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Verification of the silica deficiency hypothesis based on biogeochemical trends in the aquatic continuum of Lake Biwa-Yodo River-Seto Inland Sea, Japan.

The silica deficiency hypothesis holds that increases of still waters caused by hydraulic alterations and high nitrogen (N) and phosphorus (P) discharges enhance the growth of freshwater diatoms, which take up the dissolved silicate (DSi) supplied by natural weathering. The consequent decrease in the DSi supply to the sea is advantageous to flagellates (nonsiliceous and potentially harmful) but not to diatoms (siliceous and mostly benign) in coastal marine ecosystems. Verification of this hypothesis has been hampered by lack of relevant data, particularly in Asia. We investigated the aquatic continuum composed of Lake Biwa, the Yodo River, and the Seto Inland Sea, Japan, where the natural conditions make the silica deficiency less likely to emerge due to the inherently rich supply of DSi. The results showed that the silica was retained both in the lake and nearby the estuary. The relative dominance of diatom and flagellates could not be explained solely by the stoichiometric arguments but by the supportive discussion on the difference of their behavioral characteristics and the process nearby the estuary, where direct inputs of N and P and effluent Si enhanced diatom bloom, even though the Si/N ratio was lowered in the upstream reservoir. Thus the retention of DSi occurred in two places: in the lake and nearby the estuary, where the other N and P are loaded directly. The rate of DSi retention correlated with socio-economic changes, such as rapid economic growth in the 1960s and mitigations implemented after the 1980s. Sensitivity of this continuum to the Si processes suggests the global significance of this hypothesis.

Diatoms↗

Vectors of Leishmania braziliensis in the Petén, Guatemala.

During a 1-year study, 13 species of sand fly were collected in bite-landing collections on human attractants in Tikal, Guatemala. Using isoenzyme analysis, Leishmania braziliensis was identified among isolates from Lutzomyia ovallesi, Lu. panamensis, and Lu. ylephiletor. Lutzomyia ovallesi, Lu. shannoni, and Lu. cruciata were found with flagellates whose isoenzyme patterns matched unidentified flagellates isolated from a patient with mucosal lesions.

Animals↗

Free-living amoebae in Egypt. 1. Naegleria gruberi and Naegleria fowleri.

Two Naegleria species were isolated and identified from various water sources in Lower and Upper Egypt. Identification was based on the morphology, nuclear division and the excystation and flagellation tests. The trophic, cystic and flagellate forms of N. gruberi are larger than those of N. fowleri and the cyst of the former species has one or more pores while that of the latter species has no pores and has an outer gelatinous layer. The size and the morphological characteristics of these two free-living amoebae from Egypt were in complete agreement with those previously described for amoebae of this same genus and species endemic to other geographical areas.

Animals↗

Anopheline mosquitoes and malaria parasites in Taiwan.

Extensive taxonomic studies of the genus Anopheles were carried out in Taiwan during the first half of the 20th century. As a result, 17 species of Anopheles were identified and reported as occurring in Taiwan. However, the occurrence of two species, An. kochi and An. fluviatilis, in Taiwan is doubtful. Vector competence of the anophelines of Taiwan was also studied both in the laboratory and in the field during the same period of time, and experimental infections of 8 species of Anopheles were carried out by Anazawa in the 1920s. The results showed that all species tested were susceptible to three human plasmodia except for An. sinensis which was refractory to Plasmodium falciparum. The results of dissection of nearly 10,000 anopheline mosquitoes collected from the field by various workers suggested that An. minimus is the chief vector in Taiwan. Crithidial flagellates resembling malaria sporozoites were found in the salivary glands of 4 common species of Anopheles collected from cow-stables. These flagellates may have been misidentified as malaria sporozoites in the earlier investigations. During the first part of the operations for malaria control and eradication, anopheline mosquitoes were collected by day from 1,118 houses scattered over the island. Of the 25,656 specimens of Anopheles collected, nearly 80% were An. minimus, mostly from bedrooms. Since the highly anthropophilic and endophilic An. minimus was determined to be the chief vector of malaria in Taiwan, DDT was applied to the wall surfaces in houses for malaria control and eradication. Before spraying DDT, the rate of P. malariae infections was very high in areas where malaria is endemic. However, these infections were more quickly suppressed by DDT spraying than two other parasite species. Other details of studies on the ecology and behavior of anophelines are also presented.

