WorldWideScience

Sample records for amoeba

  1. Free-Living Amoebae Keratitis.

    Science.gov (United States)

    Pinna, Antonio; Porcu, Tiziana; Boscia, Francesco; Cano, Antonella; Erre, Giuseppe; Mattana, Antonella

    2017-07-01

    To describe the diagnostic and clinical features and treatment results in 43 consecutive patients with microbiologically proven free-living amoebae (FLA) keratitis. In this hospital-based, prospective case series, corneal scrapings from 43 patients with presumed amoebic keratitis were plated on nonnutrient agar. Amoebic isolates were identified morphologically and by the polymerase chain reaction. All patients with culture-proven FLA keratitis were treated with polyhexamethylene biguanide (PHMB) 0.02% eye drops. Forty-three corneal scrapings from 43 patients were found to be culture positive for FLA; 41 (95%) were from contact lens wearers and 2 (5%) were from noncontact lens wearers. Microscopic examination identified 4 Acanthamoeba spp, 24 Hartmannella spp, 12 vahlkampfiid amoebae, and 3 mixed infections with Hartmannella/vahlkampfiid amoebae. Morphological results were confirmed by the polymerase chain reaction. Patients with Acanthamoeba, Hartmannella, and vahlkampfiid keratitis had indistinguishable clinical features. In 38 eyes with keratitis at an early stage, treatment with PHMB 0.02% eye drops was fully successful. In 5 patients with advanced keratitis, topical PHMB 0.02% controlled the infection, but all of them developed a central corneal scar with visual deterioration. Acanthamoeba is not the only cause of amoebic keratitis, because this condition may also be caused by other FLA, such as Hartmannella and vahlkampfiid amoebae. This finding is epidemiologically interesting, suggesting a possible different geographical prevalence of the different FLA responsible for keratitis. Early diagnosis and proper antiamoebic treatment are crucial to yielding a cure.

  2. Performance of the Amoeba Distributed Operating System

    NARCIS (Netherlands)

    van Renesse, R.; van Staveren, H.; Tanenbaum, A.S.

    1989-01-01

    Amoeba is a capability‐based distributed operating system designed for high‐performance interactions between clients and servers using the well‐known RPC model. The paper starts out by describing the architecture of the Amoeba system, which is typified by specialized components such as workstations,

  3. How do amoebae swim and crawl?

    Directory of Open Access Journals (Sweden)

    Jonathan D Howe

    Full Text Available The surface behaviour of swimming amoebae was followed in cells bearing a cAR1-paGFP (cyclic AMP receptor fused to a photoactivatable-GFP construct. Sensitized amoebae were placed in a buoyant medium where they could swim toward a chemoattractant cAMP source. paGFP, activated at the cell's front, remained fairly stationary in the cell's frame as the cell advanced; the label was not swept rearwards. Similar experiments with chemotaxing cells attached to a substratum gave the same result. Furthermore, if the region around a lateral projection near a crawling cell's front is marked, the projection and the labelled cAR1 behave differently. The label spreads by diffusion but otherwise remains stationary in the cell's frame; the lateral projection moves rearwards on the cell (remaining stationary with respect to the substrate, so that it ends up outside the labelled region. Furthermore, as cAR1-GFP cells move, they occasionally do so in a remarkably straight line; this suggests they do not need to snake to move on a substratum. Previously, we suggested that the surface membrane of a moving amoeba flows from front to rear as part of a polarised membrane trafficking cycle. This could explain how swimming amoebae are able to exert a force against the medium. Our present results indicate that, in amoebae, the suggested surface flow does not exist: this implies that they swim by shape changes.

  4. Amoebas of Complex Hypersurfaces in Statistical Thermodynamics

    Energy Technology Data Exchange (ETDEWEB)

    Passare, Mikael [Stockholm University, Department of Mathematics (Sweden); Pochekutov, Dmitry, E-mail: potchekutov@gmail.com [Siberian Federal University, Institute of Core Undergraduate Programmes (Russian Federation); Tsikh, August, E-mail: atsikh@sfu-kras.ru [Siberian Federal University, Institute of Mathematics (Russian Federation)

    2013-03-15

    The amoeba of a complex hypersurface is its image under the logarithmic projection. A number of properties of algebraic hypersurface amoebas are carried over to the case of transcendental hypersurfaces. We demonstrate the potential that amoebas can bring into statistical physics by considering the problem of energy distribution in a quantum thermodynamic ensemble. The spectrum {l_brace}{epsilon}{sub k}{r_brace} Subset-Of Z{sup n} of the ensemble is assumed to be multidimensional; this leads us to the notions of multidimensional temperature and a vector of differential thermodynamic forms. Strictly speaking, in the paper we develop the multidimensional Darwin-Fowler method and give the description of the domain of admissible average values of energy for which the thermodynamic limit exists.

  5. Biodiversity of amoebae and amoebae-resisting bacteria in a drinking water treatment plant.

    Science.gov (United States)

    Thomas, Vincent; Loret, Jean-François; Jousset, Michel; Greub, Gilbert

    2008-10-01

    The complex ecology of free-living amoebae (FLA) and their role in spreading pathogenic microorganisms through water systems have recently raised considerable interest. In this study, we investigated the presence of FLA and amoebae-resisting bacteria (ARB) at various stages of a drinking water plant fed with river water. We isolated various amoebal species from the river and from several points within the plant, mostly at early steps of water treatment. Echinamoeba- and Hartmannella-related amoebae were mainly recovered in the drinking water plant whereas Acanthamoeba- and Naegleria-related amoebae were recovered from the river water and the sand filtration units. Some FLA isolates were recovered immediately after the ozonation step, thus suggesting resistance of these microorganisms to this disinfection procedure. A bacterial isolate related to Mycobacterium mucogenicum was recovered from an Echinamoeba-related amoeba isolated from ozone-treated water. Various other ARB were recovered using co-culture with axenic Acanthamoeba castellanii, including mycobacteria, legionella, Chlamydia-like organisms and various proteobacteria. Noteworthy, a new Parachlamydia acanthamoebae strain was recovered from river water and from granular activated carbon (GAC) biofilm. As amoebae mainly multiply in sand and GAC filters, optimization of filter backwash procedures probably offers a possibility to better control these protists and the risk associated with their intracellular hosts.

  6. Experiences with the Amoeba distributed operating system

    NARCIS (Netherlands)

    Tanenbaum, Andrew S.; van Renesse, Robbert; van Staveren, Hans; Sharp, Gregory J.; Mullender, Sape J.; Jansen, Jack; van Rossum, Guido

    1990-01-01

    The Amoeba project is a research effort aimed at understanding how to connect multiple computers in a seamless way [16, 17, 26, 27, 31]. The basic idea is to provide the users with the illusion of a single powerful timesharing system, when, in fact, the system is implemented on a collection of

  7. Infection and Proliferation of Giant Viruses in Amoeba Cells.

    Science.gov (United States)

    Takemura, Masaharu

    2016-01-01

    Acanthamoeba polyphaga mimivirus, the first discovered giant virus with genome size and particle size much larger than previously discovered viruses, possesses several genes for translation and CRISPER Cas system-like defense mechanism against virophages, which co-infect amoeba cells with the giant virus and which inhibit giant virus proliferation. Mimiviruses infect amoeba cells by phagocytosis and release their DNA into amoeba cytoplasm through their stargate structure. After infection, giant virion factories (VFs) form in amoeba cytoplasm, followed by DNA replication and particle formation at peripheral regions of VF. Marseilleviruses, the smallest giant viruses, infect amoeba cells by phagocytosis or endocytosis, form larger VF than Mimivirus's VF in amoeba cytoplasm, and replicate their particles. Pandoraviruses found in 2013 have the largest genome size and particle size among all viruses ever found. Pandoraviruses infect amoeba cells by phagocytosis and release their DNA into amoeba cytoplasm through their mouth-like apical pores. The proliferation of Pandoraviruses occurs along with nucleus disruption. New virions form at the periphery of the region formerly occupied by the amoeba cell nucleus.

  8. Revised Parameters for the AMOEBA Polarizable Atomic Multipole Water Model.

    Science.gov (United States)

    Laury, Marie L; Wang, Lee-Ping; Pande, Vijay S; Head-Gordon, Teresa; Ponder, Jay W

    2015-07-23

    A set of improved parameters for the AMOEBA polarizable atomic multipole water model is developed. An automated procedure, ForceBalance, is used to adjust model parameters to enforce agreement with ab initio-derived results for water clusters and experimental data for a variety of liquid phase properties across a broad temperature range. The values reported here for the new AMOEBA14 water model represent a substantial improvement over the previous AMOEBA03 model. The AMOEBA14 model accurately predicts the temperature of maximum density and qualitatively matches the experimental density curve across temperatures from 249 to 373 K. Excellent agreement is observed for the AMOEBA14 model in comparison to experimental properties as a function of temperature, including the second virial coefficient, enthalpy of vaporization, isothermal compressibility, thermal expansion coefficient, and dielectric constant. The viscosity, self-diffusion constant, and surface tension are also well reproduced. In comparison to high-level ab initio results for clusters of 2-20 water molecules, the AMOEBA14 model yields results similar to AMOEBA03 and the direct polarization iAMOEBA models. With advances in computing power, calibration data, and optimization techniques, we recommend the use of the AMOEBA14 water model for future studies employing a polarizable water model.

  9. Sucker-like structures on the pathogenic amoeba Naegleria fowleri.

    OpenAIRE

    John, D T; Cole, T B; Marciano-Cabral, F M

    1984-01-01

    Using scanning electron microscopy, we observed sucker-like structures on amoebae of 13 human isolates of Naegleria fowleri. The number of suckers per amoeba seemed to vary according to the virulence of the strain. We propose the term amoebastome to describe this unique sucker-like structure of N. fowleri.

  10. Amoeba: a distributed operating system for the 1990s

    NARCIS (Netherlands)

    Mullender, Sape J.; van Rossum, Guido; Tanenbaum, Andrew S.; van Renesse, Robbert; van Staveren, Hans

    A description is given of the Amoeba distributed operating system, which appears to users as a centralized system but has the speed, fault tolerance, security safeguards, and flexibility required for the 1990s. The Amoeba software is based on objects. Objects are managed by server processes and

  11. Amoeba: A Distributed Operating System for the 1990s

    NARCIS (Netherlands)

    Mullender, S.J.; van Rossum, G.; Tanenbaum, A.S.; van Renesse, R.; van Staveren, H.

    1990-01-01

    A description is given of the Amoeba distributed operating system, which appears to users as a centralized system but has the speed, fault tolerance, security safeguards, and flexibility required for the 1990s. The Amoeba software is based on objects. Objects are managed by server processes and

  12. Cocultivation of Legionella pneumophila and free-living amoebae.

    Science.gov (United States)

    Tyndall, R L; Domingue, E L

    1982-10-01

    Studies of the interaction of Legionella pneumophila with free-living amoebae showed that Naegleria lovaniensis and Acanthamoeba royreba could use L. pneumophila as a sole food source. However, growth of the amoebae on nonnutrient agar plates seeded with L. pneumophila was slower than growth on nonnutrient agar plates seeded with Escherichia coli. On inoculation of L. pneumophila into axenic cultures of N. lovaniensis and A. royreba, 99.9% of the L. pneumophila was destroyed within 24 h. After several weeks, however, some amoeba cultures became chronically infected and supported the growth of L. pneumophila. Amoebae exposed to L. pneumophila and containing adhered L. pneumophila, L. pneumophila antigens, or both, showed no increased pathogenic potential on intranasal inoculation of weanling mice. Similarly, L. pneumophila propagated in chronically infected amoeba cultures showed no increase in virulence on intraperitoneal inoculation of guinea pigs relative to L. pneumophila grown in yeast extract broth.

  13. Cocultivation of Legionella pneumophila and free-living amoebae

    Energy Technology Data Exchange (ETDEWEB)

    Tyndall, R.L.; Domingue, E.L.

    1982-10-01

    Studies of the interaction of Legionella pneumophila with free-living amoebae showed that Naegleria lovaniensis and Acanthamoeba royreba could use L. pneumophia as a sole food source. However, growth of the amoebae on nonnutrient agar plates seeded with L. pneumophila was slower than growth on nonnutrient agar plates seeded with Escherichia coli. On inoculation of L. pneumophila into axenic cultures of N. lovaniensis and A. roryba, 99.9% of the L. pneumophila was destroyed within 24 h. After several weeks, however, some amoeba cultures became chronically infected and supported the growth of L. pneumophila. Amoebae exposed to L. pneumophila and containing adhered L. pneumophila, L. pneumophila antigens, or both, showed no increased pathogenic potential on intranasal inoculation of weanling mice. Similarly, L. pneumophila propagated in chronically infected amoeba cultures showed no increase in virulence on intraperitoneal inoculation of guinea pigs relative to L. pneumophila grown in yeast extract broth. 20 references, 1 figure, 4 tables.

  14. Multiplication rate of amoebae related to the cultivation temperature

    Energy Technology Data Exchange (ETDEWEB)

    Sopina, V.A.

    1976-10-01

    The cultivation temperature-dependent rate of multiplication in amoebae was studied using three strains of Amoeba proteus and another amoeba strain of unknown specific and generic position. The multiplication rates are characterized by optima that vary with strains. The temperature-induced changes in the multiplication rate of a given strain are non-hereditary; they are common modifications. Hereditary interstrain differences in the multiplication rate have also been shown at constant temperatures. The range of modificational changes with increase or decrease of cultivation temperature appears to be greater than interstrain differences in multiplication rate.

  15. Buse_Francisella and free-living amoebae data sets

    Data.gov (United States)

    U.S. Environmental Protection Agency — Co-infection data in the form of colony forming units and amoeba cell counts. This dataset is associated with the following publication: Buse , H., F. Schaefer, and...

  16. Phylogeny, evolution, and taxonomy of vannellid amoebae.

    Science.gov (United States)

    Smirnov, Alexey V; Nassonova, Elena S; Chao, Ema; Cavalier-Smith, Thomas

    2007-07-01

    We sequenced 18S rRNA genes from 21 vannellid amoebae (Amoebozoa; Vannellidae), including nearly all available type cultures, and performed a comprehensive phylogenetic analysis for 57 Vannellidae sequences. The results show that species of Vannella and Platyamoeba are completely mixed and do not form distinct clades. Several very closely related species pairs exist, each with a Vannella and a Platyamoeba species differing in only a few nucleotides. Therefore, presence (Vannella) or absence (Platyamoeba) of glycostyles in the cell surface coat is an invalid generic distinction; the genera must be merged. As Vannella has priority, we formally transferred Platyamoeba species into Vannella, except for the non-vannellid P. stenopodia, here renamed Stenamoeba stenopodia gen. n. comb. n. and transferred to the family Thecamoebidae. Our trees show that Vannella glycostyles were probably easily and repeatedly evolutionarily lost. We have established a new genus Ripella, with distinct morphology and sequence signatures for Vannella platypodia and morphologically similar species that form a clearly separate clade, very distant from other Vannellidae. Vannellids form four well-separated single-genus clades: Vannella sensu stricto, Ripella, Clydonella, and Lingulamoeba. Species of the revised genus Vannella comprise four closely related, well-supported subclades: one marine and three freshwater. Here, we provide an illustrated checklist for all 40 known Vannellidae species.

  17. Pathogenic Free-Living Amoebae Isolated From Contact Lenses of Keratitis Patients

    OpenAIRE

    HAJIALILO, Elham; NIYYATI, Maryam; SOLAYMANI, Mohammad; REZAEIAN, Mostafa

    2015-01-01

    Background: Free-living amoeba (FLA)-related keratitis is a progressive infection of the cornea with poor prognosis. The present study aimed to investigates the con­tact lenses of patients with keratitis for pathogenic free-living amoebae.Methods: Overall, 62 contact lenses and their paraphernalia of patients with kerati­tis cultured and tested for the presence of free-living amoebae using morphological criteria. Unusual plates including plates containing mix amoebae and Vermamoeba were submi...

  18. Parasitic amoebae found in water bodies of Ukraine.

    Science.gov (United States)

    Patsyuk, Marina

    2017-12-01

    Two parasitic amoebian species are found in mollusks of the water bodies of Ukraine. Vahlkampfia sp. is found in hepatopankreas of Unio conus Spengler, 1793, and Acanthamoeba sp. is observed in mantle cavity of Viviparus viviparus Linnaeus, 1758. For these protist species, the mollusks are shown to be intermediate hosts where amoebae feed and reproduce. An experimental infection with Vahlkampfia sp. and Acanthamoeba sp. was not successful, no pathological changes in mollusks were observed. These amoebae are successfully cultured in fresh water and agar medium, hence we can safely consider them free-living. Copyright © 2017 Elsevier Inc. All rights reserved.

  19. The Evolution of a distributed operating system (Amoeba)

    NARCIS (Netherlands)

    van Renesse, Robbert; Tanenbaum, Andrew S.; Mullender, Sape J.

    1989-01-01

    AMOEBA is a research project to build a true distributed operating system using the object model. Under the COST11-ter MANDIS project this work was extended to cover wide-area networks. Besides describing the system, this paper discusses the successive versions in the implementation of its model,

  20. Shifts in soil testate amoeba communities associated with forest diversification.

    Science.gov (United States)

    Bobrov, Anatoly A; Zaitsev, Andrei S; Wolters, Volkmar

    2015-05-01

    We studied changes of testate amoeba communities associated with the conversion of spruce monocultures into mixed beech-fir-spruce forests in the Southern Black Forest Mountains (Germany). In this region, forest conversion is characterized by a gradual development of beech undergrowth within thinned spruce tree stands leading to multiple age continuous cover forests with a diversified litter layer. Strong shifts in the abundance of testate amoeba observed in intermediate stages levelled off to monoculture conditions again after the final stage of the conversion process had been reached. The average number of species per conversion stage (i.e., local richness) did not respond strongly to forest conversion, but the total number of species (i.e., regional richness) was considerably higher in the initial stage than in the mixed forests, due to the large number of hygrophilous species inhabiting spruce monocultures. Functional diversity of the testate amoeba community, however, significantly increased during the conversion process. This shift was closely associated with improved C and N availability as well as higher niche diversity in the continuous cover stands. Lower soil acidity in these forests coincided with a higher relative abundance of eurytopic species. Our results suggest that testate amoeba communities are much more affected by physicochemical properties of the soil than directly by litter diversity.

  1. Elemental maps of Amoeba proteus by a scanning proton microprobe

    Energy Technology Data Exchange (ETDEWEB)

    Li Minqian; Zhu Jieqing; Huang Zeqi (Shanghai Inst. of Nuclear Research, Academia Sinica, SH (China)); Zhu Jingde; Zhou Zheng; Zhou Weiying (Shanghai Inst. of Cell Biology, Academia Sinica, SH (China)); Cholewa, M.; Legge, G.J.F. (Micro Analytical Research Centre, Univ. of Melbourne, School of Physics, Parkville (Australia))

    1991-03-01

    Elemental maps for P, S, Cl, K, Ca and Zn of individual Amoeba proteus were obtained with the Melbourne scanning proton microprobe. The emphasis was put on the relationship of both distribution and concentration of Zn within the cell and the growth inhibitory effect of higher Zn concentrations in the culture medium. At a concentration of 0.04 mmol ZnCl{sub 2}. Amoeba growth was inhibited. But at a concentration of 0.0016 mmol, the Amoeba grew as well as a control grown without addition of Zn. We found that in the former (0.04 mmol) Zn concentrated three times more than in the latter (0.0016 mmol), and also that Zn was enriched much more in the nucleus and endoplasm (five to six times) than in other parts of the cell (two times). Future work along these lines may provide insight into the mechanism by which Zn affects the growth of Amoeba proteus and other cells. (orig.).

  2. A Comparison of Two Distributed Systems: Amoeba and Sprite

    NARCIS (Netherlands)

    Douglis, F.; Ousterhout, J.K.; Kaashoek, M.F.; Tanenbaum, A.S.

    This paper compares two distributed operating systems, Amoeba and Sprite. Although the systems share many goals, they diverged on two philosophical grounds: whether to emphasize a distributed computing model or traditional UNIX-style applications, and whether to use a workstation-centered model of

  3. Methodology to Implement an Amoeba Complex Object Server

    NARCIS (Netherlands)

    Teeuw, Wouter B.; Blanken, Henk

    The paper describes the methodology used in the design of a complex object server application for the Amoeba distributed operating system. It uses the top-down design that was suggested by Parnas, in which a model is turned into an implementation by gradually adding details. The paper also describes

  4. AIL - a class-oriented RPC stub generator for Amoeba

    NARCIS (Netherlands)

    G. van Rossum (Guido)

    1989-01-01

    textabstractAIL – an acronym for Amoeba Interface Language – is a class-oriented RPC stub generator, used with Amoeba’s RPC primitives. Together with Amoeba’s facilities for manipulating capabilities (bit patterns that are unforgeable references to objects maintained by servers anywhere on a

  5. The genome of the social amoeba Dictyostelium discoideum

    DEFF Research Database (Denmark)

    Eichinger, L; Pachebat, J A; Glöckner, G

    2005-01-01

    The social amoebae are exceptional in their ability to alternate between unicellular and multicellular forms. Here we describe the genome of the best-studied member of this group, Dictyostelium discoideum. The gene-dense chromosomes of this organism encode approximately 12,500 predicted proteins,...

  6. Advances in the knowledge of amphizoic amoebae infecting fish

    Czech Academy of Sciences Publication Activity Database

    Dyková, Iva; Lom, Jiří

    2004-01-01

    Roč. 51, 2/3 (2004), s. 81-97 ISSN 0015-5683 R&D Projects: GA AV ČR IAA6022202 Institutional research plan: CEZ:AV0Z6022909 Keywords : amphizoic amoebae * fish hosts * review Subject RIV: EA - Cell Biology Impact factor: 0.837, year: 2004

  7. Diagnosis of Infections Caused by Pathogenic Free-Living Amoebae

    Directory of Open Access Journals (Sweden)

    Bruno da Rocha-Azevedo

    2009-01-01

    Full Text Available Naegleria fowleri, Acanthamoeba spp., Balamuthia mandrillaris, and Sappinia sp. are pathogenic free-living amoebae. N. fowleri causes Primary Amoebic Meningoencephalitis, a rapidly fatal disease of the central nervous system, while Acanthamoeba spp. and B. mandrillaris cause chronic granulomatous encephalitis. Acanthamoeba spp. also can cause cutaneous lesions and Amoebic Keratitis, a sight-threatening infection of the cornea that is associated with contact lens use or corneal trauma. Sappinia pedata has been identified as the cause of a nonlethal case of amoebic encephalitis. In view of the potential health consequences due to infection with these amoebae, rapid diagnosis is critical for early treatment. Microscopic examination and culture of biopsy specimens, cerebral spinal fluid (CSF, and corneal scrapings have been used in the clinical laboratory. For amoebic keratitis, confocal microscopy has been used to successfully identify amoebae in corneal tissue. More recently, conventional and real-time PCR assays have been developed that are sensitive and specific for the amoebae. In addition, multiplex PCR assays are available for the rapid identification of these pathogens in biopsy tissue, CSF, and corneal specimens.

  8. Inactivation and Removal of Free-Living Amoebae.

    Science.gov (United States)

    Free-living amoebae (FLA) are ubiquitous protozoan that are predominantly harmless to humans. There are a few genera that cause disease in humans, Balamuthia, Naegleria, and Acanthamoeba. These organisms are not easily removed by physical means or inactivated by chemic...

  9. Counting Legionella cells within single amoeba host cells

    Science.gov (United States)

    Here we present the first attempt to quantify L. pneumophila cell numbers within individual amoebae hosts that may be released into engineered water systems. The maximum numbers of culturable L. pneumophila cells grown within Acanthamoeba polyphaga and Naegleria fowleri were 134...

  10. Surveillance of parasitic Legionella in surface waters by using immunomagnetic separation and amoebae enrichment.

    Science.gov (United States)

    Hsu, Tsui-Kang; Wu, Shu-Fen; Hsu, Bing-Mu; Kao, Po-Min; Tao, Chi-Wei; Shen, Shu-Min; Ji, Wen-Tsai; Huang, Wen-Chien; Fan, Cheng-Wei

    2015-01-01

    Free-living amoebae (FLA) are potential reservoirs of Legionella in aquatic environments. However, the parasitic relationship between various Legionella and amoebae remains unclear. In this study, surface water samples were gathered from two rivers for evaluating parasitic Legionella. Warmer water temperature is critical to the existence of Legionella. This result suggests that amoebae may be helpful in maintaining Legionella in natural environments because warmer temperatures could enhance parasitisation of Legionella in amoebae. We next used immunomagnetic separation (IMS) to identify extracellular Legionella and remove most free Legionella before detecting the parasitic ones in selectively enriched amoebae. Legionella pneumophila was detected in all the approaches, confirming that the pathogen is a facultative amoebae parasite. By contrast, two obligate amoebae parasites, Legionella-like amoebal pathogens (LLAPs) 8 and 9, were detected only in enriched amoebae. However, several uncultured Legionella were detected only in the extracellular samples. Because the presence of potential hosts, namely Vermamoeba vermiformis, Acanthamoeba spp. and Naegleria gruberi, was confirmed in the samples that contained intracellular Legionella, uncultured Legionella may survive independently of amoebae. Immunomagnetic separation and amoebae enrichment may have referential value for detecting parasitic Legionella in surface waters.

  11. Raman spectroscopic study on the excystation process in a single unicellular organism amoeba (Acanthamoeba polyphaga)

    Science.gov (United States)

    Lin, Yu-Chung; Perevedentseva, Elena; Cheng, Chia-Liang

    2015-05-01

    An in vivo Raman spectroscopic study of amoeba (Acanthamoeba polyphaga) is presented. The changes of the spectra during the amoeba cyst activation and excystation are analyzed. The spectra show the changes of the relative intensities of bands corresponding to protein, lipid, and carotenoid components during cyst activation. The presence of carotenoids in the amoeba is observed via characteristic Raman bands. These signals in the Raman spectra are intense in cysts but decrease in intensity with cyst activation and exhibit a correlation with the life cycle of amoeba. This work demonstrates the feasibility of using Raman spectroscopy for the detection of single amoeba microorganisms in vivo and for the analysis of the amoeba life activity. The information obtained may have implications for the estimation of epidemiological situations and for the diagnostics and prognosis of the development of amoebic inflammations.

  12. Survival of taylorellae in the environmental amoeba Acanthamoeba castellanii.

    Science.gov (United States)

    Allombert, Julie; Vianney, Anne; Laugier, Claire; Petry, Sandrine; Hébert, Laurent

    2014-03-19

    Taylorella equigenitalis is the causative agent of contagious equine metritis, a sexually-transmitted infection of Equidae characterised in infected mares by abundant mucopurulent vaginal discharge and a variable degree of vaginitis, cervicitis or endometritis, usually resulting in temporary infertility. The second species of the Taylorella genus, Taylorella asinigenitalis, is considered non-pathogenic, although mares experimentally infected with this bacterium can develop clinical signs of endometritis. To date, little is understood about the basic molecular virulence and persistence mechanisms employed by the Taylorella species. To clarify these points, we investigated whether the host-pathogen interaction model Acanthamoeba castellanii was a suitable model for studying taylorellae. We herein demonstrate that both species of the Taylorella genus are internalised by a mechanism involving the phagocytic capacity of the amoeba and are able to survive for at least one week inside the amoeba. During this one-week incubation period, taylorellae concentrations remain strikingly constant and no overt toxicity to amoeba cells was observed. This study provides the first evidence of the capacity of taylorellae to survive in a natural environment other than the mammalian genital tract, and shows that the alternative infection model, A. castellanii, constitutes a relevant alternative system to assess host-pathogen interactions of taylorellae. The survival of taylorellae inside the potential environmental reservoir A. castellanii brings new insight, fostering a broader understanding of taylorellae biology and its potential natural ecological niche.

  13. Pathogenic Free-Living Amoebae Isolated From Contact Lenses of Keratitis Patients.

    Science.gov (United States)

    Hajialilo, Elham; Niyyati, Maryam; Solaymani, Mohammad; Rezaeian, Mostafa

    2015-01-01

    Free-living amoeba (FLA)-related keratitis is a progressive infection of the cornea with poor prognosis. The present study aimed to investigates the contact lenses of patients with keratitis for pathogenic free-living amoebae. Overall, 62 contact lenses and their paraphernalia of patients with keratitis cultured and tested for the presence of free-living amoebae using morphological criteria. Unusual plates including plates containing mix amoebae and Vermamoeba were submitted to molecular analysis. Out of 62 plates, 11 revealed the outgrowth of free living amoeba of which 9 were Acanthamoeba, one plates contained mix amoebae including Acanthamoeba and Vermamoeba and one showed the presence of Vermamoeba. These two latter plates belonged to patients suffered from unilateral keratitis due to the misused of soft contact lenses. One of the patients had mix infection of Acanthamoeba (T4) and V. vermiformis meanwhile the other patient was infected with the V. vermiformis. Amoebic keratitis continues to rise in Iran and worldwide. To date, various genera of free-living amoebae such as Vermamoeba could be the causative agent of keratitis. Soft contact lens wearers are the most affected patients in the country, thus awareness of high-risk people for preventing free-living amoebae related keratitis is of utmost importance.

  14. Environmental Mycobacterium avium subsp. paratuberculosis Hosted by Free-Living Amoebae

    Directory of Open Access Journals (Sweden)

    Ascel Samba-Louaka

    2018-02-01

    Full Text Available Mycobacterium avium subsp. paratuberculosis is responsible for paratuberculosis in animals. This disease, leading to an inflammation of the gastrointestinal tract, has a high impact on animal health and an important economic burden. The environmental life cycle of M. avium subsp. paratuberculosis is poorly understood and several studies suggest that free-living amoebae (FLA might be a potential environmental host. FLA are protozoa found in water and soil that are described as reservoirs of pathogenic and non-pathogenic bacteria in the environment. Indeed, bacteria able to survive within these amoebae would survive phagocytosis from immune cells. In this study, we assessed the in vitro interactions between several strains of M. avium subsp. paratuberculosis and Acanthamoeba castellanii. The results indicate that the bacteria were able to grow within the amoeba and that they can survive for several days within their host. To explore the presence of M. avium subsp. paratuberculosis in environmental amoebae, we sampled water from farms positive for paratuberculosis. A M. avium subsp. paratuberculosis strain was detected within an environmental amoeba identified as related to the poorly described Rosculus genus. The bacterial strain was genotyped, showing that it was similar to previous infectious strains isolated from cattle. In conclusion, we described that various M. avium subsp. paratuberculosis strains were able to grow within amoebae and that these bacteria could be found on farm within amoebae isolated from the cattle environment. It validates that infected amoebae might be a reservoir and vector for the transmission of M. avium subsp. paratuberculosis.

  15. Pathogenic Free-Living Amoebae Isolated From Contact Lenses of Keratitis Patients

    Directory of Open Access Journals (Sweden)

    Elham HAJIALILO

    2015-12-01

    Full Text Available Background: Free-living amoeba (FLA-related keratitis is a progressive infection of the cornea with poor prognosis. The present study aimed to investigates the con­tact lenses of patients with keratitis for pathogenic free-living amoebae.Methods: Overall, 62 contact lenses and their paraphernalia of patients with kerati­tis cultured and tested for the presence of free-living amoebae using morphological criteria. Unusual plates including plates containing mix amoebae and Vermamoeba were submitted to molecular analysis. Results: Out of 62 plates, 11 revealed the outgrowth of free living amoeba of which 9 were Acanthamoeba, one plates contained mix amoebae including Acan­thamoeba and Vermamoeba and one showed the presence of Vermamoeba. These two latter plates belonged to patients suffered from unilateral keratitis due to the mis­used of soft contact lenses. One of the patients had mix infection of Acanthamoeba (T4 and V. vermiformis meanwhile the other patient was infected with the V. vermiformis. Conclusion: Amoebic keratitis continues to rise in Iran and worldwide. To date, various genera of free-living amoebae such as Vermamoeba could be the causative agent of keratitis. Soft contact lens wearers are the most affected patients in the country, thus awareness of high-risk people for preventing free-living amoebae re­lated keratitis is of utmost importance.

  16. Pathogenic amoebae in power-plant cooling lakes. Final report

    Energy Technology Data Exchange (ETDEWEB)

    Tyndall, R.L.; Willaert, E.; Stevens, A.R.

    1981-06-01

    Cooling waters and associated algae and sediments from four northern and four southern/western electric power plants were tested for the presence of pathogenic amoebae. Unheated control waters and algae/sediments from four northern and five southern/western sites were also tested. When comparing results from the test versus control sites, a significantly higher proportion (P less than or equal to 0.05) of the samples from the test sites were positive for thermophilic amoeba, thermophilic Naegleria and pathogenic Naegleria. The difference in number of samples positive for thermophilic Naegleria between heated and unheated waters, however, was attributable predominantly to the northern waters and algae/sediments. While two of four northern test sites yielded pathogenic Naegleria, seven of the eight isolates were obtained from one site. Seasonality effects relative to the isolation of the pathogen were also noted at this site. One pathogen was isolated from a southwestern test site. Pathogens were not isolated from any control sites. Some of the pathogenic isolates were analyzed serologically and classified as pathogenic Naegleria fowleri. Salinity, pH, conductivity, and bacteriological profiles did not obviously correlate with the presence or absence of pathogenic Naegleria. While thermal addition was significantly associated with the presence of thermophilic Naegleria (P less than or equal to 0.05), the data implicate other as yet undefined parameters associated with the presence of the pathogenic thermophile. Until further delineation of these parameters is effected, generalizations cannot be made concerning the effect of thermal impact on the growth of pathogenic amoeba in a particular cooling system.

  17. Passive versus active local microrheology in mammalian cells and amoebae

    Science.gov (United States)

    Riviere, C.; Gazeau, F.; Marion, S.; Bacri, J.-C.; Wilhelm, C.

    2004-12-01

    We compare in this paper the rotational magnetic microrheology detailed by Marion et al [18] and Wilhelm et al [19] to the passive tracking microrheology. The rotational microrheology has been designed to explore, using magnetic rotating probes, the local intracellular microenvironment of living cells in terms of viscoelasticity. Passive microrheology techniques is based on the analysis of spontaneous diffusive motions of Brownian probes. The dependence of mean square displacement (MSD) with the time then directly reflects the type of movement (sub-, hyper- or diffusive motions). Using the same intracellular probes, we performed two types of measurements (active and passive). Based on the fluctuation-dissipation theorem, one should obtain the same information from the both techniques in a thermally equilibrium system. Interestingly, our measurements differ, and the discordances directly inform on active biological processes, which add to thermally activated fluctuations in our out-of equilibrium systems. In both cell models used, mammalian Hela cells and amoebae Entamoeba Histolytica, a hyper-diffusive regime at a short time is observed, which highlights the presence of an active non-thermal driving force, acting on the probe. However, the nature of this active force in mammalian cells and amoebae is different, according to their different phenotypes. In mammalian cells active processes are governed by the transport, via molecular motors, on the microtubule network. In amoebae, which are highly motile cells free of microtubule network, the active processes are dominated by strong fluxes of cytoplasm driven by extension of pseudopodia, in random directions, leading to an amplitude of motion one order of magnitude higher than for mammalian cells. Figs 7, Refs 32.

  18. [Amoebas of the genus acanthamoeba as an etiological factor of keratitis].

    Science.gov (United States)

    Kosik-Bogacka, Danuta; Czepita, Damian; Łanocha, Natalia

    2010-01-01

    The aim of the paper was to describe the role of amoebas of the genus Acanthamoeba in the pathogenesis of keratitis. A review of literature concerning the role of amoebas of the genus Acanthamoeba in the pathogenesis of keratitis was done. Amoebas of the genus Acanthamoeba can be widely found in soil, air, fresh as well as saline water and as facultative parasites in humans. Amoebas of this genus are responsible for the creation of Acanthamoeba keratitis. In this paper the latest data concerning the epidemiology, diagnostic and treatment of Acanthamoeba keratitis have been cited and disscused. 1. Keratitis induced by amoebas of the genus Acanthamoeba are characterized as acute and difficult to treat. 2. Acanthamoeba keratitis occurs in recent years more often due to the fact that more people wear contact lenses.

  19. Free Living Amoeba Belonging to Vannella Spp. Isolated from a Hotspring in Amol City, Northern Iran

    Directory of Open Access Journals (Sweden)

    Maryam Niyyati

    2017-04-01

    Full Text Available Background: Free-living amoebae have various genera that are found in several environmental niches such as soil, freshwater, dust, seawater and hotsprings. Most of Free-living amoebae are normally harmless to humans. However, some ameoba such as Acanthamoeba and also Naegleria fowleri, Balamuthia manderillaris and Sappinia are identified as opportunistic and pathogenic amoebae that can cause eye diseases, encephalitis, and meningoencephalitis in human. Vannellidae are a family of free-living amoebae and exist mainly in soil, freshwater, and marine habitats. This amoeba is nonpathogenic for human, but can act as a Trojan horse for other pathogens such as Microsporidia. The present study reports the occurrence of Vannella spp. in a hotspring of Amol city.Materials and Methods: 22 samples were taken from hotsprings of Mazandaran province during our previous study. The plates were checked for the presence of Vannella spp. according to the specific morphological criteria. DNA extraction, PCR and sequencing was performed on the positive isolate.Results: The result showed that one plate contained fan-shaped amoebae suspected to Vannella spp. PCR analysis and sequencing was confirmed the occurrence of Vannella spp. in one sample of a hot spring of Amol, northern Iran.Conclusion: The result confirmed the presence of Vannella amoebae in the hotspring of Amol city. More studies are needed to clarify the real distribution of Vannella spp. in environmental niches and its pathogenic potential in Iran and worldwide.

  20. SPATIAL-VARIANT MORPHOLOGICAL FILTERS WITH NONLOCAL-PATCH-DISTANCE-BASED AMOEBA KERNEL FOR IMAGE DENOISING

    Directory of Open Access Journals (Sweden)

    Shuo Yang

    2015-01-01

    Full Text Available Filters of the Spatial-Variant amoeba morphology can preserve edges better, but with too much noise being left. For better denoising, this paper presents a new method to generate structuring elements for Spatially-Variant amoeba morphology.  The amoeba kernel in the proposed strategy is divided into two parts: one is the patch distance based amoeba center, and another is the geodesic distance based amoeba boundary, by which the nonlocal patch distance and local geodesic distance are both taken into consideration. Compared to traditional amoeba kernel, the new one has more stable center and its shape can be less influenced by noise in pilot image. What’s more important is that the nonlocal processing approach can induce a couple of adjoint dilation and erosion, and combinations of them can construct adaptive opening, closing, alternating sequential filters, etc. By designing the new amoeba kernel, a family of morphological filters therefore is derived. Finally, this paper presents a series of results on both synthetic and real images along with comparisons with current state-of-the-art techniques, including novel applications to medical image processing and noisy SAR image restoration.

  1. Detection and Identification of Free-living Amoeba from Environmental Water in Taiwan by PCR Method

    Science.gov (United States)

    Tsai, H. F.; Hsu, B. M.; Huang, K. H.; She, C. Y.; Kao, P. M.; Shen, S. M.; Tseng, S. F.; Chen, J. S.

    2012-04-01

    Acanthamoeba, Naegleria, Balamuthia and Hartmannella all belong to free-living amoebae that are present ubiquitously in the environment including water, soil, and air. Free-living amoebae are parasites which can infect humans and can lead to serious illness and even death. The aim of this study is to investigate the presence of free-living amoebae in aquatic environment in Taiwan, and to compare the differences between Acanthamoeba and Naegleria in diverse cultivation methods and conditions. In this study, we used molecular method by PCR amplification with specific primers to analyze the occurrence of free-living amoebae. We collected 176 samples from environmental water including drinking water treatment plants, stream water, and hot spring recreational areas in Taiwan. Based on the results of PCR, 43 water samples (24.4%) were detected positive for free-living amoebae. The most common Acanthamoeba genotype isolated from samples including T2, T4, T5, T12, and T15. N. australiensis and N. lovaniensis were also identified by molecular biology techniques. Furthermore, we found that both Acanthamoeba and Naegleria can be cultured by PYG in 30° C, but not all free-living amoebae can be isolated and enriched by using storage-cultivation method. Because of the widespread presence of Acanthamoeba and Naegleria in aquatic environments, the water quality and safety of aquatic environments should be more conscious in Taiwan and worldwide. Keywords: free-living amoebae; Acanthamoeba; Naegleria; Balamuthia; Hartmannella; PCR

  2. Genetic Variation in the Free-Living Amoeba Naegleria fowleri

    Science.gov (United States)

    Pélandakis, Michel; De Jonckheere, Johan F.; Pernin, Pierre

    1998-01-01

    In this study, 30 strains of the pathogenic free-living amoeba Naegleria fowleri were investigated by using the randomly amplified polymorphic DNA (RAPD) method. The present study confirmed our previous finding that RAPD variation is not correlated with geographical origin. In particular, Mexican strains belong to the variant previously detected in Asia, Europe, and the United States. In France, surprisingly, strains from Cattenom gave RAPD patterns identical to those of the Japanese strains. In addition, all of these strains, together with an additional French strain from Chooz, exhibited similarities to South Pacific strains. The results also confirmed the presence of numerous variants in Europe, whereas only two variants were detected in the United States. The two variants found in the United States were different from the South Pacific variants. These findings do not support the previous hypothesis concerning the origin and modes of dispersal of N. fowleri. PMID:9687460

  3. Contrasting species-environment relationships in communities of testate amoebae, bryophytes and vascular plants along the fen-bog gradient.

    Science.gov (United States)

    Lamentowicz, Mariusz; Lamentowicz, Lukasz; van der Knaap, Willem O; Gabka, Maciej; Mitchell, Edward A D

    2010-04-01

    We studied the vegetation, testate amoebae and abiotic variables (depth of the water table, pH, electrical conductivity, Ca and Mg concentrations of water extracted from mosses) along the bog to extremely rich fen gradient in sub-alpine peatlands of the Upper Engadine (Swiss Alps). Testate amoeba diversity was correlated to that of mosses but not of vascular plants. Diversity peaked in rich fen for testate amoebae and in extremely rich fen for mosses, while for testate amoebae and mosses it was lowest in bog but for vascular plants in extremely rich fen. Multiple factor and redundancy analyses (RDA) revealed a stronger correlation of testate amoebae than of vegetation to water table and hydrochemical variables and relatively strong correlation between testate amoeba and moss community data. In RDA, hydrochemical variables explained a higher proportion of the testate amoeba and moss data than water table depth. Abiotic variables explained a higher percentage of the species data for testate amoebae (30.3% or 19.5% for binary data) than for mosses (13.4%) and vascular plants (10%). These results show that (1) vascular plant, moss and testate amoeba communities respond differently to ecological gradients in peatlands and (2) testate amoebae are more strongly related than vascular plants to the abiotic factors at the mire surface. These differences are related to vertical trophic gradients and associated niche differentiation.

  4. Diversity and distribution of freshwater testate amoebae (protozoa) along latitudinal and trophic gradients in China.

    Science.gov (United States)

    Ju, Lihua; Yang, Jun; Liu, Lemian; Wilkinson, David M

    2014-11-01

    Freshwater microbial diversity is subject to multiple stressors in the Anthropocene epoch. However, the effects of climate changes and human activities on freshwater protozoa remain poorly understood. In this study, the diversity and distribution of testate amoebae from the surface sediments were investigated in 51 Chinese lakes and reservoirs along two gradients, latitude and trophic status. A total of 169 taxa belonging to 24 genera were identified, and the most diverse and dominant genera were Difflugia (78 taxa), Centropyxis (26 taxa) and Arcella (12 taxa). Our analysis revealed that biomass of testate amoebae decreased significantly along the latitudinal gradient, while Shannon-Wiener indices and species richness presented an opposite trend (P trophic status (P trophic status on diversity and distribution of testate amoebae in China. Therefore, our results provide valuable baseline data on testate amoebae and contribute to lake management and our understanding of the large-scale global patterns in microorganism diversity.

  5. Intra-amoeba multiplication induces chemotaxis and biofilm colonization and formation for Legionella.

    Directory of Open Access Journals (Sweden)

    Renaud Bigot

    Full Text Available Legionella pneumophila, a facultative intracellular bacterium, is the causative agent of legionellosis. In the environment this pathogenic bacterium colonizes the biofilms as well as amoebae, which provide a rich environment for the replication of Legionella. When seeded on pre-formed biofilms, L. pneumophila was able to establish and survive and was only found at the surface of the biofilms. Different phenotypes were observed when the L. pneumophila, used to implement pre-formed biofilms or to form mono-species biofilms, were cultivated in a laboratory culture broth or had grown intracellulary within the amoeba. Indeed, the bacteria, which developed within the amoeba, formed clusters when deposited on a solid surface. Moreover, our results demonstrate that multiplication inside the amoeba increased the capacity of L. pneumophila to produce polysaccharides and therefore enhanced its capacity to establish biofilms. Finally, it was shown that the clusters formed by L. pneumophila were probably related to the secretion of a chemotaxis molecular agent.

  6. Free-living amoebae in sediments from the Lascaux Cave in France.

    Directory of Open Access Journals (Sweden)

    Garcia-Sanchez A.M.

    2013-01-01

    Full Text Available The Lascaux Cave in France is an old karstic channel where the running waters are collected in a pool and pumped to the exterior. It is well-known that water bodies in the vicinity of humans are suspected to be reservoirs of amoebae and associated bacteria. In fact, the free-living amoebae Acanthamoeba astronyxis, Acanthamoeba castellanii, Acanthamoeba sp. and Hartmannella vermiformis were identif ied in the sediments of the cave using phylogenetic analyses and morphological traits. Lascaux Cave sediments and rock walls are wet due to a relative humidity near saturation and water condensation, and this environment and the presence of abundant bacterial communities constitute an ideal habitat for amoebae. The data suggest the need to carry out a detailed survey on all the cave compartments in order to determine the relationship between amoebae and pathogenic bacteria.

  7. Detection of Free-Living Amoebae in Some Water Sources and its Control by Ultraviolet- Radiation

    International Nuclear Information System (INIS)

    Moawed, M.A.F.; Shihab, A.

