viz. in the resting medium and at an elevated temperature. Streptomycin, dihydrostreptomycin, gentamicin, kanamycin and partially chloramphenicol exhibited mutagenic effects. Eight antibiotics eliminated chloroplasts only from growing cultures and seven antibiotics did not induce the mutation at all.

Animals↗

A new type of microscope illumination and its use in microbiology.

To overcome problems with taking photographs of motile microorganisms we used a new type of microscope illumination by means of a light-emitting diode (LED), which fully preserves the Kohler illumination principle. Oblique illumination or relief contrast after the Hostounsk, microscopy method were used for observing and photographing bacteria, flagellates and infusorians having different motion velocities with sufficient image contrast. The device, which permitted us to use short shutter time (1/30, 1/60 or 1/125 s), was successfully used in recording different motile microorganisms including their internal architecture.

Animals↗

Structure of microbial communities in Sphagnum peatlands and effect of atmospheric carbon dioxide enrichment.

Little is known about the structure of microbial communities in Sphagnum peatlands, and the potential effects of the increasing atmospheric CO2 concentration on these communities are not known. We analyzed the structure of microbial communities in five Sphagnum-dominated peatlands across Europe and their response to CO2 enrichment using miniFACE systems. After three growing seasons, Sphagnum samples were analyzed for heterotrophic bacteria, cyanobacteria, microalgae, heterotrophic flagellates, ciliates, testate amoebae, fungi, nematodes, and rotifers. Heterotrophic organisms dominated the microbial communities and together represented 78% to 97% of the total microbial biomass. Testate amoebae dominated the protozoan biomass. A canonical correspondence analysis revealed a significant correlation between the microbial community data and four environmental variables (Na+, DOC, water table depth, and DIN), reflecting continentality, hydrology, and nitrogen deposition gradients. Carbon dioxide enrichment modified the structure of microbial communities, but total microbial biomass was unaffected. The biomass of heterotrophic bacteria increased by 48%, and the biomass of testate amoebae decreased by 13%. These results contrast with the absence of overall effect on methane production or on the vegetation, but are in line with an increased below-ground vascular plant biomass at the same sites. We interpret the increase in bacterial biomass as a response to a CO2-induced enhancement of Sphagnum exudation. The causes for the decrease of testate amoebae are unclear but could indicate a top-down rather than a bottom-up control on their density.

Amoeba↗

Ribosomal DNA ITS-1 and ITS-2 sequence comparisons as a tool for predicting genetic relatedness.

The determination of the secondary structure of the internal transcribed spacer (ITS) regions separating nuclear ribosomal RNA genes of Chlorophytes has improved the fidelity of alignment of nuclear ribosomal ITS sequences from related organisms. Application of this information to sequences from green algae and plants suggested that a subset of the ITS-2 positions is relatively conserved. Organisms that can mate are identical at all of these 116 positions, or differ by at most, one nucleotide change. Here we sequenced and compared the ITS-1 and ITS-2 of 40 green flagellates in search of the nearest relative to Chlamydomonas reinhardtii. The analysis clearly revealed one unique candidate, C. incerta. Several ancillary benefits of the analysis included the identification of mislabelled cultures, the resolution of confusion concerning C. smithii, the discovery of misidentified sequences in GenBank derived from a green algal contaminant, and an overview of evolutionary relationships among the Volvocales, which is congruent with that derived from rDNA gene sequence comparisons but improves upon its resolution. The study further delineates the taxonomic level at which ITS sequences, in comparison to ribosomal gene sequences, are most useful in systematic and other studies.

Animals↗