Animals↗

[Structure and evolution of Trichomonadida].

In the article the survey of data on morphology and ultrastructure of flagellates of the order Trichomonadida is given with special attention paid to Monocercomonadidae and Trichomonadidae. The evolution within these flagellates is considered as a series of changes leading to formation of undulating membrane and reinforcement of the cytoskeleton. In both families other two evolutionary tendencies may be also observed: reduction of the flagellar apparatus (in Monocercomonadidae) and enlargement of the number of free flagella (in both families).

Animals↗

Freeze fracture and scanning electron microscope studies on the nuclear envelope and perinuclear cytomembranes (parabasal apparatus) in the protozoan, Lophomonas blattarum.

Phase contrast, scanning electron microscope (SEM), and freeze fracture studies on the parasitic flagellate, Lophomonas blattarum, have demonstrated that the endomembrane system (parabasal apparatus) is highly ordered, restricted in position to a perinuclear zone at the anterior end of the organism, and is supported in this localized cytoplasmic region by overlapping sheets or plates of microtubules, previously called the calyx and axial filament, which may participate in supporting the nucleus-endomembrane system in a restricted region of the cell. Light microscope observations, SEM and freeze fracture data provide support to previous views that the rough- and smooth-surfaced endoplasmic reticulum are interconnected and attached to the outer layer of the nuclear envelope. The continuity of these membrane systems provides an orderly and restricted packing in the perinuclear cytoplasm since other areas of the cell may become filled with yeast. These flagellates are especially adept at phagocytosis of entire yeast. In yeast-laden cells, the flagella-nucleus-parabasal body-calyx-axial filament complex may separate from the remainder of the cell and assume a motile existence for a time. The significance of the described relationships, in addition to providing efficiency in endomembrane localization, may also reflect synthesis of enzymes and proteins by the RER and packaging in the SER, both of which are continuous. Granules characteristic of glycogen are concentrated around the SER which may be involved in glycogen metabolism. Although critical information is lacking, the endomembrane system may also be involved in the synthesis and morphogenesis of lysosomes, and perhaps peroxisomes. Lophomonas thus amplifies a highly ordered spatial relationship between the nuclear envelope and the ER.

Animals↗

[Electron microscopic observations on the existence of "deflagellation tolerance" processes capable of explaining the selective isolation of vibrios in semisolid agar].

The authors identified a group of substances (flagellar depression factors--FDF) which suppress the mobility of enterobacteria (immobilization) but do not influence the Vibrio movements (tolerance). FDF do not affect the vital processes, multiplication and formation of colonies. Electron microscopic interpretation showed that, in the usual media, enterobacteria have characteristic peritrichous cilia, whereas in media with FDF they are denuded of their flagellation formation. As such a process of flagellation depletion seems to be not known, we suggest the term "deflagellation" to name it. In a completely different way, the vibrios keep their single cilium located apically and implicitly their moving ability. These behaviour differences are evident in semisolid agar with FDF. The deflagellated enterobacteria are, therefore, immobilized, and multiply, forming small punctual colonies, of 0.5-1 mm phi, whereas the vibrios tolerating FDF develop and at the same time move in the liquid agar, mass, forming large colonies, of 8-14 mm phi. The electron microscopic observations presented show the existence of a "deflagellation tolerance" process and also explain the specific typology of the colonies of vibrios and enterobacteria formed in liquid agar with FDF.

Agar↗

The affinity of the lectins Ricinus communis and Glycine maxima to carbohydrates on the cell surface of various forms of Trypanosoma cruzi and Trypanosoma rangeli, and the application of these lectins for the identification of T. cruzi in the feces of Rhodnius prolixus.