    2000-01-01

    Among the numerous free-living amoebae (FLA) of soil and water habitats, certain species belonging to two genera Acanthamoeba and Naegleria are facultative parasites of man.They cause disease called primary amoebic meningoencephalitis where history of contact with water was recorded in most of the cases especially with Naegleria species. The present work aimed to search for presence of free-living amoebae in the water and trials for its control by Ultraviolet-radiation (UV-radiation). Samples from different water sources were examined for the presence of free-living amoebae. These samples were cultured on non-nutrient agar streaked with bacteria. Amoebae were detected and identified by means of their morphological characters. Twelve positive cases of one hundred and twenty examined samples could be detected. The positive samples were exposed to different doses of UV-radiation for different times

  8. Divalent Metal Cations Potentiate the Predatory Capacity of Amoeba for Cryptococcus neoformans.

    Science.gov (United States)

    Fu, Man Shun; Casadevall, Arturo

    2018-02-01

    Among the best-studied interactions between soil phagocytic predators and a human-pathogenic fungus is that of Acanthamoeba castellanii and Cryptococcus neoformans The experimental conditions used in amoeba-fungus confrontation assays can have major effects on whether the fungus or the protozoan is ascendant in the interaction. In the presence of Mg 2+ and Ca 2+ in phosphate-buffered saline (PBS), C. neoformans was consistently killed when incubated with A. castellanii A. castellanii survived better in the presence of Mg 2+ and Ca 2+ , even when incubated with C. neoformans In the absence of Mg 2+ and Ca 2+ , C. neoformans survived when incubated with A. castellanii , and the percentage of dead amoebae was higher than when incubated without yeast cells. These results show that the presence of Mg 2+ and Ca 2+ can make a decisive contribution toward tilting the outcome of the interaction in favor of the amoeba. Of the two metals, Mg 2+ had a stronger effect than Ca 2+ The cations enhanced A. castellanii activity against C. neoformans via enhanced phagocytosis, which is the major mechanism by which amoebae kill fungal cells. We found no evidence that amoebae use extracellular killing mechanisms in their interactions with C. neoformans In summary, the presence of Mg 2+ and Ca 2+ enhanced the cell adhesion on the surfaces and the motility of the amoeba, thus increasing the chance for contact with C. neoformans and the frequency of phagocytosis. Our findings imply that the divalent cation concentration in soils could be an important variable for whether amoebae can control C. neoformans in the environment. IMPORTANCE The grazing of soil organisms by phagocytic predators such as amoebae is thought to select for traits that enable some of them to acquire the capacity for virulence in animals. Consequently, knowledge about the interactions between amoebae and soil microbes, such as pathogenic fungi, is important for understanding how virulence can emerge. We show that the

  9. Amoeba-related health risk in drinking water systems: could monitoring of amoebae be a complementary approach to current quality control strategies?

    Science.gov (United States)

    Codony, Francesc; Pérez, Leonardo Martín; Adrados, Bárbara; Agustí, Gemma; Fittipaldi, Mariana; Morató, Jordi

    2012-01-01

    Culture-based methods for fecal indicator microorganisms are the standard protocol to assess potential health risk from drinking water systems. However, these traditional fecal indicators are inappropriate surrogates for disinfection-resistant fecal pathogens and the indigenous pathogens that grow in drinking water systems. There is now a range of molecular-based methods, such as quantitative PCR, which allow detection of a variety of pathogens and alternative indicators. Hence, in addition to targeting total Escherichia coli (i.e., dead and alive) for the detection of fecal pollution, various amoebae may be suitable to indicate the potential presence of pathogenic amoeba-resisting microorganisms, such as Legionellae. Therefore, monitoring amoeba levels by quantitative PCR could be a useful tool for directly and indirectly evaluating health risk and could also be a complementary approach to current microbial quality control strategies for drinking water systems.

  10. Survival and growth of Stenotrophomonas maltophilia in free-living amoebae (FLA) and bacterial virulence properties.

    Science.gov (United States)

    Denet, Elodie; Vasselon, Valentin; Burdin, Béatrice; Nazaret, Sylvie; Favre-Bonté, Sabine

    2018-01-01

    Stenotrophomonas maltophilia is found ubiquitously in the environment and is an important emerging nosocomial pathogen. S. maltophilia has been recently described as an Amoebae-Resistant Bacteria (ARB) that exists as part of the microbiome of various free-living amoebae (FLA) from waters. Co-culture approaches with Vermamoeba vermiformis demonstrated the ability of this bacterium to resist amoebal digestion. In the present study, we assessed the survival and growth of six environmental and one clinical S. maltophilia strains within two amoebal species: Acanthamoeba castellanii and Willaertia magna. We also evaluated bacterial virulence properties using the social amoeba Dictyostelium discoideum. A co-culture approach was carried out over 96 hours and the abundance of S. maltophilia cells was measured using quantitative PCR and culture approach. The presence of bacteria inside the amoeba was confirmed using confocal microscopy. Our results showed that some S. maltophilia strains were able to multiply within both amoebae and exhibited multiplication rates up to 17.5 and 1166 for A. castellanii and W. magna, respectively. In contrast, some strains were unable to multiply in either amoeba. Out of the six environmental S. maltophilia strains tested, one was found to be virulent. Surprisingly, this strain previously isolated from a soil amoeba, Micriamoeba, was unable to infect both amoebal species tested. We further performed an assay with a mutant strain of S. maltophilia BurA1 lacking the efflux pump ebyCAB gene and found the mutant to be more virulent and more efficient for intra-amoebal multiplication. Overall, the results obtained strongly indicated that free-living amoebae could be an important ecological niche for S. maltophilia.

  11. Survival and growth of Stenotrophomonas maltophilia in free-living amoebae (FLA and bacterial virulence properties.

    Directory of Open Access Journals (Sweden)

    Elodie Denet

    Full Text Available Stenotrophomonas maltophilia is found ubiquitously in the environment and is an important emerging nosocomial pathogen. S. maltophilia has been recently described as an Amoebae-Resistant Bacteria (ARB that exists as part of the microbiome of various free-living amoebae (FLA from waters. Co-culture approaches with Vermamoeba vermiformis demonstrated the ability of this bacterium to resist amoebal digestion. In the present study, we assessed the survival and growth of six environmental and one clinical S. maltophilia strains within two amoebal species: Acanthamoeba castellanii and Willaertia magna. We also evaluated bacterial virulence properties using the social amoeba Dictyostelium discoideum. A co-culture approach was carried out over 96 hours and the abundance of S. maltophilia cells was measured using quantitative PCR and culture approach. The presence of bacteria inside the amoeba was confirmed using confocal microscopy. Our results showed that some S. maltophilia strains were able to multiply within both amoebae and exhibited multiplication rates up to 17.5 and 1166 for A. castellanii and W. magna, respectively. In contrast, some strains were unable to multiply in either amoeba. Out of the six environmental S. maltophilia strains tested, one was found to be virulent. Surprisingly, this strain previously isolated from a soil amoeba, Micriamoeba, was unable to infect both amoebal species tested. We further performed an assay with a mutant strain of S. maltophilia BurA1 lacking the efflux pump ebyCAB gene and found the mutant to be more virulent and more efficient for intra-amoebal multiplication. Overall, the results obtained strongly indicated that free-living amoebae could be an important ecological niche for S. maltophilia.

  12. Dynamics of hybrid amoeba proteus containing zoochlorellae studied using fluorescence spectroscopy

    Science.gov (United States)

    Liu, C.-H.; Fong, B. A.; Alfano, S. A., Jr.; Rakhlin, I.; Wang, W. B.; Ni, X. H.; Yang, Y. L.; Zhou, F.; Zuzolo, R. C.; Alfano, R. R.

    2011-03-01

    The microinjection of organelles, plants, particles or chemical solutions into Amoeba proteus coupled with spectroscopic analysis and observed for a period of time provides a unique new model for cancer treatment and studies. The amoeba is a eukaryote having many similar features of mammalian cells. The amoeba biochemical functions monitored spectroscopically can provide time sequence in vivo information about many metabolic transitions and metabolic exchanges between cellar organelles and substances microinjected into the amoeba. It is possible to microinject algae, plant mitochondria, drugs or carcinogenic solutions followed by recording the native fluorescence spectra of these composites. This model can be used to spectroscopically monitor the pre-metabolic transitions in developing diseased cells such as a cancer. Knowing specific metabolic transitions could offer solutions to inhibit cancer or reverse it as well as many other diseases. In the present study a simple experiment was designed to test the feasibility of this unique new model by injecting algae and chloroplasts into amoeba. The nonradiative dynamics found from these composites are evidence in terms of the emission ratios between the intensities at 337nm and 419nm; and 684nm bands. There were reductions in the metabolic and photosynthetic processes in amoebae that were microinjected with chloroplasts and zoochlorellae as well of those amoebae that ingested the algae and chloroplasts. The changes in the intensity of the emissions of the peaks indicate that the zoochlorellae lived in the amoebae for ten days. Spectral changes in intensity under the UV and 633nm wavelength excitation are from the energy transfer of DNA and RNA, protein-bound chromophores and chlorophylls present in zoochlorellae undergoing photosynthesis. The fluorescence spectroscopic probes established the biochemical interplay between the cell organelles and the algae present in the cell cytoplasm. This hybrid state is indicative

  13. Diversity and community ecology of forest epiphyte testate amoebae from European Russia.

    Science.gov (United States)

    Payne, Richard J; Belyakova, Olga; Mazei, Yuri

    2015-10-01

    Testate amoebae are an abundant group of microorganisms which make a significant contribution to the diversity of protist life. Most of the world's potential habitats for testate amoebae have been barely studied and when such places are investigated they frequently reveal novel communities and species. Here we consider the testate amoeba communities associated with boreal forest epiphytes (mosses and lichens); an environment which we argue has been under-researched. We present a dataset of 165 samples from four regions of western Russia and analyse these data in relation to micro-habitat position and selected environmental data. The testate amoebae of epiphytes are abundant but dominated by ubiquitous species. We show that there are trends toward a lower species richness and test concentration with greater elevation on the trunk and in lichens compared to mosses. There are considerable differences in community composition between sampling regions. Of all measured environmental variables only moisture content showed a significant relationship with testate amoeba community structure. Our data highlight how little is known about testate amoeba communities of this habitat and call for greater research efforts, particularly in less-studied regions and biomes. Copyright © 2015 Elsevier GmbH. All rights reserved.

  14. Behavioural differentiation induced by environmental variation when crossing a toxic zone in an amoeba

    International Nuclear Information System (INIS)

    Kunita, Itsuki; Kuroda, Shigeru; Nakagaki, Toshiyuki; Ueda, Kei-Ichi; Akita, Dai

    2017-01-01

    Organisms choose from among various courses of action in response to a wide variety of environmental conditions and the mechanism by which various behaviours are induced is an open question. Interesting behaviour was recently reported: that a unicellular organism of slime mold Physarum polycephalum known as an amoeba had multiple responses (crossing, returning, etc) when the amoeba encounters a zone with toxic levels of quinine, even under carefully controlled conditions. We here examined this elegant example in more detail to obtain insight into behavioural differentiation. We found that the statistical distribution of passage times across a quinine zone switch from unimodal to bimodal (with peaks corresponding to fast crossing and no crossing) when a periodic light stimulation to modulate a biorhythm in amoeba is applied homogeneously across the space, even under the same level of chemical stimuli. Based on a mathematical model for cell movement in amoeba, we successfully reproduced the stimulation-induced differentiation, which was observed experimentally. These dynamics may be explained by a saddle structure around a canard solution. Our results imply that the differentiation of behavioural types in amoeba is modified step-by-step via the compounding of stimulation inputs. The complex behaviour like the differentiation in amoeba may provide a basis for understanding the mechanism of behaviour selection in higher animals from an ethological perspective. (paper)

  15. Behavioural differentiation induced by environmental variation when crossing a toxic zone in an amoeba

    Science.gov (United States)

    Kunita, Itsuki; Ueda, Kei-Ichi; Akita, Dai; Kuroda, Shigeru; Nakagaki, Toshiyuki

    2017-09-01

    Organisms choose from among various courses of action in response to a wide variety of environmental conditions and the mechanism by which various behaviours are induced is an open question. Interesting behaviour was recently reported: that a unicellular organism of slime mold Physarum polycephalum known as an amoeba had multiple responses (crossing, returning, etc) when the amoeba encounters a zone with toxic levels of quinine, even under carefully controlled conditions. We here examined this elegant example in more detail to obtain insight into behavioural differentiation. We found that the statistical distribution of passage times across a quinine zone switch from unimodal to bimodal (with peaks corresponding to fast crossing and no crossing) when a periodic light stimulation to modulate a biorhythm in amoeba is applied homogeneously across the space, even under the same level of chemical stimuli. Based on a mathematical model for cell movement in amoeba, we successfully reproduced the stimulation-induced differentiation, which was observed experimentally. These dynamics may be explained by a saddle structure around a canard solution. Our results imply that the differentiation of behavioural types in amoeba is modified step-by-step via the compounding of stimulation inputs. The complex behaviour like the differentiation in amoeba may provide a basis for understanding the mechanism of behaviour selection in higher animals from an ethological perspective.

  16. Potentially pathogenic free-living amoebae in contact lenses of the asymptomatic contact lens wearers.

    Directory of Open Access Journals (Sweden)

    Maryam Niyyati

    2014-03-01

    Full Text Available Free-living amoebae (FLA including Acanthamoeba spp. and Hartmannella spp. are the causative agents of serious corneal infection especially within contact lens wearers. Thus contact lenses and their storage case could be a suitable niche for potentially pathogenic amoebae. The main objective of the present study was to evaluate the contamination of contact lenses to free living amoebae using morphological and sequencing based methods.Overall, 90 volunteers provided their contact lenses. All volunteers wore soft contact lenses. Both lenses were cultured in the same plate. Forty-eight of the volunteers were medical and dentistry student and 42 were ophthalmology attendees of hospitals in Tehran, Iran. All of the samples were inoculated to non-nutrient medium and monitored daily for the outgrowth of the amoebae. PCR and sequencing were performed using various primer pairs.Of the 90 volunteers, 9 (10% were positive for free-living amoebae outgrowth. Morphological analysis revealed that 3 isolates were belonged to Hartmannella genus according to small round cysts and 6 isolates were belonged to Acanthamoeba genus based on the star shape of endocysts. Sequencing revealed that Acanthamoeba belonged to T4, T3 and T5 genotype. Hartmannella were also belonged to vermiformis species.The presence of potentially pathogenic free living amoebae including Acanthamoeba and Hartmannella could be a high risk for people using soft contact lenses. These results revealed that improved clarification and professional recommendations for contact lens wearers is of utmost importance.

  17. Distribution of free-living amoebae in a treatment system of textile industrial wastewater.

    Science.gov (United States)

    Ramirez, Elizabeth; Robles, Esperanza; Martinez, Blanca; Ayala, Reynaldo; Sainz, Guadalupe; Martinez, Maria Elena; Gonzalez, Maria Elena

    2014-11-01

    Free-living amoebae have been found in soil, air and a variety of aquatic environments, but few studies have been conducted on industrial wastewater and none on wastewater from the textile industry. The aim of this study was to determine the presence and distribution of free-living amoebae in a biological treatment system that treats textile industrial wastewater. Samples were taken from input, aeration tank, sedimentation tank and output. Samples were centrifuged at 1200g for 15min, the sediment was seeded on non-nutritive agar with Enterobacter aerogenes (NNE) and the plates were incubated at 30 and 37°C. Free-living amoebae were present in all stages of the treatment system. The highest number of amoebic isolates was found in the aeration tank and no seasonal distribution was observed during the year. A total of 14 amoeba genera were isolated: Acanthamoeba, Echinamoeba, Korotnevella, Mayorella, Naegleria, Platyamoeba, Saccamoeba, Stachyamoeba, Thecamoeba, Vahlkampfia, Vannella, Vermamoeba, Vexillifera and Willaertia. The most frequently found amoebae were Acanthamoeba and Vermamoeba which were found in all treatment system stages. The constant presence and diversity of free-living amoebae in the treatment system were important findings due to the characteristics of the wastewater from the textile plant in terms of the residue content from colorants, fixers, carriers, surfactants, etc., used in fabric dyeing and finishing processes. The factors that determined the presence and distribution of amoebae in the activated sludge system were their capacity to form cysts, which allowed them to resist adverse conditions; food availability; an average temperature of 27-33°C; dissolved oxygen in average concentrations above 2mg/L, and pH in a range of 5.9-7.1. Copyright © 2014 Elsevier Inc. All rights reserved.

  18. Review: Occurrence of the pathogenic amoeba Naegleria fowleri in groundwater

    Science.gov (United States)

    Bright, Kelly R.; Gerba, Charles P.

    2017-06-01

    Naegleria fowleri is a thermophilic free-living amoeba found worldwide in soils and warm freshwater. It is the causative agent of primary amebic meningoencephalitis, a nearly always fatal disease afflicting mainly children and young adults. Humans are exposed to the organism via swimming, bathing, or other recreational activity during which water is forcefully inhaled into the upper nasal passages. Although many studies have looked at the occurrence of N. fowleri in surface waters, limited information is available regarding its occurrence in groundwater and geothermally heated natural waters such as hot springs. This paper reviews the current literature related to the occurrence of N. fowleri in these waters and the methods employed for its detection. Case reports of potential groundwater exposures are also included. Despite increased interest in N. fowleri in recent years due to well-publicized cases linked to drinking water, many questions still remain unanswered. For instance, why the organism persists in some water sources and not in others is not well understood. The role of biofilms in groundwater wells and plumbing in individual buildings, and the potential for warming due to climate change to expand the occurrence of the organism into new regions, are still unclear. Additional research is needed to address these questions in order to better understand the ecology of N. fowleri and the conditions that result in greater risks to bathers.

  19. Genotypic characterization of amoeba isolated from Acanthamoeba keratitis in Poland.

    Science.gov (United States)

    Derda, Monika; Solarczyk, Piotr; Cholewiński, Marcin; Hadaś, Edward

    2015-03-01

    Free-living amoebae belonging to the genus Acanthamoeba are the causative factor of many diseases. Among others, they cause Acanthamoeba keratitis (AK), a condition that usually occurs in contact lens wearers, though it is also observed in non-wearers. The number of diagnosed cases of AK increased more than eightfold during 8 years in the USA, and a proportional increase in frequency also occurred in Poland and Europe. Cases of AK are usually diagnosed late, and their therapy is difficult and rarely successful. AK is an uncommon diagnosis in Poland. The increased number of positive cases observed in our laboratory may reflect the growing at-risk population of contact lens wearers. Acanthamoeba as a genus of facultative human parasites is currently classified into 17 genotypes. Isolates belonging to seven genotypes were found to be associated with AK. One genotype in particular, T4, was found to be overrepresented in human disease. The main finding of our study is that in Poland, AK is almost always associated with the T4 genotype.

  20. Detection of the amoeba Entamoeba gingivalis in periodontal pockets

    Science.gov (United States)

    Bonner, Mark; Amard, Véronique; Bar-Pinatel, Charlotte; Charpentier, Frédéric; Chatard, Jean-Michel; Desmuyck, Yvan; Ihler, Serge; Rochet, Jean-Pierre; Roux de La Tribouille, Véronique; Saladin, Luc; Verdy, Marion; Gironès, Núria; Fresno, Manuel; Santi-Rocca, Julien

    2014-01-01

    Periodontitis is a public health issue, being one of the most prevalent diseases worldwide. However, the aetiology of the disease is still unclear: genetics of patients cannot explain the dispersed or isolated localisation of gingival pockets, while bacteria-based models are insufficient to distinguish gingivitis and periodontitis. The possible role of parasites in the establishment of periodontitis has been poorly studied until now. The aim of this project was to study a potential link between colonisation of gingival crevices by the amoeba Entamoeba gingivalis and periodontitis. In eight different dental clinics in France, samples were taken in periodontal pockets (72) or healthy sites (33), and submitted to microscopic observation and molecular identification by PCR with a new set of primers designed to specifically detect E. gingivalis. This blind sample analysis showed the strong sensitivity of PCR compared with clinical diagnosis (58/72 = 81%), and microscopy (51/65 = 78%). The results of this work show that the parasites detected by microscopy mainly – if not exclusively – belong to the species E. gingivalis and that the presence of the parasite is correlated with periodontitis. PMID:24983705

  1. Origin and diversity of testate amoebae shell composition: Example of Bullinularia indica living in Sphagnum capillifolium.

    Science.gov (United States)

    Delaine, Maxence; Bernard, Nadine; Gilbert, Daniel; Recourt, Philippe; Armynot du Châtelet, Eric

    2017-06-01

    Testate amoebae are free-living shelled protists that build a wide range of shells with various sizes, shapes, and compositions. Recent studies showed that xenosomic testate amoebae shells could be indicators of atmospheric particulate matter (PM) deposition. However, no study has yet been conducted to assess the intra-specific mineral, organic, and biologic grain diversity of a single xenosomic species in a natural undisturbed environment. This study aims at providing new information about grain selection to develop the potential use of xenosomic testate amoebae shells as bioindicators of the multiple-origin mineral/organic diversity of their proximal environment. To fulfil these objectives, we analysed the shell content of 38 Bullinularia indica individuals, a single xenosomic testate amoeba species living in Sphagnum capillifolium, by scanning electron microscope (SEM) coupled with X-ray spectroscopy. The shells exhibited high diversities of mineral, organic, and biomineral grains, which confirms their capability to recycle xenosomes. Mineral grain diversity and size of B. indica matched those of the atmospheric natural mineral PM deposited in the peatbog. Calculation of grain size sorting revealed a discrete selection of grains agglutinated by B. indica. These results are a first step towards understanding the mechanisms of particle selection by xenosomic testate amoebae in natural conditions. Copyright © 2017 Elsevier GmbH. All rights reserved.

  2. LPS-inducible factor(s) from activated macrophages mediates cytolysis of Naegleria fowleri amoebae

    Energy Technology Data Exchange (ETDEWEB)

    Cleary, S.F.; Marciano-Cabral, F.

    1986-03-01

    Soluble cytolytic factors of macrophage origin have previously been described with respect to their tumoricidal activity. The purpose of this study was to investigate the mechanism and possible factor(s) responsible for cytolysis of the amoeba Naegleria fowleri by activated peritoneal macrophages from B6C3F1 mice. Macrophages or conditioned medium (CM) from macrophage cultures were incubated with /sup 3/H-Uridine labeled amoebae. Percent specific release of label served as an index of cytolysis. Bacille Calmette-Guerin (BCG) and Corynebacterium parvum macrophages demonstrated significant cytolysis of amoebae at 24 h with an effector to target ratio of 10:1. Treatment of macrophages with inhibitors of RNA or protein synthesis blocked amoebicidal activity. Interposition of a 1 ..mu..m pore membrane between macrophages and amoebae inhibited killing. Inhibition in the presence of the membrane was overcome by stimulating the macrophages with LPS. CM from SPS-stimulated, but not unstimulated, cultures of activated macrophages was cytotoxic for amoebae. The activity was heat sensitive and was recovered from ammonium sulfate precipitation of the CM. Results indicate that amoebicidal activity is mediated by a protein(s) of macrophage origin induced by target cell contact or stimulation with LPS.

  3. Proteases from Entamoeba spp. and Pathogenic Free-Living Amoebae as Virulence Factors

    Science.gov (United States)

    Serrano-Luna, Jesús; Piña-Vázquez, Carolina; Reyes-López, Magda; Ortiz-Estrada, Guillermo

    2013-01-01

    The standard reference for pathogenic and nonpathogenic amoebae is the human parasite Entamoeba histolytica; a direct correlation between virulence and protease expression has been demonstrated for this amoeba. Traditionally, proteases are considered virulence factors, including those that produce cytopathic effects in the host or that have been implicated in manipulating the immune response. Here, we expand the scope to other amoebae, including less-pathogenic Entamoeba species and highly pathogenic free-living amoebae. In this paper, proteases that affect mucin, extracellular matrix, immune system components, and diverse tissues and cells are included, based on studies in amoebic cultures and animal models. We also include proteases used by amoebae to degrade iron-containing proteins because iron scavenger capacity is currently considered a virulence factor for pathogens. In addition, proteases that have a role in adhesion and encystation, which are essential for establishing and transmitting infection, are discussed. The study of proteases and their specific inhibitors is relevant to the search for new therapeutic targets and to increase the power of drugs used to treat the diseases caused by these complex microorganisms. PMID:23476670

  4. Detection and identification of free-living amoeba from aquatic environment in Taiwan

    Science.gov (United States)

    Jiun Tzeng, Kai; Che Tung, Min; Hsu, Bing Mu; Tsai, Hsiu Feng; Huang, Po Hsiang; Hao Huang, Kuan; Kao, Po Min; Shen, Shu Min; Chen, Jung Sheng

    2013-04-01

    Free-living amoebae including Acanthamoeba, Naegleria, Balamuthia and Hartmannella are widely distributed in water, soil, and air. They can infect humans and can lead to serious illness even death. The aim of this study is to investigate the presence of free-living amoebae from aquatic environment in Taiwan, and to compare the differences between Acanthamoeba and Naegleria in different cultivation methods and conditions. In this study, we used molecular method with specific primers by Polymerase Chain Reaction (PCR) to amplify and to analyze the occurrence of free-living amoebae in aquatic environment. We collected 92 samples from environmental water in Taiwan. The results show that 33 water samples (35.9%) and 11 water samples (12.0%) were detected positive for Acanthamoeba and Naegleria, respectively. Furthermore, both Acanthamoeba and Naegleria can be cultured by PYG in 30° C, but not all free-living amoebae can be enriched and isolated by using storage-cultivation method. Due to the presence of Acanthamoeba and Naegleria in aquatic environment, the water quality monitoring should be more conscious. Keywords: free-living amoebae; Acanthamoeba; Naegleria; Balamuthia; Hartmannella; PCR

  5. NAEGLARIA FOWLERI: THE BRAIN EATING AMOEBA OR AN ENIGMA?

    Science.gov (United States)

    Mukhtar, Fatima; Wazir, Mohammad Salim

    2015-01-01

    Naeglaria fowleri (N. fowleri), popularly known as the brain eating amoeba is the causative agent of the fulminant disease, primary amoebic meningoencephalitis (PAM). Although a rare disease, it is a threat to human health with a case fatality rate ranging from 95-99%. PAM cases have been reported from the United States of America, Australia, Europe and Asia. From 1962 to 2014, 133 people have been infected by N. fowleri in the USA, out of which only three have survived. None of the PAM cases reported in Pakistan so far has survived. This underscores the importance to identify factors, which have led to the failure in decreasing case fatality associated with N. fowleri despite major advances in medical technology, health care; and prevention and control strategies since the first reported case in 1965. We need to focus on eliciting risk factors of the disease prevalent in our part of the world, which are at variance with the developed world. A predominant number. of PAM cases in the West are reported in young males who had participated in recreational activities. However, majority of cases reported in Pakistan are also among young males but they were linked with the religious practice of ablution. What is required to better understand and hence manage this enigma is further research. Further research is to be conducted to discover potent antimicrobials, to test the effectiveness of the new transcribial device in managing PAM, and to identify host factors, which make an individual susceptible to N. fowleri. Investigation of environmental factors related to N. fowleri also needs to be done. Doing so is of paramount importance, as it will help identify the preventive strategies to be employed against N. fowleri.

  6. Late Holocene palaeohydrological changes in a Sphagnum peat bog from NW Romania based on testate amoebae

    Directory of Open Access Journals (Sweden)

    Andrei-Cosmin Diaconu

    2016-04-01

    Full Text Available This paper investigates the possibility of reconstructing the palaeohydrological changes in an active Sphagnum peat bog from north-western Romania using testate amoebae fauna and organic matter content determined by loss on ignition (LOI. In total 28 taxa of testate amoebae were identified of which 11 were frequent enough to present a remarkable ecological significance. Based on the relative abundance of these taxa nine zones were identified, crossing from very wet to dry climate conditions. The wet periods identified are characterized by taxa like Centropyxis cassis, Amphitrema flavum and Hyalosphenia papilio, while in the dry periods Difflugia pulex and Nebela militaris thrive. We showed that combining qualitative information regarding hydrological preferences with the quantitative percentage data from the fossil record it is possible to obtain information regarding major surface moisture changes from the peat bog surface. Furthermore we identified a link between distribution of testate amoebae assemblages, organic matter variation and minerogenic material.

  7. Free energy simulations with the AMOEBA polarizable force field and metadynamics on GPU platform.

    Science.gov (United States)

    Peng, Xiangda; Zhang, Yuebin; Chu, Huiying; Li, Guohui

    2016-03-05

    The free energy calculation library PLUMED has been incorporated into the OpenMM simulation toolkit, with the purpose to perform enhanced sampling MD simulations using the AMOEBA polarizable force field on GPU platform. Two examples, (I) the free energy profile of water pair separation (II) alanine dipeptide dihedral angle free energy surface in explicit solvent, are provided here to demonstrate the accuracy and efficiency of our implementation. The converged free energy profiles could be obtained within an affordable MD simulation time when the AMOEBA polarizable force field is employed. Moreover, the free energy surfaces estimated using the AMOEBA polarizable force field are in agreement with those calculated from experimental data and ab initio methods. Hence, the implementation in this work is reliable and would be utilized to study more complicated biological phenomena in both an accurate and efficient way. © 2015 Wiley Periodicals, Inc. © 2015 Wiley Periodicals, Inc.

  8. [The role of the floodplain gradient in structuring of testate amoebae communities in the Ilych River].

    Science.gov (United States)

    Mazeĭ, Iu A; Malysheva, E A; Lapteva, E M; Komarov, A A; Taskaeva, A A

    2012-01-01

    Forty-two testate amoebae taxa were identified in alluvial soils of floodplain islands in the Ilych River. Among the pedo- and eurybionts, there were aquatic rhizopods. Along the floodplain transect (willow --> meadow --> deciduous forest --> coniferous forest), the testate amoebae community changed directly. There are spatially homogeneous (low beta-diversity) testacean communities but species rich on the local level (high alpha-diversity) within forests. Within willows and meadows, communities are characterized by low alpha-diversity and high heterogeneity that leads to high gamma-diversity.

  9. Mycobacterium tuberculosis complex mycobacteria as amoeba-resistant organisms.

    Directory of Open Access Journals (Sweden)

    Felix Mba Medie

    Full Text Available BACKGROUND: Most environmental non-tuberculous mycobacteria have been demonstrated to invade amoebal trophozoites and cysts, but such relationships are largely unknown for members of the Mycobacterium tuberculosis complex. An environmental source has been proposed for the animal Mycobacterium bovis and the human Mycobacterium canettii. METHODOLOGY/PRINCIPAL FINDINGS: Using optic and electron microscopy and co-culture methods, we observed that 89±0.6% of M. canettii, 12.4±0.3% of M. tuberculosis, 11.7±2% of M. bovis and 11.2±0.5% of Mycobacterium avium control organisms were phagocytized by Acanthamoeba polyphaga, a ratio significantly higher for M. canettii (P = 0.03, correlating with the significantly larger size of M. canetti organisms (P = 0.035. The percentage of intraamoebal mycobacteria surviving into cytoplasmic vacuoles was 32±2% for M. canettii, 26±1% for M. tuberculosis, 28±2% for M. bovis and 36±2% for M. avium (P = 0.57. M. tuberculosis, M. bovis and M. avium mycobacteria were further entrapped within the double wall of <1% amoebal cysts, but no M. canettii organisms were observed in amoebal cysts. The number of intracystic mycobacteria was significantly (P = 10(-6 higher for M. avium than for the M. tuberculosis complex, and sub-culturing intracystic mycobacteria yielded significantly more (P = 0.02 M. avium organisms (34×10(4 CFU/mL than M. tuberculosis (42×10(1 CFU/mL and M. bovis (35×10(1 CFU/mL in the presence of a washing fluid free of mycobacteria. Mycobacteria survived in the cysts for up to 18 days and cysts protected M. tuberculosis organisms against mycobactericidal 5 mg/mL streptomycin and 2.5% glutaraldehyde. CONCLUSIONS/SIGNIFICANCE: These data indicate that M. tuberculosis complex organisms are amoeba-resistant organisms, as previously demonstrated for non-tuberculous, environmental mycobacteria. Intercystic survival of tuberculous mycobacteria, except for M. canettii, protect them

  10. Pathogenic amoebae in natural thermal waters of three resorts of Hidalgo, Mexico.

    Science.gov (United States)

    Rivera, F; Lares, F; Gallegos, E; Ramirez, E; Bonilla, P; Calderon, A; Martinez, J J; Rodriguez, S; Alcocer, J

    1989-12-01

    In a search for free-living amoebae, seven water samples from three thermal water bathing resorts in Tecozautla, Hidalgo, were analyzed during December 1984. The samples were concentrated by filtration and centrifugation, and inoculated later on monoxenic and axenic media. The identification of the isolates was performed by morphology and isoelectric focusing of isoenzymes and total proteins. Thirty-three strains of free-living amoebae belonging to the genera Naegleria, Acanthamoeba, and Willaertia were isolated. Twenty of these strains belonged to the Naegleria genus, 16 of them were classified as Naegleria spp., and 2 were classified as Naegleria lovaniensis. Noteworthy was the finding of two pathogenic strains of the species Naegleria australiensis. N. australiensis and N. lovaniensis may be considered good indicator organisms, since they live in the same environmental conditions as N. fowleri, the agent of primary amoebic encephalitis (PAM). On the other hand, amoebae other than Naegleria were isolated and identified as Acathamoeba castellanii (two strains), and Acanthamoeba lugdunensis (one strain), which proved to be pathogenic when tested in mice. Nine more pathogenic strains of the genus Acanthamoeba spp. were isolated together with one strain of Willaertia magna, a thermophilic nonpathogenic amoeba. The chlorination and periodical surveillance of water resorts like the one studied is recommended, in order to prevent the appearance of more cases of PAM or other human diseases associated with pathogenic Acanthamoeba spp.

  11. Presence of rotavirus and free-living amoebae in the water supplies of Karachi, Pakistan

    Science.gov (United States)

    Yousuf, Farzana Abubakar; Siddiqui, Ruqaiyyah; Khan, Naveed Ahmed

    2017-01-01

    ABSTRACT Rotavirus and pathogenic free-living amoebae are causative agents of important health problems, especially for developing countries like Pakistan where the population has limited access to clean water supplies. Here, we evaluated the prevalence of rotavirus and free-living amoebae (Acanthamoeba spp., Balamuthia mandrillaris, Naegleria fowleri) in drinking water supplies of Karachi, Pakistan. Six water filtration plants that supply drinking water to the population of Karachi were investigated. Additionally, drinking water samples from households were analyzed for the presence of rotavirus and free-living amoebae. Rotavirus was present in 35% of the water samples collected from water filtration plants; however, domestic tap water samples had a prevalence of only 5%. Out of 20 water samples from filtration plants, 13 (65%) were positive for Acanthamoeba spp., and one (5%) was positive for B. mandrillaris. Out of 20 drinking water samples collected from different areas of Karachi, 35% were positive for Acanthamoeba spp. Rotavirus was detected in 5% of the drinking water samples tested. Overall, these findings showed for the first time the presence of rotavirus, in addition to pathogenic free-living amoebae in drinking water supplies of Karachi that could be an important public health risk for the affected population. PMID:28591260

  12. Quadrulella texcalense sp nov from a Mexican desert: An unexpected new environment for hyalospheniid testate amoebae

    Czech Academy of Sciences Publication Activity Database

    Perez-Juarez, H.; Serrano-Vazquez, A.; Kosakyan, Anush; Mitchell, E.A.D.; Rivera Aguilar, V. M.; Lahr, D. J. G.; Hernandez Moreno, M. M.; Macías Cuellar, H.; Eguiarte, L. E.; Lara, E.

    2017-01-01

    Roč. 61, 17 July (2017), s. 253-264 ISSN 0932-4739 Institutional support: RVO:60077344 Keywords : Adaptation * Biological soil crust desert * Protist * Tehuacain-Cuicatlain Biosphere Reserve * Testate amoebae Subject RIV: EG - Zoology OBOR OECD: Zoology Impact factor: 2.581, year: 2016

  13. Free-living amoebae used to isolate consortia capable of degrading trichloroethylene

    Energy Technology Data Exchange (ETDEWEB)

    Tyndall, R.L.; Katz, D.S.; Little, C.D. [Oak Ridge National Lab., TN (United States); Kennedy, J.R. [Univ. of Tennessee, Knoxville, TN (United States)

    1991-12-31

    The interaction of protozoa with prokaryotes is well documented. These interactions can be either ecto- or endosymbiotic. An example of photosynthetic symbiosis is the well-defined interaction between paramecium and entrapped Chlorella. Paramecium can also form symbiotic relationships with gram-negative heterotrophic bacteria. Jeon has described an interaction between amoebae and engulfed bacteria, that eventuated into a dependency of the amoebae on the presence of the engulfed bacterium. Free-living amoebae and tetrahymena can engulf and subsequently provide the necessary niche for the replication of Legionella. Acanthamoebae trophozoites and cysts can harbor and support the replication of unidentified gram-negative bacteria. King has recently shown that bacteria associated with free-living amoebae are more resistant to toxic environments. Assuming that methylotrophic bacteria in situ are a part of a mixed community, and based on our observations that bacteria associated with protozoa may not be easily isolated by standard techniques, we attempted to use protozoa as a tool to isolate TCE-degrading bacteria.

  14. Presence of rotavirus and free-living amoebae in the water supplies of Karachi, Pakistan

    Directory of Open Access Journals (Sweden)

    Farzana Abubakar Yousuf

    Full Text Available ABSTRACT Rotavirus and pathogenic free-living amoebae are causative agents of important health problems, especially for developing countries like Pakistan where the population has limited access to clean water supplies. Here, we evaluated the prevalence of rotavirus and free-living amoebae (Acanthamoeba spp., Balamuthia mandrillaris, Naegleria fowleri in drinking water supplies of Karachi, Pakistan. Six water filtration plants that supply drinking water to the population of Karachi were investigated. Additionally, drinking water samples from households were analyzed for the presence of rotavirus and free-living amoebae. Rotavirus was present in 35% of the water samples collected from water filtration plants; however, domestic tap water samples had a prevalence of only 5%. Out of 20 water samples from filtration plants, 13 (65% were positive for Acanthamoeba spp., and one (5% was positive for B. mandrillaris. Out of 20 drinking water samples collected from different areas of Karachi, 35% were positive for Acanthamoeba spp. Rotavirus was detected in 5% of the drinking water samples tested. Overall, these findings showed for the first time the presence of rotavirus, in addition to pathogenic free-living amoebae in drinking water supplies of Karachi that could be an important public health risk for the affected population.

  15. Genome sequence of Rickettsia bellii illuminates the role of amoebae in gene exchanges between intracellular pathogens.

    Directory of Open Access Journals (Sweden)

    Hiroyuki Ogata

    2006-05-01

    Full Text Available The recently sequenced Rickettsia felis genome revealed an unexpected plasmid carrying several genes usually associated with DNA transfer, suggesting that ancestral rickettsiae might have been endowed with a conjugation apparatus. Here we present the genome sequence of Rickettsia bellii, the earliest diverging species of known rickettsiae. The 1,552,076 base pair-long chromosome does not exhibit the colinearity observed between other rickettsia genomes, and encodes a complete set of putative conjugal DNA transfer genes most similar to homologues found in Protochlamydia amoebophila UWE25, an obligate symbiont of amoebae. The genome exhibits many other genes highly similar to homologues in intracellular bacteria of amoebae. We sought and observed sex pili-like cell surface appendages for R. bellii. We also found that R. bellii very efficiently multiplies in the nucleus of eukaryotic cells and survives in the phagocytic amoeba, Acanthamoeba polyphaga. These results suggest that amoeba-like ancestral protozoa could have served as a genetic "melting pot" where the ancestors of rickettsiae and other bacteria promiscuously exchanged genes, eventually leading to their adaptation to the intracellular lifestyle within eukaryotic cells.

  16. Evaluating the use of testate amoebae for palaeohydrological reconstruction in permafrost peatlands

    DEFF Research Database (Denmark)

    Swindles, Graeme T.; Amesbury, Matthew J.; Turner, T. Edward

    2015-01-01

    The melting of high-latitude permafrost peatlands is a major concern due to a potential positive feedback on global climate change. We examine the ecology of testate amoebae in permafrost peatlands, based on sites in Sweden (similar to 200 km north of the Arctic Circle). Multivariate statistical ...

  17. Amoebae as potential environmental hosts for Mycobacterium ulcerans and other mycobacteria, but doubtful actors in Buruli ulcer epidemiology.

    Science.gov (United States)

    Gryseels, Sophie; Amissah, Diana; Durnez, Lies; Vandelannoote, Koen; Leirs, Herwig; De Jonckheere, Johan; Silva, Manuel T; Portaels, Françoise; Ablordey, Anthony; Eddyani, Miriam

    2012-01-01

    The reservoir and mode of transmission of Mycobacterium ulcerans, the causative agent of Buruli ulcer, remain unknown. Ecological, genetic and epidemiological information nonetheless suggests that M. ulcerans may reside in aquatic protozoa. We experimentally infected Acanthamoeba polyphaga with M. ulcerans and found that the bacilli were phagocytised, not digested and remained viable for the duration of the experiment. Furthermore, we collected 13 water, 90 biofilm and 45 detritus samples in both Buruli ulcer endemic and non-endemic communities in Ghana, from which we cultivated amoeboid protozoa and mycobacteria. M. ulcerans was not isolated, but other mycobacteria were as frequently isolated from intracellular as from extracellular sources, suggesting that they commonly infect amoebae in nature. We screened the samples as well as the amoeba cultures for the M. ulcerans markers IS2404, IS2606 and KR-B. IS2404 was detected in 2% of the environmental samples and in 4% of the amoeba cultures. The IS2404 positive amoeba cultures included up to 5 different protozoan species, and originated both from Buruli ulcer endemic and non-endemic communities. This is the first report of experimental infection of amoebae with M. ulcerans and of the detection of the marker IS2404 in amoeba cultures isolated from the environment. We conclude that amoeba are potential natural hosts for M. ulcerans, yet remain sceptical about their implication in the transmission of M. ulcerans to humans and their importance in the epidemiology of Buruli ulcer.

  18. Amoebae as potential environmental hosts for Mycobacterium ulcerans and other mycobacteria, but doubtful actors in Buruli ulcer epidemiology.

    Directory of Open Access Journals (Sweden)

    Sophie Gryseels

    Full Text Available The reservoir and mode of transmission of Mycobacterium ulcerans, the causative agent of Buruli ulcer, remain unknown. Ecological, genetic and epidemiological information nonetheless suggests that M. ulcerans may reside in aquatic protozoa.We experimentally infected Acanthamoeba polyphaga with M. ulcerans and found that the bacilli were phagocytised, not digested and remained viable for the duration of the experiment. Furthermore, we collected 13 water, 90 biofilm and 45 detritus samples in both Buruli ulcer endemic and non-endemic communities in Ghana, from which we cultivated amoeboid protozoa and mycobacteria. M. ulcerans was not isolated, but other mycobacteria were as frequently isolated from intracellular as from extracellular sources, suggesting that they commonly infect amoebae in nature. We screened the samples as well as the amoeba cultures for the M. ulcerans markers IS2404, IS2606 and KR-B. IS2404 was detected in 2% of the environmental samples and in 4% of the amoeba cultures. The IS2404 positive amoeba cultures included up to 5 different protozoan species, and originated both from Buruli ulcer endemic and non-endemic communities.This is the first report of experimental infection of amoebae with M. ulcerans and of the detection of the marker IS2404 in amoeba cultures isolated from the environment. We conclude that amoeba are potential natural hosts for M. ulcerans, yet remain sceptical about their implication in the transmission of M. ulcerans to humans and their importance in the epidemiology of Buruli ulcer.

  19. Seasonal abundances of naked amoebae in biofilms on shells of zebra mussels (Dreissena polymorpha) with comparative data from rock scrapings.

    Science.gov (United States)

    Bischoff, Paul J; Wetmore, Scott

    2009-01-01

    In North America, zebra mussels (Dreissena polymorpha) are notoriously known as invasive species. The abundance of naked amoebae sampled from the shells of zebra mussels was compared with abundances from rock scrapings at approximately monthly intervals for 1 year. The sites were 2 km apart along the same shoreline. No significant difference in abundance of naked amoebae (F = 1.44; P amoebae in winter, followed by peaks in early spring. The percent encysted increased from a low of 1% in the summer to 80% in early winter.

  20. [A study on the taxonomy of soil amoebas from Caspian plague foci based on an analysis of ribosomal operon sequences].

    Science.gov (United States)

    Koshel', E I; Anisimova, L V; Novichkova, L A; Vidiaeva, N A; Guseva, N P; Eroshenko, G A; Kutyrev, V V

    2015-01-01

    The results of a study on the taxonomy and quantitative abundance of free-living amoebas in soil samples from the Russian plague foci of the northwestern Caspian steppe, the Caspian sand, and the Volga-Ural steppe are presented. Amoebas of the Willaertia and Hartmanella genera, as well as representatives of myxomycetes, were isolated from samples. From these, amoebas of the Acanthamoeba genus predominated and could be as abundantas 300000 cells per 1 g of soil. Sequencing of the 18S rRNA gene region showed that Acanthamoeba from the Volga-Ural steppe focus belonged to the A. castellanii species. Phylogenetic analysis confirmed that amoebas from two other Caspian foci belonged to the species of Acanthamoeba spp.

  1. Survival of Campylobacter jejuni in co-culture with Acanthamoeba castellanii: role of amoeba-mediated depletion of dissolved oxygen

    DEFF Research Database (Denmark)

    Bui, Thanh Xuan; Winding, Anne; Qvortrup, Klaus

    2012-01-01

    of intracellular C. jejuni in A. castellanii 5 h post gentamicin treatment at 25°C. Conversely, we found that A. castellanii promoted the extracellular growth of C. jejuni in co-cultures at 37°C in aerobic conditions. This growth-promoting effect did not require amoebae – bacteria contact. The growth rates......, the dissolved oxygen levels of co-cultures with or without amoebae – bacteria contact were much lower than those observed with culture media or with C. jejuni alone incubated in aerobic conditions, and were comparable with levels obtained after 24 h of growth of C. jejuni under microaerophilic conditions. Our...... observed with or without contact with amoeba were similar to those observed when C. jejuni was grown in microaerophilic conditions. Preconditioned media prepared with live or dead amoebae cultivated with or without C. jejuni did not promote the growth of C. jejuni in aerobic conditions. Interestingly...

  2. Presence of pathogenic amoebae in power plant cooling waters. Final report, October 15, 1977-September 30, 1979. [Naegleria fowleri

    Energy Technology Data Exchange (ETDEWEB)

    Tyndall, R.L.; Willaert, E.; Stevens, A.R.

    1981-03-01

    Cooling-water-associated algae and sediments from five northern and five southern or western electric power plants were tested for the presence of pathogenic amoebae. In addition, water algae and sediments from five northern and five southern/western sites not associated with power plants were tested. There was a significant correlation at northern power plants between the presence of thermophilic, pathogenic amoebae in cooling waters and thermal additions. Presence of the pathogenic did not correlate with salinity, pH, conductivity, or a variety of various chemical components of the cooling waters. Selected pathogenic isolates were tested serologically and were classified as Naegleria fowleri. Although thermal additions were shown to be contributing factor in predisposing cooling waters to the growth of pathogenic amoebae, the data suggest the involvement of other currently undefined parameters associated with the presence of the pathogenic amoebae. 35 refs., 21 tabs.

  3. Detection and identification of free-living amoeba from aquatic environment in different seasons in Taiwan

    Science.gov (United States)

    Tzeng, K.; Hsu, B.; Tsai, H.; Huang, P.; Tsai, J.; Kao, P.; Huang, K.; Chen, J.