Flagellates of Trypanosoma cruzi (stock Molino 1), obtained from the intestine of experimentally infected Rhodnius prolixus, grown in cellular or acellular culture, as well as from the blood of infected mice, were examined by a direct fluorescence test using the lectins RCA (Ricinus communis-120) and SBA (soy bean agglutinin; Glycine maxima), conjugated with fluorescein isothiocyanate, for the detection of beta-D-galactose and alpha,beta-N-acetyl-D-galactosamine on the membranes of the flagellates. The same reactions were carried out using Trypanosoma rangeli (stock San Agustin), obtained from the intestine, hemo-lymph or salivary glands of experimentally infected R. prolixus, as well as from cultures and from the blood of experimentally infected CFW mice. The results indicate that the membrane of T. rangeli in the salivary glands of the vector contains beta-D-galactose, but that this sugar is absent from all other developmental stages of this trypanosome. All stages of intestinal and cultured. T. cruzi presented positive reactions with RCA-FITC and SBA-FITC. The high specificity of this technique makes it useful for the examination of R. prolixus, previously used in xenodiagnosis of Chagas' disease and for the examination of intradomiciliary or sylvatic vectors in epidemiological surveys in areas where T. cruzi and T. rangeli coexist. Formaldehyde fixed samples can be examined months later and false reports due to T. rangeli can be avoided.

Animals↗

UV-induced growth inhibition in the trypanosomatid Crithidia fasciculata.

The effects of short ultraviolet (253 nm) radiation on the growth of the trypanosomatid flagellate Crithidia fasciculata were studied in liquid medium and nutrient agar plates. Cell duplication was completely inhibited after exposure of the flagellates to doses equal to or higher than 50 J/m2. The UV-induced lag period was dose-dependent. Survival was reduced to 1% after exposure of the parasites to 200 J/m2. Ultrastructural changes after the lag period were studied by transmission electron microscopy. Changes of the kinetoplast network structure and sometimes of the mitochondrial matrix were observed. The existence of DNA repair mechanisms in this protozoan is discussed.

Animals↗

Reptiles and their importance in the epidemiology of leishmaniasis.

Promastigote flagellates have been isolated from various species of lizard and from some other reptiles. It is known that sandflies readily feed upon lizards and it has been thought that reptiles could be a reservoir for mammalian leishmaniasis. A feature of reptilian infections is the extreme scarcity of parasites in blood smears and in tissue impression smears but isolations may readily be made in culture media. The intradermal inoculation of promastigote cultures from lizards into mammals and man induces a positive leishmanial response and gives rise to long-lasting dermal knots from which living parasites can be recovered for periods of several months.Associations between promastigotes and lizards in parts of the USSR, particularly in the Turkmenian SSR, and experimental work in the USSR on the transmission of promastigote strains to a variety of mammals, including gerbils, mice, monkeys, and man, are reviewed. The author accepts the generally held view that although promastigote flagellates of reptiles are important in an evolutionary context, having probably given rise to the mammalian leishmanias, present-day strains must be considered nonpathogenic for mammals.

Animals↗

Infection of the colon of the rhesus monkey by spiral-shaped organisms.

Intestinal spirochetosis, an infection of the mucosa by spiral-shaped organisms, was studied in clinically normal rhesus monkeys (Macaca mulatta) by histology, transmission and scanning electron microscopy. The incidence of intestinal spirochetosis was 42% in 221 monkeys. Spiral organisms stained with hematoxylin and eosin (HE) appeared as a broad basophilic haze on the colonic surface and were strongly positive by the Warthin-Starry stain. Spiral-shaped bacteria include two structurally different organisms: spirochetes and flagellated microbes. They intimately populated the brush border of the surface of the epithelium of the large intestine. They were absent in the crypts and in the small intestine. Infection by spirochetes produced no alteration of cytocomponents of the underlying host structures. Spirochetes and flagellates infrequently penetrated beyond the brush border into the epithelial cytoplasm and also into the lamina propria. Even in cases where invasion was documented, no inflammatory response was found.

Animals↗

Infection of the colon of the rhesus monkey by spiral-shaped organisms.

Intestinal spirochetosis, an infection of the mucosa by spiral-shaped organisms, was studied in clinically normal rhesus monkeys (Macaca mulatta) by histology, transmission and scanning electron microscopy. The incidence of intestinal spirochetosis was 42% in 221 monkeys. Spiral organisms stained with hematoxylin and eosin (HE) appeared as a broad basophilic haze on the colonic surface and were strongly positive by the Warthin-Starry stain. Spiral-shaped bacteria include two structurally different organisms: spirochetes and flagellated microbes. They intimately populated the brush border of the surface of the epithelium of the large intestine. They were absent in the crypts and in the small intestine. Infection by spirochetes produced no alteration of cytocomponents of the underlying host structures. Spirochetes and flagellates infrequently penetrated beyond the brush border into the epithelial cytoplasm and also into the lamina propria. Even in cases where invasion was documented, no inflammatory response was found.

Animals↗

Appearance of heat shock proteins during the induction of multiple flagella in Naegleria gruberi.

A heat shock to amebae of the amebo-flagellate Naegleria gruberi during differentiation into swimming flagellates results in the induction of heat shock proteins as well as multiple flagella. The principal heat shock proteins migrate on sodium dodecyl sulfate-polyacrylamide gels with apparent molecular weights of 96,000, 77,000, 70,000, and 68,000. These proteins are synthesized preferentially when cells at 25 degrees C are shifted to temperatures above 32 degrees C. The maximal incorporation of methionine into heat shock proteins occurs at 38.2 degrees C, the temperature at which maximal induction of multiple flagella has been reported. Synthesis of heat shock proteins requires new transcription as judged by the ability of actinomycin D to inhibit their synthesis during the first 15 min of heat shock but not thereafter. Although heat shock can induce multiple flagella only when applied during a restricted interval, heat shock proteins are induced at any time cells are shifted to 38.2 degrees C. The response to heat shock of the Naegleria heat shock proteins resembles that of Drosophila heat shock proteins, but the two groups of proteins differ in both size and number. Naegleria heat shock proteins are, however, strikingly similar in size to a group of heat-induced proteins found in chick embryo fibroblast, mouse L, and BHK cells.

Amoeba↗

Leishmaniasis in the lowlands of Bolivia. Entomological studies on sandflies of the "Valle del Sacta". Tropical Carrasco of the Department of Cochabamba.

Up to 1987, through the LEISHBOL project (CEE-STD1) studies on sandflies were performed in the rain forests of Yapacani, a region of the Department of Santa Cruz. In another set of studies, also carried out within the LEISHBOLPE project (CEE-STD2), sandflies were studied in the rain forests of the "Valle del Sacta, Prov. Carrasco" of the Departament of Cochabamba, to determine the composition of the species, its density, the anthropophilic species, and its infection by flagellates. 18 species were identified, from those collected in the Valle del Sacta, one of which was a new species now being described as Lutzomyia lerayi n-sp. Only 7 species are common with those identified in Yapacani, in spite of the proximity and of the apparent similarity of the biotopes. Lu. shawi was the only species captured practically year round, and highest densities were identified in October with 7.6 "females/hour/shannon trap". The lowest monthly density occurred in July with "0.2 female/hour/shannon trap." In relation to the altitudinal density of Lu. shawi, it was higher at 5 meters (5.5 females/night/CDC in October) than at 1 meter (2.3 females/night/CDC-trap in November) above ground level. The following species was detected as anthropophilic: Lu. shawi, Lu. amazonensis, Lu. evangelistai and Lu. sherlocki. 4.5% (7 of 154) of Lu. shawi were found to be infected by flagellates in suprapiloric position. The composition of the sandfly species of Valle del Sacta is different than that of Yacapani. The most important species of Valle del Sacta is Lu. shawi, instead of Lu. yucumensis of Yapacani.

Altitude↗

Interaction of Listeria species with human cell monolayers.

In foodborne listeriosis, the first step of infection must be attachment to, and invasion of, the gastrointestinal epithelium by virulent Listeria monocytogenes. Virulence factors affecting this invasion are only now being determined. We examined the interaction of L. monocytogenes, serotypes 4b and 1/2a strains with "smooth" and "rough" characteristics. In addition, flagellated and non-flagellated isogenic strains altered by transposon mutagenesis were examined to study the effect of flagellae on virulence. Strains of Listeria ivanovii and Listeria innocua were also studied. L. ivanovii demonstrated the greatest degree of internalization and L. innocua, the least. L. monocytogenes strains were intermediate and there were no significant differences between "rough" and "smooth" isolates. The presence or absence of flagellae did not effect cell association in this model. Minor differences between cell lines for the same strains were demonstrated. Variations in cell association for differing inocula temperatures (4 degrees, 22 degrees, 37 degrees) were also noted for all strains. The interaction of Listeria species with human cell monolayers appears to be complex. Hemolysin clearly play a significant role but other virulence factors may also be important.

Bacterial Adhesion↗

An animal model of foodborne Listeria monocytogenes virulence: effect of alterations in local and systemic immunity on invasive infection.