    2013-12-01

    Free-living amoeba includes Acanthamoeba and Naegleria, which are widely distributed in water and soil. Human infection with free-living amoeba leads to serious illness, even lethal. For example, central nervous system infection will cause amoebic meningoencephalitis, and infections will cause amoebic keratitis. The presence of free-living amoeba in environment water can be used as a water quality indicator in ecosystem assessment. In Taiwan, reservoirs are indispensable because of the water source are limited by the steep terrain and the short river flow. Therefore, we need to pay more attention in the quality control of reservoirs water. The aims of this study are to investigate the presence of free-living amoeba in Taiwan reservoirs, and to compare the differences among seasons. At last, the identification and genotyping of Acanthamoeba and Naegleria are investigated. In this study, we use polymerase chain reaction with specific primers to analyze the presence of free-living amoeba in aquatic environment. We collected total 60 samples from reservoirs in Taiwan. The water samples are divided into two parts for both direct concentration method and culture method. The results show the different detection rates among seasons. For Acanthamoeba, the detection rates were 28.3% (17 of 60 water samples), 21.7% (13 of 60 water samples) and 8.3% (5 of 60 water samples) in autumn, winter and spring, respectively. For Naegleria, the detection rates were 6.7% (4 of 60 water samples), 0% (0 of 60 water samples) and 0% (0 of 60 water samples) were detected positive in autumn, winter and spring, respectively. Sequence analysis showed that the major genotypes in Acanthamoeba were T3, T4, T10 and T11 in autumn, T2, T4 and T10 in winter, T4 in spring. Due to the presences of Acanthamoeba and Naegleria in reservoirs, we should pay more attention in water quality monitoring to prevent the potential risks of diseases. Keywords: free-living amoeba, Acanthamoeba, Naegleria, polymerase

  4. Evidence of test detachment in Astrorhiza limicola and two consequential synonyms: Amoeba gigantea and Megamoebomyxa argillobia (Foraminiferida)

    DEFF Research Database (Denmark)

    Cedhagen, Tomas; Tendal, Ole S.

    1989-01-01

    Laboratory observations and experiments demonstrate that the naked rhizopods Amoeba gigantea SANDAHL, 1857 and Megamoebomyxa argillo~ia NYHOLM, 1950, and the foraminifers Astrorhiza arenifera STSCHEDRlNA, 1946, A. sabulifera STSCHEDRINA, 1946 and A. arctlca STSCHEDRINA, 1958 are synonyms of Astro......Laboratory observations and experiments demonstrate that the naked rhizopods Amoeba gigantea SANDAHL, 1857 and Megamoebomyxa argillo~ia NYHOLM, 1950, and the foraminifers Astrorhiza arenifera STSCHEDRlNA, 1946, A. sabulifera STSCHEDRINA, 1946 and A. arctlca STSCHEDRINA, 1958 are synonyms...

  5. Seasonal distribution of air-borne pathogenic and free-living amoebae in Mexico City and its suburbs

    Energy Technology Data Exchange (ETDEWEB)

    Rivera, F.; Bonilla, P.; Ramirez, E.; Calderon, A.; Gallegos, E.; Rodriguez, S.; Ortiz, R.; Hernandez, D.; Rivera, V. (ENEP-Iztacala, Tlalnepantla (Mexico). Lab. of Environmental Microbiology)

    1994-03-01

    A survey was carried out over a one-year period to isolate amoebae suspended in the air of Mexico City and its suburbs. Sampling stations were placed at the four cardinal points of the metropolitan area. Selective media were used to culture the amoebae isolated. Specialized taxonomic keys and physical and physiological tests were used for identification, and a statistical analysis was performed to determine the correlations between physico-chemical and biological parameters. 108 strains were isolated, of which 19 were pathogenic via intracerebral inoculation and 9 via intranasal inoculation. Species of the genera Acanthamoeba, Vahlkampfia and Hartmannella were most abundant. Acanthamoeba polyphaga showed the highest abundance. Several times during the period of the study SO[sub 2], O[sub 3], CO NO and NO[sub 2] exceeded the permissible levels established by the Mexican government. The ability of amoebae to form cysts and cyst size were important factors for their presence, survival, abundance and diversity in the atmosphere. The main source of air-borne amoebae was the soil. Factors that favored the incidence and diversity of the isolates were wind speed and direction, low relative humidity, generation of frequent dust-storms, resuspension of amoebae by vehicular traffic, proximity to garbage dumps and large extensions of bare soil. Soil cover was a factor associated with a reduction in the incidence and diversity of the aerial amoebae. This study demonstrates that there are viable cysts of amoebae in the atmosphere of Mexico City, that may have potential importance in the case of certain kinds of human allergies and diseases. Further research is needed to find out the aerial presence of viable cysts of obligatory, amphizoic or opportunistic amoebic parasites, and to clarify the qualitative and quantitative effects of the local meteorological and physico-chemical environment on the free-living amoebae present in the atmosphere. 48 refs., 11 figs., 4 tabs.

  6. Free-Living Amoebae as Hosts for and Vectors of Intracellular Microorganisms with Public Health Significance

    Science.gov (United States)

    Balczun, Carsten; Scheid, Patrick L.

    2017-01-01

    Free-living amoebae (FLA) are parasites within both humans and animals causing a wide range of symptoms and act as hosts of, and vehicles for phylogenetically diverse microorganisms, called endocytobionts. The interaction of the FLA with sympatric microorganisms leads to an exceptional diversity within FLA. Some of these bacteria, viruses, and even eukaryotes, can live and replicate intracellularly within the FLA. This relationship provides protection to the microorganisms from external interventions and a dispersal mechanism across various habitats. Among those intracellularly-replicating or -residing organisms there are obligate and facultative pathogenic microorganisms affecting the health of humans or animals and are therefore of interest to Public Health Authorities. Mimiviruses, Pandoraviruses, and Pithoviruses are examples for interesting viral endocytobionts within FLA. Future research is expected to reveal further endocytobionts within free-living amoebae and other protozoa through co-cultivation studies, genomic, transcriptomic, and proteomic analyses. PMID:28368313

  7. Reconstruction of active regular motion in amoeba extract: dynamic cooperation between sol and gel states.

    Directory of Open Access Journals (Sweden)

    Yukinori Nishigami

    Full Text Available Amoeboid locomotion is one of the typical modes of biological cell migration. Cytoplasmic sol-gel conversion of an actomyosin system is thought to play an important role in locomotion. However, the mechanisms underlying sol-gel conversion, including trigger, signal, and regulating factors, remain unclear. We developed a novel model system in which an actomyosin fraction moves like an amoeba in a cytoplasmic extract. Rheological study of this model system revealed that the actomyosin fraction exhibits shear banding: the sol-gel state of actomyosin can be regulated by shear rate or mechanical force. Furthermore, study of the living cell indicated that the shear-banding property also causes sol-gel conversion with the same order of magnitude as that of shear rate. Our results suggest that the inherent sol-gel transition property plays an essential role in the self-regulation of autonomous translational motion in amoeba.

  8. An intracellular replication niche for Vibrio cholerae in the amoeba Acanthamoeba castellanii.

    Science.gov (United States)

    Van der Henst, Charles; Scrignari, Tiziana; Maclachlan, Catherine; Blokesch, Melanie

    2016-04-01

    Vibrio cholerae is a human pathogen and the causative agent of cholera. The persistence of this bacterium in aquatic environments is a key epidemiological concern, as cholera is transmitted through contaminated water. Predatory protists, such as amoebae, are major regulators of bacterial populations in such environments. Therefore, we investigated the interaction between V. cholerae and the amoeba Acanthamoeba castellanii at the single-cell level. We observed that V. cholerae can resist intracellular killing. The non-digested bacteria were either released or, alternatively, established a replication niche within the contractile vacuole of A. castellanii. V. cholerae was maintained within this compartment even upon encystment. The pathogen ultimately returned to its aquatic habitat through lysis of A. castellanii, a process that was dependent on the production of extracellular polysaccharide by the pathogen. This study reinforces the concept that V. cholerae is a facultative intracellular bacterium and describes a new host-pathogen interaction.

  9. [Free-living amoebae: analytical methods for water and biofilm quality control].

    Science.gov (United States)

    Bonadonna, L; Lacchetti, I; Paradiso, R

    2006-01-01

    Free-living amoebae are ubiquitous. Some species can cause infections in humans and it is known that they can ingest and protect many microrganisms, such as species belonging to Legionella genus. Till now in Italy the environmental surveillance is still scarce because of both the lack of appropriate and promptly practical methods and drawbacks due to matrices characteristics. In the present study simple techniques were investigated and evaluated to detect and observe free-living amoebae in good water quality and biofilm samples. The membrane filtration technique for water analysis and the direct presence/absence technique for biofilm were recognized as workable methods, easily to be used also by laboratories with little skills and equipments.

  10. Engineering an artificial amoeba propelled by nanoparticle-triggered actin polymerization

    Energy Technology Data Exchange (ETDEWEB)

    Yi Jinsoo; Schmidt, Jacob; Chien Aichi; Montemagno, Carlo D [Department of Bioengineering, University of California Los Angeles, 420 Westwood Plaza, 7523 Boelter Hall, Los Angeles, CA 90095-1600 (United States)], E-mail: montemcd@ucmail.uc.edu

    2009-02-25

    We have engineered an amoeba system combining nanofabricated inorganic materials with biological components, capable of propelling itself via actin polymerization. The nanofabricated materials have a mechanism similar to the locomotion of the Listeria monocytogenes, food poisoning bacteria. The propulsive force generation utilizes nanoparticles made from nickel and gold functionalized with the Listeria monocytogenes transmembrane protein, ActA. These Listeria-mimic nanoparticles were in concert with actin, actin binding proteins, ATP (adenosine triphosphate) and encapsulated within a lipid vesicle. This system is an artificial cell, such as a vesicle, where artificial nanobacteria and actin polymerization machinery are used in driving force generators inside the cell. The assembled structure was observed to crawl on a glass surface analogously to an amoeba, with the speed of the movement dependent on the amount of actin monomers and ATP present.

  11. Interactions Between Testate Amoebae and Saprotrophic Microfungi in a Scots Pine Litter Microcosm

    Czech Academy of Sciences Publication Activity Database

    Vohník, Martin; Burdíková, Zuzana; Vyhnal, Aleš; Koukol, O.

    2011-01-01

    Roč. 61, č. 3 (2011), s. 660-668 ISSN 0095-3628 R&D Projects: GA ČR GP206/09/P340; GA ČR(CZ) GD204/09/H084 Institutional research plan: CEZ:AV0Z50110509; CEZ:AV0Z60050516 Keywords : testate amoebae * saprotrophic fungi * litter Subject RIV: EF - Botanics Impact factor: 2.912, year: 2011

  12. Amoebae, Giant Viruses, and Virophages Make Up a Complex, Multilayered Threesome

    OpenAIRE

    Diesend, Jan; Kruse, Janis; Hagedorn, Monica; Hammann, Christian

    2018-01-01

    Viral infection had not been observed for amoebae, until the Acanthamoeba polyphaga mimivirus (APMV) was discovered in 2003. APMV belongs to the nucleocytoplasmatic large DNA virus (NCLDV) family and infects not only A. polyphaga, but also other professional phagocytes. Here, we review the Megavirales to give an overview of the current members of the Mimi- and Marseilleviridae families and their structural features during amoebal infection. We summarize the different steps of their infection ...

  13. An Amoeba/Zoozanthellae Consortium as a Model System for Animal/Algal Symbiosis

    Science.gov (United States)

    1991-06-18

    of the 9 isolates of Symbiodinium anthopleura which contained swimming small flagellates resistant to the antobiotics and too motile to be engulfed by...was used as a tool for 9 cies. The resistance of these unicellular algae to the amoe- cleaning these algae from digestible contaminating 0 bal... resistant to the antibiotics and the external contaminants but did ne, kill the algae protected tOO motile to be engulfed by the amoebae. inside the amoebal

  14. Water quality bioassay using selected protozoa. I. [Paramecium candatum; Amoeba proteus; Euglena gracilis

    Energy Technology Data Exchange (ETDEWEB)

    Mills, W.L.

    1976-01-01

    The suitability of certain species of protozoa as indicators of water quality has been determined. Experiments were conducted under laboratory conditions to standardize a bioassay procedure for water quality using either Paramecium caudatum, Amoeba proteus, or Euglena gracilis as the indicator organism. The bioassay, which consists of exposing the organisms to a known concentration of pollutant under laboratory conditions, followed by microscopic observation to establish the time of death, affords a reliable, convenient and inexpensive way to monitor for water quality.

  15. Ecology of Testate Amoebae in the Komořany Ponds in the Vltava Basin

    Czech Academy of Sciences Publication Activity Database

    Burdíková, Zuzana; Čapek, Martin; Švindrych, Zdeněk; Gryndler, M.; Kubínová, Lucie; Holcová, K.

    2012-01-01

    Roč. 64, č. 1 (2012), s. 117-130 ISSN 0095-3628 R&D Projects: GA MŠk(CZ) LC06063; GA ČR(CZ) GAP108/11/0794 Institutional research plan: CEZ:AV0Z50110509 Keywords : ecology * freshwater ecosystem * testate amoeba e * seasonal variability Subject RIV: CF - Physical ; Theoretical Chemistry Impact factor: 3.277, year: 2012

  16. Phospholipids trigger Cryptococcus neoformans capsular enlargement during interactions with amoebae and macrophages.

    Directory of Open Access Journals (Sweden)

    Cara J Chrisman

    2011-05-01

    Full Text Available A remarkable aspect of the interaction of Cryptococcus neoformans with mammalian hosts is a consistent increase in capsule volume. Given that many aspects of the interaction of C. neoformans with macrophages are also observed with amoebae, we hypothesized that the capsule enlargement phenomenon also had a protozoan parallel. Incubation of C. neoformans with Acanthamoeba castellanii resulted in C. neoformans capsular enlargement. The phenomenon required contact between fungal and protozoan cells but did not require amoeba viability. Analysis of amoebae extracts showed that the likely stimuli for capsule enlargement were protozoan polar lipids. Extracts from macrophages and mammalian serum also triggered cryptococcal capsular enlargement. C. neoformans capsule enlargement required expression of fungal phospholipase B, but not phospholipase C. Purified phospholipids, in particular, phosphatidylcholine, and derived molecules triggered capsular enlargement with the subsequent formation of giant cells. These results implicate phospholipids as a trigger for both C. neoformans capsule enlargement in vivo and exopolysaccharide production. The observation that the incubation of C. neoformans with phospholipids led to the formation of giant cells provides the means to generate these enigmatic cells in vitro. Protozoan- or mammalian-derived polar lipids could represent a danger signal for C. neoformans that triggers capsular enlargement as a non-specific defense mechanism against potential predatory cells. Hence, phospholipids are the first host-derived molecules identified to trigger capsular enlargement. The parallels apparent in the capsular response of C. neoformans to both amoebae and macrophages provide additional support for the notion that certain aspects of cryptococcal virulence emerged as a consequence of environmental interactions with other microorganisms such as protists.

  17. Environmental Free-Living Amoebae Isolated from Soil in Khon Kaen, Thailand, Antagonize Burkholderia pseudomallei.

    Directory of Open Access Journals (Sweden)

    Parumon Noinarin

    Full Text Available Presence of Burkholderia pseudomallei in soil and water is correlated with endemicity of melioidosis in Southeast Asia and northern Australia. Several biological and physico-chemical factors have been shown to influence persistence of B. pseudomallei in the environment of endemic areas. This study was the first to evaluate the interaction of B. pseudomallei with soil amoebae isolated from B. pseudomallei-positive soil site in Khon Kaen, Thailand. Four species of amoebae, Paravahlkampfia ustiana, Acanthamoeba sp., Naegleria pagei, and isolate A-ST39-E1, were isolated, cultured and identified based on morphology, movement and 18S rRNA gene sequence. Co-cultivation combined with a kanamycin-protection assay of B. pseudomallei with these amoebae at MOI 20 at 30°C were evaluated during 0-6 h using the plate count technique on Ashdown's agar. The fate of intracellular B. pseudomallei in these amoebae was also monitored by confocal laser scanning microscopy (CLSM observation of the CellTracker™ Orange-B. pseudomallei stained cells. The results demonstrated the ability of P. ustiana, Acanthamoeba sp. and isolate A-ST39-E1 to graze B. pseudomallei. However, the number of internalized B. pseudomallei substantially decreased and the bacterial cells disappeared during the observation period, suggesting they had been digested. We found that B. pseudomallei promoted the growth of Acanthamoeba sp. and isolate A-ST39-E1 in co-cultures at MOI 100 at 30°C, 24 h. These findings indicated that P. ustiana, Acanthamoeba sp. and isolate A-ST39-E1 may prey upon B. pseudomallei rather than representing potential environmental reservoirs in which the bacteria can persist.

  18. Detection of antibody against limax amoebae by means of the indirect haemagglutination test.

    Science.gov (United States)

    Cerva, L

    1977-01-01

    A detailed description is given of the procuedure of the indirect haemagglutination test employed in the detection of specific antibodies against pathogenic amoebae of the genera Naegleria and Acanthamoeba. The preservation of all ingredients obtained by lyophilisation provide for a standard pattern of the test. The results of the test with experimentally prepared immune rabbit sera demonstrate an antigenic uniformity of the various isolates of Naegleria fowleri, and their marked difference from Acanthamoeba.

  19. Presence of Legionella and Free-Living Amoebae in Composts and Bioaerosols from Composting Facilities

    OpenAIRE

    Conza, Lisa; Pagani, Simona Casati; Gaia, Valeria

    2013-01-01

    Several species of Legionella cause Legionnaires' disease (LD). Infection may occur through inhalation of Legionella or amoebal vesicles. The reservoirs of Legionella are water, soil, potting soil and compost. Some species of free-living amoebae (FLA) that are naturally present in water and soil were described as hosts for Legionella. This study aimed to understand whether or not the composting facilities could be sources of community-acquired Legionella infections after development of bioaer...

  20. Pathogenic waterborne free-living amoebae: An update from selected Southeast Asian countries.

    Science.gov (United States)

    Abdul Majid, Mohamad Azlan; Mahboob, Tooba; Mong, Brandon G J; Jaturas, Narong; Richard, Reena Leeba; Tian-Chye, Tan; Phimphila, Anusorn; Mahaphonh, Panomphanh; Aye, Kyaw Nyein; Aung, Wai Lynn; Chuah, Joon; Ziegler, Alan D; Yasiri, Atipat; Sawangjaroen, Nongyao; Lim, Yvonne A L; Nissapatorn, Veeranoot

    2017-01-01

    Data on the distribution of free-living amoebae is still lacking especially in Southeast Asian region. The aquatic environment revealed a high occurrence of free-living amoebae (FLA) due to its suitable condition and availability of food source, which subsequently causes infection to humans. A total of 94 water samples consisted of both treated and untreated from Laos (31), Myanmar (42), and Singapore (21) were investigated for the presence of pathogenic FLA. Each water sample was filtered and cultured onto non-nutrient agar seeded with live suspension of Escherichia coli and incubated at room temperature. Morphological identification was conducted for both trophozoites and cysts via microscopic stains (Giemsa and immunofluorescence). The presence of Naegleria-like structures was the most frequently encountered in both treated and untreated water samples, followed by Acanthamoeba-like and Vermamoeba-like features. To identify the pathogenic isolates, species-specific primer sets were applied for molecular identification of Acanthamoeba, Naegleria, and Vermamoeba. The pathogenic species of Acanthamoeba lenticulata and A. triangularis were detected from untreated water samples, while Vermamoeba vermiformis was found in both treated and untreated water samples. Our results suggested that poor water quality as well as inadequate maintenance and treatment might be the cause of this alarming problem since chlorine disinfection is ineffective in eradicating these amoebas in treated water samples. Regular monitoring and examination of water qualities are necessary in order to control the growth, hence, further preventing the widespread of FLA infections among the public.

  1. Enhanced survival but not amplification of Francisella spp. in the presence of free-living amoebae.

    Science.gov (United States)

    Buse, Helen Y; Schaefer, Frank W; Rice, Eugene W

    2017-03-01

    Transmission of Francisella tularensis, the etiologic agent of tularemia, has been associated with various water sources. Survival of many waterborne pathogens within free-living amoeba (FLA) is well documented; however, the role of amoebae in the environmental persistence of F. tularensis is unclear. In this study, axenic FLA cultures of Acanthamoeba castellanii, Acanthamoeba polyphaga, and Vermamoeba vermiformis were each inoculated with virulent strains of F. tularensis (Types A and B), the attenuated live vaccine strain, and Francisella novicida. Experimental parameters included low and high multiplicity of infection and incubation temperatures of 25 and 30 °C for 0-10 days. Francisella spp. survival was enhanced by the presence of FLA; however, bacterial growth and protozoa infectivity were not observed. In contrast, co-infections of A. polyphaga and Legionella pneumophila, used as an amoeba pathogen control, resulted in bacterial proliferation, cytopathic effects, and amoebal lysis. Collectively, even though short-term incubation with FLA was beneficial, the long-term effects on Francisella survival are unknown, especially given the expenditure of available amoebal derived nutrients and the fastidious nature of Francisella spp. These factors have clear implications for the role of FLA in Francisella environmental persistence.

  2. Exploring Virulence Determinants of Filamentous Fungal Pathogens through Interactions with Soil Amoebae.

    Science.gov (United States)

    Novohradská, Silvia; Ferling, Iuliia; Hillmann, Falk

    2017-01-01

    Infections with filamentous fungi are common to all animals, but attention is rising especially due to the increasing incidence and high mortality rates observed in immunocompromised human individuals. Here, Aspergillus fumigatus and other members of its genus are the leading causative agents. Attributes like their saprophytic life-style in various ecological niches coupled with nutritional flexibility and a broad host range have fostered the hypothesis that environmental predators could have been the actual target for some of their virulence determinants. In this mini review, we have merged the recent findings focused on the potential dual-use of fungal defense strategies against innate immune cells and soil amoebae as natural phagocytes. Well-established virulence attributes like the melanized surface of fungal conidia or their capacity to produce toxic secondary metabolites have also been found to be protective against the model amoeba Dictyostelium discoideum . Some of the recent advances during interaction studies with human cells have further promoted the adaptation of other amoeba infection models, including the wide-spread generalist Acanthamoeba castellanii , or less prominent representatives like Vermamoeba vermiformis . We further highlight prospects and limits of these natural phagocyte models with regard to the infection biology of filamentous fungi and in comparison to the phagocytes of the innate immune system.

  3. Bacteriolytic activities of the free-living soil amoebae, Acanthamoeba castellanii, Acanthamoeba polyphaga and Hartmannella vermiformis.

    Science.gov (United States)

    Weekers, P H; Engelberts, A M; Vogels, G D

    1995-10-01

    Bacteriolytic activities of axenically grown free-living soil amoebae Acanthamoeba castellanii, Acanthamoeba polyphaga and Hartmannella vermiformis towards various Gram-positive and Gram-negative bacteria were determined. A spectrophotometric assay revealed that the specific bacteriolytic activities of both Acanthamoeba species were higher as those of the three Hartmannella strains. Bacillus megaterium, Bacillus subtilis, Chromatium vinosum, Micrococcus luteus and Pseudomonas fluorescens were more easily lysed than the other bacteria tested. Agrobacterium tumefaciens, Klebsiella aerogenes and Serratia marcescens were hardly affected at all by the amoebal bacteriolytic activities. Among the Gram-negative bacteria we observed differences in lysis sensitivity while the Gram-positive bacteria tested were sensitive to lysis. Isoelectric focusing (IEF) gel-electrophoresis in the pH range 3-10 was performed to separate the bacteriolytic isoenzymes of amoebae. Bacteriolytic patterns were shown by using an activity assay in which lysis bands were formed in the agar/bacteria gel-overlay. The activity assay revealed remarkable differences in typical banding patterns for bacteriolytic activities among amoebae. Distinct differences between typical pI points of bacteriolytic activities in Acanthamoeba and Hartmannella were shown. Bacteriolytic activities of Hartmannella were more pronounced and observed in the isoelectric points (pI) range of 4.0-9.3 while for Acanthamoeba the range was pI 4.5-8.9.

  4. Elimination of free-living amoebae in fresh water with pulsed electric fields.

    Science.gov (United States)

    Vernhes, M C; Benichou, A; Pernin, P; Cabanes, P A; Teissié, J

    2002-08-01

    This study investigates the effects of pulsed electric fields on the inactivation of trophozoite form of Naegleria lovaniensis Ar9M-1 in batch and flow processes, systematically examining the lethal effect of field strength, pulse duration, number of pulses, and pulse frequency. Our results show that amoebae eradication is modulated by pulse parameters, composition of the pulsing medium, and physiological state of the cells. Cell survival is not related to the energy delivered to the cell suspension during the electrical treatment. For a given energy a strong field applied for a short cumulative pulse duration affects viability more than a weak field with a long cumulative pulsation. We also determine the optimal electrical conditions to obtain an inactivation rate higher than 95% while using the least energy. Flow processes allow to treat large-scale volumes. Our results show that the most efficient flow process for amoeba eradication requires a field parallel to the flow. Pulsed electric fields are a new and attractive method for inactivating amoebae in large volumes of fresh water.

  5. [Isolation of Legionella and free-living amoebae at hot spring spas in Kanagawa, Japan].

    Science.gov (United States)

    Kuroki, T; Yagita, K; Yabuuchi, E; Agata, K; Ishima, T; Katsube, Y; Endo, T

    1998-10-01

    Microbiological contamination of hot spring bath water is a public health concern. A province-wide survey was carried out to determine the extent and distribution of both Legionella and free-living amoebae contamination. Among 30 samples of hot spring bath from 12 sites in Kanagawa, Japan, L. pneumophila was detected in 21 water samples from 11 sites, ranging from 10(1)-10(3) CFU/100 ml. Serogroups 3, 5 and 6 of L. pneumophila were predominantly isolated from the samples. Naegleria (46.7%), Platyamoeba (33.3%), Acanthamoeba (10.0%) and 2 other genera of free-living amoebae were detected in 22 samples from 11 sites. One or more genera of host amoebae of Legionella occurred in 17 samples (56.7%) from 9 sites. Another thing to be noted is that 13 water samples contained N. lovaniensis. Although N. lovaniensis is nonpathogenic, it is considered an indicator organism for places that are suitable for the growth of N. fowleri, a causative agent of primary amoebic meningoencephalitis in man.

  6. Isolation of free-living amoebae from sarein hot springs in ardebil province, iran.

    Science.gov (United States)

    Badirzadeh, A; Niyyati, M; Babaei, Z; Amini, H; Badirzadeh, H; Rezaeian, M

    2011-06-01

    Free-living amoebae (FLA) are a group of ubiquitous protozoan, which are distributed in the natural and artificial environment sources. The main aim of the current study was to identify the presence of FLA in the recreational hot springs of Sarein in Ardebil Province of Iran. Seven recreational hot springs were selected in Sarein City and 28 water samples (four from each hot spring) were collected using 500 ml sterile plastic bottles during three month. Filtration of water samples was performed, and culture was done in non-nutrient agar medium enriched with Escherichia coli. Identification of the FLA was based on morphological criteria of cysts and trophozoites. Genotype identification of Acanthamoeba positive samples were also performed using sequencing based method. Overall, 12 out of 28 (42.9%) samples were positive for FLA which Acanthamoeba and Vahlkampfiid amoebae were found in one (3.6%) and 11 (39.3%) samples, respectively. Sequence analysis of the single isolate of Acanthamoeba revealed potentially pathogenic T(4) genotype corresponding to A. castellanii. Contamination of hot springs to FLA, such as Acanthamoeba T(4) genotype (A. castellanii) and Vahlkampfiid amoebae, could present a sanitary risk for high risk people, and health authorities must be aware of FLA presence.

  7. Calcium and initial surface binding phase of pinocytosis in Amoeba proteus

    Energy Technology Data Exchange (ETDEWEB)

    Prusch, R.D.

    1986-08-01

    The uptake of membrane-bound solute and external medium by bulk-phase pinocytosis in Amoeba proteus is influenced by the level of Ca/sup 2 +/ in the external medium. Increasing external Ca/sup 2 +/ to approx.10/sup -4/ M increases pinocytotic intensity, while increases in Ca/sup 2 +/ above this level decrease the intensity of pinocytosis. The initial interaction of pinocytotic inducers and Ca/sup +2/ at the surface of A moeba proteus was therefore examined. Alcain blue and Na/sup +/, both inducers of pinocytosis, differ in the manner with which they associate with the amoeba surface, suggesting the possibility of different pinocytosis-inducing sites on the amoeba surface. Low levels of external Ca/sup 2 +/ in the range of 3 x 10/sup -5/ to 4.5 x 10/sup -4/ M increase the amount of cationic inducer associated with the cell surface while, at the same time, decreasing anion association with the cell surface. It is suggested that Ca/sup 2 +/ influences ion association with the cell surface by controlling the availability of negative surface sites, which in turn influences pinocytotic intensity. Surface binding of Na/sup +/, Ca/sup 2 +/ and Cl/sup -/ was determined by adding /sup 22/Na, /sup 45/Ca or /sup 36/Cl.

  8. Genome-wide identification of pathogenicity factors of the free-living amoeba Naegleria fowleri.

    Science.gov (United States)

    Zysset-Burri, Denise C; Müller, Norbert; Beuret, Christian; Heller, Manfred; Schürch, Nadia; Gottstein, Bruno; Wittwer, Matthias

    2014-06-19

    The free-living amoeba Naegleria fowleri is the causative agent of the rapidly progressing and typically fatal primary amoebic meningoencephalitis (PAM) in humans. Despite the devastating nature of this disease, which results in > 97% mortality, knowledge of the pathogenic mechanisms of the amoeba is incomplete. This work presents a comparative proteomic approach based on an experimental model in which the pathogenic potential of N. fowleri trophozoites is influenced by the compositions of different media. As a scaffold for proteomic analysis, we sequenced the genome and transcriptome of N. fowleri. Since the sequence similarity of the recently published genome of Naegleria gruberi was far lower than the close taxonomic relationship of these species would suggest, a de novo sequencing approach was chosen. After excluding cell regulatory mechanisms originating from different media compositions, we identified 22 proteins with a potential role in the pathogenesis of PAM. Functional annotation of these proteins revealed, that the membrane is the major location where the amoeba exerts its pathogenic potential, possibly involving actin-dependent processes such as intracellular trafficking via vesicles. This study describes for the first time the 30 Mb-genome and the transcriptome sequence of N. fowleri and provides the basis for the further definition of effective intervention strategies against the rare but highly fatal form of amoebic meningoencephalitis.

  9. Exploring Virulence Determinants of Filamentous Fungal Pathogens through Interactions with Soil Amoebae

    Directory of Open Access Journals (Sweden)

    Silvia Novohradská

    2017-12-01

    Full Text Available Infections with filamentous fungi are common to all animals, but attention is rising especially due to the increasing incidence and high mortality rates observed in immunocompromised human individuals. Here, Aspergillus fumigatus and other members of its genus are the leading causative agents. Attributes like their saprophytic life-style in various ecological niches coupled with nutritional flexibility and a broad host range have fostered the hypothesis that environmental predators could have been the actual target for some of their virulence determinants. In this mini review, we have merged the recent findings focused on the potential dual-use of fungal defense strategies against innate immune cells and soil amoebae as natural phagocytes. Well-established virulence attributes like the melanized surface of fungal conidia or their capacity to produce toxic secondary metabolites have also been found to be protective against the model amoeba Dictyostelium discoideum. Some of the recent advances during interaction studies with human cells have further promoted the adaptation of other amoeba infection models, including the wide-spread generalist Acanthamoeba castellanii, or less prominent representatives like Vermamoeba vermiformis. We further highlight prospects and limits of these natural phagocyte models with regard to the infection biology of filamentous fungi and in comparison to the phagocytes of the innate immune system.

  10. Isolation of Free-Living Amoebae from Sarein Hot Springs in Ardebil Province, Iran

    Directory of Open Access Journals (Sweden)

    A Badirzadeh

    2011-06-01

    Full Text Available Background: Free-living amoebae (FLA are a group of ubiquitous protozoan, which are distrib­uted in the natural and artificial environment sources. The main aim of the current study was to identify the presence of FLA in the recreational hot springs of Sarein in Ardebil Province of Iran.Methods: Seven recreational hot springs were selected in Sarein City and 28 water samples (four from each hot spring were collected using 500 ml sterile plastic bottles during three month. Filtra­tion of water samples was performed, and culture was done in non-nutrient agar medium enriched with Escherichia coli. Identification of the FLA was based on morphological criteria of cysts and trophozoites. Genotype identification of Acanthamoeba positive samples were also per­formed using sequencing based method.Results: Overall, 12 out of 28 (42.9% samples were positive for FLA which Acanthamoeba and Vahlkampfiid amoebae were found in one (3.6% and 11 (39.3% samples, respectively. Se­quence analysis of the single isolate of Acanthamoeba revealed potentially pathogenic T4 geno­type corresponding to A. castellanii.Conclusion: Contamination of hot springs to FLA, such as Acanthamoeba T4 genotype (A. castel­lanii and Vahlkampfiid amoebae, could present a sanitary risk for high risk people, and health authorities must be aware of FLA presence.

  11. The potential influence of short-term environmental variability on the composition of testate amoeba communities in Sphagnum peatlands.

    Science.gov (United States)

    Sullivan, Maura E; Booth, Robert K

    2011-07-01

    Testate amoebae are a group of moisture-sensitive, shell-producing protozoa that have been widely used as indicators of changes in mean water-table depth within oligotrophic peatlands. However, short-term environmental variability (i.e., sub-annual) also probably influences community composition. The objective of this study was to assess the potential influence of short-term environmental variability on the composition of testate amoeba communities in Sphagnum-dominated peatlands. Testate amoebae and environmental conditions, including hourly measurements of relative humidity within the upper centimeter of the peatland surface, were examined throughout the 2008 growing season at 72 microsites within 11 peatlands of Pennsylvania and Wisconsin, USA. Relationships among testate amoeba communities, vegetation, depth to water table, pH, and an index of short-term environmental variability (EVI), were examined using nonmetric multidimensional scaling and correlation analysis. Results suggest that EVI influences testate amoeba communities, with some taxa more abundant under highly variable conditions (e.g., Arcella discoides, Difflugia pulex, and Hyalosphenia subflava) and others more abundant when environmental conditions at the peatland surface were relatively stable (e.g., Archerella flavum and Bullinularia indica). The magnitude of environmental variability experienced at the peatland surface appears to be primarily controlled by vegetation composition and density. In particular, sites with dense Sphagnum cover had lower EVI values than sites with loose-growing Sphagnum or vegetation dominated by vascular plants and/or non-Sphagnum bryophytes. Our results suggest that more environmental information may be inferred from testate amoebae than previously recognized. Knowledge of relationships between testate amoebae and short-term environmental variability should lead to more detailed and refined environmental inferences.

  12. Genetic structure of a morphological species within the amoeba genus Korotnevella (Amoebozoa: Discosea), revealed by the analysis of two genes.

    Science.gov (United States)

    Zlatogursky, Vasily V; Kudryavtsev, Alexander; Udalov, Ilya A; Bondarenko, Natalya; Pawlowski, Jan; Smirnov, Alexey

    2016-10-01

    Amoebae of the genus Korotnevella are covered with scales, the structure of which is believed to be species-specific and allows distinguishing species reliably at the morphological level. We studied members of this genus in order to assess the genetic structure of the local populations of amoebae. For the present study we isolated nine freshwater strains of Korotnevella, belonging to three species, from two locations in North-Western Russia. In order to obtain data on the population structure of these amoebae, we identified all isolates based on the light-microscopic morphology and scale structure and investigated both inter-strain and intra-strain polymorphism of Cox I and 18S rRNA genes. Results show that both genes provide congruent patterns of population structure. The Cox I gene appears to be more reliable DNA barcode while the 18S rRNA gene shows an interesting pattern of polymorphism, which may represent phylotypes of amoebae. Local population of amoebae in every studied species consists of a number of genetic lineages (phylotypes), some shared between the populations while others are unique to a local habitat. Copyright © 2016. Published by Elsevier GmbH.

  13. Hunting for agile prey: trophic specialisation in leptophryid amoebae (Vampyrellida, Rhizaria) revealed by two novel predators of planktonic algae.

    Science.gov (United States)

    Hess, Sebastian

    2017-09-01

    Vampyrellid amoebae (Vampyrellida, Rhizaria) are widespread in freshwater, marine and terrestrial ecosystems and consume a wide range of eukaryotes, e.g. algae, fungi and micrometazoa. Environmental sequences indicate that only a small fraction of their genetic diversity is phenotypically characterised, emphasising the need to further explore unknown vampyrellids and their interactions with prey organisms. This study tests the prey range specificity of three vampyrellid amoebae with 49 strains of three common groups of freshwater algae (Zygnematophyceae, Euglenophyceae and Volvocales), and documents specific interactions by time-lapse microscopy. Two of the amoebae, here introduced as the novel genera Arachnomyxa and Planctomyxa based on morphology and SSU rRNA gene comparisons, display a complementary prey range and consume motile algae, namely Volvocales and Euglenophyceae, respectively. This reveals the existence of specialised 'plankton feeders' in the vampyrellid family Leptophryidae, contrasting with the strikingly broad prey range of Leptophrys vorax. The distinct autecological characteristics found in this group of morphologically rather indistinct amoebae contribute to our knowledge about the vastly understudied vampyrellid amoebae. Furthermore, time-lapse observations suggest that euglenoid movements exerted by the sluggish species of the 'Euglena deses group' as a reaction to vampyrellid contact may serve as an effective defence against microbial predators. © FEMS 2017. All rights reserved. For permissions, please e-mail: journals.permissions@oup.com.

  14. Survival of Campylobacter jejuni in co-culture with Acanthamoeba castellanii: role of amoeba-mediated depletion of dissolved oxygen.

    Science.gov (United States)

    Bui, Xuan Thanh; Winding, Anne; Qvortrup, Klaus; Wolff, Anders; Bang, Dang Duong; Creuzenet, Carole

    2012-08-01

    Campylobacter jejuni is a major cause of infectious diarrhoea worldwide but relatively little is known about its ecology. In this study, we examined its interactions with Acanthamoeba castellanii, a protozoan suspected to serve as a reservoir for bacterial pathogens. We observed rapid degradation of intracellular C.jejuni in A.castellanii 5 h post gentamicin treatment at 25°C. Conversely, we found that A.castellanii promoted the extracellular growth of C.jejuni in co-cultures at 37°C in aerobic conditions. This growth-promoting effect did not require amoebae - bacteria contact. The growth rates observed with or without contact with amoeba were similar to those observed when C.jejuni was grown in microaerophilic conditions. Preconditioned media prepared with live or dead amoebae cultivated with or without C.jejuni did not promote the growth of C.jejuni in aerobic conditions. Interestingly, the dissolved oxygen levels of co-cultures with or without amoebae - bacteria contact were much lower than those observed with culture media or with C.jejuni alone incubated in aerobic conditions, and were comparable with levels obtained after 24 h of growth of C.jejuni under microaerophilic conditions. Our studies identified the depletion of dissolved oxygen by A.castellanii as the major contributor for the observed amoeba-mediated growth enhancement. © 2011 Society for Applied Microbiology and Blackwell Publishing Ltd.

  15. Control of Branchionus sp. and Amoeba sp. in cultures of Arthrospira sp. Control de Branchionus sp. y Amoeba sp. en cultivos de Arthrospira sp.

    Directory of Open Access Journals (Sweden)

    Carlos Méndez

    2012-09-01

    Full Text Available Cultivation of cyanobacterium Arthrospira sp. has been developed in many countries for the production of proteins, pigments and other compounds. Outdoor mass cultures are often affected by biological contamination, drastically reducing productivity as far as bringing death. This study evaluates the control of Branchionus sp. and Amoeba sp. with two chemical compounds: urea (U and ammonium bicarbonate (AB, in laboratory conditions and outdoor mass culture of Arthrospira sp. The lethal concentration 100 (LC100 at 24 h for Branchionus sp. and Amoeba sp. determined was of 60-80 mg L-1 (U and 100-150 mg L-1 (AB. The average effective inhibition concentration for 50% of the population (IC50 in Arthrospira sp., after 72 h, was 80 mg L-1 (U and 150 mg L-1 (AB. The application of doses of 60 mg L-1 (U or 100 mg L-1 (AB in the outdoor mass culture of this contaminated microalga, completely inhibited grazing and did not affect the growth of Arthrospira sp. but rather promoted rapid recovery of algal density at levels prior to infestation. These compounds provided an economical and effective control of predators in cultures of Arthrospira sp.El cultivo de la cianobacteria Arthrospira sp. ha sido desarrollado en muchos países para la obtención de proteínas, pigmentos y otros compuestos. Cultivo que a nivel industrial se ve afectado frecuentemente por contaminación biológica, reduciendo drásticamente la productividad hasta causar la muerte. Este estudio evalúa el control de Branchionus sp. y de Amoeba sp. con dos compuestos químicos, la urea (U y bicarbonato de amonio (AB en cultivos de Arthrospira sp. La concentración letal 100 (LC100 determinada a las 24 h para Branchionus sp. y Amoeba sp. fue de 60-80 mg L-1 (U y 100-150 mg L-1 (AB. La concentración media de inhibición efectiva, después de 72 h, para el 50% de la población (IC50 en Arthrospira fue de 80 mg L-1 (U y 150 mg L-1 (AB. La aplicación de dosis de 60 mg L-1 (U ó 100 mg L-1 (AB en

  16. Investigating the role of free-living amoebae as a reservoir for Mycobacterium ulcerans.

    Science.gov (United States)

    Amissah, Nana Ama; Gryseels, Sophie; Tobias, Nicholas J; Ravadgar, Bahram; Suzuki, Mitsuko; Vandelannoote, Koen; Durnez, Lies; Leirs, Herwig; Stinear, Timothy P; Portaels, Françoise; Ablordey, Anthony; Eddyani, Miriam

    2014-09-01

    The reservoir and mode of transmission of Mycobacterium ulcerans, the causative agent of Buruli ulcer, still remain a mystery. It has been suggested that M. ulcerans persists with difficulty as a free-living organism due to its natural fragility and inability to withstand exposure to direct sunlight, and thus probably persists within a protective host environment. We investigated the role of free-living amoebae as a reservoir of M. ulcerans by screening the bacterium in free-living amoebae (FLA) cultures isolated from environmental specimens using real-time PCR. We also followed the survival of M. ulcerans expressing green fluorescence protein (GFP) in Acanthameoba castellanii by flow cytometry and observed the infected cells using confocal and transmission electron microscopy for four weeks in vitro. IS2404 was detected by quantitative PCR in 4.64% of FLA cultures isolated from water, biofilms, detritus and aerosols. While we could not isolate M. ulcerans, 23 other species of mycobacteria were cultivated from inside FLA and/or other phagocytic microorganisms. Laboratory experiments with GFP-expressing M. ulcerans in A. castellani trophozoites for 28 days indicated the bacteria did not replicate inside amoebae, but they could remain viable at low levels in cysts. Transmission electron microscopy of infected A. castellani confirmed the presence of bacteria within both trophozoite vacuoles and cysts. There was no correlation of BU notification rate with detection of the IS2404 in FLA (r = 0.07, n = 539, p = 0.127). This study shows that FLA in the environment are positive for the M. ulcerans insertion sequence IS2404. However, the detection frequency and signal strength of IS2404 positive amoabae was low and no link with the occurrence of BU was observed. We conclude that FLA may host M. ulcerans at low levels in the environment without being directly involved in the transmission to humans.

  17. Coexistence of free-living amoebae and bacteria in selected South African hospital water distribution systems.

    Science.gov (United States)

    Muchesa, P; Leifels, M; Jurzik, L; Hoorzook, K B; Barnard, T G; Bartie, C

    2017-01-01

    Pathogenic free-living amoebae (FLA), such as Naegleria fowleri, Balamuthia mandrillaris and Acanthamoeba species isolated from aquatic environments have been implicated in central nervous system, eye and skin human infections. They also allow the survival, growth and transmission of bacteria such as Legionella, Mycobacteria and Vibrio species in water systems. The purpose of this study was to investigate the co-occurrence of potentially pathogenic FLA and their associated bacteria in hospital water networks in Johannesburg, South Africa. A total of 178 water (n = 95) and swab (n = 83) samples were collected from two hospital water distribution systems. FLA were isolated using the amoebal enrichment technique and identified using PCR and 18S rDNA sequencing. Amoebae potentially containing intra-amoebal bacteria were lysed and cultured on blood agar plates. Bacterial isolates were characterized using the VITEK®2 compact System. Free-living amoebae were isolated from 77 (43.3 %) of the samples. Using microscopy, PCR and 18S rRNA sequencing, Acanthamoeba spp. (T3 and T20 genotypes), Vermamoeba vermiformis and Naegleria gruberi specie were identified. The Acanthamoeba T3 and T20 genotypes have been implicated in eye and central nervous system infections. The most commonly detected bacterial species were Serratia marcescens, Stenotrophomonas maltophilia, Delftia acidovorans, Sphingomonas paucimobilis and Comamonas testosteroni. These nosocomial pathogenic bacteria are associated with systematic blood, respiratory tract, the urinary tract, surgical wounds and soft tissues infections. The detection of FLA and their associated opportunistic bacteria in the hospital water systems point out to a potential health risk to immune-compromised individuals.

  18. Computational identification of putative miRNAs and their target genes in pathogenic amoeba Naegleria fowleri.

    Science.gov (United States)

    Padmashree, Dyavegowda; Swamy, Narayanaswamy Ramachandra

    2015-01-01

    Naegleria fowleri is a parasitic unicellular free living eukaryotic amoeba. The parasite spreads through contaminated water and causes primary amoebic meningoencephalitis (PAM). Therefore, it is of interest to understand its molecular pathogenesis. Hence, we analyzed the parasite genome for miRNAs (microRNAs) that are non-coding, single stranded RNA molecules. We identified 245 miRNAs using computational methods in N. fowleri, of which five miRNAs are conserved. The predicted miRNA targets were analyzed by using miRanda (software) and further studied the functions by subsequently annotating using AmiGo (a gene ontology web tool).

  19. Confocal Laser Scanning Microscopy and Two Photon Excitation Microscopy as Tools to Study Testate Amoebae

    Czech Academy of Sciences Publication Activity Database

    Burdíková, Zuzana; Čapek, Martin; Ostašov, Pavel; Mitchell, E.A.D.; Machač, Jiří; Kubínová, Lucie

    2010-01-01

    Roč. 16, Suppl.2 (2010), s. 1142-1143 ISSN 1431-9276. [Microscopy and Microanalysis 2010. Portland, 01.08.2010-05.08.2010] R&D Projects: GA MŠk(CZ) LC06063; GA ČR(CZ) GA102/08/0691; GA ČR(CZ) GA304/09/0733 Institutional research plan: CEZ:AV0Z50110509; CEZ:AV0Z60050516 Keywords : testate amoeba e * confocal microscopy * two-photon microscopy Subject RIV: BH - Optics, Masers, Lasers Impact factor: 2.179, year: 2010

  20. Effects of Grazing by the Free-Living Soil Amoebae Acanthamoeba castellanii, Acanthamoeba polyphaga, and Hartmannella vermiformis on Various Bacteria

    Science.gov (United States)

    Weekers, Peter H. H.; Bodelier, Paul L. E.; Wijen, John P. H.; Vogels, Godfried D.

    1993-01-01

    Cultures of 10 different bacteria were used to serve as food sources for axenically grown Acanthamoeba castellanii, Acanthamoeba polyphaga, and Hartmannella vermiformis. The nonpigmented enterobacteriaceae Escherichia coli K-12 and Klebsiella aerogenes appeared to be excellent feed to all three amoebae. Hardly any growth or ammonium production was observed in tests with Chromatium vinosum and Serratia marcescens, which share the presence of pigmented compounds. Distinct differences in net ammonium production were detected and were correlated to the amoebal growth yield. In general, growth of amoebae and ammonium production increased in the order A. polyphaga, A. castellanii, and H. vermiformis. PMID:16349000

  1. Effects of Naphthalene on DNA and RNA quantity in Amoeba proteus by using confocal laser scanning microscope

    Directory of Open Access Journals (Sweden)

    Khwanmuni, J.

    2006-01-01

    Full Text Available Effects of Naphthalene which is a carcinogen on changes of DNA and RNA quantities were studied with acridine orange stained cells under a confocal laser scanning microscope. It was found that DNA and RNA in amoebae nucleus and cytoplasm, reared in 0 (control, 3 and 8.85 mg/l (24h-LD50 at 0 and 12 h. showed a statistically significant difference (p<0.05. The more naphthalene concentrations and larger incubation periods had greater effects on DNA and RNA decreases in amoebae nucleus and cytoplasm.

  2. SSU rRNA reveals a sequential increase in shell complexity among the euglyphid testate amoebae (Rhizaria: Euglyphida)

    DEFF Research Database (Denmark)

    Lara, Enrique; Heger, Thierry J; Mitchell, Edward A D

    2007-01-01

    The existing data on the molecular phylogeny of filose testate amoebae from order Euglyphida has revealed contradictions between traditional morphological classification and SSU rRNA phylogeny and, moreover, the position of several important genera remained unknown. We therefore carried out a study...... aiming to fill several important gaps and better understand the relationships among the main euglyphid testate amoebae and the evolutionary steps that led to the present diversity at a higher level. We obtained new SSU rRNA sequences from five genera and seven species. This new phylogeny obtained shows...

  3. Old lineages in a new ecosystem: diversification of arcellinid amoebae (Amoebozoa and peatland mosses.

    Directory of Open Access Journals (Sweden)

    Omar Fiz-Palacios

    Full Text Available Arcellinid testate amoebae (Amoebozoa form a group of free-living microbial eukaryotes with one of the oldest fossil records known, yet several aspects of their evolutionary history remain poorly understood. Arcellinids occur in a range of terrestrial, freshwater and even brackish habitats; however, many arcellinid morphospecies such as Hyalosphenia papilio are particularly abundant in Sphagnum-dominated peatlands, a relatively new ecosystem that appeared during the diversification of Sphagnum species in the Miocene (5-20 Myr ago. Here, we reconstruct divergence times in arcellinid testate amoebae after selecting several fossils for clock calibrations and then infer whether or not arcellinids followed a pattern of diversification that parallels the pattern described for Sphagnum. We found that the diversification of core arcellinids occurred during the Phanerozoic, which is congruent with most arcellinid fossils but not with the oldest known amoebozoan fossil (i.e. at ca. 662 or ca. 750 Myr. Overall, Sphagnum and the Hyalospheniidae exhibit different patterns of diversification. However, an extensive molecular phylogenetic analysis of distinct clades within H. papilio species complex demonstrated a correlation between the recent diversification of H. papilio, the recent diversification of Sphagnum mosses, and the establishment of peatlands.

  4. Isolation and molecular characterization of free-living amoebae from different water sources in Italy.

    Science.gov (United States)

    Montalbano Di Filippo, Margherita; Santoro, Maristella; Lovreglio, Piero; Monno, Rosa; Capolongo, Carmen; Calia, Carla; Fumarola, Luciana; D'Alfonso, Rossella; Berrilli, Federica; Di Cave, David

    2015-03-24

    Free-living amoebae (FLA) are protozoa ubiquitous in Nature, isolated from a variety of environments worldwide. In addition to their natural distribution, some species have been found to be pathogenic to humans. In the present study a survey was conducted in order to evaluate the presence and to characterize at molecular level the isolates of amoebic organisms collected from different water sources in Italy. A total of 160 water samples were analyzed by culture and microscopic examination. FLA were found in 46 (28.7%) of the investigated water samples. Groundwater, well waters, and ornamental fountain waters were the sources with higher prevalence rates (85.7%, 50.0%, and 45.9%, respectively). Identification of FLA species/genotypes, based on the 18S rDNA regions, allowed to identify 18 (39.1%) Acanthamoeba isolates (genotypes T4 and T15) and 21 (45.6%) Vermamoeba vermiformis isolates. Other FLA species, including Vahlkampfia sp. and Naegleria spp., previously reported in Italy, were not recovered. The occurrence of potentially pathogenic free-living amoebae in habitats related to human population, as reported in the present study, supports the relevance of FLA as a potential health threat to humans.

  5. Two distinct sensing pathways allow recognition of Klebsiella pneumoniae by Dictyostelium amoebae.

    Science.gov (United States)

    Lima, Wanessa C; Balestrino, Damien; Forestier, Christiane; Cosson, Pierre

    2014-03-01

    Recognition of bacteria by metazoans is mediated by receptors that recognize different types of microorganisms and elicit specific cellular responses. The soil amoebae Dictyostelium discoideum feeds upon a variable mixture of environmental bacteria, and it is expected to recognize and adapt to various food sources. To date, however, no bacteria-sensing mechanisms have been described. In this study, we isolated a Dictyostelium mutant (fspA KO) unable to grow in the presence of non-capsulated Klebsiella pneumoniae bacteria, but growing as efficiently as wild-type cells in the presence of other bacteria, such as Bacillus subtilis. fspA KO cells were also unable to respond to K. pneumoniae and more specifically to bacterially secreted folate in a chemokinetic assay, while they responded readily to B. subtilis. Remarkably, both WT and fspA KO cells were able to grow in the presence of capsulated LM21 K. pneumoniae, and responded to purified capsule, indicating that capsule recognition may represent an alternative, FspA-independent mechanism for K. pneumoniae sensing. When LM21 capsule synthesis genes were deleted, growth and chemokinetic response were lost for fspA KO cells, but not for WT cells. Altogether, these results indicate that Dictyostelium amoebae use specific recognition mechanisms to respond to different K. pneumoniae elements. © 2013 John Wiley & Sons Ltd.

  6. Evidence of parasexual activity in "asexual amoebae" Cochliopodium spp. (Amoebozoa): extensive cellular and nuclear fusion.

    Science.gov (United States)

    Tekle, Yonas I; Anderson, O Roger; Lecky, Ariel F

    2014-09-01

    The majority of microbial eukaryotes have long been considered asexual, though new evidence indicates sex, or sexual-like (parasexual) behaviors that deviate from the usual union of two gametes, among other variant aspects. Over a dozen amoebozoans are implicated to have sexual stages. However, the exact mechanism by which sex occurs in these lineages remains elusive. This is mainly due to the diverse quality and cryptic nature of their life cycle. In this study we present evidence of some previously unreported aspects of the life cycle of an amoeba, Cochliopodium, that undergoes unusual intraspecific interactions using light microscopy and immunocytochemistry. Similar to other amoebozoans, Cochliopodium, is considered asexual with no published reports of sex or parasexuality. We also investigated environmental conditions that govern the observed intraspecific interactions. Both light microscopic and immunocytochemistry evidence demonstrates Cochliopodium undergoes cellular fusion (plasmogamy) and nuclear fusion (karyogamy). Large plasmodia eventually undergo karyogamy and contain large fused, polyploid, nuclei. These are observed to fragment, subsequently, by karyotomy (nuclear fission) and cytoplasmic fission to yield uninucleated amoebae. This process could lead to a non-meiotic, parasexual exchange of chromosomes in Cochliopodium. These findings strongly suggest that Cochliopodium is involved in parasexual activity and should no longer be considered strictly asexual. Copyright © 2014 Elsevier GmbH. All rights reserved.

  7. Development of a nested PCR assay to detect the pathogenic free-living amoeba Naegleria fowleri.

    Science.gov (United States)

    Réveiller, Fabienne L; Cabanes, Pierre-André; Marciano-Cabral, Francine

    2002-05-01

    Naegleria fowleri is the causative agent of primary amoebic meningoencephalitis, a fatal disease of the central nervous system that is acquired while swimming or diving in freshwater. A cDNA clone designated Mp2C15 obtained from N. fowleri was used as a probe to distinguish N. fowleri from other free-living amoebae. The Mp2C15 probe hybridized to genomic DNA from pathogenic N. fowleri and antigenically related non-pathogenic N. lovaniensis. Mp2C15 was digested with the restriction enzyme XbaI, resulting in two fragments, Mp2C15.G and Mp2C15.P. Four species of Naegleria and four species of Acanthamoeba were examined for reactivity with Mp2C15.P. Mp2C15.P was specific for N. fowleri and was used in the development of a nested PCR assay which is capable of detecting as little as 5 pg of N. fowleri DNA or five intact N. fowleri amoebae. In summary, a rapid, sensitive, and specific assay for the detection of N. fowleri was developed.

  8. Archaeological occurrences and historical review of the human amoeba, Entamoeba histolytica, over the past 6000years.

    Science.gov (United States)

    Le Bailly, Matthieu; Maicher, Céline; Dufour, Benjamin

    2016-08-01

    Understanding parasite history and the evolution of host/parasite relationships is one of the most important aspects of paleoparasitology. Within the framework of this research topic, this paper focuses on the human pathogenic amoeba, Entamoeba histolytica. The compilation of all the available archaeological data concerning this parasite leads to a first glimpse of the history of this parasite of current medical importance. Paleoparasitological investigation into this parasite uses immunological techniques and shows that the modern strain of E. histolytica has been present in Western Europe since at least the Neolithic period (3700yearsBCE), and could have originated in the Old World. The appearance of the modern amoeba strain in the pre-Columbian Americas and the Middle East around the 12th century CE gives rise to hypotheses as to how human migrations (Atlantic or Pacific routes) contributed to the diffusion of this pathogen, resulting in its current distribution. This compilation proves that parasites are valuable proxies for studying past human and animal migrations, and should be given more consideration in the future. Copyright © 2016 Elsevier B.V. All rights reserved.

  9. Serologic prevalence of amoeba-associated microorganisms in intensive care unit pneumonia patients.

    Directory of Open Access Journals (Sweden)

    Sabri Bousbia

    Full Text Available BACKGROUND: Patients admitted to intensive care units are frequently exposed to pathogenic microorganisms present in their environment. Exposure to these microbes may lead to the development of hospital-acquired infections that complicate the illness and may be fatal. Amoeba-associated microorganisms (AAMs are frequently isolated from hospital water networks and are reported to be associated to cases of community and hospital-acquired pneumonia. METHODOLOGY/PRINCIPAL FINDINGS: We used a multiplexed immunofluorescence assay to test for the presence of antibodies against AAMs in sera of intensive care unit (ICU pneumonia patients and compared to patients at the admission to the ICU (controls. Our results show that some AAMs may be more frequently detected in patients who had hospital-acquired pneumonia than in controls, whereas other AAMs are ubiquitously detected. However, ICU patients seem to exhibit increasing immune response to AAMs when the ICU stay is prolonged. Moreover, concomitant antibodies responses against seven different microorganisms (5 Rhizobiales, Balneatrix alpica, and Mimivirus were observed in the serum of patients that had a prolonged ICU stay. CONCLUSIONS/SIGNIFICANCE: Our work partially confirms the results of previous studies, which show that ICU patients would be exposed to water amoeba-associated microorganisms, and provides information about the magnitude of AAM infection in ICU patients, especially patients that have a prolonged ICU stay. However, the incidence of this exposure on the development of pneumonia remains to assess.

  10. Elucidation of the Role of 3-Hydroxy Fatty Acids in Cryptococcus-amoeba Interactions

    Directory of Open Access Journals (Sweden)

    Olihile M. Sebolai

    2017-04-01

    Full Text Available We previously reported that 3-hydroxy fatty acids promoted the survival of cryptococcal cells when acted upon by amoebae. To expand on this, the current study sought to explain how these molecules may protect cells. Our data suggest that 3-hydroxy fatty acids may subvert the internalization of cryptococcal cells via suppression of the levels of a fetuin A-like amoebal protein, which may be important for enhancing phagocytosis. Additionally, we show that an acapsular strain (that is devoid of 3-hydroxy fatty acids was protected against the effects of hydrogen peroxide when exogenous 3-hydroxy fatty acids were present, but not in the absence of 3-hydroxy fatty acids. A similar response profile was noted when a strain with a capsule was challenged with hydrogen peroxide. We also show that cryptococcal cells that naturally produce 3-hydroxy fatty acids were more resistant to the effects of amoebapore (an amoeba-specific hydrolytic enzyme, compared to cells that do not produce these molecules. Taken together, our findings suggest that 3-hydroxy fatty acids possess an anti-phagocytic activity that may be expressed when cells interact with macrophages. This may allow the yeast cells to evade immuno-processing.

  11. Spontaneous occurrence of antibodies against pathogenic amoebae of the limax group in domestic animals.

    Science.gov (United States)

    Cerva, L

    1981-01-01

    Sera of 1 218 animals were examined using indirect haemagglutination reaction with antigens of Naegleria fowleri and Acanthamoeba culbertsoni. Rabbit sera (214 samples) gave negative reaction with both antigens. Bovine sera (593 samples) reacted positively in 1.5%, particularly with the antigen of A. culbertsoni. Out of the 411 pigs examined 12% were positive with one or both of the antigens. In one of the pig farms of a modern type the positivity with one or both of the antigens. In one of the pig farms of a modern type the positivity was as much as 32.2%. The titres of positive animal sera have never been higher than 1 : 320. The amoebae of the limax group do not seem to be involved in the pathogenesis of diseases of cattle and rabbits. A sporadic occurrence of peracute cases is possible. In case of a sudden death, animals with symptoms of encephalitis or pneumonia should be examined for the presence of amoebae. Hypersensitivity should be considered as a possible cause of pig diseases.

  12. Isolation and Molecular Characterization of Free-Living Amoebae from Different Water Sources in Italy

    Directory of Open Access Journals (Sweden)

    Margherita Montalbano Di Filippo

    2015-03-01

    Full Text Available Free-living amoebae (FLA are protozoa ubiquitous in Nature, isolated from a variety of environments worldwide. In addition to their natural distribution, some species have been found to be pathogenic to humans. In the present study a survey was conducted in order to evaluate the presence and to characterize at molecular level the isolates of amoebic organisms collected from different water sources in Italy. A total of 160 water samples were analyzed by culture and microscopic examination. FLA were found in 46 (28.7% of the investigated water samples. Groundwater, well waters, and ornamental fountain waters were the sources with higher prevalence rates (85.7%, 50.0%, and 45.9%, respectively. Identification of FLA species/genotypes, based on the 18S rDNA regions, allowed to identify 18 (39.1% Acanthamoeba isolates (genotypes T4 and T15 and 21 (45.6% Vermamoeba vermiformis isolates. Other FLA species, including Vahlkampfia sp. and Naegleria spp., previously reported in Italy, were not recovered. The occurrence of potentially pathogenic free-living amoebae in habitats related to human population, as reported in the present study, supports the relevance of FLA as a potential health threat to humans.

  13. Contrasting diversity of testate amoebae communities in Sphagnum and brown-moss dominated patches in relation to shell counts.

    Science.gov (United States)

    Lizoňová, Zuzana; Horsák, Michal

    2017-04-01

    Ecological studies of peatland testate amoebae are generally based on totals of 150 individuals per sample. However, the suitability of this standard has never been assessed for alkaline habitats such as spring fens. We explored the differences in testate amoeba diversity between Sphagnum and brown-moss microhabitats at a mire site with a highly diversified moss layer which reflects the small-scale heterogeneity in groundwater chemistry. Relationships between sampling efficiency and sample completeness were explored using individual-based species accumulation curves and the effort required to gain an extra species was assessed. Testate amoeba diversity differed substantially between microhabitats, with brown mosses hosting on average twice as many species and requiring greater shell totals to reach comparable sample analysis efficiency as for Sphagnum. Thus, for samples from alkaline conditions an increase in shell totals would be required and even an overall doubling up to 300 individuals might be considered for reliable community description. Our small-scale data are likely not robust enough to provide an ultimate solution for the optimization of shell totals. However, the results proved that testate amoebae communities from acidic and alkaline environments differ sharply in both species richness and composition and they might call for different methodological approaches. Copyright © 2017 Elsevier GmbH. All rights reserved.

  14. Shedding light on microbial dark matter: a TM6 bacterium as natural endosymbiont of a free-living amoeba.

    Science.gov (United States)

    Delafont, Vincent; Samba-Louaka, Ascel; Bouchon, Didier; Moulin, Laurent; Héchard, Yann

    2015-12-01

    The TM6 phylum belongs to the so-called microbial dark matter that gathers uncultivated bacteria detected only via DNA sequencing. Recently, the genome sequence of a TM6 bacterium (TM6SC1) has led to suggest that this bacterium would adopt an endosymbiotic life. In the present paper, free-living amoebae bearing a TM6 strain were isolated from a water network. The amoebae were identified as Vermamoeba vermiformis and the presence of a TM6 strain was detected by polymerase chain reaction and microscopy. The partial sequence of its 16S rRNA gene showed this strain to be closely related to the sequenced TM6SC1 strain. These bacteria displayed a pyriform shape and were found within V. vermiformis. Therefore, these bacteria were named Vermiphilus pyriformis. Interactions studies showed that V. pyriformis was highly infectious and that its relation with V. vermiformis was specific and highly stable. Finally, it was found that V. pyriformis inhibited the encystment of V. vermiformis. Overall, this study describes for the first time an endosymbiotic relationship between a TM6 bacterium and a free-living amoeba in the environment. It suggests that other bacteria of the TM6 phylum might also be endosymbiotic bacteria and may be found in other free-living amoebae or other organisms. © 2015 Society for Applied Microbiology and John Wiley & Sons Ltd.

  15. A free-living amoeba with unusual pattern of mitochondrial structure isolated from Atlantic salmon, Salmo salar L

    Czech Academy of Sciences Publication Activity Database

    Dyková, Iva; Veverková, Marie; Fiala, Ivan; Macháčková, Blanka

    2002-01-01

    Roč. 41, - (2002), s. 415-419 ISSN 0065-1583 R&D Projects: GA AV ČR IAA6022202 Institutional research plan: CEZ:AV0Z6022909 Keywords : free-living amoeba * mitochondrial cristae * Salmo salar Subject RIV: EA - Cell Biology Impact factor: 0.446, year: 2002

  16. Quantitative detection of the free-living amoeba Hartmannella vermiformis in surface water by using real-time PCR

    NARCIS (Netherlands)

    Kuiper, M.W.; Valster, R.M.; Wullings, B.A.; Boonstra, H.; Smidt, H.; Kooij, van der D.

    2006-01-01

    A real-time PCR-based method targeting the 18S rRNA gene was developed for the quantitative detection of Hartmannella vermiformis, a free-living amoeba which is a potential host for Legionella pneumophila in warm water systems and cooling towers. The detection specificity was validated using genomic

  17. Effect of Common Drinking Water Disinfectants, Chlorine and Heat, on Free Legionella and Amoebae-Associated Legionella

    Science.gov (United States)

    Cervero-Aragó, Sílvia; Rodríguez-Martínez, Sarah; Puertas-Bennasar, Antoni; Araujo, Rosa M.

    2015-01-01

    Chlorine and thermal treatments are the most commonly used procedures to control and prevent Legionella proliferation in drinking water systems of large buildings. However, cases of legionellosis still occur in facilities with treated water. The purpose of this work was to model the effect of temperature and free chlorine applied in similar exposure conditions as in drinking water systems on five Legionella spp. strains and two amoebal strains of the genera Acanthamoeba. Inactivation models obtained were used to determine the effectiveness of the treatments applied which resulted more effective against Legionella than Acanthamoeba, especially those in cystic stages. Furthermore, to determine the influence of the relationship between L. pneumophila and Acanthamoeba spp. on the treatment effectiveness, inactivation models of the bacteria-associated amoeba were also constructed and compared to the models obtained for the free living bacteria state. The Legionella-amoeba association did not change the inactivation models, but it reduced the effectiveness of the treatments applied. Remarkably, at the lowest free chlorine concentration, 0.5 mg L-1, as well as at the lowest temperatures, 50°C and 55°C, the influence of the Legionella-amoeba associate state was the strongest in reducing the effectiveness of the treatments compared to the free Legionella state. Therefore, the association established between L. pneumophila and amoebae in the water systems indicate an increased health risk in proximal areas of the system (close to the tap) where lower free chlorine concentrations and lower temperatures are commonly observed. PMID:26241039

  18. Differential growth of Legionella pneumophila strains within a range of amoebae at various temperatures associated with in-premise plumbing

    Science.gov (United States)

    The potential effect of in-premise plumbing temperatures (24, 32, 37 and 41 °C) on the growth of five different L. pneumophila strains within free-living amoebae (Acanthamoeba polyphaga, Hartmannella vermiformis and Naegleria fowleri) was examined. Compared to controls only fed E...

  19. Effect of UV irradiation (253.7 nm) on free Legionella and Legionella associated with its amoebae hosts.

    Science.gov (United States)

    Cervero-Aragó, Sílvia; Sommer, Regina; Araujo, Rosa M

    2014-12-15

    Water systems are the primary reservoir for Legionella spp., where the bacteria live in association with other microorganisms, such as free-living amoebae. A wide range of disinfection treatments have been studied to control and prevent Legionella colonization but few of them were performed considering its relation with protozoa. In this study, the effectiveness of UV irradiation (253.7 nm) using low-pressure lamps was investigated as a disinfection method for Legionella and amoebae under controlled laboratory conditions. UV treatments were applied to 5 strains of Legionella spp., 4 strains of free-living amoeba of the genera Acanthamoeba and Vermamoeba, treating separately trophozoites and cysts, and to two different co-cultures of Legionella pneumophila with the Acanthamoeba strains. No significant differences in the UV inactivation behavior were observed among Legionella strains tested which were 3 logs reduced for fluences around 45 J/m(2). UV irradiation was less effective against free-living amoebae; which in some cases required up to 990 J/m(2) to obtain the same population reduction. UV treatment was more effective against trophozoites compared to cysts; moreover, inactivation patterns were clearly different between the genus Acanthamoeba and Vermamoeba. For the first time data about Vermamoeba vermiformis UV inactivation has been reported in a study. Finally, the results showed that the association of L. pneumophila with free-living amoebae decreases the effectiveness of UV irradiation against the bacteria in a range of 1.5-2 fold. That fact demonstrates that the relations established between different microorganisms in the water systems can modify the effectiveness of the UV treatments applied. Copyright © 2014 Elsevier Ltd. All rights reserved.

  20. Cheating does not explain selective differences at high and low relatedness in a social amoeba

    Directory of Open Access Journals (Sweden)

    Queller David C

    2010-03-01

    Full Text Available Abstract Background Altruism can be favored by high relatedness among interactants. We tested the effect of relatedness in experimental populations of the social amoeba Dictyostelium discoideum, where altruism occurs in a starvation-induced social stage when some amoebae die to form a stalk that lifts the fertile spores above the soil facilitating dispersal. The single cells that aggregate during the social stage can be genetically diverse, which can lead to conflict over spore and stalk allocation. We mixed eight genetically distinct wild isolates and maintained twelve replicated populations at a high and a low relatedness treatment. After one and ten social generations we assessed the strain composition of the populations. We expected that some strains would be out-competed in both treatments. In addition, we expected that low relatedness might allow the persistence of social cheaters as it provides opportunity to exploit other strains. Results We found that at high relatedness a single clone prevailed in all twelve populations. At low relatedness three clones predominated in all twelve populations. Interestingly, exploitation of some clones by others in the social stage did not explain the results. When we mixed each winner against the pool of five losers, the winner did not prevail in the spores because all contributed fairly to the stalk and spores. Furthermore, the dominant clone at high-relatedness was not cheated by the other two that persisted at low relatedness. A combination of high spore production and short unicellular stage most successfully explained the three successful clones at low relatedness, but not why one of them fared better at high relatedness. Differences in density did not account for the results, as the clones did not differ in vegetative growth rates nor did they change the growth rates over relevant densities. Conclusions These results suggest that social competition and something beyond solitary growth differences

  1. A temporary flagellate (mastigote) stage in the vahlkampfiid amoeba Willaertia magna and its possible evolutionary significance.

    Science.gov (United States)

    Robinson, B S; Christy, P E; De Jonckheere, J F

    1989-01-01

    A temporary flagellate (mastigote) stage has been observed in several isolates of the vahlkampfiid amoeba Willaertia magna. In an Australian isolate studied in detail, flagellates appeared synchronously, although later than in Naegleria fowleri or N. lovaniensis under similar conditions (half-maximal time, t50 = 168 min at 37 degrees C). The flagellates initially have four flagella and lack a cytostome, but undergo several successive divisions, the first of them synchronous, resulting in progressive reduction in cell volume. New flagella appear during and after division, and the number of flagella in daughter cells of later divisions is rather variable. Comparison of these observations with descriptions of other amoeboflagellates confirms that Willaertia is a valid genus. A likely sequence of morphological changes in the evolution of Willaertia and Naegleria from a hypothetical ancestral vahlkampfiid is proposed.

  2. The Laredo strain and other 'Entamoeba histolytica-like' amoebae are Entamoeba moshkovskii.

    Science.gov (United States)

    Clark, C G; Diamond, L S

    1991-05-01

    A small number of Entamoeba isolates from humans, the best known of which is the 'Laredo' strain, have the ability to grow at room temperature. This peculiarity, along with other characteristics, distinguishes the strains from the human pathogen E. histolytica despite their being morphologically inseparable. In contrast, these 'E. histolytica-like' strains share several features with E. moshkovskii, which is most frequently isolated from polluted water. To examine the taxonomic relationships among these morphologically similar organisms, we have used polymerase chain reaction amplification of the small subunit ribosomal RNA gene combined with restriction fragment length polymorphism analysis, 'riboprinting'. The results clearly show that the 'E. histolytica-like' amoebae are indeed strains of E. moshkovskii, and not closely related to E. histolytica.

  3. Bioaccumulation of pathogenic bacteria and amoeba by zebra mussels and their presence in watercourses.

    Science.gov (United States)

    Mosteo, R; Goñi, P; Miguel, N; Abadías, J; Valero, P; Ormad, M P

    2016-01-01

    Dreissena polymorpha (the zebra mussel) has been invading freshwater bodies in Europe since the beginning of the nineteenth century. Filter-feeding organisms can accumulate and concentrate both chemical and biological contaminants in their tissues. Therefore, zebra mussels are recognized as indicators of freshwater quality. In this work, the capacity of the zebra mussel to accumulate human pathogenic bacteria and protozoa has been evaluated and the sanitary risk associated with their presence in surface water has also been assessed. The results show a good correlation between the pathogenic bacteria concentration in zebra mussels and in watercourses. Zebra mussels could therefore be used as an indicator of biological contamination. The bacteria (Escherichia coli, Enterococcus spp., Pseudomonas spp., and Salmonella spp.) and parasites (Cryptosporidium oocysts and free-living amoebae) detected in these mussels reflect a potential sanitary risk in water.

  4. Contribution to the problem of the so-called nonpathogenic amoebae in the intestine of man.

    Science.gov (United States)

    Cerva, L; Kliment, V

    1978-01-01

    Among 10,418 patients of a Prague hospital, a plain infection with intestinal parasitic protozoans was identified in 1,319 persons (12.7%). Of these, 3.5% were infested with Giardia intestinalis, 0.3% with Entamoeba histolytica forma minuta, 5.7% with Endolimax nana. We evaluated the frequency of findings of protozoans in various clinical diagnoses. A statistically significant increase in frequency was recorded for E. nana in diagnoses of eosinophilia, giardiasis, amoebiasis and helminthiasis. A slight increase above the average was found for Entamoeba coli in diagnoses of giardiasis and helminthiasis. Most cases of infection with Entamoeba histolytica were associated with a stay abroad. No increase in the frequency of these protozoans was recorded for patients with diarrhea. An analysis of the results indicated that a nonpathogenic amoeba might participate in the origin of intestinal disorders in man.

  5. Examination of thermally polluted water for free living amoebae and testing for their possible pathogenic properties

    Energy Technology Data Exchange (ETDEWEB)

    Janitschke, K.; Lichy, S.; Westphal, C.

    1982-05-01

    Water and mud samples were collected from canals and rivers which were adjacent to outlets discharging warm water of 3 power plants in Berlin. Downstream samples from 1 bathing resort were also collected. Free living amoebae were isolated from 138 water and 69 mud samples. From these respectively 156 and 73 strains could be cultured and were administered intranasally to mice for pathogenicity tests. Two Acanthamoeba strains from water and 7 from mud could be reisolated from mouse brain and or lungs, although no pathological disorders could be observed. Five Naegleria strains were negative in mouse inoculation tests. Four Acanthamoeba strains which were positive in mice were cultured at + 45 degrees C; no cytopathogenic effects were observed in tissue cultures. Acanthamoeba infective for mice could also be isolated from samples at low water temperatures. Further investigations have to show, whether changes in virulence of amoebic strains are of significance and therefore for epidemiology and pathogenicity in man.

  6. Reaching an entrepreneurial management system of amoebas. A qualitative insight into the European experiences

    Directory of Open Access Journals (Sweden)

    Urban Wiesław

    2016-03-01

    Full Text Available The aim of the study is an assessment of the Amoeba Management System (AMS introduction advancements in some European companies. The study takes the practically focused research approach. The approaches, achievements and phases whilst introducing the AMS principles by companies are observed and critically assessed. First insight into the challenges of AMS introduction is taken basing on critical study of the literature output. The scientific studies and managerial publications are taken into consideration. The empirical part of the study is based on the qualitative approach. A multiple case study methodology is employed. The research objects are three companies, one of them operates in Sweden, the next two in Poland. Each of them have different experiences in AMS implementation, they also manifest different management styles and habits. The study demonstrates that AMS is a very prospective management methodology which can support companies in employees commitment during their journey towards operational excellence. The analysis results show different motivations for AMS introduction as well as different development paths, these are harmonized with different management styles in companies and culture occurring in countries. The study is particularly valuable because this is one of the first empirical investigations of AMS implementation in European companies. In the field of theory the study proposes the four level scale for amoebas system maturity. This scale allows to classify companies following AMS principles and, at the same time, this scale is also the kind of path of AMS implementation. The study points out basic tools for companies which support AMS implementation. These tools are already known in management literature, but experience of investigated companies shows that they are fundamental for successful AMS implementation.

  7. Prevalence of potentially pathogenic free-living amoebae from Acanthamoeba and Naegleria genera in non-hospital, public, internal environments from the city of Santos, Brazil

    Directory of Open Access Journals (Sweden)

    Lais Helena Teixeira

    Full Text Available Acanthamoeba and Naegleria species are free-living amoebae (FLA found in a large variety of natural habitats. The prevalence of such amoebae was determined from dust samples taken from public non-hospital internal environments with good standards of cleanliness from two campuses of the same University in the city of Santos (SP, Brazil, and where young and apparently healthy people circulate. The frequency of free-living amoebae in both campuseswas 39% and 17% respectively, with predominance of the genus Acanthamoeba. On the campus with a much larger number of circulating individuals, the observed frequency of free-living amoebae was 2.29 times larger (P< 0.00005. Two trophozoite forms of Naegleria fowleri, are the only species of this genus known to cause primary amoebian meningoencephalitis, a rare and non-opportunistic infection. We assume that the high frequency of these organisms in different internal locations represents some kind of public health risk.

  8. Sterol biosynthesis via cycloartenol and other biochemical features related to photosynthetic phyla in the amoeba Naegleria lovaniensis and Naegleria gruberi.

    Science.gov (United States)

    Raederstorff, D; Rohmer, M

    1987-04-15

    The sterols and sterol precursors of two amoebae of the genus Naegleria, Naegleria lovaniensis and Naegleria gruberi were investigated. Cycloartenol, the sterol precursor in photosynthetic organisms, is present in both amoebae. In N. lovaniesis, it is accompanied by lanosterol and parkeol, as well as by the 24,25-dihydro derivatives of these triterpenes. One of the most striking features of these amoebae is the accumulation of 4 alpha-methylsterols which are present in similar amounts as those of 4,4-desmethylsterols (3-5 mg/g, dry weight). 4 alpha-Methylergosta-7,22-dienol was identified as a new compound. Ergosterol was the major 4,4-desmethylsterol, accompanied by small amounts of C27 and other C28 sterols. Treatment of N. lovaniensis with fenpropimorph modified the sterol pattern of this amoeba and inhibited its growth. This fungicide, known to inhibit steps of sterol biosynthesis in fungi and plants, induced the disappearance of 4 alpha-methyl-delta 7-sterols and the appearance of the unusual delta 6,8,22-ergostatrienol as in A. polyphaga. These results might be explained by a partial inhibition of the delta 8----delta 7 isomerase, the small amounts of delta 7-sterols formed being converted into ergosterol which is still present in fenpropimorph-exposed cells. De novo sterol biosynthesis in N. lovaniensis was shown by incorporation of [1-14C]acetate into sterols and sterol precursors, especially cycloartenol. Lanosterol and parkeol were not significantly labelled. Furthermore, [3-3H]squalene epoxide was efficiently cyclized by a cell-free system of this amoeba into cycloartenol, and again no significant radioactivity was detected in lanosterol and parkeol. This shows that cycloartenol, the sterol precursor in plants and algae, is also the sterol precursor in Naegleria species, and that these amoebae, like A. polyphaga, are related by some biosynthetic pathways to photosynthetic phyla. Lanosterol, the sterol precursor in non-photosynthetic phyla (animal and

  9. First Record of Potentially Pathogenic Amoeba Vermamoeba vermiformis (Lobosea: Gymnamoebia) Isolated from a Freshwater of Dokdo Island in the East Sea, Korea

    OpenAIRE

    Jong Soo Park

    2016-01-01

    Vermamoeba vermiformis is a very important free-living amoeba for human health in association with Legionnaires’ disease and keratitis. This interesting amoeba was firstly isolated from a freshwater of Dokdo (island), which was historically used for drinking water. Trophozoites and cyst forms of V. vermiformis strain MG1 are very similar to previous reported species. Trophozoites of V. vermiformis strain MG1 showed cylindrical shape with prominent anterior hyaline region. The average ratio of...

  10. Growth regulation of Legionella Pneumophila in biofilms and amoebae; Wachstumsregulation von Legionella Pneumophila in Biofilmen und Amoeben

    Energy Technology Data Exchange (ETDEWEB)

    Hilbi, H.

    2006-07-01

    This final report for the Swiss Federal Office of Energy (SFOE) presents the results of studies made on the regulation of the growth of Legionella Pneumophila bacteria in biofilms and amoebae. In a first project, the formation of biofilms by Legionella Pneumophila bacteria was analysed in static and dynamic systems using a complex growth medium. Under static and dynamic clinical and environmental conditions, the adherence of the biofilms on polystyrene tissue was studied. This was also examined under dynamic flow conditions. In a second part of the project, the regulation of growth of Legionella Pneumophila in amoebae was examined in that changes were made to the genome of the bacteria. The importance of the work for the de-activation of Legionella Pneumophila bacteria in biofilms is noted in the conclusions of the report.

  11. Investigation of Heterotrophic Bacteria, Legionella and Free-Living Amoeba in Cooling Tower Samples by FISH and Culture Methods

    OpenAIRE

    Zeybek, Zuhal; Dogruoz Gungor, Nihal; Turetgen, Irfan

    2018-01-01

    Themicroorganisms living in the cooling towers water can affect both human healththrough inhalation of aerosolized water as well as industrial processes. Inorder to analyse such man-made water systems, microbiological tests that cangive results in a short time are needed. In this study, the presence ofheterotrophic bacteria, Legionella bacteria and free - living amoeba, FLA,including Acanthamoeba, in cooling-tower water and biofilm samples wereinvestigated using two different methods, fluores...

  12. Development of a high- versus low-pathogenicity model of the free-living amoeba Naegleria fowleri.

    Science.gov (United States)

    Burri, Denise C; Gottstein, Bruno; Zumkehr, Béatrice; Hemphill, Andrew; Schürch, Nadia; Wittwer, Matthias; Müller, Norbert

    2012-10-01

    Species in the genus Naegleria are free-living amoebae of the soil and warm fresh water. Although around 30 species have been recognized, Naegleria fowleri is the only one that causes primary amoebic meningoencephalitis (PAM) in humans. PAM is an acute and fast progressing disease affecting the central nervous system. Most of the patients die within 1-2 weeks of exposure to the infectious water source. The fact that N. fowleri causes such fast progressing and highly lethal infections has opened many questions regarding the relevant pathogenicity factors of the amoeba. In order to investigate the pathogenesis of N. fowleri under defined experimental conditions, we developed a novel high- versus low-pathogenicity model for this pathogen. We showed that the composition of the axenic growth media influenced growth behaviour and morphology, as well as in vitro cytotoxicity and in vivo pathogenicity of N. fowleri. Trophozoites maintained in Nelson's medium were highly pathogenic for mice, demonstrated rapid in vitro proliferation, characteristic expression of surface membrane vesicles and a small cell diameter, and killed target mouse fibroblasts by both contact-dependent and -independent destruction. In contrast, N. fowleri cultured in PYNFH medium exhibited a low pathogenicity, slower growth, increased cell size and contact-dependent target cell destruction. However, cultivation of the amoeba in PYNFH medium supplemented with liver hydrolysate (LH) resulted in trophozoites that were highly pathogenic in mice, and demonstrated an intermediate proliferation rate in vitro, diminished cell diameter and contact-dependent target cell destruction. Thus, in this model, the presence of LH resulted in increased proliferation of trophozoites in vitro and enhanced pathogenicity of N. fowleri in mice. However, neither in vitro cytotoxicity mechanisms nor the presence of membrane vesicles on the surface correlated with the pathologic potential of the amoeba. This indicated that the

  13. Growth-supporting activity for Legionella pneumophila in tap water cultures and implication of hartmannellid amoebae as growth factors.

    Science.gov (United States)

    Wadowsky, R M; Butler, L J; Cook, M K; Verma, S M; Paul, M A; Fields, B S; Keleti, G; Sykora, J L; Yee, R B

    1988-11-01

    Photosynthetic cyanobacteria, heterotrophic bacteria, free-living amoebae, and ciliated protozoa may support growth of Legionella pneumophila. Studies were done with two tap water cultures (WS1 and WS2) containing L. pneumophila and associated microbiota to characterize growth-supporting activity and assess the relative importance of the microbiota in supporting multiplication of L. pneumophila. The water cultures were incubated in the dark at 35 degrees C. The growth-supporting factor(s) was separated from each culture by filtration through 1-micron-pore-size membrane filters. The retentate was then suspended in sterile tap water. Multiplication of L. pneumophila occurred when both the retentate suspension and the filtrate from either culture were inoculated into sterile tap water. L. pneumophila did not multiply in tap water inoculated with only the filtrate, even though filtration did not reduce the concentration of L. pneumophila or heterotrophic bacteria in either culture. Growth-supporting activity of the retentate suspension from WS1 was inactivated at 60 degrees C but unaffected at 0, 25, and 45 degrees C after 30-min incubations. Filtration experiments indicated that the growth-supporting factor(s) in WS1 was 2 to 5 micron in diameter. Ciliated protozoa were not detected in either culture. Hartmannellid amoebae were conclusively demonstrated in WS2 but not in WS1. L. pneumophila multiplied in tap water inoculated with the amoebae (10(3)/ml) and the 1-micron filtrate of WS2. No multiplication occurred in tap water inoculated with the filtrate only. Growth-supporting activity for L. pneumophila may be present in plumbing systems; hartmannellid amoebae appear to be important determinants of multiplication of L. pneumophila in some tap water cultures.

  14. The Phanerozoic diversification of silica-cycling testate amoebae and its possible links to changes in terrestrial ecosystems

    Science.gov (United States)

    Bosak, Tanja; Lara, Enrique; Mitchell, Edward A.D.

    2015-01-01

    The terrestrial cycling of Si is thought to have a large influence on the terrestrial and marine primary production, as well as the coupled biogeochemical cycles of Si and C. Biomineralization of silica is widespread among terrestrial eukaryotes such as plants, soil diatoms, freshwater sponges, silicifying flagellates and testate amoebae. Two major groups of testate (shelled) amoebae, arcellinids and euglyphids, produce their own silica particles to construct shells. The two are unrelated phylogenetically and acquired biomineralizing capabilities independently. Hyalosphenids, a group within arcellinids, are predators of euglyphids. We demonstrate that hyalosphenids can construct shells using silica scales mineralized by the euglyphids. Parsimony analyses of the current hyalosphenid phylogeny indicate that the ability to “steal” euglyphid scales is most likely ancestral in hyalosphenids, implying that euglyphids should be older than hyalosphenids. However, exactly when euglyphids arose is uncertain. Current fossil record contains unambiguous euglyphid fossils that are as old as 50 million years, but older fossils are scarce and difficult to interpret. Poor taxon sampling of euglyphids has also prevented the development of molecular clocks. Here, we present a novel molecular clock reconstruction for arcellinids and consider the uncertainties due to various previously used calibration points. The new molecular clock puts the origin of hyalosphenids in the early Carboniferous (∼370 mya). Notably, this estimate coincides with the widespread colonization of land by Si-accumulating plants, suggesting possible links between the evolution of Arcellinid testate amoebae and the expansion of terrestrial habitats rich in organic matter and bioavailable Si. PMID:26734499

  15. Surveillance and evaluation of the infection risk of free-living amoebae and Legionella in different aquatic environments.

    Science.gov (United States)

    Ji, Wen-Tsai; Hsu, Bing-Mu; Chang, Tien-Yu; Hsu, Tsui-Kang; Kao, Po-Min; Huang, Kuan-Hao; Tsai, Shiou-Feng; Huang, Yu-Li; Fan, Cheng-Wei

    2014-11-15

    Free-living amoebae (FLA) are ubiquitous in various aquatic environments. Several amoebae species are pathogenic and host other pathogens such as Legionella, but the presence of FLA and its parasites as well as the related infection risk are not well known. In this study, the presence of pathogenic FLA and Legionella in various water bodies was investigated. Water samples were collected from a river, intake areas of drinking water treatment plants, and recreational hot spring complexes in central and southern Taiwan. A total of 140 water samples were tested for the presence of Acanthamoeba spp., Naegleria spp., Vermamoeba vermiformis, and Legionella. In addition, phylogenetic characteristics and water quality parameters were also assessed. The pathogenic genotypes of FLA included Acanthamoeba T4 and Naegleria australiensis, and both were abundant in the hot spring water. In contrast, Legionella pneumophila was detected in different aquatic environments. Among the FLA assessed, V. vermiformis was most likely to coexist with Legionella spp. The total bacteria level was associated with the presence of FLA and Legionella especially in hot spring water. Taken together, FLA contamination in recreational hot springs and drinking water source warrants more attention on potential legionellosis and amoebae infections. Copyright © 2014. Published by Elsevier B.V.

  16. Detection and Molecular Characterization of Potentially Pathogenic Free-living Amoebae from Water Sources in Kish Island, Southern Iran.

    Science.gov (United States)

    Niyyati, Maryam; Lasgerdi, Zohreh; Lorenzo-Morales, Jacob

    2015-01-01

    Amoebic keratitis, a sight-threatening corneal infection, mainly occurs in contact lens wearers who wash their eyes with tap water. The present research was conducted to identify the occurrence of potentially pathogenic free-living amoebae (FLA) in tap water sources on Kish Island, a tourist region in Iran. Amoebae were detected using a culture-enriched method and by polymerase chain reaction (PCR)/sequencing of the diagnostic fragment 3 region of the 18S rRNA gene of Acanthamoeba. In the case of other free-living amoebae species, PCR/sequencing analysis of the 18S rDNA was conducted. Results of this study showed the presence of Acanthamoeba belonging to T3, T4, T5, and T11 genotypes in tap water sources. Additionally, Vermamoebae vermiformis was detected in three water samples. This is the first report of the Acanthamoeba genotypes T3, T4, T5, and T11 and V. vermiformis species in tap water sources in a tourist region in Iran.

  17. Pathogenic free-living amoebae in a closed-loop power plant; risk assessment and risk management

    Energy Technology Data Exchange (ETDEWEB)

    Cabanes, P. [Electricite de France (EDF), 75 - Paris (France). Service Etudes Medicales; Pringuez, E.; Siclet, F.; Khalanski, M. [Electricite de France (EDF), 75 - Paris (France). Direction des Etudes et Recherches; Bard, D. [CEA/Fontenay-aux-Roses, Inst. de Protection et de Surete Nucleaire (IPSN), 92 (France); Pernin, P. [Faculte de Pharmacie de Lyon, 69 (France)

    1998-07-01

    Since 1980, the water used for cooling in French power plants has been tested for pathogenic amoebae, especially Naegleria fowleri, the causative agent of Primary Amoebic Meningoencephalitis (PAM), a rapid fatal disease of the central nervous system. The replacement of brass condensers by stainless steel condensers resulted in increased Naegleria fowleri development, to a density of as much as 3000 l{sup -1} in the Dampierre power plant cooling water. Downstream from Dampierre, the maximum detected density of this amoeba during the summer of 1995 was 80 l{sup -1}, at low river flow. The replacement of a second condenser in 1996 at the same power plant was expected to double the amoebae concentration in the river. The hypothetical PAM risk for swimmers was then predicted to be 10{sup -4} per swim. To reduce the risk continuous chlorination of the closed-loop cooling system was implemented at a free residual chlorine level in the range of 0.3-0.5 mg.l{sup -1}. Naegleria fowleri concentrations decreased immediately and thereafter remained under 4 l{sup -1}. Total residual chlorine and chlorinated organic compounds were also monitored in the evaluation of the environmental impact of this preventive action. (authors)

  18. The Phanerozoic diversification of silica-cycling testate amoebae and its possible links to changes in terrestrial ecosystems

    Directory of Open Access Journals (Sweden)

    Daniel J.G. Lahr

    2015-09-01

    Full Text Available The terrestrial cycling of Si is thought to have a large influence on the terrestrial and marine primary production, as well as the coupled biogeochemical cycles of Si and C. Biomineralization of silica is widespread among terrestrial eukaryotes such as plants, soil diatoms, freshwater sponges, silicifying flagellates and testate amoebae. Two major groups of testate (shelled amoebae, arcellinids and euglyphids, produce their own silica particles to construct shells. The two are unrelated phylogenetically and acquired biomineralizing capabilities independently. Hyalosphenids, a group within arcellinids, are predators of euglyphids. We demonstrate that hyalosphenids can construct shells using silica scales mineralized by the euglyphids. Parsimony analyses of the current hyalosphenid phylogeny indicate that the ability to “steal” euglyphid scales is most likely ancestral in hyalosphenids, implying that euglyphids should be older than hyalosphenids. However, exactly when euglyphids arose is uncertain. Current fossil record contains unambiguous euglyphid fossils that are as old as 50 million years, but older fossils are scarce and difficult to interpret. Poor taxon sampling of euglyphids has also prevented the development of molecular clocks. Here, we present a novel molecular clock reconstruction for arcellinids and consider the uncertainties due to various previously used calibration points. The new molecular clock puts the origin of hyalosphenids in the early Carboniferous (∼370 mya. Notably, this estimate coincides with the widespread colonization of land by Si-accumulating plants, suggesting possible links between the evolution of Arcellinid testate amoebae and the expansion of terrestrial habitats rich in organic matter and bioavailable Si.