Development of foodborne listeriosis is dependent on an interplay between organism-specific virulence factors and host susceptibility. Gastric inoculation of Sprague-Dawley rats was used as a model to explore Listeria-specific virulence and host susceptibility. Gastric inoculation leads to invasive infection with "smooth" hemolytic Listeria monocytogenes but not with "rough" L. monocytogenes or other Listeria species. Infection is dose-dependent with an ID50 of 10(6) virulent Listeria monocytogenes. In these experiments, the ID50 was not altered by pregnancy but invasive infection led to abnormal reproductive outcomes including stillbirth and reabsorption of fetuses. Immunosuppression by cyclosporin A led to more prolonged infection but did not alter the ID50. Manipulation of intestinal flora with antibiotics suggested increased rates of infection with antibiotics that decreased anaerobic flora. Growth of virulent Listeria in milk at varying temperatures did not enhance virulence. No differences in invasive potential of flagellated vs. non-flagellated L. monocytogenes were found. Oral models of invasive Listeria monocytogenes infection provide a useful tool for studying organism virulence and host susceptibility.

Animals↗

Evaluation of Extracellular, High-affinity beta-N-acetylglucosaminidase Measurements from Freshwater Lakes: An Enzyme Assay to Estimate Protistan Grazing on Bacteria and Picocyanobacteria

Protistan community grazing rates upon both bacterioplankton and autotrophic picoplankton were estimated using fluorescently-labeled prey and by measurement of extracellular hydrolysis of 4-methylumbelliferyl (MUF) beta-N-acetylglucosaminide in a eutrophic reservoir and an oligo-mesotrophic lake during phytoplankton blooms. In addition, enzyme methods were optimized in bacterivorous flagellate cultures by two enzyme assays, based on fluorometric detection of protistan digestive activity, which were compared and calibrated independently against flagellate bacterivory. Enzymatic hydrolyses of MUF beta-N,N',N''-triacetylchitotriose and MUF beta-N-acetylglucosaminide were measured in cell-free (sonicated) and whole-cell (unsonicated) samples. The hydrolysis of both substrates, using the whole-cell enzyme assay at in situ pH, was correlated significantly with total grazing rate of Bodo saltans. Thus the whole-cell enzyme assay with MUF beta-N-acetylglucosaminide was used for freshwater samples. High-affinity (Km < 1 μmol l-1) and low-affinity (Km > 100μmol l-1) enzymes were distinguished kinetically in most samples from both systems studied. Activities (Vmax) of the high-affinity enzyme varied from 0.24 to 1.43 nmol l-1 h-1. Protistan community grazing on bacterioplankton was in the range of 0.15-1.36 μg C l-1 h-1 both for lake and reservoir, the differences being observed in grazing on picocyanobacteria (lake, 0.03-0.22 μg C l-1 h-1; reservoir, 0.35-1.56 μg C l-1 h-1). The enzyme activities were correlated significantly with the protistan grazing both on bacterioplakton (rs=0.62, P<0.001) and total procaryotic picoplankton (the sum of organic carbon grazed from bacteria and picocyanobacteria, rs=0.73, P<0.001) in the eutrophic reservoir. Weaker relationships (rs=0.42) with a lower slope were found for the oligo-mesotrophic lake. Ingestion rate studies are time-consuming and the digestive enzyme assay with MUF beta-N-acetylglucosaminide presents a rapid alternative for estimating total protistan prokaryotic picoplanktivory in freshwaters.

Journal Article↗

Leishmania major: amastigote formation in cell-free media.

The life cycle of Leishmania consists of two distinct developmental stages: the amastigote, which is the ovoid non-flagellated form found in the vertebrate host, and the promastigote, which is an elongated flagellated form found in the gut of an infected sandfly. Following its injection into the vertebrate host by the sandfly vector, the promastigote transforms into an amastigote after entering the host macrophage. The environmental cues inducing this transformation are not fully understood. Attempts to axenically develop and cultivate amastigotes from different Leishmania species have indicated that species and sometimes even strains of the same species vary in their requirements for this process (Pan et al., 1993). The majority of the available published data on transformation pertains to New World Leishmania and provides evidence that elevation in the incubation temperature and/or acidic pH can in some cases induce amastigote formation from promastigotes.

Animals↗