  19. Competition between Naegleria fowleri and Free Living Amoeba Colonizing Laboratory Scale and Operational Drinking Water Distribution Systems.

    Science.gov (United States)

    Miller, Haylea C; Wylie, Jason T; Kaksonen, Anna H; Sutton, David; Puzon, Geoffrey J

    2018-03-06

    Free living amoebae (FLA), including pathogenic Naegleria fowleri, can colonize and grow within pipe wall biofilms of drinking water distribution systems (DWDSs). Studies on the interactions between various FLA species in biofilms are limited. Understanding the interaction between FLA and the broader biofilm ecology could help better predict DWDS susceptibility to N. fowleri colonization. The aim of this study was to determine if N. fowleri and other FLAs ( Naegleria, Vermamoeba, Willaertia, and Vahlkampfia spp.) cocolonize DWDS biofilm. FLAs commonly isolated from DWDSs ( N. fowleri, V. vermiformis, and N. lovaniensis) were introduced into laboratory-scale biomonitors to determine the impact of these amoebae on N. fowleri's presence and viability. Over 18 months, a single viable amoebae ( N. fowleri, N. lovaniensis, or V. vermiformis) was detected in each biofilm sample, with the exception of N. lovaniensis and N. fowleri, which briefly cocolonized biofilm following their coinoculation. The analysis of biofilm and bulk water samples from operational DWDSs revealed a similar lack of cocolonization with a single FLA detected in 99% ( n = 242) of samples. Interestingly, various Naegleria spp. did colonize the same DWDS locations but at different times. This knowledge furthers the understanding of ecological factors which enable N. fowleri to colonize and survive within operational DWDSs and could aid water utilities to control its occurrence.

  20. Pathogenic free-living amoebae in a closed-loop power plant; risk assessment and risk management

    International Nuclear Information System (INIS)

    Cabanes, P.; Pernin, P.

    1998-01-01

    Since 1980, the water used for cooling in French power plants has been tested for pathogenic amoebae, especially Naegleria fowleri, the causative agent of Primary Amoebic Meningoencephalitis (PAM), a rapid fatal disease of the central nervous system. The replacement of brass condensers by stainless steel condensers resulted in increased Naegleria fowleri development, to a density of as much as 3000 l -1 in the Dampierre power plant cooling water. Downstream from Dampierre, the maximum detected density of this amoeba during the summer of 1995 was 80 l -1 , at low river flow. The replacement of a second condenser in 1996 at the same power plant was expected to double the amoebae concentration in the river. The hypothetical PAM risk for swimmers was then predicted to be 10 -4 per swim. To reduce the risk continuous chlorination of the closed-loop cooling system was implemented at a free residual chlorine level in the range of 0.3-0.5 mg.l -1 . Naegleria fowleri concentrations decreased immediately and thereafter remained under 4 l -1 . Total residual chlorine and chlorinated organic compounds were also monitored in the evaluation of the environmental impact of this preventive action. (authors)

  1. Detection and Molecular Characterization of Potentially Pathogenic Free-living Amoebae from Water Sources in Kish Island, Southern Iran

    Directory of Open Access Journals (Sweden)

    Maryam Niyyati

    2015-01-01

    Full Text Available Amoebic keratitis, a sight-threatening corneal infection, mainly occurs in contact lens wearers who wash their eyes with tap water. The present research was conducted to identify the occurrence of potentially pathogenic free-living amoebae (FLA in tap water sources on Kish Island, a tourist region in Iran. Amoebae were detected using a culture-enriched method and by polymerase chain reaction (PCR/sequencing of the diagnostic fragment 3 region of the 18S rRNA gene of Acanthamoeba . In the case of other free-living amoebae species, PCR/sequencing analysis of the 18S rDNA was conducted. Results of this study showed the presence of Acanthamoeba belonging to T3, T4, T5, and T11 genotypes in tap water sources. Additionally, Vermamoebae vermiformis was detected in three water samples. This is the first report of the Acanthamoeba genotypes T3, T4, T5, and T11 and V. vermiformis species in tap water sources in a tourist region in Iran.

  2. High relatedness in a social amoeba: the role of kin-discriminatory segregation.

    Science.gov (United States)

    Gilbert, Owen M; Strassmann, Joan E; Queller, David C

    2012-07-07

    A major challenge for social theory is to explain the importance of kin discrimination for the evolution of altruism. One way to assess the importance of kin discrimination is to test its effects on increasing relatedness within groups. The social amoeba Dictyostelium discoideum aggregates to form a fruiting body composed of dead stalk and live spores. Previous studies of a natural population showed that where D. discoideum occurs in the soil, multiple clones are often found in the same small soil samples. However, actual fruiting bodies usually contain only one clone. We here performed experiments to gauge the effect of kin-discriminatory segregation on increasing relatedness. We mixed co-occurring clones from this population using a relatedness level found in small soil samples. We found a lower proportion of uniclonal fruiting bodies and a lower level of relatedness compared with natural fruiting bodies. We found that the amount of relatedness increase attributable to kin-discriminatory segregation was small. These findings suggest a relatively minor influence of kin-discriminatory segregation on relatedness in D. discoideum. We discuss our results comparing with the results of previous studies, including those of wild clones and laboratory mutants. We ask why wild clones of D. discoideum exhibit a low degree of kin-discriminatory segregation, and what alternative factors might account for high relatedness in D. discoideum.

  3. Campylobacter jejuni Actively Invades the Amoeba Acanthamoeba polyphaga and Survives within Non Digestive Vacuoles

    Science.gov (United States)

    Olofsson, Jenny; Axelsson-Olsson, Diana; Brudin, Lars; Olsen, Björn; Ellström, Patrik

    2013-01-01

    The Gram-negative bacterium Campylobacter jejuni is able to enter, survive and multiply within the free living amoeba Acanthamoeba polyphaga, but the molecular mechanisms behind these events are still unclear. We have studied the uptake and intracellular trafficking of viable and heat killed bacterial cells of the C. jejuni strain 81–176 in A. polyphaga. We found that viable bacteria associated with a substantially higher proportion of Acanthamoeba trophozoites than heat killed bacteria. Furthermore, the kinetics of internalization, the total number of internalized bacteria as well as the intracellular localization of internalized C. jejuni were dramatically influenced by bacterial viability. Viable bacteria were internalized at a high rate already after 1 h of co-incubation and were observed in small vacuoles tightly surrounding the bacteria. In contrast, internalization of heat killed C. jejuni was low at early time points and did not peak until 96 h. These cells were gathered in large spacious vacuoles that were part of the degradative pathway as determined by the uptake of fluorescently labeled dextran. The amount of heat killed bacteria internalized by A. polyphaga did never reach the maximal amount of internalized viable bacteria. These results suggest that the uptake and intracellular survival of C. jejuni in A. polyphaga is bacterially induced. PMID:24223169

  4. Free-living amoebae (FLA) co-occurring with legionellae in industrial waters☆

    Science.gov (United States)

    Scheikl, Ute; Sommer, Regina; Kirschner, Alexander; Rameder, Alexandra; Schrammel, Barbara; Zweimüller, Irene; Wesner, Wolfgang; Hinker, Manfred; Walochnik, Julia

    2014-01-01

    Legionella pneumophila is known as the causative agent of Legionnaires’ disease and free-living amoebae (FLA) can serve as vehicles for legionellae. The aim of this study was to screen industrial waters for the occurrence of FLA and their co-occurrence with legionellae. A total of 201 water samples, including 129 cooling waters and 72 process waters, and 30 cooling lubricants were included in the study. Treated waters were screened periodically, pre and post treatment. Altogether, 72.6% of the water samples were positive for FLA, acanthamoebae being most prevalent (in 23.9% of the samples) followed by Vermamoeba vermiformis (19.4%). Only one cooling lubricant was positive (Acanthamoeba genotype T4). Legionella spp. were detected in 34.8% of the water samples and in 15% in high concentrations (>1000 CFU/100 ml). Altogether, 81.4% of the Legionella-positive samples were positive for FLA by standard methods. By applying a highly sensitive nested PCR to a representative set of random samples it was revealed that Legionella spp. always co-occurred with Acanthamoeba spp. Although the addition of disinfectants did influence amoebal density and diversity, treated waters showed no difference concerning FLA in the interphases of disinfection. It appears that FLA can re-colonize treated waters within a short period of time. PMID:25062389

  5. Ophthalmology hospital wards contamination to pathogenic free living Amoebae in Iran.

    Science.gov (United States)

    Lasjerdi, Zohreh; Niyyati, Maryam; Lorenzo-Morales, Jacob; Haghighi, Ali; Taghipour, Niloofar

    2015-09-01

    The present study was conducted to determine the occurrence of potentially pathogenic free-living amoeba in ophthalmology wards in reference hospitals in Iran. Since an increasing number of Acanthamoeba Keratitis cases after eye surgery and eye trauma have been recently observed in this country, it could be possible that the disinfection procedures undertaken in the clinical setting may not have a good hygiene and disinfection procedures, hence the aim of this study. Therefore, 42 dust and biofilm samples were collected from different areas of ophthalmology wards and checked for the presence of FLA using morphological criteria, PCR based analysis and DNA sequencing. Of the 42 samples from dust and biofilm sources, 18(42.86%) isolates were found to contain FLA and 12(92.3%) isolates belonged to Acanthamoeba T4 genotype. Isolation of the pathogenic genotype T4 from medical instruments, including slit lamp in corneal wards, may be a threat for patients undergoing eye surgery in these wards. Other FLA isolated in this study included Acanthamoeba genotype T5, Vahlkampfia sp, Naegleria australiensis, Vermamoeba vermiformis and Echinamoeba exudans. To our knowledge, this is the first report of the presence of potentially pathogenic FLA in ophthalmology wards in Iran. Improved disinfection methods and monitoring of hospitals ward are thus necessary in this area in order to minimize the risk of infection in patients.

  6. Free-living amoebae (FLA) co-occurring with legionellae in industrial waters.

    Science.gov (United States)

    Scheikl, Ute; Sommer, Regina; Kirschner, Alexander; Rameder, Alexandra; Schrammel, Barbara; Zweimüller, Irene; Wesner, Wolfgang; Hinker, Manfred; Walochnik, Julia

    2014-08-01

    Legionella pneumophila is known as the causative agent of Legionnaires' disease and free-living amoebae (FLA) can serve as vehicles for legionellae. The aim of this study was to screen industrial waters for the occurrence of FLA and their co-occurrence with legionellae. A total of 201 water samples, including 129 cooling waters and 72 process waters, and 30 cooling lubricants were included in the study. Treated waters were screened periodically, pre and post treatment. Altogether, 72.6% of the water samples were positive for FLA, acanthamoebae being most prevalent (in 23.9% of the samples) followed by Vermamoeba vermiformis (19.4%). Only one cooling lubricant was positive (Acanthamoeba genotype T4). Legionella spp. were detected in 34.8% of the water samples and in 15% in high concentrations (>1000 CFU/100 ml). Altogether, 81.4% of the Legionella-positive samples were positive for FLA by standard methods. By applying a highly sensitive nested PCR to a representative set of random samples it was revealed that Legionella spp. always co-occurred with Acanthamoeba spp. Although the addition of disinfectants did influence amoebal density and diversity, treated waters showed no difference concerning FLA in the interphases of disinfection. It appears that FLA can re-colonize treated waters within a short period of time. Copyright © 2014 The Authors. Published by Elsevier GmbH.. All rights reserved.

  7. Planktonic free-living amoebae susceptibility to dental unit waterlines disinfectants.

    Science.gov (United States)

    Costa, Damien; Bossard, Valentin; Brunet, Kévin; Fradin, Benjamin; Imbert, Christine

    2017-11-30

    A high diversity of microorganisms is encountered inside dental unit waterlines (DUWL). Among those the presence of free-living amoebae (FLA) appears currently underestimated, although human infections may occur due to contact with FLA-contaminated water during dental cares. In order to limit microbial DUWL contamination, disinfectants are provided by dental unit manufacturer, however, with limited documentation on their activities against FLA. The aim of this study was to evaluate the efficiency of three commercial DUWL disinfectants: the Calbenium© (Airel, Champigny-sur-Marne, France), the Oxygenal 6© (Kavo, Biberach, Germany) and the Sterispray© (Gammasonic, Billom, France), against two FLA species, i.e. Acanthamoeba castellanii and Vermamoeba vermiformis alone or co-cultured with Pseudomonas aeruginosa and Candida albicans at concentrations ranging from 0% to 5% (v/v). Results showed varied efficacies of disinfectants: the Oxygenal 6© did not exhibit FLA killing activity, while the Sterispray© and the Calbenium© displayed concentration- and species-dependent activities with a maximum eradication rates of 100% and 86%, and 79% and 97% for A. castellani and V. vermiformis, respectively. None of the disinfectants were able to totally eradicate FLA at concentrations recommended by manufacturers. Present results highlight unsatisfactory anti-FLA activities of 3 DUWL disinfectant preparations advocating deeper investigation of antimicrobial spectra of commercial disinfectants in use for DUWL maintenance. © FEMS 2017. All rights reserved. For permissions, please e-mail: journals.permissions@oup.com.

  8. Lateral gene exchanges shape the genomes of amoeba-resisting microorganisms

    Directory of Open Access Journals (Sweden)

    Claire eBertelli

    2012-08-01

    Full Text Available Based on Darwin’s concept of the tree of life, vertical inheritance was thought to be dominant, and mutations, deletions and duplication were streaming the genomes of living organisms. In the current genomic era, increasing data indicated that both vertical and lateral gene inheritance interact in space and time to trigger genome evolution, particularly among microorganisms sharing a given ecological niche. As a paradigm to their diversity and their survival in a variety of cell types, intracellular microorganisms, and notably intracellular bacteria, were considered as less prone to lateral genetic exchanges. Such specialized microorganisms generally have a smaller gene repertoire because they do rely on their host’s factors for some basic regulatory and metabolic functions. Here we review events of lateral gene transfer (LGT that illustrate the genetic exchanges among intra-amoebal microorganisms or between the microorganism and its amoebal host. We tentatively investigate the functions of laterally transferred genes in the light of the interaction with their host as they should confer a selective advantage and success to the amoeba-resisting microorganisms.

  9. Comparative recoveries of Naegleria fowleri amoebae from seeded river water by filtration and centrifugation.

    Science.gov (United States)

    Pernin, P; Pélandakis, M; Rouby, Y; Faure, A; Siclet, F

    1998-03-01

    Detection of pathogenic Naegleria fowleri in environmental water samples, which is necessary for the prevention of primary amoebic meningoencephalitis, generally requires concentrating the samples. Two concentration techniques, filtration and centrifugation, were used to study the recovery of N. fowleri, in vegetative or cystic form, that had been mixed with the two other thermotolerant Naegleria species, N. lovaniensis and N. australiensis. Counting of amoebae was performed by the most probable number method on 10 water replicates of 100 ml and 10 ml each. With both concentration methods, recovery was better for cysts than for trophozoites (53% +/- 21% versus 5% +/- 5% by filtration and 57% +/- 25% versus 22% +/- 5% by centrifugation). The recovery of Naegleria trophozoites by filtration was very low, and centrifugation was significantly better than filtration in recovery of Naegleria trophozoites (22% +/- 5% versus 5% +/- 5%; P 0.7). Although the recovery of cysts of N. fowleri obtained by filtration (51% +/- 24%) appeared higher than that by centrifugation (36% +/- 23%), the difference was not significant (P > 0.1). Both concentration methods have highly variable recovery rates, making accurate quantification of low concentrations (< 100/liter) of N. fowleri in the environment difficult.

  10. Amoebae, Giant Viruses, and Virophages Make Up a Complex, Multilayered Threesome.

    Science.gov (United States)

    Diesend, Jan; Kruse, Janis; Hagedorn, Monica; Hammann, Christian

    2017-01-01

    Viral infection had not been observed for amoebae, until the Acanthamoeba polyphaga mimivirus (APMV) was discovered in 2003. APMV belongs to the nucleocytoplasmatic large DNA virus (NCLDV) family and infects not only A. polyphaga , but also other professional phagocytes. Here, we review the Megavirales to give an overview of the current members of the Mimi - and Marseilleviridae families and their structural features during amoebal infection. We summarize the different steps of their infection cycle in A. polyphaga and Acanthamoeba castellani . Furthermore, we dive into the emerging field of virophages, which parasitize upon viral factories of the Megavirales family. The discovery of virophages in 2008 and research in recent years revealed an increasingly complex network of interactions between cell, giant virus, and virophage. Virophages seem to be highly abundant in the environment and occupy the same niches as the Mimiviridae and their hosts. Establishment of metagenomic and co-culture approaches rapidly increased the number of detected virophages over the recent years. Genetic interaction of cell and virophage might constitute a potent defense machinery against giant viruses and seems to be important for survival of the infected cell during mimivirus infections. Nonetheless, the molecular events during co-infection and the interactions of cell, giant virus, and virophage have not been elucidated, yet. However, the genetic interactions of these three, suggest an intricate, multilayered network during amoebal (co-)infections. Understanding these interactions could elucidate molecular events essential for proper viral factory activity and could implicate new ways of treating viruses that form viral factories.

  11. Comparative Recoveries of Naegleria fowleri Amoebae from Seeded River Water by Filtration and Centrifugation

    Science.gov (United States)

    Pernin, P.; Pélandakis, M.; Rouby, Y.; Faure, A.; Siclet, F.

    1998-01-01

    Detection of pathogenic Naegleria fowleri in environmental water samples, which is necessary for the prevention of primary amoebic meningoencephalitis, generally requires concentrating the samples. Two concentration techniques, filtration and centrifugation, were used to study the recovery of N. fowleri, in vegetative or cystic form, that had been mixed with the two other thermotolerant Naegleria species, N. lovaniensis and N. australiensis. Counting of amoebae was performed by the most probable number method on 10 water replicates of 100 ml and 10 ml each. With both concentration methods, recovery was better for cysts than for trophozoites (53% ± 21% versus 5% ± 5% by filtration and 57% ± 25% versus 22% ± 5% by centrifugation). The recovery of Naegleria trophozoites by filtration was very low, and centrifugation was significantly better than filtration in recovery of Naegleria trophozoites (22% ± 5% versus 5% ± 5%; P 0.7). Although the recovery of cysts of N. fowleri obtained by filtration (51% ± 24%) appeared higher than that by centrifugation (36% ± 23%), the difference was not significant (P > 0.1). Both concentration methods have highly variable recovery rates, making accurate quantification of low concentrations (fowleri in the environment difficult. PMID:9501435

  12. Quadrulella texcalense sp. nov. from a Mexican desert: An unexpected new environment for hyalospheniid testate amoebae.

    Science.gov (United States)

    Pérez-Juárez, Horacio; Serrano-Vázquez, Angélica; Kosakyan, Anush; Mitchell, Edward A D; Rivera Aguilar, Víctor M; Lahr, Daniel J G; Hernández Moreno, Mayra M; Cuellar, Humberto Macías; Eguiarte, Luis E; Lara, Enrique

    2017-10-01

    Quadrulella (Amoebozoa, Arcellinida, Hyalospheniidae) is a genus of testate amoebae with unmistakable morphology, which secretes characteristic square plates to reinforce the test. They are mainly known from fens and freshwater habitats and have never been documented in deserts. We describe a new species, Quadrulella texcalense, from biological soil crusts in the intertropical desert of Tehuacán (state of Puebla, Mexico). Quadrulella texcalense occurred only at altitudes between 2140 and 2221m.a.s.l., together with the bryophyte genera Pseudocrossidium, Weissia, Bryum, Didymodon, Neohyophyla and Aloina. The soil was extremely dry (moisture of 1.97-2.6%), which contrasts sharply with previous reports for the Quadrulella genus. Single cell mitochondrial cytochrome oxidase I (COI) barcoding of thirteen isolated cells showed an important morphological variability despite having all the same COI barcode sequence. Quadrulella texcalense was placed in a tree containing other Hyalsopheniidae, including a newly barcoded South African species, Q. elegans. Q. texcalense unambiguously branched within genus Quadrulella in a compact clade but with a long branch, suggesting accelerated evolution due to a transition towards a new environment and/or under-sampling. Copyright © 2017 Elsevier GmbH. All rights reserved.

  13. Extracellular matrix dynamics and functions in the social amoeba Dictyostelium: A critical review.

    Science.gov (United States)

    Huber, Robert J; O'Day, Danton H

    2017-01-01

    The extracellular matrix (ECM) is a dynamic complex of glycoproteins, proteoglycans, carbohydrates, and collagen that serves as an interface between mammalian cells and their extracellular environment. Essential for normal cellular homeostasis, physiology, and events that occur during development, it is also a key functionary in a number of human diseases including cancer. The social amoeba Dictyostelium discoideum secretes an ECM during multicellular development that regulates multicellularity, cell motility, cell differentiation, and morphogenesis, and provides structural support and protective layers to the resulting differentiated cell types. Proteolytic processing within the Dictyostelium ECM leads to specific bioactive factors that regulate cell motility and differentiation. Here we review the structure and functions of the Dictyostelium ECM and its role in regulating multicellular development. The questions and challenges that remain and how they can be answered are also discussed. The Dictyostelium ECM shares many of the features of mammalian and plant ECM, and thus presents an excellent system for studying the structure and function of the ECM. As a genetically tractable model organism, Dictyostelium offers the potential to further elucidate ECM functions, and to possibly reveal previously unknown roles for the ECM. Copyright © 2016 Elsevier B.V. All rights reserved.

  14. Campylobacter jejuni actively invades the amoeba Acanthamoeba polyphaga and survives within non digestive vacuoles.

    Directory of Open Access Journals (Sweden)

    Jenny Olofsson

    Full Text Available The Gram-negative bacterium Campylobacter jejuni is able to enter, survive and multiply within the free living amoeba Acanthamoeba polyphaga, but the molecular mechanisms behind these events are still unclear. We have studied the uptake and intracellular trafficking of viable and heat killed bacterial cells of the C. jejuni strain 81-176 in A. polyphaga. We found that viable bacteria associated with a substantially higher proportion of Acanthamoeba trophozoites than heat killed bacteria. Furthermore, the kinetics of internalization, the total number of internalized bacteria as well as the intracellular localization of internalized C. jejuni were dramatically influenced by bacterial viability. Viable bacteria were internalized at a high rate already after 1 h of co-incubation and were observed in small vacuoles tightly surrounding the bacteria. In contrast, internalization of heat killed C. jejuni was low at early time points and did not peak until 96 h. These cells were gathered in large spacious vacuoles that were part of the degradative pathway as determined by the uptake of fluorescently labeled dextran. The amount of heat killed bacteria internalized by A. polyphaga did never reach the maximal amount of internalized viable bacteria. These results suggest that the uptake and intracellular survival of C. jejuni in A. polyphaga is bacterially induced.

  15. Isolation and Identification of Free-Living Amoebae from Tap Water in Sivas, Turkey

    Directory of Open Access Journals (Sweden)

    Kübra Açıkalın Coşkun

    2013-01-01

    Full Text Available The present work focuses on a local survey of free-living amoebae (FLA that cause opportunistic and nonopportunistic infections in humans. Determining the prevalence of FLA in water sources can shine a light on the need to prevent FLA related illnesses. A total of 150 samples of tap water were collected from six districts of Sivas province. The samples were filtered and seeded on nonnutrient agar containing Escherichia coli spread. Thirty-three (22% out of 150 samples were found to be positive for FLA. The FLA were identified by morphology and by PCR using 18S rDNA gene. The morphological analysis and partial sequencing of the 18S rDNA gene revealed the presence of three different species, Acanthamoeba castellanii, Acanthamoeba polyphaga, and Hartmannella vermiformis. Naegleria fowleri, Balamuthia mandrillaris, or Sappinia sp. was not isolated during the study. All A. castellanii and A. polyphaga sequence types were found to be genotype T4 that contains most of the pathogenic Acanthamoeba strains. The results indicated the occurrence and distribution of FLA species in tap water in these localities of Sivas, Turkey. Furthermore, the presence of temperature tolerant Acanthamoeba genotype T4 in tap water in the region must be taken into account for health risks.

  16. Influence of climate and geography on the occurrence of Legionella and amoebae in composting facilities.

    Science.gov (United States)

    Conza, Lisa; Casati Pagani, Simona; Gaia, Valeria

    2014-11-24

    The incidence of Legionnaires' disease (LD) in southern Switzerland is three times higher than in northern Switzerland. Climatic and geographic factors may be potential causes for this difference.We studied the prevalence of Legionella and free-living amoebae (FLA) in compost and bioaerosol in two Swiss regions to understand the role of climate and geography in the transmission of LD. We also tried to investigate whether or not compost storage duration would influence the composition of Legionella and FLA communities. A larger proportion of compost heaps in facilities from southern Switzerland harbor more diverse Legionella compared to the north (P=0.0146). FLA were isolated from composts in northern facilities at slightly higher rates (88.2% vs. 69.2%) and at lower rates from bioaerosols (6.3% vs. 13%) than in southern Switzerland. The diversity of FLA was higher in northern than in southern Switzerland (80% vs. 65%). A general decrease in the presence and variety of species was observed with decreasing compost storage time length. A discriminant model showed that values of vapour pressure, relative humidity and temperature distinguish the two regions, which were also characterised by different contamination rates by FLA and Legionella. The duration of outdoor storage may favour contamination of the compost by Legionella, and may increase the number and isolation of Legionella naturally occurring in compost. The climate in the south seems to favour higher Legionella contamination of compost heaps: this could explain the higher incidence of LD in southern Switzerland.

  17. Flourish or flush: effects of simulated extreme rainfall events on Sphagnum-dwelling testate amoebae in a subarctic bog (Abisko, Sweden).

    Science.gov (United States)

    Tsyganov, Andrey N; Keuper, Frida; Aerts, Rien; Beyens, Louis

    2013-01-01

    Extreme precipitation events are recognised as important drivers of ecosystem responses to climate change and can considerably affect high-latitude ombrotrophic bogs. Therefore, understanding the relationships between increased rainfall and the biotic components of these ecosystems is necessary for an estimation of climate change impacts. We studied overall effects of increased magnitude, intensity and frequency of rainfall on assemblages of Sphagnum-dwelling testate amoebae in a field climate manipulation experiment located in a relatively dry subarctic bog (Abisko, Sweden). The effects of the treatment were estimated using abundance, species diversity and structure of living and empty shell assemblages of testate amoebae in living and decaying layers of Sphagnum. Our results show that increased rainfall reduced the mean abundance and species richness of living testate amoebae. Besides, the treatment affected species structure of both living and empty shell assemblages, reducing proportions of hydrophilous species. The effects are counterintuitive as increased precipitation-related substrate moisture was expected to have opposite effects on testate amoeba assemblages in relatively dry biotopes. Therefore, we conclude that other rainfall-related factors such as increased infiltration rates and frequency of environmental disturbances can also affect testate amoeba assemblages in Sphagnum and that hydrophilous species are particularly sensitive to variation in these environmental variables.

  18. Effect of thermal additions on the density and distribution of thermophilic amoebae and pathogenic Naegleria fowleri in a newly created cooling lake

    Energy Technology Data Exchange (ETDEWEB)

    Tyndall, R.L.; Ironside, K.S.; Metler, P.L.; Tan, E.L. (Univ. of Tennessee, Knoxville (USA)); Hazen, T.C.; Fliermans, C.B. (E.I. du Pont de Nemours and Co., Inc., Aiken, SC (USA))

    1989-03-01

    Pathogenic Naegleria fowleri is the causative agent of fatal human amoebic meningoencephalitis. The protozoan is ubiquitous in nature, and its presence is enhanced by thermal additions. In this investigation, water and sediments from a newly created cooling lake were quantitatively analyzed for the presence of thermophilic amoebae, thermophilic Naegleria spp., and the pathogen Naegleria fowleri. During periods of thermal additions, the concentrations of thermophilic amoebae and thermophilic Naegleria spp. increased as much as 5 orders of magnitude, and the concentration of the pathogen N. fowleri increased as much as 2 orders of magnitude. Concentrations of amoebae returned to prior thermal perturbation levels within 30 to 60 days after cessation of thermal additions. Increases in the thermophilic amoeba concentrations were noted in Savannah River oxbows downriver from the Savannah River plant discharge streams as compared with oxbows upriver from the discharges. Concentrations of thermophilic amoebae and thermophilic Naegleria spp. correlated significantly with temperature and conductivity. Air samples taken proximal to the lade during periods of thermal addition showed no evidence of thermophilic Naegleria spp. Isoenzyme patterns of the N. fowleri isolated from the cooling lake were identical to patterns of N. fowleri isolated from other sites in the United States and Belgium.

  19. Social amoebae mating types do not invest unequally in sexual offspring.

    Science.gov (United States)

    Douglas, T E; Queller, D C; Strassmann, J E

    2017-05-01

    Unequal investment by different sexes in their progeny is common and includes differential investment in the zygote and differential care of the young. The social amoeba Dictyostelium discoideum has a sexual stage in which isogamous cells of any two of the three mating types fuse to form a zygote which then attracts hundreds of other cells to the macrocyst. The latter cells are cannibalized and so make no genetic contribution to reproduction. Previous literature suggests that this sacrifice may be induced in cells of one mating type by cells of another, resulting in a higher than expected production of macrocysts when the inducing type is rare and giving a reproductive advantage to this social cheat. We tested this hypothesis in eight trios of field-collected clones of each of the three D. discoideum mating types by measuring macrocyst production at different pairwise frequencies. We found evidence that supported differential contribution in only two of the 24 clone pairs, so this pattern is rare and clone-specific. In general, we did not reject the hypothesis that the mating types contribute cells relative to their proportion in the population. We also found a significant quadratic relationship between partner frequency and macrocyst production, suggesting that when one clone is rare, macrocyst production is limited by partner availability. We were also unable to replicate previous findings that macrocyst production could be induced in the absence of a compatible mating partner. Overall, mating type-specific differential investment during sex is unlikely in microbial eukaryotes like D. discoideum. © 2017 European Society For Evolutionary Biology. Journal of Evolutionary Biology © 2017 European Society For Evolutionary Biology.

  20. Exposure to synthetic gray water inhibits amoeba encystation and alters expression of Legionella pneumophila virulence genes.

    Science.gov (United States)

    Buse, Helen Y; Lu, Jingrang; Ashbolt, Nicholas J

    2015-01-01

    Water conservation efforts have focused on gray water (GW) usage, especially for applications that do not require potable water quality. However, there is a need to better understand environmental pathogens and their free-living amoeba (FLA) hosts within GW, given their growth potential in stored gray water. Using synthetic gray water (sGW) we examined three strains of the water-based pathogen Legionella pneumophila and its FLA hosts Acanthamoeba polyphaga, A. castellanii, and Vermamoeba vermiformis. Exposure to sGW for 72 h resulted in significant inhibition (P vermiformis (1 versus 92%), suggesting sGW induced maintenance of the actively feeding trophozoite form. During sGW exposure, L. pneumophila culturability decreased as early as 5 h (1.3 to 2.9 log10 CFU, P < 0.001) compared to controls (Δ0 to 0.1 log10 CFU) with flow cytometric analysis revealing immediate changes in membrane permeability. Furthermore, reverse transcription-quantitative PCR was performed on total RNA isolated from L. pneumophila cells at 0 to 48 h after sGW incubation, and genes associated with virulence (gacA, lirR, csrA, pla, and sidF), the type IV secretion system (lvrB and lvrE), and metabolism (ccmF and lolA) were all shown to be differentially expressed. These results suggest that conditions within GW may promote interactions between water-based pathogens and FLA hosts, through amoebal encystment inhibition and alteration of bacterial gene expression, thus warranting further exploration into FLA and L. pneumophila behavior in GW systems. Copyright © 2015, American Society for Microbiology. All Rights Reserved.

  1. Arcellacea (testate amoebae) as bio-indicators of road salt contamination in lakes.

    Science.gov (United States)

    Roe, Helen M; Patterson, R Timothy

    2014-08-01

    Winter deicing operations occur extensively in mid- to high-latitude metropolitan regions around the world and result in a significant reduction in road accidents. Deicing salts can, however, pose a major threat to water quality and aquatic organisms. In this paper, we examine the utility of Arcellacea (testate amoebae) for monitoring lakes that have become contaminated by winter deicing salts, particularly sodium chloride. We analysed 50 sediment samples and salt-related water property variables (chloride concentrations; conductivity) from 15 lakes in the Greater Toronto Area and adjacent areas of southern Ontario, Canada. The sampled lakes included lakes in proximity to major highways and suburban roads and control lakes in forested settings away from road influences. Samples from the most contaminated lakes, with chloride concentrations in excess of 400 mg/l and conductivities of >800 μS/cm, were dominated by species typically found in brackish and/or inhospitable lake environments and by lower faunal diversities (lowest Shannon diversity index values) than samples with lower readings. Q-R-mode cluster analysis and detrended correspondence analysis (DCA) resulted in the recognition of four assemblage groupings. These reflect varying levels of salt contamination in the study lakes, along with other local influences, including nutrient loading. The response to nutrients can, however, be isolated if the planktic eutrophic indicator species Cucurbitella tricuspis is removed from the counts. The findings show that the group has considerable potential for biomonitoring in salt-contaminated lakes, and their presence in lake sediment cores may provide significant insights into long-term benthic community health, which is integral for remedial efforts.

  2. Presence of Legionella and Free-Living Amoebae in Composts and Bioaerosols from Composting Facilities

    Science.gov (United States)

    Conza, Lisa; Pagani, Simona Casati; Gaia, Valeria

    2013-01-01

    Several species of Legionella cause Legionnaires’ disease (LD). Infection may occur through inhalation of Legionella or amoebal vesicles. The reservoirs of Legionella are water, soil, potting soil and compost. Some species of free-living amoebae (FLA) that are naturally present in water and soil were described as hosts for Legionella. This study aimed to understand whether or not the composting facilities could be sources of community-acquired Legionella infections after development of bioaerosols containing Legionella or FLA. We looked for the presence of Legionella (by co-culture) and FLA (by culture) in composts and bioaerosols collected at four composting facilities located in southern Switzerland. We investigated the association between the presence of Legionella and compost and air parameters and presence of FLA. Legionella spp. (including L. pneumophila) were detected in 69.3% (61/88) of the composts and FLA (mainly Acanthamoeba, Vermamoeba, Naegleria and Stenamoeba) in 92.0% (81/88). L. pneumophila and L. bozemanii were most frequently isolated. FLA as potential host for Legionella spp. were isolated from 40.9% (36/88) of the composts in all facilities. In Legionella-positive samples the temperature of compost was significantly lower (P = 0.012) than in Legionella-negative samples. Of 47 bioaerosol samples, 19.1% (9/47) were positive for FLA and 10.6% (5/47) for L. pneumophila. Composts (62.8%) were positive for Legionella and FLA contemporaneously, but both microorganisms were never detected simultaneously in bioaerosols. Compost can release bioaerosol containing FLA or Legionella and could represent a source of infection of community-acquired Legionella infections for workers and nearby residents. PMID:23844174

  3. An expanded phylogeny of social amoebas (Dictyostelia shows increasing diversity and new morphological patterns

    Directory of Open Access Journals (Sweden)

    Stephenson Steven L

    2011-03-01

    Full Text Available Abstract Background Social Amoebae or Dictyostelia are eukaryotic microbes with a unique life cycle consisting of both uni- and multicellular stages. They have long fascinated molecular, developmental and evolutionary biologists, and Dictyostelium discoideum is now one of the most widely studied eukaryotic microbial models. The first molecular phylogeny of Dictyostelia included most of the species known at the time and suggested an extremely deep taxon with a molecular depth roughly equivalent to Metazoa. The group was also shown to consist of four major clades, none of which correspond to traditional genera. Potential morphological justification was identified for three of the four major groups, on the basis of which tentative names were assigned. Results Over the past four years, the Mycetozoan Global Biodiversity Survey has identified many new isolates that appear to be new species of Dictyostelia, along with numerous isolates of previously described species. We have determined 18S ribosomal RNA gene sequences for all of these new isolates. Phylogenetic analyses of these data show at least 50 new species, and these arise from throughout the dictyostelid tree breaking up many previously isolated long branches. The resulting tree now shows eight well-supported major groups instead of the original four. The new species also expand the known morphological diversity of the previously established four major groups, violating nearly all previously suggested deep morphological patterns. Conclusions A greatly expanded phylogeny of Dictyostelia now shows even greater morphological plasticity at deep taxonomic levels. In fact, there now seem to be no obvious deep evolutionary trends across the group. However at a finer level, patterns in morphological character evolution are beginning to emerge. These results also suggest that there is a far greater diversity of Dictyostelia yet to be discovered, including novel morphologies.

  4. Disruption of MDCK cell tight junctions by the free-living amoeba Naegleria fowleri.

    Science.gov (United States)

    Shibayama, Mineko; Martínez-Castillo, Moisés; Silva-Olivares, Angélica; Galindo-Gómez, Silvia; Navarro-García, Fernando; Escobar-Herrera, Jaime; Sabanero, Myrna; Tsutsumi, Víctor; Serrano-Luna, Jesús

    2013-02-01

    Naegleria fowleri is the aetiological agent of primary amoebic meningoencephalitis. This parasite invades its host by penetrating the olfactory mucosa. However, the mechanism of epithelium penetration is not well understood. In the present study, we evaluated the effect of N. fowleri trophozoites and the non-pathogenic Naegleria gruberi on Madin-Darby canine kidney (MDCK) tight junction proteins, including claudin-1, occludin and ZO-1, as well as on the actin cytoskeleton. Trophozoites from each of the free-living amoeba species were co-cultured with MDCK cells in a 1 : 1 ratio for 1, 3, 6 or 10 h. Light microscopy revealed that N. fowleri caused morphological changes as early as 3 h post-infection in an epithelial MDCK monolayer. Confocal microscopy analysis revealed that after 10 h of co-culture, N. fowleri trophozoites induced epithelial cell damage, which was characterized by changes in the actin apical ring and disruption of the ZO-1 and claudin-1 proteins but not occludin. Western blot assays revealed gradual degradation of ZO-1 and claudin-1 as early as 3 h post-infection. Likewise, there was a drop in transepithelial electrical resistance that resulted in increased epithelial permeability and facilitated the invasion of N. fowleri trophozoites by a paracellular route. In contrast, N. gruberi did not induce alterations in MDCK cells even at 10 h post-infection. Based on these results, we suggest that N. fowleri trophozoites disrupt epithelial monolayers, which could enable their penetration of the olfactory epithelium and subsequent invasion of the central nervous system.

  5. LAMARCKIAN EVOLUTION OF THE GIANT MIMIVIRUS IN ALLOPATRIC LABORATORY CULTURE ON AMOEBAE

    Directory of Open Access Journals (Sweden)

    Philippe eColson

    2012-07-01

    Full Text Available Acanthamoeba polyphaga Mimivirus has been subcultured 150 times on germ-free amoebae. This allopatric niche is very different from that found in the natural environment, where the virus is in competition with many other organisms. In this experiment, substantial gene variability and loss occurred concurrently with the emergence of phenotypically different viruses. We sought to quantify the respective roles of Lamarckian and Darwinian evolution during this experiment. We postulated that the Mimivirus genes that were down-regulated at the beginning of the allopatric laboratory culture and inactivated after 150 passages experienced Lamarckian evolution because phenotypic modifications preceded genotypic modifications, whereas we considered that genes that were highly transcribed in the new niche but were later inactivated obeyed Darwinian rules. We used the total transcript abundances and sequences described for the genes of Mimivirus at the beginning of its laboratory life and after 150 passages in allopatric culture on Acanthamoeba spp. We found a statistically significant positive correlation between the level of gene expression at the beginning of the culture and gene inactivation during the 150 passages. In particular, the mean transcript abundance at baseline was significantly lower for inactivated genes than for unchanged genes (165±589 vs. 470±1,625; p<1e-3, and the mean transcript levels during the replication cycle of Mimivirus M1 were up to 8.5-fold lower for inactivated genes than for unchanged genes. In addition, proteins tended to be less frequently identified from purified virions in their early life in allopatric laboratory culture if they were encoded by variable genes than if they were encoded by conserved genes (9% vs. 15%; p= 0.062. Finally, Lamarckian evolution represented the evolutionary process encountered by 63% of the inactivated genes. Such observations may be explained by the lower level of DNA repair of useless genes.

  6. Amoebae, Giant Viruses, and Virophages Make Up a Complex, Multilayered Threesome

    Directory of Open Access Journals (Sweden)

    Jan Diesend

    2018-01-01

    Full Text Available Viral infection had not been observed for amoebae, until the Acanthamoeba polyphaga mimivirus (APMV was discovered in 2003. APMV belongs to the nucleocytoplasmatic large DNA virus (NCLDV family and infects not only A. polyphaga, but also other professional phagocytes. Here, we review the Megavirales to give an overview of the current members of the Mimi- and Marseilleviridae families and their structural features during amoebal infection. We summarize the different steps of their infection cycle in A. polyphaga and Acanthamoeba castellani. Furthermore, we dive into the emerging field of virophages, which parasitize upon viral factories of the Megavirales family. The discovery of virophages in 2008 and research in recent years revealed an increasingly complex network of interactions between cell, giant virus, and virophage. Virophages seem to be highly abundant in the environment and occupy the same niches as the Mimiviridae and their hosts. Establishment of metagenomic and co-culture approaches rapidly increased the number of detected virophages over the recent years. Genetic interaction of cell and virophage might constitute a potent defense machinery against giant viruses and seems to be important for survival of the infected cell during mimivirus infections. Nonetheless, the molecular events during co-infection and the interactions of cell, giant virus, and virophage have not been elucidated, yet. However, the genetic interactions of these three, suggest an intricate, multilayered network during amoebal (co-infections. Understanding these interactions could elucidate molecular events essential for proper viral factory activity and could implicate new ways of treating viruses that form viral factories.

  7. Presence of Legionella and free-living Amoebae in composts and bioaerosols from composting facilities.

    Directory of Open Access Journals (Sweden)

    Lisa Conza

    Full Text Available Several species of Legionella cause Legionnaires' disease (LD. Infection may occur through inhalation of Legionella or amoebal vesicles. The reservoirs of Legionella are water, soil, potting soil and compost. Some species of free-living amoebae (FLA that are naturally present in water and soil were described as hosts for Legionella. This study aimed to understand whether or not the composting facilities could be sources of community-acquired Legionella infections after development of bioaerosols containing Legionella or FLA. We looked for the presence of Legionella (by co-culture and FLA (by culture in composts and bioaerosols collected at four composting facilities located in southern Switzerland. We investigated the association between the presence of Legionella and compost and air parameters and presence of FLA. Legionella spp. (including L. pneumophila were detected in 69.3% (61/88 of the composts and FLA (mainly Acanthamoeba, Vermamoeba, Naegleria and Stenamoeba in 92.0% (81/88. L. pneumophila and L. bozemanii were most frequently isolated. FLA as potential host for Legionella spp. were isolated from 40.9% (36/88 of the composts in all facilities. In Legionella-positive samples the temperature of compost was significantly lower (P = 0.012 than in Legionella-negative samples. Of 47 bioaerosol samples, 19.1% (9/47 were positive for FLA and 10.6% (5/47 for L. pneumophila. Composts (62.8% were positive for Legionella and FLA contemporaneously, but both microorganisms were never detected simultaneously in bioaerosols. Compost can release bioaerosol containing FLA or Legionella and could represent a source of infection of community-acquired Legionella infections for workers and nearby residents.

  8. Presence of Legionella and free-living Amoebae in composts and bioaerosols from composting facilities.

    Science.gov (United States)

    Conza, Lisa; Pagani, Simona Casati; Gaia, Valeria

    2013-01-01

    Several species of Legionella cause Legionnaires' disease (LD). Infection may occur through inhalation of Legionella or amoebal vesicles. The reservoirs of Legionella are water, soil, potting soil and compost. Some species of free-living amoebae (FLA) that are naturally present in water and soil were described as hosts for Legionella. This study aimed to understand whether or not the composting facilities could be sources of community-acquired Legionella infections after development of bioaerosols containing Legionella or FLA. We looked for the presence of Legionella (by co-culture) and FLA (by culture) in composts and bioaerosols collected at four composting facilities located in southern Switzerland. We investigated the association between the presence of Legionella and compost and air parameters and presence of FLA. Legionella spp. (including L. pneumophila) were detected in 69.3% (61/88) of the composts and FLA (mainly Acanthamoeba, Vermamoeba, Naegleria and Stenamoeba) in 92.0% (81/88). L. pneumophila and L. bozemanii were most frequently isolated. FLA as potential host for Legionella spp. were isolated from 40.9% (36/88) of the composts in all facilities. In Legionella-positive samples the temperature of compost was significantly lower (P = 0.012) than in Legionella-negative samples. Of 47 bioaerosol samples, 19.1% (9/47) were positive for FLA and 10.6% (5/47) for L. pneumophila. Composts (62.8%) were positive for Legionella and FLA contemporaneously, but both microorganisms were never detected simultaneously in bioaerosols. Compost can release bioaerosol containing FLA or Legionella and could represent a source of infection of community-acquired Legionella infections for workers and nearby residents.

  9. Evidence of transfer by conjugation of type IV secretion system genes between Bartonella species and Rhizobium radiobacter in amoeba.

    Directory of Open Access Journals (Sweden)

    Watcharee Saisongkorh

    Full Text Available BACKGROUND: Bartonella species cospeciate with mammals and live within erythrocytes. Even in these specific niches, it has been recently suggested by bioinformatic analysis of full genome sequences that Lateral Gene Transfer (LGT may occur but this has never been demonstrated biologically. Here we describe the sequence of the B. rattaustraliani (AUST/NH4(T circular plasmid (pNH4 that encodes the tra cluster of the Type IV secretion system (T4SS and we eventually provide evidence that Bartonella species may conjugate and exchange this plasmid inside amoeba. PRINCIPAL FINDINGS: The T4SS of pNH4 is critical for intracellular viability of bacterial pathogens, exhibits bioinformatic evidence of LGT among bacteria living in phagocytic protists. For instance, 3 out of 4 T4SS encoding genes from pNH4 appear to be closely related to Rhizobiales, suggesting that gene exchange occurs between intracellular bacteria from mammals (bartonellae and plants (Rhizobiales. We show that B. rattaustraliani and Rhizobium radiobacter both survived within the amoeba Acanthamoeba polyphaga and can conjugate together. Our findings further support the hypothesis that tra genes might also move into and out of bacterial communities by conjugation, which might be the primary means of genomic evolution for intracellular adaptation by cross-talk of interchangeable genes between Bartonella species and plant pathogens. CONCLUSIONS: Based on this, we speculate that amoeba favor the transfer of genes as phagocytic protists, which allows for intraphagocytic survival and, as a consequence, promotes the creation of potential pathogenic organisms.

  10. Study of interactions between metal ions and protein model compounds by energy decomposition analyses and the AMOEBA force field

    Science.gov (United States)

    Jing, Zhifeng; Qi, Rui; Liu, Chengwen; Ren, Pengyu

    2017-10-01

    The interactions between metal ions and proteins are ubiquitous in biology. The selective binding of metal ions has a variety of regulatory functions. Therefore, there is a need to understand the mechanism of protein-ion binding. The interactions involving metal ions are complicated in nature, where short-range charge-penetration, charge transfer, polarization, and many-body effects all contribute significantly, and a quantitative description of all these interactions is lacking. In addition, it is unclear how well current polarizable force fields can capture these energy terms and whether these polarization models are good enough to describe the many-body effects. In this work, two energy decomposition methods, absolutely localized molecular orbitals and symmetry-adapted perturbation theory, were utilized to study the interactions between Mg2+/Ca2+ and model compounds for amino acids. Comparison of individual interaction components revealed that while there are significant charge-penetration and charge-transfer effects in Ca complexes, these effects can be captured by the van der Waals (vdW) term in the AMOEBA force field. The electrostatic interaction in Mg complexes is well described by AMOEBA since the charge penetration is small, but the distance-dependent polarization energy is problematic. Many-body effects were shown to be important for protein-ion binding. In the absence of many-body effects, highly charged binding pockets will be over-stabilized, and the pockets will always favor Mg and thus lose selectivity. Therefore, many-body effects must be incorporated in the force field in order to predict the structure and energetics of metalloproteins. Also, the many-body effects of charge transfer in Ca complexes were found to be non-negligible. The absorption of charge-transfer energy into the additive vdW term was a main source of error for the AMOEBA many-body interaction energies.

  11. Quantitative detection and differentiation of free-living amoeba species using SYBR green-based real-time PCR melting curve analysis.

    Science.gov (United States)

    Behets, Jonas; Declerck, Priscilla; Delaedt, Yasmine; Verelst, Lieve; Ollevier, Frans

    2006-12-01

    Real-time polymerase chain reaction melting curve analysis (MCA) allows differentiation of several free-living amoebae species. Distinctive characteristics were found for Naegleria fowleri, N. lovaniensis, N. australiensis, N. gruberi, Hartmanella vermiformis, and Willaertia magna. Species specificity of the amplicons was confirmed using agarose gel electrophoresis and sequence-based approaches. Amplification efficiency ranged from 91% to 98%, indicating the quantitative potential of the assay. This MCA approach can be used for quantitative detection of free-living amoebae after cultivation but also as a culture-independent detection method.

  12. Paravannella minima n. g. n. sp. (Discosea, Vannellidae) and distinction of the genera in the vannellid amoebae.

    Science.gov (United States)

    Kudryavtsev, Alexander

    2014-06-01

    Paravannella minima n. g. n. sp. (Amoebozoa, Vannellidae) isolated from a freshwater aquarium, possesses all light-microscopic and ultrastructural characteristics of the genus Vannella, being one of the smallest species among the vannellid amoebae (cell size during locomotion usually between 4.5 and 10μm). At the same time, sequence analysis of the genes encoding for nuclear SSU rRNA, actin and mitochondrial Cox1 shows this species as the earliest-branching vannellid that appears to be sister to the rest of this clade. This is correlated with the presence of some plesiomorphic characters; in particular, the secondary structure of the hypervariable helices E23-1-E23-7 in the studied species is shared with Vannella and most of the genera of Dactylopodida. The cell coat structure of the studied species corresponds to the hypothesis that vannellid amoebae were ancestrally enclosed in a cell coat consisting of pentagonal glycostyles that have undergone multiple independent losses in various clades of Vannellidae. Copyright © 2013 Elsevier GmbH. All rights reserved.

  13. Ptolemeba n. gen., a novel genus of hartmannellid amoebae (Tubulinea, Amoebozoa); with an emphasis on the taxonomy of Saccamoeba.

    Science.gov (United States)

    Watson, Pamela M; Sorrell, Stephanie C; Brown, Matthew W

    2014-01-01

    Hartmannellid amoebae are an unnatural assemblage of amoeboid organisms that are morphologically difficult to discern from one another. In molecular phylogenetic trees of the nuclear-encoded small subunit rDNA, they occupy at least five lineages within Tubulinea, a well-supported clade in Amoebozoa. The polyphyletic nature of the hartmannellids has led to many taxonomic problems, in particular paraphyletic genera. Recent taxonomic revisions have alleviated some of the problems. However, the genus Saccamoeba is paraphyletic and is still in need of revision as it currently occupies two distinct lineages. Here, we report a new clade on the tree of Tubulinea, which we infer represents a novel genus that we name Ptolemeba n. gen. This genus subsumes a clade of hartmannellid amoebae that were previously considered in the genus Saccamoeba, but whose mitochondrial morphology is distinct from Saccamoeba. In accordance with previous research, we formalize the clade as distinct from Saccamoeba. Transmission electron microscopy of our isolates illustrate that both molecularly discrete species can be further differentiated by their unique mitochondrial cristal morphology. © 2014 The Author(s) Journal of Eukaryotic Microbiology © 2014 International Society of Protistologists.

  14. The reconstruction of late Holocene depth-to-water-table based on testate amoebae in an eastern Australian mire

    Science.gov (United States)

    Zheng, X.; Money, S.; Hope, G.

    2017-12-01

    There are relatively few quantitative palaeo-hydrological records available in eastern Australia, and those that are available, for example from dendroclimatology and the reconstruction of lake level, are often relatively short or have a relatively coarse temporal resolution (e.g. Wilkins et al. 2013; Palmer et al. 2015). Testate amoebae, a widely used hydrological proxy in the Northern Hemisphere, were used here to reconstruct depth to water table (DWT) at Snowy Flat, which is a Sphagnum-Richea-Empodismahigh altitude (1618 m asl) shrub bog in the Australian Capital Territory, Australia. Testate amoebae were quantified in a Snowy Flat core representing 4,200 cal Y BP and the community composition was used to reconstruct DWT based on our recently established transfer functions. Results from three different types of transfer functions (Fig. 1) consistently show there was a decreasing DWT (wetter) period centred on about 3350 cal Y BP, a trend towards increased dryness from about 3300 to 2200 cal Y BP and a distinctly drier period 850 to 700 cal Y BP which was immediately followed by a wetter period from 700 to 500 cal Y BP. We discuss these episodes and trends in relation to the drivers of climatic variability in this region and in particular, by comparing our results with other south-eastern Australia records, comment on the history of the southern annular mode.

  15. Loss of Cln3 impacts protein secretion in the social amoeba Dictyostelium.

    Science.gov (United States)

    Huber, Robert J

    2017-07-01

    Neuronal ceroid lipofuscinosis (NCL), also referred to as Batten disease, is the most common form of childhood neurodegeneration. Mutations in CLN3 cause the most prevalent subtype of the disease, which manifests during early childhood and is currently untreatable. The precise function of the CLN3 protein is still not known, which has inhibited the development of targeted therapies. In the social amoeba Dictyostelium discoideum, loss of the CLN3 homolog, Cln3, reduces adhesion during early development, which delays streaming and aggregation. The results of the present study indicate that this phenotype may be at least partly due to aberrant protein secretion in cln3 - cells. It is well-established that Cln3 localizes primarily to the contractile vacuole (CV) system in Dictyostelium, and to a lesser extent, compartments of the endocytic pathway. Intriguingly, the CV system has been linked to the secretion of proteins that do not contain a signal peptide for secretion (i.e., unconventional protein secretion). Proteins that do contain a signal peptide are secreted via a conventional mechanism involving the endoplasmic reticulum, transport through the Golgi, and secretion via vesicle release. In this study, Cln3 was observed to co-localize with the Golgi marker wheat germ agglutinin suggesting that Cln3 participates in both secretion mechanisms. Chimeras of wild-type (WT) and cln3 - cells displayed delayed streaming and aggregation, and interestingly, cln3 - cells starved in conditioned media (CM) harvested from starving WT cells showed near normal timing of streaming and aggregation suggesting aberrant protein secretion in Cln3-deficient cells. Based on these observations, LC-MS/MS was used to reveal the protein content of CM from starved cells (mass spectrometry data are available via ProteomeXchange with identifier PXD004897). A total of 450 proteins were detected in WT and cln3 - CM, of which 3 were absent in cln3 - CM. Moreover, 12 proteins that were present in

  16. Micriamoeba tesseris nov. gen. nov. sp.: a new taxon of free-living small-sized Amoebae non-permissive to virulent Legionellae.

    Science.gov (United States)

    Atlan, Danièle; Coupat-Goutaland, Bénédicte; Risler, Arnaud; Reyrolle, Monique; Souchon, Maud; Briolay, Jérôme; Jarraud, Sophie; Doublet, Patricia; Pélandakis, Michel

    2012-11-01

    Investigation of soil amoebae in 11 cooling towers allowed us to isolate a major unknown small-sized amoeba population (SZA). However, SZA did not appear to be specific to cooling tower ecosystems since they are also a major amoeba population found in muds isolated from different points of a water treatment plant. The SSU-rDNA sequences from SZA strains did not match any known database sequences, suggesting that SZA constitutes a new amoeba taxon. We isolated and further described one of the SZA that we named Micriamoeba tesseris. The phylogenetic analyses showed that Micriamoeba tesseris belongs to the Amebozoa and branched together with genus Echinamoeba+Vermamoeba vermiformis. Phylogenetic analyses within the Micriamoeba group distinguished different subgroups of Micriamoeba strains according to their origin, i.e. cooling tower or mud. Although Micriamoeba are able to feed on viable E. coli cells, they do not uptake virulent Legionella pneumophila strains, thus enabling them to avoid infection by Legionella. Consequently, Micriamoeba is not directly involved in L. pneumophila multiplication. However, an indirect role of Micriamoeba in Legionella risk is discussed. Copyright © 2012 Elsevier GmbH. All rights reserved.

  17. First Evidence for Diploidy and Genetic Recombination in Free-Living Amoebae of the Genus Naegleria on the Basis of Electrophoretic Variation

    OpenAIRE

    Cariou, Marie Louise; Pernin, Pierre

    1987-01-01

    Electrophoretic variation for 15 enzyme-coding genes was studied in various Naegleria (Rhizopoda, Vahlkampfiidae) species. The occurrence of complex banding patterns provided the first evidence of a diploid structure of the genome of these amoebae. The putative loci identified were found not to be linked and the genotypic distribution suggested chromosomal recombination for one species (Naegleria lovaniensis).

  18. First evidence for diploidy and genetic recombination in free-living amoebae of the genus naegleria on the basis of electrophoretic variation.

    Science.gov (United States)

    Cariou, M L; Pernin, P

    1987-02-01

    Electrophoretic variation for 15 enzyme-coding genes was studied in various Naegleria (Rhizopoda, Vahlkampfiidae) species. The occurrence of complex banding patterns provided the first evidence of a diploid structure of the genome of these amoebae. The putative loci identified were found not to be linked and the genotypic distribution suggested chromosomal recombination for one species (Naegleria lovaniensis).

  19. Isolamento de amebas de vida livre a partir de água mineral engarrafada Isolation of free-living amoebae from bottled mineral water

    Directory of Open Access Journals (Sweden)

    Homero Coutinho Salazar

    1982-10-01

    Full Text Available Foram isoladas amebas dos gêneros Vahlkampfia, Glaeseria, Acanthamoeba, Filamoeba, Amoeba, Platyamoeba e Hartmanella de dez diferentes marcas de água mineral engarrafadas, servidas no Rio de Janeiro, RJ (Brasil. Somente uma das marcas foi negativa para protozoários, enquanto que de todas as outras foram isoladas amebas de vida livre. O significado desses resultados é discutido como um indicador de qualidade da água mineral usada comercialmente, destacando-se o fato de que alguns desses microorganismos são potencialmente patogênicos.Amoebae of the genera Vahlkampfia, Glaeseria, Acanthamoeba, Filamoeba, Amoeba, Platyamoeba and Hartmanella were isolated from ten different brands of bottled mineral water used in Rio de Janeiro. Of these brands, only one was negative after an incubation period of twenty days, while the others were all positive for free living amoebae. The results are discussed as an indicator of the quality of commercially used mineral water and it is stressed that some of these microorganisms are potentially pathogenic.

  20. Use of an Amoeba proteus model for in vitro cytotoxicity testing in phytochemical research. Application to Euphorbia hirta extracts.

    Science.gov (United States)

    Duez, P; Livaditis, A; Guissou, P I; Sawadogo, M; Hanocq, M

    1991-09-01

    Amoeba proteus is proposed as a low-cost multi-purpose biochemical tool for screening and standardizing cytotoxic plant extracts with possible application in the laboratories of developing countries. Advantages and limitations of this test are examined and different mathematical treatments (probit analysis versus curve fitting to Von Bertalanffy and Hill functions) are investigated. Known anti-cancer (doxorubicin, daunorubicin, dacarbazine, 5-fluorouracil) and antiparasitic (emetine, dehydroemetine, metronidazole, cucurbitine, chloroquine) drugs were tested using this method and only metronidazole appeared inactive. Application of this model to Euphorbia hirta established that a 100 degrees C aqueous extraction of fresh aerial parts allows efficient extraction of active constituents and that drying the plant material before extraction considerably reduces activity.

  1. The abundant free-living amoeba, Acanthamoeba polyphaga, increases the survival of Campylobacter jejuni in milk and orange juice

    Directory of Open Access Journals (Sweden)

    Jenny Olofsson

    2015-09-01

    Full Text Available Background: Campylobacter jejuni is a common cause of human bacterial diarrhea in most parts of the world. Most C. jejuni infections are acquired from contaminated poultry, milk, and water. Due to health care costs and human suffering, it is important to identify all possible sources of infection. Unpasteurized milk has been associated with several outbreaks of C. jejuni infection. Campylobacter has been identified on fresh fruit, and other gastrointestinal pathogens such as Salmonella, E. coli O157:H7 and Cryptosporidium have been involved in fruit juice outbreaks. C. jejuni is sensitive to the acidic environment of fruit juice, but co-cultures with the amoeba, Acanthamoeba polyphaga, have previously been shown to protect C. jejuni at low pH. Methods: To study the influence of A. polyphaga on the survival of C. jejuni in milk and juice, the bacteria were incubated in the two products at room temperature and at 4°C with the following treatments: A C. jejuni preincubated with A. polyphaga before the addition of product, B C. jejuni mixed with A. polyphaga after the addition of product, and C C. jejuni in product without A. polyphaga. Bacterial survival was assessed by colony counts on blood agar plates. Results: Co-culture with A. polyphaga prolonged the C. jejuni survival both in milk and juice. The effect of co-culture was most pronounced in juice stored at room temperature. On the other hand, A. polyphaga did not have any effect on C. jejuni survival during pasteurization of milk or orange juice, indicating that this is a good method for eliminating C. jejuni in these products. Conclusion: Amoebae-associated C. jejuni in milk and juice might cause C. jejuni infections.

  2. Coexistence of Legionella pneumophila Bacteria and Free-Living Amoebae in Lakes Serving as a Cooling System of a Power Plant.

    Science.gov (United States)

    Zbikowska, Elżbieta; Kletkiewicz, Hanna; Walczak, Maciej; Burkowska, Aleksandra

    2014-01-01

    The study was aimed at determining whether potentially pathogenic free-living amoebae (FLA) and Legionella pneumophila can be found in lakes serving as a natural cooling system of a power plant. Water samples were collected from five lakes forming the cooling system of the power plants Pątnów and Konin (Poland). The numbers of investigated organisms were determined with the use of a very sensitive molecular method-fluorescence in situ hybridization (FISH). The result of the present study shows that thermally altered aquatic environments provide perfect conditions for the growth of L. pneumophila and amoebae. The bacteria were identified in the biofilm throughout the entire research period and in the subsurface water layer in July and August. Hartmanella sp. and/or Naegleria fowleri were identified in the biofilm throughout the entire research period.

  3. Molecular identification of waterborne free living amoebae (Acanthamoeba, Naegleria and Vermamoeba) isolated from municipal drinking water and environmental sources, Semnan province, north half of Iran.

    Science.gov (United States)

    Javanmard, Ehsan; Niyyati, Maryam; Lorenzo-Morales, Jacob; Lasjerdi, Zohreh; Behniafar, Hamed; Mirjalali, Hamed

    2017-12-01

    The present study tested 80 samples of municipal, geothermal and recreational water samples for the occurrence of waterborne free living amoebae (FLA) including Acanthamoeba, Balamuthia mandrillaris, Vahlkampfiids and Vermamoeba in Semnan province, North half of Iran. Four sets of primers including JDP1,2 primers, ITS1,2 primers (Vahlkampfiids), 16S rRNABal primers (Balamuthia mandrillaris) and NA1,2 primers (Vermamoeba) were used to confirm the morphological identification. From the 80 water samples tested in the present study, 16 (20%) were positive for the outgrowth of free living amoebae based on the morphological page key. Out of the 34 municipal water samples, 7 (20.6%) were positive for outgrowth of Free living amoeba, belonging to Vermamoeba, Naegleria and Acanthamoeba using molecular tools. Three out of the six investigated hot springs were also contaminated with Naegleria spp. Sequencing of the ITS1,2 region of the Vahlkampfiid isolates revealed the highest homology with N. gruberi (2 isolates), N. australiensis (1 isolate) and N. pagei (3 isolates). This is the first report of N. gruberi in the country. Using morphological and molecular analysis, Balamuthia mandrillaris was undetected in all the water samples. The present study further confirmed the occurrence of potentially pathogenic waterborne free living amoebae in habitats with high human activity. It is of utmost importance that more studies are conducted to evaluate the niches of B. mandrillaris and N. fowleri in Iran and worldwide. Such investigations regarding the relevance of FLA as a hazard to humans, should be brought to the notice of the health authorities. Copyright © 2017. Published by Elsevier Inc.

  4. Occurrence and molecular characterization of free-living amoeba species (Acanthamoeba, Hartmannella, and Saccamoeba limax) in various surface water resources of Iran.

    Science.gov (United States)

    Mahmoudi, Mohammad Reza; Rahmati, Behnaz; Seyedpour, Seyed Hosssen; Karanis, Panagiotis

    2015-12-01

    This study was conducted to determine the presence and molecular identity of Acanthamoeba species in the surface water resources of four provinces in Iran, namely Guilan, Mazandaran (North of Iran), Alborz, and Tehran (capital city), using culture- and molecular-based methods. During March to November 2014, 49 surface water samples were collected from environmental water sources-the distinct surface waters of Guilan, Mazandaran, Alborz, and Tehran provinces, in Iran. For the isolation of Acanthamoeba species, approximately 500 ml of the water samples were filtered through a cellulose nitrate membrane with a pore size of 0.45 μ. The filter was transferred onto non-nutrient agar plates seeded with Gram-negative bacteria (Escherichia coli) as a food source. The presence of Acanthamoeba was confirmed by the genus-specific primer pair JDP1 and 2, and/or NA primers were used to identify Acanthamoeba and certain other free-living amoebae. In total, 38 out of 49 samples were positive by culture and/or PCR for Acanthamoeba and other free-living amoebae from all three provinces. By sequencing the positive isolates, the strains were shown to belong to Acanthamoeba (16 isolates belonged to T4 and 2 isolates belonged to T5), Hartmannella vermiformis (3/24), and Saccamoeba limax (2/24). The T4 and T5 genotypes were detected in Guilan and Mazandaran provinces. Two isolates from Guilan and Tehran provinces belonged to S. limax, and H. vermiformis was detected in Guilan province. The results of this study highlight the need to pay more attention to free-living amoebae, as human activity was observed in all of the localities from which these samples were taken. These surface waters can be potential sources for the distribution and transmission of pathogenic Acanthamoeba in the study areas, and free-living amoebas (FLA) (particularly the Acanthamoeba species) can serve as hosts for and vehicles of various microorganisms.

  5. First Record of Potentially Pathogenic Amoeba Vermamoeba vermiformis (Lobosea: Gymnamoebia Isolated from a Freshwater of Dokdo Island in the East Sea, Korea

    Directory of Open Access Journals (Sweden)

    Jong Soo Park

    2016-01-01

    Full Text Available Vermamoeba vermiformis is a very important free-living amoeba for human health in association with Legionnaires’ disease and keratitis. This interesting amoeba was firstly isolated from a freshwater of Dokdo (island, which was historically used for drinking water. Trophozoites and cyst forms of V. vermiformis strain MG1 are very similar to previous reported species. Trophozoites of V. vermiformis strain MG1 showed cylindrical shape with prominent anterior hyaline region. The average ratio of length and width was about 6.5. Typically, cysts of the strain MG1 showed a spherical or slightly ovoidal shape with smooth wall, and lacked cyst pores. Some cysts had crenulatewalled ectocyst, which was separated from endocyst wall. Further, 18S rRNA gene sequence of V. vermiformis strain MG1 showed very high similarity to other V. vermiformis species (99.4%-99.9% identity. Molecular phylogenetic analysis based on 18S rRNA gene sequences clearly confirmed that the isolate was one strain of V. vermiformis with maximum bootstrap value (maximum likelihood: 100% and Bayesian posterior probability of 1. Thus, the freshwater of Dokdo in Korea could harbor potentially pathogenic amoeba that may cause diseases in humans.

  6. Changes in Structure and Functioning of Protist (Testate Amoebae) Communities Due to Conversion of Lowland Rainforest into Rubber and Oil Palm Plantations.

    Science.gov (United States)

    Krashevska, Valentyna; Klarner, Bernhard; Widyastuti, Rahayu; Maraun, Mark; Scheu, Stefan

    2016-01-01

    Large areas of tropical rainforest are being converted to agricultural and plantation land uses, but little is known of biodiversity and ecological functioning under these replacement land uses. We investigated the effects of conversion of rainforest into jungle rubber, intensive rubber and oil palm plantations on testate amoebae, diverse and functionally important protists in litter and soil. Living testate amoebae species richness, density and biomass were all lower in replacement land uses than in rainforest, with the impact being more pronounced in litter than in soil. Similar abundances of species of high and low trophic level in rainforest suggest that trophic interactions are more balanced, with a high number of functionally redundant species, than in rubber and oil palm. In contrast, plantations had a low density of high trophic level species indicating losses of functions. This was particularly so in oil palm plantations. In addition, the relative density of species with siliceous shells was >50% lower in the litter layer of oil palm and rubber compared to rainforest and jungle rubber. This difference suggests that rainforest conversion changes biogenic silicon pools and increases silicon losses. Overall, the lower species richness, density and biomass in plantations than in rainforest, and the changes in the functional composition of the testate amoebae community, indicate detrimental effects of rainforest conversion on the structure and functioning of microbial food webs.

  7. Isolamento de amebas de vida livre potencialmente patogênicas em poeira de hospitais Isolation of potencially pathogenic free-living amoebas in hospital dust

    Directory of Open Access Journals (Sweden)

    Maria Aparecida da Silva

    2003-04-01

    Full Text Available OBJETIVO: Observar a ocorrência de amebas de vida livre dos gêneros Acanthamoeba e Naegleria em amostras de poeira coletadas em hospitais. MÉTODOS: Foram coletadas 132 amostras de poeira em dois hospitais do município de Presidente Prudente, São Paulo. Os locais da coleta foram: Unidade de Terapia Intensiva, Centro Cirúrgico, Isolamento de Moléstias Infecciosas, Berçário, Emergência e Cozinha. As amostras foram semeadas em três meios de cultura: meio de ágar não nutriente com Escherichia coli, meio de ágar infusão de soja e microcultivo em meio de Pavlova modificado por Giazzi. As amebas isoladas foram identificadas segundo critérios morfológicos. RESULTADOS: O índice geral de positividade para amebas de vida livre, potencialmente patogênicas, dos gêneros Acanthamoeba e Naegleria, foi de 45,5%, sendo positivas 41,6% das amostras de poeira coletadas no hospital universitário e 50% no hospital estadual. Obtiveram-se 45,5% de positividade do gênero Acanthamoeba e 3,8% para amebas do gênero Naegleria. CONCLUSÕES: As amebas de vida livre, potencialmente patogênicas, estavam presentes em todos os ambientes estudados dos dois hospitais, sendo que as espécies do gênero Acanthamoeba foram as isoladas com maior freqüência.OBJECTIVE: To evaluate the occurrence of free-living amoebas of the genera Acanthamoeba and Naegleria is dust samples colleted in two hospitals. METHODS: One-hundred and thirty-two dust samples were collected in two hospitals in Brazil. Hospital collection sites were the following: intensive care unit, operation rooms, nursery, kitchen, emergency and infectious diseases isolation room . The isolation of the amoebas was performed in three culture media: non-nutrient agar inoculated with Escherichia coli, soy agar, and microculture in Giazzi-modified Pavlova's medium. The amoebas were identified according to morphological criteria. RESULTS: Amoebas of the genera Acanthamoeba and Naegleria were found in 45.5% of

  8. Detection of viable Helicobacter pylori inside free-living amoebae in wastewater and drinking water samples from Eastern Spain.

    Science.gov (United States)

    Moreno-Mesonero, Laura; Moreno, Yolanda; Alonso, José Luis; Ferrús, M Antonia

    2017-10-01

    Helicobacter pylori is one of the most concerning emerging waterborne pathogens. It has been suggested that it could survive in water inside free-living amoebae (FLA), but nobody has studied this relationship in the environment yet. Thus, we aimed to detect viable H. pylori cells from inside FLA in water samples. Sixty-nine wastewater and 31 drinking water samples were collected. FLA were purified and identified by PCR and sequencing. For exclusively detecting H. pylori inside FLA, samples were exposed to sodium hypochlorite and assayed by specific PMA-qPCR, DVC-FISH and culture. FLA were detected in 38.7% of drinking water and 79.7% of wastewater samples, even after disinfection. In wastewater, Acanthamoeba spp. and members of the family Vahlkampfiidae were identified. In drinking water, Acanthamoeba spp. and Echinamoeba and/or Vermamoeba were present. In 39 (58.2%) FLA-positive samples, H. pylori was detected by PMA-qPCR. After DVC-FISH, 21 (31.3%) samples harboured viable H. pylori internalized cells. H. pylori was cultured from 10 wastewater samples. To our knowledge, this is the first report that demonstrates that H. pylori can survive inside FLA in drinking water and wastewater, strongly supporting the hypothesis that FLA could play an important role in the transmission of H. pylori to humans. © 2017 Society for Applied Microbiology and John Wiley & Sons Ltd.

  9. The TOM Complex of Amoebozoans: the Cases of the Amoeba Acanthamoeba castellanii and the Slime Mold Dictyostelium discoideum.

    Science.gov (United States)

    Wojtkowska, Małgorzata; Buczek, Dorota; Stobienia, Olgierd; Karachitos, Andonis; Antoniewicz, Monika; Slocinska, Małgorzata; Makałowski, Wojciech; Kmita, Hanna

    2015-07-01

    Protein import into mitochondria requires a wide variety of proteins, forming complexes in both mitochondrial membranes. The TOM complex (translocase of the outer membrane) is responsible for decoding of targeting signals, translocation of imported proteins across or into the outer membrane, and their subsequent sorting. Thus the TOM complex is regarded as the main gate into mitochondria for imported proteins. Available data indicate that mitochondria of representative organisms from across the major phylogenetic lineages of eukaryotes differ in subunit organization of the TOM complex. The subunit organization of the TOM complex in the Amoebozoa is still elusive, so we decided to investigate its organization in the soil amoeba Acanthamoeba castellanii and the slime mold Dictyostelium discoideum. They represent two major subclades of the Amoebozoa: the Lobosa and Conosa, respectively. Our results confirm the presence of Tom70, Tom40 and Tom7 in the A. castellanii and D. discoideum TOM complex, while the presence of Tom22 and Tom20 is less supported. Interestingly, the Tom proteins display the highest similarity to Opisthokonta cognate proteins, with the exception of Tom40. Thus representatives of two major subclades of the Amoebozoa appear to be similar in organization of the TOM complex, despite differences in their lifestyle. Copyright © 2015 The Authors. Published by Elsevier GmbH.. All rights reserved.

  10. A simple method for the cryopreservation of free-living amoebae belonging to the genera Naegleria and Acanthamoeba.

    Science.gov (United States)

    Kilvington, S; White, D

    1991-06-21

    The cryopreservation of free-living amoebae of the genera Naegleria and Acanthamoeba from axenic or monoxenic culture with Escherichia coli was achieved using a two-step cooling process in laboratory freezers and storage in liquid nitrogen. Trophozoites were suspended in axenic culture medium and dimethylsulphoxide (DMSO) added to a final concentration of 5%. Ampoules were placed directly at -20°C for 60 min, followed by a further 60 min at -70°C and then stored in liquid nitrogen. On rapid thawing of the ampoules at 37°C, recovery rates with standard errors from three separate experiments performed in triplicate were: N. fowleri 8% ± 3%, N. lovaniensis 12% ± 3%, N. gruberi 25% ± 6% and A. polyphaga 34% ± 5.5% from axenic cultures, and21% ± 7%, 31% ± 10%, 44% ± 11%, 49% ± 9%, respectively from monoxenic cultures. Although not optimal, the method is proposed as a simple and reproducible means of cryopreserving Naegleria and Acanthamoeba strains. Copyright © 1991 Gustav Fischer Verlag · Stuttgart · Jena · New York. Published by Elsevier GmbH.. All rights reserved.

  11. Brain-Eating Amoebae: Silver Nanoparticle Conjugation Enhanced Efficacy of Anti-Amoebic Drugs against Naegleria fowleri.

    Science.gov (United States)

    Rajendran, Kavitha; Anwar, Ayaz; Khan, Naveed Ahmed; Siddiqui, Ruqaiyyah

    2017-12-20

    The overall aim of this study was to determine whether conjugation with silver nanoparticles enhances effects of available drugs against primary amoebic meningoencephalitis due to Naegleria fowleri. Amphotericin B, Nystatin, and Fluconazole were conjugated with silver nanoparticles, and synthesis was confirmed using UV-visible spectrophotometry. Atomic force microscopy determined their size in range of 20-100 nm. To determine amoebicidal effects, N. fowleri were incubated with drugs-conjugated silver nanoparticles, silver nanoparticles alone, and drugs alone. The findings revealed that silver nanoparticles conjugation significantly enhanced antiamoebic effects of Nystatin and Amphotericin B but not Fluconazole at micromolar concentrations, compared with the drugs alone. For the first time, our findings showed that silver nanoparticle conjugation enhances efficacy of antiamoebic drugs against N. fowleri. Given the rarity of the disease and challenges in developing new drugs, it is hoped that modifying existing drugs to enhance their antiamoebic effects is a useful avenue that holds promise in improving the treatment of brain-eating amoebae infection due to N. fowleri.

  12. Sphagnum-dwelling testate amoebae in subarctic bogs are more sensitive to soil warming in the growing season than in winter: the results of eight-year field climate manipulations.

    Science.gov (United States)

    Tsyganov, Andrey N; Aerts, Rien; Nijs, Ivan; Cornelissen, Johannes H C; Beyens, Louis

    2012-05-01

    Sphagnum-dwelling testate amoebae are widely used in paleoclimate reconstructions as a proxy for climate-induced changes in bogs. However, the sensitivity of proxies to seasonal climate components is an important issue when interpreting proxy records. Here, we studied the effects of summer warming, winter snow addition solely and winter snow addition together with spring warming on testate amoeba assemblages after eight years of experimental field climate manipulations. All manipulations were accomplished using open top chambers in a dry blanket bog located in the sub-Arctic (Abisko, Sweden). We estimated sensitivity of abundance, diversity and assemblage structure of living and empty shell assemblages of testate amoebae in the living and decaying layers of Sphagnum. Our results show that, in a sub-arctic climate, testate amoebae are more sensitive to climate changes in the growing season than in winter. Summer warming reduced species richness and shifted assemblage composition towards predominance of xerophilous species for the living and empty shell assemblages in both layers. The higher soil temperatures during the growing season also decreased abundance of empty shells in both layers hinting at a possible increase in their decomposition rates. Thus, although possible effects of climate changes on preservation of empty shells should always be taken into account, species diversity and structure of testate amoeba assemblages in dry subarctic bogs are sensitive proxies for climatic changes during the growing season. Copyright © 2011 Elsevier GmbH. All rights reserved.

  13. Whole genome sequencing of mutation accumulation lines reveals a low mutation rate in the social amoeba Dictyostelium discoideum.

    Directory of Open Access Journals (Sweden)

    Gerda Saxer

    Full Text Available Spontaneous mutations play a central role in evolution. Despite their importance, mutation rates are some of the most elusive parameters to measure in evolutionary biology. The combination of mutation accumulation (MA experiments and whole-genome sequencing now makes it possible to estimate mutation rates by directly observing new mutations at the molecular level across the whole genome. We performed an MA experiment with the social amoeba Dictyostelium discoideum and sequenced the genomes of three randomly chosen lines using high-throughput sequencing to estimate the spontaneous mutation rate in this model organism. The mitochondrial mutation rate of 6.76×10(-9, with a Poisson confidence interval of 4.1×10(-9 - 9.5×10(-9, per nucleotide per generation is slightly lower than estimates for other taxa. The mutation rate estimate for the nuclear DNA of 2.9×10(-11, with a Poisson confidence interval ranging from 7.4×10(-13 to 1.6×10(-10, is the lowest reported for any eukaryote. These results are consistent with low microsatellite mutation rates previously observed in D. discoideum and low levels of genetic variation observed in wild D. discoideum populations. In addition, D. discoideum has been shown to be quite resistant to DNA damage, which suggests an efficient DNA-repair mechanism that could be an adaptation to life in soil and frequent exposure to intracellular and extracellular mutagenic compounds. The social aspect of the life cycle of D. discoideum and a large portion of the genome under relaxed selection during vegetative growth could also select for a low mutation rate. This hypothesis is supported by a significantly lower mutation rate per cell division in multicellular eukaryotes compared with unicellular eukaryotes.

  14. Isolation and identification of pathogenic free-living amoeba from surface and tap water of Shiraz City using morphological and molecular methods.

    Science.gov (United States)

    Armand, B; Motazedian, M H; Asgari, Q

    2016-01-01

    Free-living amoebae (FLA) are the most abundant and widely distributed protozoa in the environment. An investigation was conducted to determine the presence of free-living amoebae (FLA), Acanthamoeba and Vermamoeba in waterfronts of parks and squares and tap water of Shiraz City, Iran. FLA are considered pathogenic for human. These ubiquitous organisms have been isolated from different environments such as water, soil, and air. Eighty-two water samples were collected from different places of Shiraz City during the summer of 2013. All samples were processed in Dept. of Parasitology and Mycology, Shiraz University of Medical Sciences, Fars, Iran. Samples were screened for FLA and identified by morphological characters in the cultures, PCR amplification targeting specific genes for each genus and sequencing determined frequent species and genotypes base on NCBI database. Overall, 48 samples were positive for Acanthamoeba and Vermamoeba in non-nutrient agar culture based on morphological characteristics. The PCR examination was done successfully. Sequencing results were revealed T4 (62.96 %) genotypes as the most common genotype of Acanthamoeba in the Shiraz water sources. In addition, T5 (33.33 %) and T15 (3.71 %) were isolated from water supplies. Vermamoeba vermiformis was known the dominant species from this genus. The high frequency of Acanthamoeba spp. and Vermamoeba in different environmental water sources of Shiraz is an alert for the public health related to water sources. The result highlights a need for taking more attention to water supplies in order to prevent illnesses related to free-living amoebae.

  15. Multiple Legionella pneumophila Type II secretion substrates, including a novel protein, contribute to differential infection of the amoebae Acanthamoeba castellanii, Hartmannella vermiformis, and Naegleria lovaniensis.

    Science.gov (United States)

    Tyson, Jessica Y; Pearce, Meghan M; Vargas, Paloma; Bagchi, Sreya; Mulhern, Brendan J; Cianciotto, Nicholas P

    2013-05-01

    Type II protein secretion (T2S) by Legionella pneumophila is required for intracellular infection of host cells, including macrophages and the amoebae Acanthamoeba castellanii and Hartmannella vermiformis. Previous proteomic analysis revealed that T2S by L. pneumophila 130b mediates the export of >25 proteins, including several that appeared to be novel. Following confirmation that they are unlike known proteins, T2S substrates NttA, NttB, and LegP were targeted for mutation. nttA mutants were impaired for intracellular multiplication in A. castellanii but not H. vermiformis or macrophages, suggesting that novel exoproteins which are specific to Legionella are especially important for infection. Because the importance of NttA was host cell dependent, we examined a panel of T2S substrate mutants that had not been tested before in more than one amoeba. As a result, RNase SrnA, acyltransferase PlaC, and metalloprotease ProA all proved to be required for optimal intracellular multiplication in H. vermiformis but not A. castellanii. Further examination of an lspF mutant lacking the T2S apparatus documented that T2S is also critical for infection of the amoeba Naegleria lovaniensis. Mutants lacking SrnA, PlaC, or ProA, but not those deficient for NttA, were defective in N. lovaniensis. Based upon analysis of a double mutant lacking PlaC and ProA, the role of ProA in H. vermiformis was connected to its ability to activate PlaC, whereas in N. lovaniensis, ProA appeared to have multiple functions. Together, these data document that the T2S system exports multiple effectors, including a novel one, which contribute in different ways to the broad host range of L. pneumophila.

  16. Ubiquiter circovirus sequences raise challenges in laboratory diagnosis: the case of honey bee and bee mite, reptiles, and free living amoebae.

    Science.gov (United States)

    Marton, Szilvia; Ihász, Katalin; Lengyel, György; Farkas, Szilvia L; Dán, Ádám; Paulus, Petra; Bányai, Krisztián; Fehér, Enikő

    2015-03-01

    Circoviruses of pigs and birds are established pathogens, however, the exact role of other, recently described circoviruses and circovirus-like viruses remains to be elucidated. The aim of this study was the detection of circoviruses in neglected host species, including honey bees, exotic reptiles and free-living amoebae by widely used broad-spectrum polymerase chain reaction (PCR) assays specific for the replication initiation protein coding gene of these viruses. The majority of sequences obtained from honey bees were highly similar to canine and porcine circoviruses, or, were distantly related to dragonfly cycloviruses. Other rep sequences detected in some honey bees, reptiles and amoebae showed similarities to various rep sequences deposited in the GenBank. Back-to-back PCR primers designed for the amplification of whole viral genomes failed to work that suggested the existence of integrated rep-like elements in many samples. Rolling circle amplification and exonuclease treatment confirmed the absence of small circular DNA genomes in the specimens analysed. In case of honey bees Varroa mite DNA contamination might be a source of the identified endogenous rep-like elements. The reptile and amoebae rep-like sequences were nearly identical with each other and with sequences detected in chimpanzee feces raising the possibility that detection of novel or unusual rep-like elements in some host species might originate from the microbial community of the host. Our results indicate that attention is needed when broad-spectrum rep gene specific polymerase chain reaction is chosen for laboratory diagnosis of circovirus infections.

  17. Defects in the synthetic pathway prevent DIF-1 mediated stalk lineage specification cascade in the non-differentiating social amoeba, Acytostelium subglobosum.

    Science.gov (United States)

    Mohri, Kurato; Hata, Takashi; Kikuchi, Haruhisa; Oshima, Yoshiteru; Urushihara, Hideko

    2014-05-29

    Separation of somatic cells from germ-line cells is a crucial event for multicellular organisms, but how this step was achieved during evolution remains elusive. In Dictyostelium discoideum and many other dictyostelid species, solitary amoebae gather and form a multicellular fruiting body in which germ-line spores and somatic stalk cells differentiate, whereas in Acytostelium subglobosum, acellular stalks form and all aggregated amoebae become spores. In this study, because most D. discoideum genes known to be required for stalk cell differentiation have homologs in A. subglobosum, we inferred functional variations in these genes and examined conservation of the stalk cell specification cascade of D. discoideum mediated by the polyketide differentiation-inducing factor-1 (DIF-1) in A. subglobosum. Through heterologous expression of A. subglobosum orthologs of DIF-1 biosynthesis genes in D. discoideum, we confirmed that two of the three genes were functional equivalents, while DIF-methyltransferase (As-dmtA) involved at the final step of DIF-1 synthesis was not. In fact, DIF-1 activity was undetectable in A. subglobosum lysates and amoebae of this species were not responsive to DIF-1, suggesting a lack of DIF-1 production in this species. On the other hand, the molecular function of an A. subglobosum ortholog of DIF-1 responsive transcription factor was equivalent with that of D. discoideum and inhibition of polyketide synthesis caused developmental arrest in A. subglobosum, which could not be rescued by DIF-1 addition. These results suggest that non-DIF-1 polyketide cascades involving downstream transcription factors are required for fruiting body development of A. subglobosum. © 2014. Published by The Company of Biologists Ltd.

  18. An evolutionarily significant unicellular strategy in response to starvation in Dictyostelium social amoebae [v2; ref status: indexed, http://f1000r.es/4kb

    Directory of Open Access Journals (Sweden)

    Darja Dubravcic

    2014-12-01

    Full Text Available The social amoeba Dictyostelium discoideum is widely studied for its multicellular development program as a response to starvation. Aggregates of up to 106 cells form fruiting bodies containing (i dormant spores (~80% that can persist for months in the absence of nutrients, and (ii dead stalk cells (~20% that promote the dispersion of the spores towards nutrient-rich areas. It is often overlooked that not all cells aggregate upon starvation. Using a new quantitative approach based on time-lapse fluorescence microscopy and a low ratio of reporting cells, we have quantified this fraction of non-aggregating cells. In realistic starvation conditions, up to 15% of cells do not aggregate, which makes this third cell fate a significant component of the population-level response of social amoebae to starvation. Non-aggregating cells have an advantage over cells in aggregates since they resume growth earlier upon arrival of new nutrients, but have a shorter lifespan under prolonged starvation. We find that phenotypic heterogeneities linked to cell nutritional state bias the representation of cells in the aggregating vs. non-aggregating fractions, and thus affect population partitioning. Next, we report that the fraction of non-aggregating cells depends on genetic factors that regulate the timing of starvation, signal sensing efficiency and aggregation efficiency. In addition, interactions between clones in mixtures of non-isogenic cells affect the partitioning of each clone into both fractions. We further build a numerical model to test the evolutionary significance of the non-aggregating cell fraction. The partitioning of cells into aggregating and non-aggregating fractions is optimal in fluctuating environments with an unpredictable duration of starvation periods. Our study highlights the unicellular component of the response of social amoebae to starvation, and thus extends its evolutionary and ecological framework.

  19. Could the canopy structure of bryophytes serve as an indicator of microbial biodiversity? A test for testate amoebae and microcrustaceans from a subtropical cloud forest in Dominican Republic.

    Science.gov (United States)

    Acosta-Mercado, D; Cancel-Morales, N; Chinea, J D; Santos-Flores, C J; De Jesús, I Sastre

    2012-07-01

    The mechanisms that ultimately regulate the diversity of microbial eukaryotic communities in bryophyte ecosystems remain a contentious topic in microbial ecology. Although there is robust consensus that abiotic factors, such as water chemistry of the bryophyte and pH, explain a significant proportion of protist and microcrustacean diversity, there is no systematic assessment of the role of bryophyte habitat complexity on such prominent microbial groups. Water-holding capacity is correlated with bryophyte morphology and canopy structure. Similarly, canopy structure explains biodiversity dynamics of the macrobiota suggesting that canopy structure may also be a potential parameter for understanding microbial diversity. Canopy roughness of the dominant bryophyte species within the Bahoruco Cloud Forest, Cachote, Dominican Republic, concomitant with their associated diversity of testate amoebae and microcrustaceans was estimated to determine whether canopy structure could be added to the list of factors explaining microbial biodiversity in bryophytes. We hypothesized that smooth (with high moisture content) canopies will have higher species richness, density, and biomass of testate amoebae and higher richness and density of microcrustaceans than rough (desiccation-prone) canopies. For testate amoebae, we found 83 morphospecies with relative low abundances. Species richness and density differed among bryophytes with different bryophyte canopy structures and based on non-metric multidimensional scaling, canopy roughness explained 25% of the variation in species composition although not as predicted. Acroporium pungens (low roughness, LR) had the lowest species richness (2 ± 0.61 SD per gram dry weight bryophyte), and density (2.1 ± 0.61 SD individual per gram of dry weight bryophyte); whereas Thuidium urceolatum (high roughness) had the highest richness (24 ± 10.82 SD) and density (94 ± 64.30 SD). The fact that the bryophyte with the highest roughness had the highest

  20. Molecular Detection of Legionella spp. and their associations with Mycobacterium spp., Pseudomonas aeruginosa and amoeba hosts in a drinking water distribution system

    Science.gov (United States)

    Quantity of Legionella spp., Mycobacterium spp., Acanthamoeba,Vermamoeba vermiformis and Pseudomonas aeruginosa were estimated using qPCR methods.This dataset is associated with the following publication:Lu , J., I. Struewing, E. Vereen, A.E. Kirby, K. Levy, C. Moe, and N. Ashbolt. Molecular detection of Legionella spp. and their associations with Mycobacterium spp., Pseudomonas aeruginosa and amoeba hosts in a drinking water distribution system (Journal Article). JOURNAL OF APPLIED MICROBIOLOGY. Blackwell Publishing, Malden, MA, USA, 120(2): 509-521, (2016).

  1. Squamamoeba japonica n. g. n. sp. (Amoebozoa): a deep-sea amoeba from the Sea of Japan with a novel cell coat structure.

    Science.gov (United States)

    Kudryavtsev, Alexander; Pawlowski, Jan

    2013-01-01

    Squamamoeba japonica n. g. n. sp. was isolated and described from marine bottom sediments collected at a depth of ca. 2700 m in the Sea of Japan. Trophic amoebae of this species are elongated and flattened, with a wide anterior hyaloplasm producing numerous ventral subpseudopodia for adhesion to the substratum. The cell coat consists of flat oval scales tightly packed together to form a continuous layer separated from the plasma membrane. Amoebae can form cytoplasmic projections protruding through the scale layer and having tips covered only with the plasma membrane. Small subunit ribosomal RNA gene phylogeny shows that S. japonica forms a long branch in the amoebozoan tree, robustly grouping with the marine strain 'Pessonella' sp. PRA-29. Morphological data available for the latter, although scarce, give additional support for the relatedness of both species. The resulting clade comprising the two taxa shows no close relationships to other Amoebozoa and seems to be a novel lineage that developed an ability to temporarily liberate local areas of the plasma membrane from the cell coat independently from Himatismenida, Trichosida, Pellitida and Dermamoeba. Copyright © 2012 Elsevier GmbH. All rights reserved.

  2. MicroRNAs in Amoebozoa: deep sequencing of the small RNA population in the social amoeba Dictyostelium discoideum reveals developmentally regulated microRNAs.

    Science.gov (United States)

    Avesson, Lotta; Reimegård, Johan; Wagner, E Gerhart H; Söderbom, Fredrik

    2012-10-01

    The RNA interference machinery has served as a guardian of eukaryotic genomes since the divergence from prokaryotes. Although the basic components have a shared origin, silencing pathways directed by small RNAs have evolved in diverse directions in different eukaryotic lineages. Micro (mi)RNAs regulate protein-coding genes and play vital roles in plants and animals, but less is known about their functions in other organisms. Here, we report, for the first time, deep sequencing of small RNAs from the social amoeba Dictyostelium discoideum. RNA from growing single-cell amoebae as well as from two multicellular developmental stages was sequenced. Computational analyses combined with experimental data reveal the expression of miRNAs, several of them exhibiting distinct expression patterns during development. To our knowledge, this is the first report of miRNAs in the Amoebozoa supergroup. We also show that overexpressed miRNA precursors generate miRNAs and, in most cases, miRNA* sequences, whose biogenesis is dependent on the Dicer-like protein DrnB, further supporting the presence of miRNAs in D. discoideum. In addition, we find miRNAs processed from hairpin structures originating from an intron as well as from a class of repetitive elements. We believe that these repetitive elements are sources for newly evolved miRNAs.

  3. Protein migration from transplanted nuclei in Amoeba proteus. I. The relation to the cell cycle and RNA migration, as studied by autoradiography

    Energy Technology Data Exchange (ETDEWEB)

    Mills, K.I.; Bell, L.G.

    1982-11-01

    Autoradiography has been used to examine the migration of proteins from a radioactivity labelled amoeba nucleus following transplantation into an unlabelled homophasic amoeba. Nuclei were transferred at three times in the cell cycle coinciding with DNA synthesis (4 h post-division); a peak of RNA synthesis (25 h); and a relative lull in synthetic activity (43 h). Six amino acids were added individually to the culture medium to label the nuclear proteins. Migration of the proteins from the donor nucleui and least with proteins labelled with the basic amino acids. All amino acids exhibited the greatest extent of migration following the 25-h transfers, i.e., coinciding with a peak of RNA synthesis at 26-27.5 h. Actinomycin D (actD) inhibition of RNA synthesis reduced, but did not eliminate the extent of protein migration from the transplanted nucleus, thus indicating the existence of two classes of migratory proteins. Firstly, proteins, associated with RNA transport, which migrated mainly into the host cytoplasm. The second class migrated into the host nucleus from the transplanted nucleus, irrespective of RNA synthesis. The shuttling character of the latter class of proteins is consistent with a role of regulation of nuclear activity.

  4. Response of sphagnum peatland testate amoebae to a 1-year transplantation experiment along an artificial hydrological gradient.

    Science.gov (United States)

    Marcisz, Katarzyna; Fournier, Bertrand; Gilbert, Daniel; Lamentowicz, Mariusz; Mitchell, Edward A D

    2014-05-01

    Peatland testate amoebae (TA) are well-established bioindicators for depth to water table (DWT), but effects of hydrological changes on TA communities have never been tested experimentally. We tested this in a field experiment by placing Sphagnum carpets (15 cm diameter) collected in hummock, lawn and pool microsites (origin) at three local conditions (dry, moist and wet) using trenches dug in a peatland. One series of samples was seeded with microorganism extract from all microsites. TA community were analysed at T0: 8-2008, T1: 5-2009 and T2: 8-2009. We analysed the data using conditional inference trees, principal response curves (PRC) and DWT inferred from TA communities using a transfer function used for paleoecological reconstruction. Density declined from T0 to T1 and then increased sharply by T2. Species richness, Simpson diversity and Simpson evenness were lower at T2 than at T0 and T1. Seeded communities had higher species richness in pool samples at T0. Pool samples tended to have higher density, lower species richness, Simpson diversity and Simpson Evenness than hummock and/or lawn samples until T1. In the PRC, the effect of origin was significant at T0 and T1, but the effect faded away by T2. Seeding effect was strongest at T1 and lowest vanished by T2. Local condition effect was strong but not in line with the wetness gradient at T1 but started to reflect it by T2. Likewise, TA-inferred DWT started to match the experimental conditions by T2, but more so in hummock and lawn samples than in pool samples. This study confirmed that TA responds to hydrological changes over a 1-year period. However, sensitivity of TA to hydrological fluctuations, and thus the accuracy of inferred DWT changes, was habitat specific, pool TA communities being least responsive to environmental changes. Lawns and hummocks may be thus better suited than pools for paleoecological reconstructions. This, however, contrasts with the higher prediction error and species' tolerance for

  5. Sphagnum-dwelling testate amoebae in subarctic bogs are more sensitive to soil warming in the growing season than in winter: the results of eight-year field climate manipulations.

    NARCIS (Netherlands)

    Tsyganov, A.N.; Aerts, R.; Nijs, I.; Cornelissen, J.H.C.; Beyens, L.

    2012-01-01

    Sphagnum-dwelling testate amoebae are widely used in paleoclimate reconstructions as a proxy for climate-induced changes in bogs. However, the sensitivity of proxies to seasonal climate components is an important issue when interpreting proxy records. Here, we studied the effects of summer warming,

  6. The Bionic Amoeba.

    Science.gov (United States)

    Wright, Emmett L.

    1979-01-01

    A demonstration is described that encourages students to engage in inquiry in biology. Using chemicals and an overhead projector, the instructor can simulate a living organism projected onto a screen. The reaction can aid students in defining the characteristics of life. (SA)

  7. [i]Legionella spp[/i]., amoebae and not-fermenting Gram negative bacteria in an Italian university hospital water system

    Directory of Open Access Journals (Sweden)

    Pasqualina Laganà

    2014-09-01

    Full Text Available [b]Introduction. [/b]In hospital and other health care facilities, contamination of water systems by potentially infectious microorganisms, such as bacteria, viruses and protozoa, is a source of nosocomial infections, which may originate fromcolonization of water pipes, cooling towers, spa pools, taps, showers and water supplies. [b]Objective. [/b]The study focuses on the occurrence of [i]Legionella spp.[/i], free-living amoebae and non-fermenting Gram-negative microorganisms in a University hospital water system located in the town of Messina (Sicily, Italy, which had never been examined previously. Materials and Methods. From January 2008 – March 2009, hot tap water samples were collected from 10 wards.[i] Legionella spp[/i]. recovered on selective culture medium were identified by microagglutination latex test; free-living amoebae were cultured using [i]Escherichia coli [/i]as a food source. Non-fermenting Gram negative microorganisms were identified by API 20 NE strips. [b]Results.[/b] [i]Legionella spp.[/i] were found in 33.33% of the samples. [i]L. pneumophila[/i] serogroup 1 was recovered from the Laboratory Diagnostic and Anaesthesia-Neurology Wards, with a peak of 3.5 × 10[sup]4[/sup] cfu/L in May 2008. [i]L. pneumophila[/i] serogroups 2–14 were found in the Othorhinolaryngology, Pathologic Anatomy, Paediatrics and Surgery Wards, and peaked (4 × 10[sup]4[/sup] cfu/L in April 2008. Pseudomonadaceae and Hyphomycetes were also detected. Legionella spp. were recovered from samples positive for non-pathogenic amoebae [i]Hartmannella spp[/i]. [b]Conclusion.[/b] This first study of a Messina hospital water system suggested potential health risks related to the detection of [i]Hartmannella spp[/i]., as reservoirs for[i] Legionella spp.[/i], and Pseudomonas aeruginosa, a Gram negative non-fermenting bacterium frequently causing nosocomial pneumonia. The urgent need for monitoring programmes and prevention measures to ensure hospital water

  8. Species richness of testate amoebae in different environments from the upper Paraná river floodplain (PR/MS - doi: 10.4025/actascibiolsci.v33i3.7261 Species richness of testate amoebae in different environments from the upper Paraná river floodplain (PR/MS - doi: 10.4025/actascibiolsci.v33i3.7261

    Directory of Open Access Journals (Sweden)

    Fábio Amodêo Lansac-Tôha

    2011-07-01

    Full Text Available This study evaluated the species richness of testate amoebae in the plankton from different environments of the upper Paraná river floodplain. Samplings were performed at subsurface of pelagic region from twelve environments using motorized pump and plankton net (68 µm, during four hydrological periods. We identified 67 taxa, distributed in seven families and Arcellidae, Difflugiidae and Centropyxidae were the most representative families. Higher values of species richness were observed in the lakes (connected and isolated during the flood pulses. Centropyxis aculeata, Difflugia gramem and D. pseudogramem were frequent throughout the study period. Seasonal variability of species in the channels and isolated lakes was evidenced by beta diversity. Besides that, in the rivers, extreme changes in species composition were verified during the high-water period. Our results highlight the importance of the present study to improve the knowledge about the diversity and geographic distribution of these organisms in Brazil and emphasize the importance of current flow in the displacement of testate amoebae from their preferred habitats, marginal vegetation and sediment.This study evaluated the species richness of testate amoebae in the plankton from different environments of the upper Paraná river floodplain. Samplings were performed at subsurface of pelagic region from twelve environments using motorized pump and plankton net (68 µm, during four hydrological periods. We identified 67 taxa, distributed in seven families and Arcellidae, Difflugiidae and Centropyxidae were the most representative families. Higher values of species richness were observed in the lakes (connected and isolated during the flood pulses. Centropyxis aculeata, Difflugia gramem and D. pseudogramem were frequent throughout the study period. Seasonal variability of species in the channels and isolated lakes was evidenced by beta diversity. Besides that, in the rivers, extreme

  9. Detection of the free living amoeba Naegleria fowleri by using conventional and real-time PCR based on a single copy DNA sequence.

    Science.gov (United States)

    Régoudis, Estelle; Pélandakis, Michel

    2016-02-01

    The amoeba-flagellate Naegleria fowleri is a causative agent of primary amoebic meningoencephalitis (PAM). This thermophilic species occurs worldwide and tends to proliferate in warm aquatic environment. The PAM cases remain rare but this infection is mostly fatal. Here, we describe a single copy region which has been cloned and sequenced, and was used for both conventional and real-time PCR. Targeting a single-copy DNA sequence allows to directly quantify the N. fowleri cells. The real-time PCR results give a detection limit of 1 copy per reaction with high reproducibility without the need of a Taqman probe. This procedure is of interest as compared to other procedures which are mostly based on the detection of multi-copy DNA associated with a Taqman probe. Copyright © 2015 Elsevier Inc. All rights reserved.

  10. Mycobacterium bovis hosted by free-living-amoebae permits their long-term persistence survival outside of host mammalian cells and remain capable of transmitting disease to mice.

    Science.gov (United States)

    Sanchez-Hidalgo, Andrea; Obregón-Henao, Andrés; Wheat, William H; Jackson, Mary; Gonzalez-Juarrero, Mercedes

    2017-10-01

    Bovine tuberculosis (TB) is a zoonotic disease caused by Mycobacterium bovis. Despite intensive TB control campaigns, there are sporadic outbreaks of bovine TB in regions declared TB free. It is unclear how M. bovis is able to survive in the environment for long periods of time. We hypothesized that Free-living amoebae (FLA), as ubiquitous inhabitants of soil and water, may act as long-term reservoirs of M. bovis in the environment. In our model, M. bovis would be taken up by amoebal trophozoites, which are the actively feeding, replicating and mobile form of FLA. Upon exposure to hostile environmental conditions, infected FLA will encyst and provide an intracellular niche allowing their M. bovis cargo to persist for extended periods of time. Here, we show that five FLA species (Acanthamoeba polyphaga, Acanthamoeba castellanii, Acanthamoeba lenticulata, Vermamoeba vermiformis and Dictyostellium discoideum) are permissive to M. bovis infection and that the M. bovis bacilli may survive within the cysts of four of these species for over 60 days. We further show that exposure of M. bovis-infected trophozoites and cysts to Balb/c mice leads to pulmonary TB. This work describes for the first time that FLA carrying M. bovis can transmit TB. © 2017 Society for Applied Microbiology and John Wiley & Sons Ltd.

  11. Loss of Cln3 Function in the Social Amoeba Dictyostelium discoideum Causes Pleiotropic Effects That Are Rescued by Human CLN3

    Science.gov (United States)

    Huber, Robert J.

    2014-01-01

    The neuronal ceroid lipofuscinoses (NCL) are a group of inherited, severe neurodegenerative disorders also known as Batten disease. Juvenile NCL (JNCL) is caused by recessive loss-of-function mutations in CLN3, which encodes a transmembrane protein that regulates endocytic pathway trafficking, though its primary function is not yet known. The social amoeba Dictyostelium discoideum is increasingly utilized for neurological disease research and is particularly suited for investigation of protein function in trafficking. Therefore, here we establish new overexpression and knockout Dictyostelium cell lines for JNCL research. Dictyostelium Cln3 fused to GFP localized to the contractile vacuole system and to compartments of the endocytic pathway. cln3− cells displayed increased rates of proliferation and an associated reduction in the extracellular levels and cleavage of the autocrine proliferation repressor, AprA. Mid- and late development of cln3− cells was precocious and cln3− slugs displayed increased migration. Expression of either Dictyostelium Cln3 or human CLN3 in cln3− cells suppressed the precocious development and aberrant slug migration, which were also suppressed by calcium chelation. Taken together, our results show that Cln3 is a pleiotropic protein that negatively regulates proliferation and development in Dictyostelium. This new model system, which allows for the study of Cln3 function in both single cells and a multicellular organism, together with the observation that expression of human CLN3 restores abnormalities in Dictyostelium cln3− cells, strongly supports the use of this new model for JNCL research. PMID:25330233

  12. Loss of Cln3 function in the social amoeba Dictyostelium discoideum causes pleiotropic effects that are rescued by human CLN3.

    Directory of Open Access Journals (Sweden)

    Robert J Huber

    Full Text Available The neuronal ceroid lipofuscinoses (NCL are a group of inherited, severe neurodegenerative disorders also known as Batten disease. Juvenile NCL (JNCL is caused by recessive loss-of-function mutations in CLN3, which encodes a transmembrane protein that regulates endocytic pathway trafficking, though its primary function is not yet known. The social amoeba Dictyostelium discoideum is increasingly utilized for neurological disease research and is particularly suited for investigation of protein function in trafficking. Therefore, here we establish new overexpression and knockout Dictyostelium cell lines for JNCL research. Dictyostelium Cln3 fused to GFP localized to the contractile vacuole system and to compartments of the endocytic pathway. cln3- cells displayed increased rates of proliferation and an associated reduction in the extracellular levels and cleavage of the autocrine proliferation repressor, AprA. Mid- and late development of cln3- cells was precocious and cln3- slugs displayed increased migration. Expression of either Dictyostelium Cln3 or human CLN3 in cln3- cells suppressed the precocious development and aberrant slug migration, which were also suppressed by calcium chelation. Taken together, our results show that Cln3 is a pleiotropic protein that negatively regulates proliferation and development in Dictyostelium. This new model system, which allows for the study of Cln3 function in both single cells and a multicellular organism, together with the observation that expression of human CLN3 restores abnormalities in Dictyostelium cln3- cells, strongly supports the use of this new model for JNCL research.

  13. Free-living amoebae (FLA: detection, morphological and molecular identification of Acanthamoeba genus in the hydraulic system of an haemodialysis unit in Tunisia

    Directory of Open Access Journals (Sweden)

    Dendana F.

    2008-06-01

    Full Text Available The free-living amoebae (FLA are ubiquitous and opportunistic protozoa. They can induce human and animal diseases. The aim of our study was to detect the FLA and Acanthamoeba genus in the hydraulic system of an hemodialysis unit. It was a prospective study of 46 water samples. The first collect (23 was before cleaning and after the haemodialysis sessions and the second (23 after cleaning and before the hemodialysis sessions. Results: the morphological study enabled us to detect morphotypic diversity. The predominant morphotypes were the acanthopodial forms (29%. At the entrance of hemodialysis unit there were acanthopodial (44% and monotactic (25% forms; at the outlet, acanthopodial and fan-shaped forms (25% each. In addition, Acanthamoeba genus was present in 39% (1st collect and 18% (2nd collect. The amplification of the FLA 18S rDNA gene was negative in only one sample localized in the last stage of water treatment unit (WTU. The amplification of the 18S rDNA (ASA.A1 Acanthamoeba gene was positive in15 samples. Conclusions: we noted that, in the hemodialysis unit, the purification techniques used in the WTU were effective, but there is a problem of water stagnation in the drain, which constitutes an appropriate condition for the biofilms formation. It is then necessary to use a filter with a low porosity (0.2 μm at the entrance of the hemodialysis unit and if possible to change the drain

  14. Antibiotic-resistant heterotrophic plate count bacteria and amoeba-resistant bacteria in aquifers of the Mooi River, North West province, South Africa.

    Science.gov (United States)

    Carstens, Alewyn; Bartie, Catheleen; Dennis, Rainier; Bezuidenhout, Carlos

    2014-12-01

    Groundwater in the Mooi River catchment is prone to mining, agricultural, municipal and septic tank pollution. In this study physico-chemical and microbiological parameters were determined using appropriate methods. Bacterial isolates were identified by 16S rRNA sequencing (heterotrophic plate count (HPC) bacteria and amoeba-resistant bacteria (ARB)) and multiplex polymerase chain reaction (Escherichia coli). Antibiotic resistance tests were also performed. Physico-chemical parameters were generally within target water quality ranges for drinking water. HPC bacteria ranged between 10(5) and 10(7) colony-forming units (cfu)/ml. E. coli were enumerated from Trimpark, School and Cemetery. The Blaauwbank borehole was negative for faecal streptococci. Pseudomonas spp. were most abundant in the bulk water. Opportunistic pathogens isolated included Pseudomonas aeruginosa, Acinetobacter, Aeromonas, Alcaligenes, Flavobacterium, Bacillus cereus and Mycobacterium spp. Varying patterns of antibiotic resistance were observed. Most HPC bacterial isolates were resistant to cephalothin and/or amoxicillin and a few were resistant to erythromycin and streptomycin. Pseudomonas spp. was also the most abundant ARB. Other ARBs included Alcaligenes faecalis, Ochrobactrum sp. and Achromobacter sp. ARBs were resistant to streptomycin, chloramphenicol, cephalothin, and/or amoxicillin compared to HPCs. The presence of E. coli and ARB in these groundwater sources indicates potential human health risks. These risks should be further investigated and quantified, and groundwater should be treated before use.

  15. The novel Legionella pneumophila type II secretion substrate NttC contributes to infection of amoebae Hartmannella vermiformis and Willaertia magna.

    Science.gov (United States)

    Tyson, Jessica Y; Vargas, Paloma; Cianciotto, Nicholas P

    2014-12-01

    The type II protein secretion (T2S) system of Legionella pneumophila secretes over 25 proteins, including novel proteins that have no similarity to proteins of known function. T2S is also critical for the ability of L. pneumophila to grow within its natural amoebal hosts, including Acanthamoeba castellanii, Hartmannella vermiformis and Naegleria lovaniensis. Thus, T2S has an important role in the natural history of legionnaires' disease. Our previous work demonstrated that the novel T2S substrate NttA promotes intracellular infection of A. castellanii, whereas the secreted RNase SrnA, acyltransferase PlaC, and metalloprotease ProA all promote infection of H. vermiformis and N. lovaniensis. In this study, we determined that another novel T2S substrate that is specific to Legionella, designated NttC, is unique in being required for intracellular infection of H. vermiformis but not for infection of N. lovaniensis or A. castellanii. Expanding our repertoire of amoebal hosts, we determined that Willaertia magna is susceptible to infection by L. pneumophila strains 130b, Philadelphia-1 and Paris. Furthermore, T2S and, more specifically, NttA, NttC and PlaC were required for infection of W. magna. Taken together, these data demonstrate that the T2S system of L. pneumophila is critical for infection of at least four types of aquatic amoebae and that the importance of the individual T2S substrates varies in a host cell-specific fashion. Finally, it is now clear that novel T2S-dependent proteins that are specific to the genus Legionella are particularly important for L. pneumophila infection of key, environmental hosts. © 2014 The Authors.

  16. Molecular Detection of Legionella spp. and their associations with Mycobacterium spp., Pseudomonas aeruginosa and amoeba hosts in a drinking water distribution system.

    Science.gov (United States)

    Lu, J; Struewing, I; Vereen, E; Kirby, A E; Levy, K; Moe, C; Ashbolt, N

    2016-02-01

    This study investigated waterborne opportunistic pathogens (OPs) including potential hosts, and evaluated the use of Legionella spp. for indicating microbial water quality for OPs within a full-scale operating drinking water distribution system (DWDS). To investigate the occurrence of specific microbial pathogens within a major city DWDS we examined large volume (90 l drinking water) ultrafiltration (UF) concentrates collected from six sites between February, 2012 and June, 2013. The detection frequency and concentration estimates by qPCR were: Legionella spp. (57%/85 cell equivalent, CE l(-1) ), Mycobacterium spp. (88%/324 CE l(-1) ), Pseudomonas aeruginosa (24%/2 CE l(-1) ), Vermamoeba vermiformis (24%/2 CE l(-1) ) and Acanthamoeba spp. (42%/5 cyst equivalent, CE l(-1) ). There was no detection of the following microorganisms: human faecal indicator Bacteroides (HF183), Salmonella enterica, Campylobacter spp., Escherichia coli O157:H7, Giardia intestinalis, Cryptosporidium spp. or Naegleria fowleri. There were significant correlations between the qPCR signals of Legionella spp. and Mycobacterium spp., and their potential hosts V. vermiformis and Acanthamoeba spp. Sequencing of Legionella spp. demonstrated limited diversity, with most sequences coming from two dominant groups, of which the larger dominant group was an unidentified species. Other known species including Legionella pneumophila were detected, but at low frequency. The densities of Legionella spp. and Mycobacterium spp. were generally higher (17 and 324 folds, respectively) for distal sites relative to the entry point to the DWDS. Legionella spp. occurred, had significant growth and were strongly associated with free-living amoebae (FLA) and Mycobacterium spp., suggesting that Legionella spp. could provide a useful DWDS monitoring role to indicate potential conditions for non-faecal OPs. The results provide insight into microbial pathogen detection that may aid in the monitoring of microbial water

  17. Prevalence of pathogenic free-living amoeba and other protozoa in natural and communal piped tap water from Queen Elizabeth protected area, Uganda.

    Science.gov (United States)

    Sente, Celsus; Erume, Joseph; Naigaga, Irene; Mulindwa, Julius; Ochwo, Sylvester; Magambo, Phillip Kimuda; Namara, Benigna Gabriela; Kato, Charles Drago; Sebyatika, George; Muwonge, Kevin; Ocaido, Michael

    2016-08-03

    Pathogenic water dwelling protozoa such as Acanthamoeba spp., Hartmannella spp., Naegleria spp., Cryptosporidium spp. and Giardia spp. are often responsible for devastating illnesses especially in children and immunocompromised individuals, yet their presence and prevalence in certain environment in sub-Saharan Africa is still unknown to most researchers, public health officials and medical practitioners. The objective of this study was to establish the presence and prevalence of pathogenic free-living amoeba (FLA), Cryptosporidium and Giardia in Queen Elizabeth Protected Area (QEPA). Samples were collected from communal taps and natural water sites in QEPA. Physical water parameters were measured in situ. The samples were processed to detect the presence of FLA trophozoites by xenic cultivation, Cryptosporidium oocysts by Ziehl-Neelsen stain and Giardia cysts by Zinc Sulphate floatation technique. Parasites were observed microscopically, identified, counted and recorded. For FLA, genomic DNA was extracted for amplification and sequencing. Both natural and tap water sources were contaminated with FLA, Cryptosporidium spp. and Giardia spp. All protozoan parasites were more abundant in the colder rainy season except for Harmannella spp. and Naegleria spp. which occurred more in the warmer months. The prevalence of all parasites was higher in tap water than in natural water samples. There was a strong negative correlation between the presence of Acanthamoeba spp., Hartmannella spp., Cryptosporidium spp. and Giardia spp. with Dissolved Oxygen (DO) (P protozoa that could possibly be the cause of a number of silent morbidities and mortalities among rural households in QEPA. This implies that water used by communities in QEPA is of poor quality and predisposes them to a variety of protozoan infections including the FLA whose public health importance was never reported, thus necessitating adoption of proper water safety measures.

  18. Holocene palaeohydrological history of the Tǎul Muced peat bog (Northern Carpathians, Romania) based on testate amoebae (Protozoa) and plant macrofossils

    Science.gov (United States)

    Cosmin Diaconu, Andrei; Feurdean, Angelica; Lamentowicz, Mariusz; Gałka, Mariusz; Tanţǎu, Ioan

    2016-04-01

    Knowledge of past local vs. regional hydro-climate variability is a priority in climate research. This is because ecosystems and human depend on local climatic conditions and the magnitude of these climate changes is more variable at local and regional rather than at global scales. Ombrotrophic bogs are highly suitable for hydro-climate reconstructions as they are entirely dependent on the water from precipitation. We used stratigraphy, radiocarbon dating, testate amoebae (TA) and plant macrofossils on a peat profile from an ombrotrophic bog (Tǎul Muced) located in the Biosphere Reserve of the Rodna National Park Romania. We performed quantitative reconstruction of the depth to water table (DWT) and pH over the last 8000 years in a continental area of CE Europe. We identified six main stages in the development of the bog based on changes in TA assemblages in time. Wet conditions and pH between 2 and 4.5 were recorded between 4600-2750 and 1300-400 cal. yr BP, by the occurrence of Archerella flavum, Amphitrema wrightianum and Hyalosphenia papilio. This was associated to a local vegetation primarily composed of Sphagnum magellanicum and S. angustifolium. Dry stages and pH of 2.5 to 5 were inferred between 7550-4600, 2750-1300 and -50 cal. yr BP, by the dominance of Nebela militaris, Difflugia pulex and Phryganella acropodia. These overall dry conditions were also connected with increased abundance of Eriophorum vaginatum. The period between 400 and -50 cal. yr BP was characterized by a rapid shift from dry to wet conditions on the surface of the bog. Vegetation shifted from Sphagnum magellanicum to Sphagnum russowii dominated community. Our reconstruction remains in relatively good agreement with other palaeohydrological records from Central Eastern Europe. However, it shows contrasting conditions to others particularly with records from NW Europe. The valuable information regarding bog hydrology offered by our record puts an accent on the need of more regional TA

  19. Species richness of testate amoebae in different environments from the upper Paraná river floodplain (PR/MS) = Riqueza de amebas testáceas em diferentes ambientes da planície de inundação do alto rio Paraná (PR/MS)

    OpenAIRE

    Deise de Morais Costa; Geziele Mucio Alves; Luiz Felipe Machado Velho; Fábio Amodêo Lansac-Tôha

    2011-01-01

    This study evaluated the species richness of testate amoebae in the plankton from different environments of the upper Parana river floodplain. Samplings were performed at subsurface of pelagic region from twelve environments using motorized pump and plankton net (68 ƒÝm), during four hydrological periods. We identified 67 taxa, distributed in seven families and Arcellidae, Difflugiidae and Centropyxidae were the most representative families. Higher values of species richness were observed in ...

  20. Giant viruses of amoebas: an update

    Directory of Open Access Journals (Sweden)

    Sarah eAherfi

    2016-03-01

    Full Text Available During the 12 past years, five new or putative virus families encompassing several members, namely Mimiviridae, Marseilleviridae, pandoraviruses, faustoviruses, and virophages were described. In addition, Pithovirus sibericum and Mollivirus sibericum represent type strains of putative new giant virus families. All these viruses were isolated using amoebal coculture methods. These giant viruses were linked by phylogenomic analyses to other large DNA viruses. They were then proposed to be classified in a new viral order, the Megavirales, on the basis of their common origin, as shown by a set of ancestral genes encoding key viral functions, a common virion architecture, and shared major biological features including replication inside cytoplasmic factories. Megavirales is increasingly demonstrated to stand in the tree of life aside Bacteria, Archaea and Eukarya, and the megavirus ancestor is suspected to be as ancient as cellular ancestors. In addition, giant amoebal viruses are visible under a light microscope and display many phenotypic and genomic features not found in other viruses, while they share other characteristics with parasitic microbes. Moreoever, these organisms appear to be common inhabitants of our biosphere, and mimiviruses and marseilleviruses were isolated from human samples and associated to diseases. In the present review, we describe the main features and recent findings on these giant amoebal viruses and virophages.

  1. The multicellularity genes of dictyostelid social amoebas

    Science.gov (United States)

    Glöckner, Gernot; Lawal, Hajara M.; Felder, Marius; Singh, Reema; Singer, Gail; Weijer, Cornelis J.; Schaap, Pauline

    2016-01-01

    The evolution of multicellularity enabled specialization of cells, but required novel signalling mechanisms for regulating cell differentiation. Early multicellular organisms are mostly extinct and the origins of these mechanisms are unknown. Here using comparative genome and transcriptome analysis across eight uni- and multicellular amoebozoan genomes, we find that 80% of proteins essential for the development of multicellular Dictyostelia are already present in their unicellular relatives. This set is enriched in cytosolic and nuclear proteins, and protein kinases. The remaining 20%, unique to Dictyostelia, mostly consists of extracellularly exposed and secreted proteins, with roles in sensing and recognition, while several genes for synthesis of signals that induce cell-type specialization were acquired by lateral gene transfer. Across Dictyostelia, changes in gene expression correspond more strongly with phenotypic innovation than changes in protein functional domains. We conclude that the transition to multicellularity required novel signals and sensors rather than novel signal processing mechanisms. PMID:27357338

  2. Efecto de la estructuración por macrófitas y por recursos alimentarios en la distribución horizontal de tecamebas y rotíferos en un lago andino patagónico Effect of macrophytes and food resources on the horizontal distribution of testate amoebae and rotifers in an Andean-Patagonian lake

    Directory of Open Access Journals (Sweden)

    MARCELA BASTIDAS-NAVARRO

    2007-09-01

    con el aumento de la abundancia de la cianofita Coelosphaerium kuetzingianum. Las diferencias señaladas indican que los recursos alimentarios serían un factor determinante en la distribución de especies de tecamebas y rotíferos en el lago Escondido. Para estos zoopláncteres de pequeño tamaño las macrófitas litorales brindarían alimento al favorecer el incremento del fitoplancton de redThe presence of macrophytes in the littoral zone of lakes produces particular conditions including higher resource availability for consumers. For this reason, the littoral zone is generally the area with the highest diversity of lakes and rivers. In this work we studied the horizontal distribution of testate amoebae and rotifers in Lago Escondido (Argentina in relation to food resources availability. The study was carried out along a north-south transect that includes the littoral and the pelagic zone of the lake. Phytoplankton and zooplankton were sampled during summer and spring (2001-2003 in five sampling stations: one pelagic and four littoral. Rotifers and testate amoebae, as well as phytoplanktonic algae abundance and biomass were estimated. Food resources were classified as nanoplankton ( 20 μm and the biovolume of these fractions varied significantly within the pelagic and littoral zones of the lake. Nanoplankon dominated the pelagic zone and was mainly composed by nanoflagellates while net phytoplankton prevailed in the littoral zone and was composed by diatoms, cyanophytes and chlorophytes. The highest number of species and diversity of testate amoebae and rotifers were observed in the littoral stations; nevertheless, no significant differences between the zones colonized by different macrophytes were observed. The CCA analysis showed four different groups. Keratella cochlearis, Synchaeta spp., Polyarthra vulgaris and Collotheca mutabilis characterized the pelagic samples and were related with a high abundance of nanoplankton. On the other hand, rotifers like

  3. Species richness and geographic distribution of testate amoebae (Rhizopoda in Brazilian freshwater environments = Riqueza de espécies e distribuição geográfica de amebas testáceas (Rhizopoda em ambientes aquáticos continentais brasileiros

    Directory of Open Access Journals (Sweden)

    Fábio Amodêo Lansac-Tôha

    2007-04-01

    Full Text Available The present study aimed to analyze the species richness of testate amoebae, as well as to describe their geographic distribution and different habitat types in Brazilian freshwater environments. Until now, 346 infrageneric taxa have been recorded, belonging to 13 families and 40 genera. In the Center-West region, 267 taxa were recorded; 188 taxa in the Southeast; 129 taxa in the South, 53 taxa in the North; and 18 taxa in Northeast region. A total of 282 taxa were recorded in plankton; 80 taxa in aquatic macrophytes; 81 taxa insediment; and 73 taxa in moss/sphagnum. The results regarding testate amoebae species richness are not yet conclusive, given that most research on these organisms was carried out in central, southeastern and southern Brazil. The higher number of taxa observed in plankton may be due to the fact that most studies on testate amoebae in Brazil had been carried out in the planktonic compartment, including reservoirs, floodplain lagoons, channels, tributaries and rivers. In addition, the majority of studies with sediment samples were conducted in estuaries or coastal lagoons, where salinity is a restricting factor for the occurrence of these organisms.Este trabalho visa realizar um levantamento da riqueza de espécies de amebas testáceas, bem como descrever a distribuição geográfica e a distribuição em diferentes tipos de habitats desses organismos em ambientes aquáticos continentais brasileiros. Até o momento, são registrados 346 táxons infragenéricos, pertencentes a 13 famílias e 40 gêneros. São registrados 267 táxons na região centro-oeste, 188táxons na região sudeste, 129 táxons na região sul, 53 táxons na região norte e 18 táxons na região nordeste. São registrados 282 táxons no plâncton; 131 táxons em macrófitas aquáticas, 81 táxonsno sedimento e 73 táxons em musgos/esfágnos. Os resultados sobre a riqueza de tecamebas não é ainda conclusiva, visto que a maioria das pesquisas foi conduzida

  4. Species richness of testate amoebae in different environments from the upper Paraná river floodplain (PR/MS = Riqueza de amebas testáceas em diferentes ambientes da planície de inundação do alto rio Paraná (PR/MS

    Directory of Open Access Journals (Sweden)

    Deise de Morais Costa

    2011-07-01

    Full Text Available This study evaluated the species richness of testate amoebae in the plankton from different environments of the upper Parana river floodplain. Samplings were performed at subsurface of pelagic region from twelve environments using motorized pump and plankton net (68 ƒÝm, during four hydrological periods. We identified 67 taxa, distributed in seven families and Arcellidae, Difflugiidae and Centropyxidae were the most representative families. Higher values of species richness were observed in the lakes (connected and isolated during the flood pulses. Centropyxis aculeata, Difflugia gramem and D.pseudogramem were frequent throughout the study period. Seasonal variability of species in the channels and isolated lakes was evidenced by beta diversity. Besides that, in the rivers, extreme changes in species composition were verified during the high-water period. Our results highlight the importance of the present study to improve the knowledge about the diversity and geographic distribution of these organisms in Brazil and emphasize the importance of current flow in the displacement of testate amoebae from their preferredhabitats, marginal vegetation and sediment.Este estudo objetivou avaliar a riqueza de taxons de amebas testaceas no plancton de diferentes ambientes da planicie de inundacao do alto rio Parana. Foram amostrados 12 pontos da regiao pelagica em diferentes ambientes (rios, canais e lagoas e em quatro periodos hidrologicos. Foram identificados 67 taxons, distribuidos em sete familias. Arcellidae, Difflugiidae e Centropyxidae foram as familiasmais especiosas. Nas lagoas (abertas e fechadas, durante os pulsos de inundacao, foram observados os maiores valores para a riqueza de especies. Centropyxis aculeata, Difflugia gramem e D. pseudogramem foram frequentes durante todo o periodo estudado. Os dados obtidos pela diversidade beta evidenciaram a variabilidade sazonal das especies nos canais e lagoas fechadas. Nos rios, as alteracoes

  5. The Amoeba Distributed Operating System - A Status Report

    NARCIS (Netherlands)

    Tanenbaum, A.S.; Kaashoek, M.F.; van Renesse, R.; Bal, H.E.

    1991-01-01

    As the price of CPU chips continues to fall rapidly, it will soon be economically feasible to build computer systems containing a large number of processors. The question of how this computing power should be organized, and what kind of operating system is appropriate then arises. Our research

  6. Phylogeny and Systematics of Leptomyxid Amoebae (Amoebozoa, Tubulinea, Leptomyxida)

    NARCIS (Netherlands)

    Smirnov, Alexey; Nassonova, Elena; Geisen, Stefan; Bonkowski, Michael; Kudryavtsev, Alexander; Berney, Cedric; Glotova, Anna; Bondarenko, Natalya; Dyková, Iva; Mrva, Martin; Fahrni, Jose; Pawlowski, Jan

    2017-01-01

    We describe four new species of Flabellula, Leptomyxa and Rhizamoeba and publish new SSU rRNA gene and actin gene sequences of leptomyxids. Using these data we provide the most comprehensive SSU phylogeny of leptomyxids to date. Based on the analyses of morphological data and results of the SSU rRNA

  7. Phylogeny and Systematics of Leptomyxid Amoebae (Amoebozoa, Tubulinea, Leptomyxida).

    Science.gov (United States)

    Smirnov, Alexey; Nassonova, Elena; Geisen, Stefan; Bonkowski, Michael; Kudryavtsev, Alexander; Berney, Cedric; Glotova, Anna; Bondarenko, Natalya; Dyková, Iva; Mrva, Martin; Fahrni, Jose; Pawlowski, Jan

    2017-04-01

    We describe four new species of Flabellula, Leptomyxa and Rhizamoeba and publish new SSU rRNA gene and actin gene sequences of leptomyxids. Using these data we provide the most comprehensive SSU phylogeny of leptomyxids to date. Based on the analyses of morphological data and results of the SSU rRNA gene phylogeny we suggest changes in the systematics of the order Leptomyxida (Amoebozoa: Lobosa: Tubulinea). We propose to merge the genera Flabellula and Paraflabellula (the genus Flabellula remains valid by priority rule). The genus Rhizamoeba is evidently polyphyletic in all phylogenetic trees; we suggest retaining the generic name Rhizamoeba for the group unifying R. saxonica, R.matisi n. sp. and R. polyura, the latter remains the type species of the genus Rhizamoeba. Based on molecular and morphological evidence we move all remaining Rhizamoeba species to the genus Leptomyxa. New family Rhizamoebidae is established here in order to avoid paraphyly of the family Leptomyxidae. With the suggested changes both molecular and morphological systems of the order Leptomyxida are now fully congruent to each other. Copyright © 2016 Elsevier GmbH. All rights reserved.

  8. Free-living amoebae isolated in the Central African Republic ...

    African Journals Online (AJOL)

    Bathing sites and cerebrospinal fluid from patients died of bacterial meningitis untagged were explored by culture and PCR and the amplicons were sequenced which allowed to characterize the species found. Only species of the genus Tetramitus, namely T. Entericus, T. waccamawensis and T.sp similar to those already ...

  9. Amoebas and Euglenas and Paramecia...Oh, My!

    Science.gov (United States)

    Farenga, Stephen; Joyce, Beverly

    2000-01-01

    Describes a partnership involving fifth grade students, preservice teachers, a classroom teacher, and a university faculty member in introducing students to the basic anatomy of a cell using three-dimensional models, analogies, and hands-on/minds-on activities. (ASK)

  10. An Overview of the Amoeba Distributed Operating System

    NARCIS (Netherlands)

    Tanenbaum, Andrew S.; Mullender, Sape J.

    As hardware prices continue to drop rapidly, building large computer systems by interconnecting substantial numbers of microcomputers becomes increasingly attractive. Many techniques for interconnecting the hardware, such as Ethernet [Metcalfe and Boggs, 1976], ring nets [Farber and Larson, 1972],

  11. Human infections caused by free-living amoebae

    Directory of Open Access Journals (Sweden)

    Katarzyna Król-Turmińska

    2017-05-01

    Amoebic diseases are difficult to diagnose which leads to delayed treatment, and result in a high mortality rate. Considering those issues, there is an urgent need to draw more attention to this type of diseases.

  12. Regulation of dynamic cyclic nucleotide signalling in social amoebas

    NARCIS (Netherlands)

    Weening, Karin Esther

    2010-01-01

    Over the past years, much information has been gathered about the importance of the individual key components that control cAMP signalling in eukaryotes and prokaryotes. In mammalians, several different classes of ACs produce cAMP and much is known about their mechanism of stimulation and function.

  13. Between a Pod and a Hard Test: The Deep Evolution of Amoebae

    Czech Academy of Sciences Publication Activity Database

    Kang, S.; Tice, A. K.; Spiegel, F. W.; Silberman, J. D.; Pánek, T.; Čepička, I.; Kostka, Martin; Kosakyan, A.; Alcantara, D. M. C.; Roger, A. J.; Shadwick, L.; Smirnov, A.; Kudryavtsev, A.; Lahr, D. J. G.; Brown, M. W.

    2017-01-01

    Roč. 34, č. 9 (2017), s. 2258-2270 ISSN 0737-4038 Institutional support: RVO:60077344 Keywords : phylogenomics * transcriptomes * Amoebozoa * reductive evolution * phylotranscriptomics Subject RIV: EB - Genetics ; Molecular Biology OBOR OECD: Biology (theoretical, mathematical, thermal, cryobiology, biological rhythm), Evolutionary biology Impact factor: 6.202, year: 2016

  14. Environmental Mycobacterium avium subsp. paratuberculosis hosted by free-living amoebae

    Science.gov (United States)

    Mycobacterium avium subsp. paratuberculosis is responsible for paratuberculosis in animals. This disease, leading to an inflammation of the gastrointestinal tract, has a high impact on animal health and an important economic burden. The environmental life cycle of Mycobacterium avium subsp. paratube...

  15. Pseudomonas aeruginosa uses type III secretion system to kill biofilm-associated amoebae

    DEFF Research Database (Denmark)

    Matz, Carsten; Moreno, Ana Maria; Alhede, Morten

    2008-01-01

    Bacteria and protozoa coexist in a wide range of biofilm communities of natural, technical and medical importance. Generally, this interaction is characterized by the extensive grazing activity of protozoa on bacterial prey populations. We hypothesized that the close spatial coexistence in biofil...

  16. Faustovirus, an asfarvirus-related new lineage of giant viruses infecting amoebae.

    Science.gov (United States)

    Reteno, Dorine Gaëlle; Benamar, Samia; Khalil, Jacques Bou; Andreani, Julien; Armstrong, Nicholas; Klose, Thomas; Rossmann, Michael; Colson, Philippe; Raoult, Didier; La Scola, Bernard

    2015-07-01

    Giant viruses are protist-associated viruses belonging to the proposed order Megavirales; almost all have been isolated from Acanthamoeba spp. Their isolation in humans suggests that they are part of the human virome. Using a high-throughput strategy to isolate new giant viruses from their original protozoan hosts, we obtained eight isolates of a new giant viral lineage from Vermamoeba vermiformis, the most common free-living protist found in human environments. This new lineage was proposed to be the faustovirus lineage. The prototype member, faustovirus E12, forms icosahedral virions of ≈ 200 nm that are devoid of fibrils and that encapsidate a 466-kbp genome encoding 451 predicted proteins. Of these, 164 are found in the virion. Phylogenetic analysis of the core viral genes showed that faustovirus is distantly related to the mammalian pathogen African swine fever virus, but it encodes ≈ 3 times more mosaic gene complements. About two-thirds of these genes do not show significant similarity to genes encoding any known proteins. These findings show that expanding the panel of protists to discover new giant viruses is a fruitful strategy. By using Vermamoeba, a protist living in humans and their environment, we isolated eight strains of a new giant virus that we named faustovirus. The genomes of these strains were sequenced, and their sequences showed that faustoviruses are related to but different from the vertebrate pathogen African swine fever virus (ASFV), which belongs to the family Asfarviridae. Moreover, the faustovirus gene repertoire is ≈ 3 times larger than that of ASFV and comprises approximately two-thirds ORFans (open reading frames [ORFs] with no detectable homology to other ORFs in a database).

  17. Between a Pod and a Hard Test: The Deep Evolution of Amoebae.

    Science.gov (United States)

    Kang, Seungho; Tice, Alexander K; Spiegel, Frederick W; Silberman, Jeffrey D; Pánek, Tomáš; Cepicka, Ivan; Kostka, Martin; Kosakyan, Anush; Alcântara, Daniel M C; Roger, Andrew J; Shadwick, Lora L; Smirnov, Alexey; Kudryavtsev, Alexander; Lahr, Daniel J G; Brown, Matthew W

    2017-09-01

    Amoebozoa is the eukaryotic supergroup sister to Obazoa, the lineage that contains the animals and Fungi, as well as their protistan relatives, and the breviate and apusomonad flagellates. Amoebozoa is extraordinarily diverse, encompassing important model organisms and significant pathogens. Although amoebozoans are integral to global nutrient cycles and present in nearly all environments, they remain vastly understudied. We present a robust phylogeny of Amoebozoa based on broad representative set of taxa in a phylogenomic framework (325 genes). By sampling 61 taxa using culture-based and single-cell transcriptomics, our analyses show two major clades of Amoebozoa, Discosea, and Tevosa. This phylogeny refutes previous studies in major respects. Our results support the hypothesis that the last common ancestor of Amoebozoa was sexual and flagellated, it also may have had the ability to disperse propagules from a sporocarp-type fruiting body. Overall, the main macroevolutionary patterns in Amoebozoa appear to result from the parallel losses of homologous characters of a multiphase life cycle that included flagella, sex, and sporocarps rather than independent acquisition of convergent features. © The Author 2017. Published by Oxford University Press on behalf of the Society for Molecular Biology and Evolution.

  18. Monitoring for the Presence of Parasitic Protozoa and Free-living Amoebae in Drinking Water Plants

    Directory of Open Access Journals (Sweden)

    Amany Saad Amer.

    2012-07-01

    Full Text Available Contamination of drinking water by microorganisms represents a major human health hazard in many parts of the world. The main objective of drinking water treatment is to provide microbiologically safe drinking water. The conventional drinking water treatment and disinfection has proved to be one of the major public health advances in modern times. A number of processes; namely water treatment, disinfection and changes influence the quality of drinking water delivered to the customer’s tap during transport of treated water via the distribution system. At least 325 water-associated outbreaks of parasitic protozoan disease have reported. In this study, drinking water from treatment plants evaluated for the presence of parasitic protozoa. Water samples collected from two main points: (a outlet of the water treatment plants (b distribution system at different distances from the water treatment plants. Protozoa were concentrated from each water sample by adsorption and accumulation on the nitrocellulose membrane filters (0.45 μm pore size and detected by conventional staining methods.

  19. Deficiency of huntingtin has pleiotropic effects in the social amoeba Dictyostelium discoideum.

    Directory of Open Access Journals (Sweden)

    Michael A Myre

    2011-04-01

    Full Text Available Huntingtin is a large HEAT repeat protein first identified in humans, where a polyglutamine tract expansion near the amino terminus causes a gain-of-function mechanism that leads to selective neuronal loss in Huntington's disease (HD. Genetic evidence in humans and knock-in mouse models suggests that this gain-of-function involves an increase or deregulation of some aspect of huntingtin's normal function(s, which remains poorly understood. As huntingtin shows evolutionary conservation, a powerful approach to discovering its normal biochemical role(s is to study the effects caused by its deficiency in a model organism with a short life-cycle that comprises both cellular and multicellular developmental stages. To facilitate studies aimed at detailed knowledge of huntingtin's normal function(s, we generated a null mutant of hd, the HD ortholog in Dictyostelium discoideum. Dictyostelium cells lacking endogenous huntingtin were viable but during development did not exhibit the typical polarized morphology of Dictyostelium cells, streamed poorly to form aggregates by accretion rather than chemotaxis, showed disorganized F-actin staining, exhibited extreme sensitivity to hypoosmotic stress, and failed to form EDTA-resistant cell-cell contacts. Surprisingly, chemotactic streaming could be rescued in the presence of the bivalent cations Ca(2+ or Mg(2+ but not pulses of cAMP. Although hd(- cells completed development, it was delayed and proceeded asynchronously, producing small fruiting bodies with round, defective spores that germinated spontaneously within a glassy sorus. When developed as chimeras with wild-type cells, hd(- cells failed to populate the pre-spore region of the slug. In Dictyostelium, huntingtin deficiency is compatible with survival of the organism but renders cells sensitive to low osmolarity, which produces pleiotropic cell autonomous defects that affect cAMP signaling and as a consequence development. Thus, Dictyostelium provides a novel haploid organism model for genetic, cell biological, and biochemical studies to delineate the functions of the HD protein.

  20. Strains of Uronema marinum (Scuticociliatia) co-isolated with amoebae of the genus Neoparamoeba.

    Science.gov (United States)

    Dyková, Iva; Tyml, Tomás; Kostka, Martin; Pecková, Hana

    2010-02-24

    Mixed infections of histophagous ciliates and Neoparamoeba spp. Page, 1987 were diagnosed in gill tissue of farmed turbot Psetta maxima (synonym: Scophthalmus maximus) and Atlantic salmon Salmo salar during a study of amoebic gill disease. Ciliates co-isolated from lesions grossly visible on gills of both fish hosts and from 2 species of red algae Lithophyllum racemus and Palmaria palmata were characterized morphologically and by using molecular markers. Sequences of small subunit (SSU) rDNA were determined for 6 strains of ciliates isolated from hosts collected in geographically distant localities. Of these, sequences of 5 strains revealed a surprisingly high level of similarity and identified the corresponding strains with Uronema marinum Lynn et Small, 1997. Thus, the set of environmental sequences of U. marinum available in the GenBank database to date was supplemented with the first sequences of potentially histophagous strains. On the basis of SSU rDNA, the 6th strain, also isolated from affected fish gills, was identified as Aristerostoma sp.

  1. Strains of Uronema marinum (Scuticociliatia) co-isolated with amoebae of the genus Neoparamoeba

    Czech Academy of Sciences Publication Activity Database

    Dyková, Iva; Tyml, Tomáš; Kostka, Martin; Pecková, Hana

    2010-01-01

    Roč. 89, č. 1 (2010), s. 71-77 ISSN 0177-5103 R&D Projects: GA ČR GA524/09/0137; GA MŠk LC522 Institutional research plan: CEZ:AV0Z60220518 Keywords : Uronema marinum * SSU rDNA * histophagous strains Subject RIV: GJ - Animal Vermins ; Diseases, Veterinary Medicine Impact factor: 1.572, year: 2010

  2. Axenic mass cultivation of the free-living soil amoeba, Acanthamoeba castellanii in a laboratory fermentor.

    Science.gov (United States)

    Weekers, P H; Wijen, J P; Lomans, B P; Vogels, G D

    1996-05-01

    Axenic mass cultivation of Acanthamoeba castellanii in laboratory fermentors (141) yielded after 20 days approximately 3 g cells (wet weight). After a short lag phase amoebal cell numbers increased exponentially to a maximum of 3.5 x 10(5) cells per ml until cell death occurred after 20 days. Optical density and protein concentrations revealed identical patterns. During amoebal growth only 12-19% of the initially added glucose (100 mM) as sole carbon source was used. Large amounts of ammonia (1 g in 10.51 culture volume) were excreted into the medium which subsequently raised the pH from 6.6 to 7.7, and from 6.6 to 6.8 in 2 and 20 mM buffered media, respectively. Growth inhibition and cell death could not be explained by a depletion of glucose or oxygen limitations during growth. The production of ammonia had a growth inhibitory effect, however, the sudden termination of the exponential growth phase and cell death could not be explained by the toxic influence of ammonia only.

  3. Biochemical identification and phylogenetic relationships in free-living amoebas of the genus Naegleria.

    Science.gov (United States)

    Pernin, P; Cariou, M L; Jacquier, A

    1985-11-01

    Using isoelectric focusing, the zymograms of 23 pathogenic and nonpathogenic Naegleria strains were studied for the activity of 16 enzymes. Certain enzymes (lactate dehydrogenase, L-threonine dehydrogenase, superoxide dismutase, acid phosphatase, malic enzyme, and leucine aminopeptidase) proved particularly useful from a practical point of view as they allow easy and reliable identification of pathogenic N. fowleri and N. australiensis as well as nonpathogenic N. lovaniensis strains. Genetic interpretation of these zymograms gave estimates of genetic distances that largely confirmed the taxonomic position of the Naegleria species. In addition, the genetic data suggest that there are two main phylogenetic groups in the genus Naegleria.

  4. A genetic approach to species criteria in the amoeba genus Naegleria using allozyme electrophoresis.

    Science.gov (United States)

    Adams, M; Andrews, R H; Robinson, B; Christy, P; Baverstock, P R; Dobson, P J; Blackler, S J

    1989-12-01

    The present study employs allozyme electrophoresis to characterize and inter-relate 61 isolates of Naegleria. Diploidy was confirmed, with heterozygotes observed at 29 of the 33 loci established and in all but two isolates. With a single exception, isolates clustered at two levels of similarity, either below 21% or above 52%. It is argued that such a major discontinuity provides a sound biological basis for a species concept in Naegleria. On this basis the present species-level taxonomy does not reflect the genetic diversity of the genus. The study recognized 18 genetic groups of species rank. The subspecies N. australiensis italica deserves specific rank; additional thermophilic species not closely related to N. fowleri and N. lovaniensis are recognized; and N. gruberi as currently conceived is a complex of 10 species, at least five of which are represented in the formal culture collections. Most species are genetically too different for relationships to be elucidated by allozyme electrophoresis, supporting the view that some of the times of divergence within the genus are extremely ancient.

  5. Identification of Naegleria fowleri in fresh isolates of environmental amoebae using a staphylococcal coagglutination technique.

    Science.gov (United States)

    Kilvington, S; White, D G

    1986-01-01

    A coagglutination test using Staphylococcus aureus Cowan strain 1 sensitized with specific antiserum successfully identified Naegleria fowleri in axenic and monoxenic culture by detection of a heat stable antigen released into the culture medium during growth. Strong coagglutination, visible to the naked eye, occurred within 30 sec when the sensitized reagent was mixed on a slide with medium from N. fowleri cultures. N. lovaniensis gave a weak cross-reaction which developed over one min clearly distinguishable from that of N.fowleri.N.gruberi and N. australiensis gave no reaction. The technique is proposed as part of a rapid and economical scheme for detecting N. fowleri in environmental samples very soon after primary isolation, with distinct advantages over conventional methods. Further development of the technique to detect other Naegleria spp. and the early diagnosis of primary amoebic meningoencephalitis are suggested as future applications.

  6. Did terrestrial diversification of amoebas (amoebozoa) occur in synchrony with land plants?

    Science.gov (United States)

    Fiz-Palacios, Omar; Romeralo, Maria; Ahmadzadeh, Afsaneh; Weststrand, Stina; Ahlberg, Per Erik; Baldauf, Sandra

    2013-01-01

    Evolution of lineage diversification through time is an active area of research where much progress has been made in the last decade. Contrary to the situation in animals and plants little is known about how diversification rates have evolved in most major groups of protist. This is mainly due to uncertainty about phylogenetic relationships, scarcity of the protist fossil record and the unknown diversity within these lineages. We have analyzed the evolutionary history of the supergroup Amoebozoa over the last 1000 million years using molecular dating and species number estimates. After an origin in the marine environment we have dated the colonization of terrestrial habitats by three distinct lineages of Amoebozoa: Dictyostelia, Myxogastria and Arcellinida. The common ancestor of the two sister taxa, Dictyostelia and Myxogastria, appears to have existed before the colonization of land by plants. In contrast Arcellinida seems to have diversify in synchrony with land plant radiation, and more specifically with that of mosses. Detection of acceleration of diversification rates in Myxogastria and Arcellinida points to a co-evolution within the terrestrial habitats, where land plants and the amoebozoans may have interacted during the evolution of these new ecosystems.

  7. Did terrestrial diversification of amoebas (amoebozoa occur in synchrony with land plants?

    Directory of Open Access Journals (Sweden)

    Omar Fiz-Palacios

    Full Text Available Evolution of lineage diversification through time is an active area of research where much progress has been made in the last decade. Contrary to the situation in animals and plants little is known about how diversification rates have evolved in most major groups of protist. This is mainly due to uncertainty about phylogenetic relationships, scarcity of the protist fossil record and the unknown diversity within these lineages. We have analyzed the evolutionary history of the supergroup Amoebozoa over the last 1000 million years using molecular dating and species number estimates. After an origin in the marine environment we have dated the colonization of terrestrial habitats by three distinct lineages of Amoebozoa: Dictyostelia, Myxogastria and Arcellinida. The common ancestor of the two sister taxa, Dictyostelia and Myxogastria, appears to have existed before the colonization of land by plants. In contrast Arcellinida seems to have diversify in synchrony with land plant radiation, and more specifically with that of mosses. Detection of acceleration of diversification rates in Myxogastria and Arcellinida points to a co-evolution within the terrestrial habitats, where land plants and the amoebozoans may have interacted during the evolution of these new ecosystems.

  8. Living together: The marine amoeba Thecamoeba hilla Schaeffer, 1926 and its endosymbiont Labyrinthula sp

    Czech Academy of Sciences Publication Activity Database

    Dyková, Iva; Fiala, Ivan; Dvořáková, Helena; Pecková, Hana

    2008-01-01

    Roč. 44, č. 4 (2008), s. 308-316 ISSN 0932-4739 R&D Projects: GA MŠk LC522 Institutional research plan: CEZ:AV0Z60220518 Keywords : Thecamoeba hilla * Labyrinthula sp. * Symbiotic association Subject RIV: GJ - Animal Vermins ; Diseases, Veterinary Medicine Impact factor: 1.000, year: 2008

  9. FYVE-dependent endosomal targeting of an arrestin-related protein in amoeba.

    Directory of Open Access Journals (Sweden)

    Dorian Guetta

    Full Text Available BACKGROUND: Visual and β-arrestins are scaffolding proteins involved in the regulation of receptor-dependent intracellular signaling and their trafficking. The arrestin superfamilly includes several arrestin domain-containing proteins and the structurally related protein Vps26. In Dictyostelium discoideum, the arrestin-domain containing proteins form a family of six members, namely AdcA to -F. In contrast to canonical arrestins, Dictyostelium Adc proteins show a more complex architecture, as they possess, in addition to the arrestin core, other domains, such as C2, FYVE, LIM, MIT and SAM, which potentially mediate selective interactions with either lipids or proteins. METHODOLOGY AND PRINCIPAL FINDINGS: A detailed analysis of AdcA has been performed. AdcA extends on both sides of the arrestin core, in particular by a FYVE domain which mediates selective interactions with PI(3P, as disclosed by intrinsic fluorescence measurements and lipid overlay assays. Localization studies showed an enrichment of tagged- and endogenous AdcA on the rim of early macropinosomes and phagosomes. This vesicular distribution relies on a functional FYVE domain. Our data also show that the arrestin core binds the ADP-ribosylation factor ArfA, the unique amoebal Arf member, in its GDP-bound conformation. SIGNIFICANCE: This work describes one of the 6 arrestin domain-containing proteins of Dictyostelium, a novel and atypical member of the arrestin clan. It provides the basis for a better understanding of arrestin-related protein involvement in trafficking processes and for further studies on the expanding roles of arrestins in eukaryotes.

  10. Propensity of heavier halides for the water/vapor interface revisited using the Amoeba force field

    Czech Academy of Sciences Publication Activity Database

    Tůma, L.; Jeníček, D.; Jungwirth, Pavel

    2005-01-01

    Roč. 411, - (2005), s. 70-74 ISSN 0009-2614 R&D Projects: GA MŠk(CZ) LC512 Grant - others:NSF(US) CHE0431312; NSF(US) CHE0209719 Institutional research plan: CEZ:AV0Z4055905 Keywords : halide anions * water/vapor interface * molecular dynamics Subject RIV: CF - Physical ; Theoretical Chemistry Impact factor: 2.438, year: 2005

  11. Marine amoebae with cytoplasmic and perinuclear symbionts deeply branching in the Gammaproteobacteria

    Czech Academy of Sciences Publication Activity Database

    Schulz, F.; Tyml, Tomáš; Pizzetti, I.; Dyková, I.; Fazi, S.; Kostka, Martin; Horn, M.

    2015-01-01

    Roč. 5, 25 August (2015), č. článku 13381. ISSN 2045-2322 R&D Projects: GA ČR(CZ) GBP505/12/G112 Institutional support: RVO:60077344 Keywords : free- living amebas * bacterial endosymbionts * phylogenetic inference * Paramecium caudatum * Naegleria * gene expression * environment * chlamydiae * diversity * pathogens Subject RIV: EG - Zoology Impact factor: 5.228, year: 2015

  12. Variation, sex, and social cooperation: molecular population genetics of the social amoeba Dictyostelium discoideum.

    Directory of Open Access Journals (Sweden)

    Jonathan M Flowers

    2010-07-01

    Full Text Available Dictyostelium discoideum is a eukaryotic microbial model system for multicellular development, cell-cell signaling, and social behavior. Key models of social evolution require an understanding of genetic relationships between individuals across the genome or possibly at specific genes, but the nature of variation within D. discoideum is largely unknown. We re-sequenced 137 gene fragments in wild North American strains of D. discoideum and examined the levels and patterns of nucleotide variation in this social microbial species. We observe surprisingly low levels of nucleotide variation in D. discoideum across these strains, with a mean nucleotide diversity (pi of 0.08%, and no strong population stratification among North American strains. We also do not find any clear relationship between nucleotide divergence between strains and levels of social dominance and kin discrimination. Kin discrimination experiments, however, show that strains collected from the same location show greater ability to distinguish self from non-self than do strains from different geographic areas. This suggests that a greater ability to recognize self versus non-self may arise among strains that are more likely to encounter each other in nature, which would lead to preferential formation of fruiting bodies with clonemates and may prevent the evolution of cheating behaviors within D. discoideum populations. Finally, despite the fact that sex has rarely been observed in this species, we document a rapid decay of linkage disequilibrium between SNPs, the presence of recombinant genotypes among natural strains, and high estimates of the population recombination parameter rho. The SNP data indicate that recombination is widespread within D. discoideum and that sex as a form of social interaction is likely to be an important aspect of the life cycle.

  13. Monitoring for the Presence of Parasitic Protozoa and Free-living Amoebae in Drinking Water Plants

    OpenAIRE

    Amany Saad Amer.

    2012-01-01

    Contamination of drinking water by microorganisms represents a major human health hazard in many parts of the world. The main objective of drinking water treatment is to provide microbiologically safe drinking water. The conventional drinking water treatment and disinfection has proved to be one of the major public health advances in modern times. A number of processes; namely water treatment, disinfection and changes influence the quality of drinking water delivered to the customer’s tap dur...

  14. Lemons, Mealworms, and the Giant Amoeba: A Eureka Trip through School Science.

    Science.gov (United States)

    McCormack, Alan J.

    1980-01-01

    Outlines teaching methods that emphasize the use of the right hemisphere of the brain. Argues that most productive thinking requires both modes of thinking and that any good elementary science program should include activities designed to develop both modes. (Author/IRT)

  15. Genome sequencing reveals metabolic and cellular interdependence in an amoeba-kinetoplastid symbiosis

    Czech Academy of Sciences Publication Activity Database

    Tanifuji, G.; Cenci, U.; Moog, D.; Dean, S.; Nakayama, T.; David, Vojtěch; Fiala, Ivan; Curtis, B.A.; Sibbald, S. J.; Onodera, N. T.; Colp, M.; Flegontov, Pavel; Johnson-MacKinnon, J.; McPhee, M.; Inagaki, Y.; Hashimoto, T.; Kelly, S.; Gull, K.; Lukeš, Julius; Archibald, J.M.

    2017-01-01

    Roč. 7, SEP 15 (2017), č. článku 11688. ISSN 2045-2322 R&D Projects: GA ČR(CZ) GA14-23986S; GA MŠk LL1601 Institutional support: RVO:60077344 Keywords : trypanosoma-brucei reveals * hidden markov model * neoparamoeba-pemaquidensis * gill disease * phylogenetic analyses * ichthyobodo-necator * gene prediction * host control * evolution * proteomics Subject RIV: EB - Gene tics ; Molecular Biology OBOR OECD: Biochemistry and molecular biology Impact factor: 4.259, year: 2016

  16. Heterolobosean amoebae from Arctic and Antarctic extremes: 18 novel strains of Allovahlkampfia, Vahlkampfia and Naegleria

    Czech Academy of Sciences Publication Activity Database

    Tyml, Tomáš; Skulinová, K.; Kavan, J.; Ditrich, O.; Kostka, M.; Dyková, I.

    2016-01-01

    Roč. 56, October (2016), s. 119-133 ISSN 0932-4739 R&D Projects: GA ČR(CZ) GBP505/12/G112 Institutional support: RVO:60077344 Keywords : Arctic * Antarctic * Heterolobosea * Molecular taxonomy * Tetramitia Subject RIV: GJ - Animal Vermins ; Diseases, Veterinary Medicine Impact factor: 2.581, year: 2016

  17. Therapeutic agents and biocides for ocular infections by free-living amoebae of Acanthamoeba genus.

    Science.gov (United States)

    Carrijo-Carvalho, Linda Christian; Sant'ana, Viviane Peracini; Foronda, Annette Silva; de Freitas, Denise; de Souza Carvalho, Fabio Ramos

    Acanthamoeba keratitis is a sight-threatening infectious disease. Resistance of the cystic form of the protozoan to biocides and the potential toxicity of chemical compounds to corneal cells are the main concerns related to long-term treatment with the clinically available ophthalmic drugs. Currently, a limited number of recognized antimicrobial agents are available to treat ocular amoebic infections. Topical application of biguanide and diamidine antiseptic solutions is the first-line therapy. We consider the current challenges when treating Acanthamoeba keratitis and review the chemical properties, toxicities, and mechanisms of action of the available biocides. Antimicrobial therapy using anti-inflammatory drugs is controversial, and aspects related to this topic are discussed. Finally, we offer our perspective on potential improvement of the effectiveness and safety of therapeutic profiles, with the focus on the quality of life and the advancement of individualized medicine. Copyright © 2016 Elsevier Inc. All rights reserved.

  18. Effects of temporally persistent ant nests of soil protozoan communities and the abundance of morphological types of amoeba

    Science.gov (United States)

    We compared soil protozoan communities near ant nests with soil protozoans in reference soils 5m from the edge of any mounds. We sampled three species of Chihuahuan Desert ants that construct nests that persist for more than a decade: a seed harvester, Pogonomymex rugosus, a liquid feeding honey-po...

  19. Ménage-a-trois: The amoeba Nuclearia sp. from Lake Zurich with its ecto- and endosymbiotic bacteria

    Czech Academy of Sciences Publication Activity Database

    Dirren, S.; Salcher, Michaela M.; Blom, J. F.; Schweikert, M.; Posch, T.

    2014-01-01

    Roč. 165, č. 5 (2014), s. 745-758 ISSN 1434-4610 Institutional support: RVO:60077344 Keywords : bacteria-protist symbioses * ectosymbionts * endosymbionts * feeding * Nucleariidae * Paucibacter toxinivorans Subject RIV: DA - Hydrology ; Limnology Impact factor: 3.045, year: 2014

  20. Genetic structure of natural populations of the free-living amoeba, Naegleria lovaniensis. Evidence for sexual reproduction.

    Science.gov (United States)

    Pernin, P; Ataya, A; Cariou, M L

    1992-02-01

    The genetic structure of two populations of Naegleria lovaniensis, comprising 71 isolates collected from the same local geographical area was investigated by isoenzyme analysis. Allelic variation at seven polymorphic enzymatic loci allowed identification of 45 distinctive genotype associations. Analysis of single locus variation reveals that most of them are close to Hardy-Weinberg equilibrium, which indicates segregation and free recombination between alleles. The recovery of a relatively high number of distinct genotypic associations (most of them being unique), and the absence of linkage disequilibrium between genotypes at the different loci also support the existence of recombination. Although we have no idea about the process involved, the results clearly indicate that genetic exchanges occur, at least occasionally, in natural populations of N. lovaniensis.

  1. Molecular differentiation of Entamoeba histolytica and Entamoeba dispar from Tunisian food handlers with amoeba infection initially diagnosed by microscopy

    Directory of Open Access Journals (Sweden)

    Ben Ayed S.

    2008-03-01

    Full Text Available The purpose of the study was to obtain more reliable epidemiological data concerning Entamoeba (E. histolytica infection in Tunisian food handlers using established molecular tools able to differentiate E. histolytica from E. dispar. From 2002 to 2005, 4,266 fresh stools specimens received in the setting of the National program of food handlers’ control were analysed by optical microscopy. Twelve (2.8 ‰ were positive for the presence of four nuclei cysts identified as E. histolytica/E. dispar. Extraction of DNA from the 12 samples, followed by specific amplifications of E. histolytica and E. dispar SSU rDNA, showed that 11 samples (92% were positive for E. dispar and negative for E. histolytica. Sequencing analysis of 8 PCR products permitted to verify the results obtained with conventional PCR. The remaining sample was negative by PCR amplifying E. histolytica DNA or E. dispar DNA specifically, although it did not show any inhibition. It probably contains protozoan cysts genetically distinct from these two species but morphological similar. Estimation of relative proportions between E. histolytica and E. dispar in cyst carriers showed that all explored individuals harboured the non pathogenic E. dispar strains. This result highlights the need of use in this population of complementary tests that allow specific diagnosis and obviate unnecessary chemotherapy.

  2. Contributions to the knowledge of a new disease caused by an amoeba in ongrowing Senegalese sole, Solea senegalensis (Kaup 1858)

    OpenAIRE

    Constenla Matalobos, María

    2013-01-01

    Esta patología se caracteriza por la presencia de protuberancias que se evidencian a través de la piel de los peces afectados. Estas lesiones se corresponden con nódulos en el tejido muscular, que muestran un aspecto de absceso. Lesiones similares a éstas también se han detectado en riñón, corazón, hígado y tracto digestivo, lo que nos permite definir la enfermedad como sistémica. Las secciones histológicas de los nódulos revelaron un extenso núcleo compuesto mayoritariamente por tejido necró...

  3. Laboratory diagnosis of primary amoebic meningo-encephalitis and methods for the detection of limax amoebae in the environment.

    Science.gov (United States)

    Cerva, L

    1980-01-01

    Laboratory examination methods and processes recommended for the diagnosis of primary amoebic meningo-encephalitis and detection of Naegleria fowleri in the environment are summarized. The most simple methods suitable for diagnostic laboratories in the sphere of medical parasitology have been chosen.

  4. Amoebae as Potential Environmental Hosts for Mycobacterium ulcerans and Other Mycobacteria, but Doubtful Actos in Buruli Ulcer Epidemiology

    DEFF Research Database (Denmark)

    Gryseels, Sophie; Amissah, Diana; Durnez, Lies

    2012-01-01

    experimentally infected Acanthamoeba polyphaga with M. ulcerans and found that the bacilli were phagocytised, not digested and remained viable for the duration of the experiment. Furthermore, we collected 13 water, 90 biofilm and 45 detritus samples in both Buruli ulcer endemic and non-endemic communities...

  5. Two new species of the genus Stenamoeba (Discosea, Longamoebia): cytoplasmic MTOC is present in one more amoebae lineage.

    Science.gov (United States)

    Geisen, Stefan; Weinert, Jan; Kudryavtsev, Alexander; Glotova, Anna; Bonkowski, Michael; Smirnov, Alexey

    2014-04-01

    Two new species of the recently described genus Stenamoeba, named S. berchidia and S. sardiniensis were isolated from a single soil sample on Sardinia, Italy. Both share morphological features characteristic to Stenamoeba and form in phylogenetic analyses together with other Stenamoeba spp. a highly supported clade within the family Thecamoebidae. The ultrastructural investigation of Stenamoeba sardiniensis revealed the presence of cytoplasmic microtubule-organizing centers (MTOCs), located close to one of several dictyosomes found inside the cell. This is the first report of cytoplasmic MTOCs among Thecamoebidae. The presence of MTOCs is now shown in five of nine orders comprising the class Discosea and potentially could be a phylogenetic marker in this group. We re-isolated Stenamoeba limacina from German soils. This strain shows a similar morphology and an almost complete SSU rDNA sequence identity with the type strain of S. limacina originating from gills of fishes, collected in Czech Republic. Copyright © 2014 Elsevier GmbH. All rights reserved.

  6. Temperature-dependent changes in plasma-membrane lipid order and the phagocytotic activity of the amoeba Acanthamoeba castellanii are closely correlated.

    Science.gov (United States)

    Avery, S V; Lloyd, D; Harwood, J L

    1995-12-15

    The relationship between temperature-dependent changes in phagocytotic activity of Acanthamoeba castellanii and the fatty acid composition and physical properties of plasma membrane-enriched fractions were determined in cultures acclimated to 30 degrees C and 15 degrees C. Chilling (from 30 degrees C to 15 degrees C) had a very pronounced short-term inhibitory effect on phagocytosis only in stationary-phase cultures, which displayed a low degree of fatty acid unsaturation. A subsequent increase in phagocytosis by these cells was correlated with a low-temperature-induced increase in fatty acid unsaturation (shown previously [Jones, Lloyd and Harwood (1993) Biochem. J. 296, 183-188] to be due to n-6 desaturase induction). Plasma membrane-enriched fractions from 15 degrees C-acclimated cells also showed a marked increase in the relative proportion of polyunsaturated fatty acids. Steady-state fluorescence depolarization studies, using the membrane probe diphenylhexatriene, revealed increases in plasma membrane order with decreasing assay temperature. Over the upper assay-temperature range (25-40 degrees C), fluorescence anisotropy values were higher in membranes from 30 degrees C-acclimated cells; a 3.3 degrees C relative displacement of plots indicated that temperature-induced changes in membrane lipid composition compensated for approx. 22% of the ordering effect of low temperature. Changes in the temperature-dependence of fluorescence anisotropy, possibly corresponding to lateral phase separations or alterations in other bulk physical properties of membranes, occurred between 20 and 25 degrees C in membranes from 30 degrees C-acclimated cells and between 15 and 20 degrees C in membranes from 15 degrees C-acclimated cells. Fluorescence anisotropy plots were superimposed at assay temperatures between 5 and 15 degrees C. Short-term phagocytotic rates in whole cells decreased with assay temperature. Arrhenius discontinuities in rates of phagocytosis occurred at approx. 25.0 degrees C and 17.5 degrees C in 30 degrees C- and 15 degrees C-acclimated cells respectively, and in each case were thus within the temperature ranges of slope-change in the corresponding fluorescence anisotropy plots. The results show a direct correlation between plasma membrane fatty acid unsaturation, membrane physical properties and phagocytotic activity in A. castellanii. Therefore, a specific integrated physiological process has been correlated with fatty acid desaturase induction for the first time.

  7. Nuclearia pattersoni sp. n. (Filosea), a new species of amphizoic amoeba isolated from gills of roach (Rutilus rutilus), and its rickettsial endosymbiont

    Czech Academy of Sciences Publication Activity Database

    Dyková, Iva; Veverková, Marie; Fiala, Ivan; Macháčková, Blanka; Pecková, Hana

    2003-01-01

    Roč. 50, č. 3 (2003), s. 161-170 ISSN 0015-5683 R&D Projects: GA AV ČR IAA6022202 Institutional research plan: CEZ:AV0Z6022909; CEZ:MSM 123100003 Keywords : Filosea * Nuclearia pattersoni * rickettsial endosymbiont Subject RIV: EA - Cell Biology Impact factor: 0.469, year: 2003

  8. Molecular Detection of Legionella spp. and their associations with Mycobacterium spp., Pseudomonas aeruginosa and amoeba hosts in a drinking water distribution system

    Data.gov (United States)

    U.S. Environmental Protection Agency — Quantity of Legionella spp., Mycobacterium spp., Acanthamoeba,Vermamoeba vermiformis and Pseudomonas aeruginosa were estimated using qPCR methods. This dataset is...

  9. Use of Multiplex PCR and PCR Restriction Enzyme Analysis for Detection and Exploration of the Variability in the Free-Living Amoeba Naegleria in the Environment

    Science.gov (United States)

    Pélandakis, Michel; Pernin, Pierre

    2002-01-01

    A multiplex PCR was developed to simultaneously detect Naegleria fowleri and other Naegleria species in the environment. Multiplex PCR was also capable of identifying N. fowleri isolates with internal transcribed spacers of different sizes. In addition, restriction fragment length polymorphism analysis of the PCR product distinguished the main thermophilic Naegleria species from the sampling sites. PMID:11916734

  10. Checklist of the soil and moss testate amoebae (Protozoa, Rhizopoda) from the National Nature Reserve Voděradské bučiny (Czech Republic)

    Czech Academy of Sciences Publication Activity Database

    Balík, Vladimír

    2001-01-01

    Roč. 170, 1-4 (2001), s. 91-104 ISSN 0139-9543 R&D Projects: GA AV ČR IAA6066702 Institutional research plan: CEZ:AV0Z6066911 Keywords : Testacea * faunistic * Voděradské bučiny National Nature Reserve Subject RIV: EH - Ecology, Behaviour

  11. Bacterivory: a novel dual role for thraustochytrids in the sea

    Digital Repository Service at National Institute of Oceanography (India)

    Raghukumar, S.

    -forming motile bacterium, isolated 166 S. Raghukumar: Bacterivory of thraustochytrids Figs. 1 to 9. Thraustochytrium striatum. Formation of amoebae. Figs. 1 to 3 Liberation of limax amoebae from sporangia. Duration of sequence = 8 min. Figs. 4 to 7... Transformation of a group of veg- etative cells into amoebae. Duration of sequence = 13 min. Figs. 8 and 9 Transformation of a large vegetative cell into an amoeba. Arrow indicates the same cell. Scale bar = 10 gm from Mandovi estuary, Goa, was introduced...

  12. Recurrent amoebic gill infestation in rainbow trout cultured in a semiclosed water recirculation system

    Science.gov (United States)

    Noble, A.C.; Herman, R.L.; Noga, E.J.; Bullock, G.L.

    1997-01-01

    Five lots of commercially purchased juvenile rainbow trout Oncorhynchus mykiss (17-44 g) stocked in a continuous-production water recirculation system became infested with gilt amoebae. The amoebae were introduced into the recirculation system, as evidenced by their presence on gills of fish held in quarantine tanks. Based on their morphology, as seen in histological sections and by electron microscopy, the amoebae appeared to be more closely related to the family Cochliopodiidae than to other taxa of free living amoebae. Attempts to culture the amoebae in different media, at different temperatures of incubation, and in fish cell culture were not successful. Initial treatment of the recirculation system with formalin at 167 parts per million (ppm) for 1 h eliminated amoebae from the gills. Subsequent treatments of the entire system with formalin at 50-167 ppm reduced the intensity of further infestations.

  13. Survival of polioviruses and echoviruses in Acanthamoeba castellanii cultivated in vitro.

    Science.gov (United States)

    Danes, L; Cerva, L

    1981-01-01

    Cultures of Acanthamoeba castellanii, kept at room temperature in sterile Bacto-Casitone (Difco) medium, were artificially infected with vaccination poliovirus strains type 1 and type 3 and with echovirus type 4 and echovirus type 30. No remarkable virus cumulation was observed on the surface or inside amoeba cells during the observation period of 21 days. Amoeba-adsorbed viruses were impossible to remove by repeated washings. Virus neutralization with specific antisera showed that enteroviruses were most probably present only on amoeba surfaces. In contrast to amoeba-free virus suspension, echoviruses bound to amoebae and their cell pulp persisted even after 52 to 75 days. However, the tested amoeba species played only the role of a solids-like carrier in this survival of echoviruses.

  14. Compositions and methods for pathogen transport

    Energy Technology Data Exchange (ETDEWEB)

    El-Etr, Sahar; Farquar, George R.

    2016-01-26

    This disclosure provides a method for transporting a pathogen under ambient conditions, by culturing the pathogen with an amoeba under conditions that favor the incorporation of the pathogen into a trophozoite, starving the amoeba until it encysts, then culturing under conditions that favor conversion of the amoeba back to a trophozoite. In one aspect, the conditions that favor incorporation of the pathogen into the cyst of the amoeba comprises contacting the pathogen with the amoeba in an iron rich environment. Virus and/or bacteria are pathogens that can be transported by the disclosed method. Amoeba that are useful in the disclosed methods include, without limitation Acanthamoeba castellanii, Hartmannella vermiformis and Naegleria gruberi. The disclosed methods have utility in: transporting pathogens from military field hospitals and clinics to the laboratory; transporting pathogens from global satellite laboratories to clinical laboratories; long term storage of pathogens; enriching contaminated patient samples for pathogens of interest; biosurveillance and detection efforts.

  15. Susceptibility of pathogenic and nonpathogenic Naegleria spp. to complement-mediated lysis.

    OpenAIRE

    Whiteman, L Y; Marciano-Cabral, F

    1987-01-01

    The susceptibility of four species of Naegleria amoebae to complement-mediated lysis was determined. The amoebicidal activity of normal human serum (NHS) and normal guinea pig serum (NGPS) for Naegleria amoebae was measured by an in vitro cytotoxicity assay. Release of radioactivity from amoebae labeled with [3H]uridine and visual observation with a compound microscope were used as indices of lysis. Highly pathogenic mouse-passaged N. fowleri was less susceptible to the lytic effects of NHS a...

  16. Effects of Mannose on Pathogenesis of Acanthamoeba castellanii

    OpenAIRE

    Yoo, Kyung-Tae; Jung, Suk-Yul

    2012-01-01

    Acanthamoeba spp. are single-celled protozoan organisms that are widely distributed in the environment. In this study, to understand functional roles of a mannose-binding protein (MBP), Acanthamoeba castellanii was treated with methyl-alpha-D-mannopyranoside (mannose), and adhesion and cytotoxicity of the amoeba were analyzed. In addition, to understand the association of MBP for amoeba phagocytosis, phagocytosis assay was analyzed using non-pathogenic bacterium, Escherichia coli K12. Amoebae...

  17. Acanthamoeba polyphaga-enhanced growth of Mycobacterium smegmatis.

    Directory of Open Access Journals (Sweden)

    Otmane Lamrabet

    Full Text Available BACKGROUND: Mycobacterium smegmatis is a rapidly-growing mycobacterium causing rare opportunistic infections in human patients. It is present in soil and water environments where free-living amoeba also reside, but data regarding M. smegmatis-amoeba relationships have been contradictory from mycobacteria destruction to mycobacteria survival. METHODOLOGY/PRINCIPAL FINDINGS: Using optic and electron microscopy and culture-based microbial enumeration we investigated the ability of M. smegmatis mc(2 155, M. smegmatis ATCC 19420(T and M. smegmatis ATCC 27204 organisms to survive into Acanthamoeba polyphaga trophozoites and cysts. We observed that M. smegmatis mycobacteria penetrated and survived in A. polyphaga trophozoites over five-day co-culture resulting in amoeba lysis and the release of viable M. smegmatis mycobacteria without amoebal cyst formation. We further observed that amoeba-co-culture, and lysed amoeba and supernatant and pellet, significantly increased five-day growth of the three tested M. smegmatis strains, including a four-fold increase in intra-amoebal growth. CONCLUSIONS/SIGNIFICANCE: Amoebal co-culture increases the growth of M. smegmatis resulting in amoeba killing by replicating M. smegmatis mycobacteria. This amoeba-M. smegmatis co-culture system illustrates an unusual paradigm in the mycobacteria-amoeba interactions as mycobacteria have been mainly regarded as amoeba-resistant organisms. Using these model organisms, this co-culture system could be used as a simple and rapid model to probe mycobacterial factors implicated in the intracellular growth of mycobacteria.

  18. Acanthamoeba containing endosymbiotic chlamydia isolated from hospital environments and its potential role in inflammatory exacerbation.

    Science.gov (United States)

    Fukumoto, Tatsuya; Matsuo, Junji; Okubo, Torahiko; Nakamura, Shinji; Miyamoto, Kentaro; Oka, Kentaro; Takahashi, Motomichi; Akizawa, Kouji; Shibuya, Hitoshi; Shimizu, Chikara; Yamaguchi, Hiroyuki

    2016-12-15

    Environmental chlamydiae belonging to the Parachlamydiaceae are obligate intracellular bacteria that infect Acanthamoeba, a free-living amoeba, and are a risk for hospital-acquired pneumonia. However, whether amoebae harboring environmental chlamydiae actually survive in hospital environments is unknown. We therefore isolated living amoebae with symbiotic chlamydiae from hospital environments. One hundred smear samples were collected from Hokkaido University Hospital, Sapporo, Japan; 50 in winter (February to March, 2012) and 50 in summer (August, 2012), and used for the study. Acanthamoebae were isolated from the smear samples, and endosymbiotic chlamydial traits were assessed by infectivity, cytokine induction, and draft genomic analysis. From these, 23 amoebae were enriched on agar plates spread with heat-killed Escherichia coli. Amoeba prevalence was greater in the summer-collected samples (15/30, 50%) than those of the winter season (8/30, 26.7%), possibly indicating a seasonal variation (p = 0.096). Morphological assessment of cysts revealed 21 amoebae (21/23, 91%) to be Acanthamoeba, and cultures in PYG medium were established for 11 of these amoebae. Three amoebae contained environmental chlamydiae; however, only one amoeba (Acanthamoeba T4) with an environmental chlamydia (Protochlamydia W-9) was shown the infectious ability to Acanthamoeba C3 (reference amoebae). While Protochlamydia W-9 could infect C3 amoeba, it failed to replicate in immortal human epithelial, although exposure of HEp-2 cells to living bacteria induced the proinflammatory cytokine, IL-8. Comparative genome analysis with KEGG revealed similar genomic features compared with other Protochlamydia genomes (UWE25 and R18), except for a lack of genes encoding the type IV secretion system. Interestingly, resistance genes associated with several antibiotics and toxic compounds were identified. These findings are the first demonstration of the distribution in a hospital of a living

  19. Legionella pneumophila: Virulent and Avirulent Interaction with Acanthamoeba castellanii.

    Science.gov (United States)

    1993-08-01

    enzymatically active site of P. aeruginosa elastase. Additionally, elastase inhibitors and related thermolysins were shown to inhibit L. pneumophila protease...legionellae by an amoeba and human monocyte-like cell line (88). Treatment of Hartmannella veriformis with cytocholasin D, an inhibitor of microfilament...18 dependent phagocytosis, did not inhibit intracellular legionellae growth. Alternately, methylamine, an inhibitor of pinocytosis, inhibited amoeba

  20. What Entamoeba histolytica and Giardia lamblia tell us about the ...

    Indian Academy of Sciences (India)

    However, our studies have shown that amoebae and giardia contain splicoeosomal introns, ras-family signal-transduction proteins, ATP-binding casettes (ABC)-family drug transporters, Golgi, and a mitochondrion-derived organelle (amoebae only). These results suggest that most of the organelles of higher eukaryotes ...

  1. Heterogeneity in the genus Allovahlkampfia and the description of the new genus Parafumarolamoeba (Vahlkampfiidae; Heterolobosea)

    NARCIS (Netherlands)

    Geisen, Stefan; Bonkowski, Michael; Zhang, Junling; De Jonckheere, Johan F.

    Abstract Heterolobosean amoebae are common and diverse members of soil protist communities. In this study, we isolated seven strains of amoebae from soil samples taken in Tibet (at high altitude), Sardinia and the Netherlands, all resembling to belong to a similar heterolobosean morphospecies.

  2. Susceptibility of pathogenic and nonpathogenic Naegleria ssp

    Energy Technology Data Exchange (ETDEWEB)

    Whiteman, L.Y.

    1988-01-01

    The susceptibility of four species of Naegleria amoebae to complement-mediated lysis was determined. The amoebicidal activity of normal human serum (NHS) and normal guinea pig serum (NGPS) for Naegleria amoebae was measured by an in vitro cytotoxicity assay. Release of radioactivity from amoebae labeled with {sup 3}H-uridine and visual observation with a compound microscope were used as indices of lysis. Susceptibility or resistance to complement-mediated lysis in vitro correlated with the in vivo pathogenic potential. Nonpathogenic Naegleria amoebae were lysed at a faster rate and at higher cell concentrations than were pathogenic amoebae. Electrophoretic analysis of NHS incubated with pathogenic or nonpathogenic Naegleria spp. demonstrated that amoebae activate the complement cascade resulting in the production of C3 and C5 complement cleavage products. Treatment with papain or trypsin for 1 h, but not with sialidase, increase the susceptibility of highly pathogenic, mouse-passaged N. fowleri to lysis. Treatment with actinomycin D, cycloheximide or various protease inhibitors for 4 h did not increase susceptibility to lysis. Neither a repair process involving de novo protein synthesis nor a complement-inactivating protease appear to account for the increase resistance of N. fowleri amoebae to complement-mediated lysis. A binding study with {sup 125}I radiolabeled C9 indicated that the terminal complement component does not remain stably bound to the membrane of pathogenic amoebae.

  3. 'Dicty dynamics': Dictyostelium motility as persistent random motion

    DEFF Research Database (Denmark)

    Li, Liang; Cox, Edward C; Flyvbjerg, Henrik

    2011-01-01

    to the amoeba's direction of motion. This motion propels the amoeba with a random periodic left–right waddle in a direction that has a long persistence time. The model fully accounts for the statistics of the experimental trajectories, including velocity power spectra and auto-correlations, non...

  4. A functional connection of Dictyostelium paracaspase with the ...

    Indian Academy of Sciences (India)

    2013-07-12

    Jul 12, 2013 ... In Dictyostelium, a single-cell haploid amoeba, starvation causes the amoeba to aggregate and ..... likely due to the loss of water from the cells. The pcp- cell line was smaller .... In the WT-AX4 and GFP-Pcp cell lines, little disassociated CVs network have seen due to the water lost by cells in the hypertonic ...

  5. Effects of mannose on pathogenesis of Acanthamoeba castellanii.

    Science.gov (United States)

    Yoo, Kyung-Tae; Jung, Suk-Yul

    2012-12-01

    Acanthamoeba spp. are single-celled protozoan organisms that are widely distributed in the environment. In this study, to understand functional roles of a mannose-binding protein (MBP), Acanthamoeba castellanii was treated with methyl-alpha-D-mannopyranoside (mannose), and adhesion and cytotoxicity of the amoeba were analyzed. In addition, to understand the association of MBP for amoeba phagocytosis, phagocytosis assay was analyzed using non-pathogenic bacterium, Escherichia coli K12. Amoebae treated with mannose for 20 cycles exhibited larger vacuoles occupying the most area of the amoebic cytoplasm in comparison with the control group amoebae and glucose-treated amoebae. Mannose-selected amoebae exhibited lower levels of binding to Chinese hamster ovary (CHO) cells. Exogenous mannose inhibited >50% inhibition of amoebae (control group) binding to CHO cells. Moreover, exogenous mannose inhibited amoebae (i.e., man-treated) binding to CHO cells by castellanii trophozoites should be severely damaged and do not well interact with a target cell via a lectin of MBP.

  6. Biology of Alkylphosphonic Acids. A Review of the Distribution, Metabolism, and Structure of Naturally Occuring Alkylphosphonic Acids.

    Science.gov (United States)

    1977-12-22

    been found in an echinoderm (Quin, 1965), a salt water crab (de Koning, 1970), the abalone (de Koning, 1966), the amoeba Acanthamoeba castellanii ...Dearborn, D.G., and Wright, P.1., 1974. Lipophosphonoglycan ofthe plasma membrane of Acanthamoeba castellanii : Isolation from whole amoebae and identificato

  7. A distributed file service based on optimistic concurrency control

    NARCIS (Netherlands)

    Mullender, Sape J.; Tanenbaum, Andrew S.

    1985-01-01

    The design of a layered file service for the Amoeba Distributed System is discussed, on top of which various applications can easily be intplemented. The bottom layer is formed by the Amoeba Block Services, responsible for implementing stable storage and repficated, highly available disk blocks. The

  8. Amoebic PI3K and PKC is required for Jurkat T cell death induced by Entamoeba histolytica.

    Science.gov (United States)

    Lee, Young Ah; Kim, Kyeong Ah; Min, Arim; Shin, Myeong Heon

    2014-08-01

    The enteric protozoan parasite Entamoeba histolytica is the causative agent of human amebiasis. During infection, adherence of E. histolytica through Gal/GalNAc lectin on the surface of the amoeba can induce caspase-3-dependent or -independent host cell death. Phosphorylinositol 3-kinase (PI3K) and protein kinase C (PKC) in E. histolytica play an important function in the adhesion, killing, or phagocytosis of target cells. In this study, we examined the role of amoebic PI3K and PKC in amoeba-induced apoptotic cell death in Jurkat T cells. When Jurkat T cells were incubated with E. histolytica trophozoites, phosphatidylserine (PS) externalization and DNA fragmentation in Jurkat cells were markedly increased compared to those of cells incubated with medium alone. However, when amoebae were pretreated with a PI3K inhibitor, wortmannin before being incubated with E. histolytica, E. histolytica-induced PS externalization and DNA fragmentation in Jurkat cells were significantly reduced compared to results for amoebae pretreated with DMSO. In addition, pretreatment of amoebae with a PKC inhibitor, staurosporine strongly inhibited Jurkat T cell death. However, E. histolytica-induced cleavage of caspase-3, -6, and -7 were not inhibited by pretreatment of amoebae with wortmannin or staurosporin. In addition, we found that amoebic PI3K and PKC have an important role on amoeba adhesion to host compartment. These results suggest that amebic PI3K and PKC activation may play an important role in caspase-independent cell death in Entamoeba-induced apoptosis.

  9. Isolation of Vermamoeba vermiformis and associated bacteria in hospital water.

    Science.gov (United States)

    Pagnier, Isabelle; Valles, Camille; Raoult, Didier; La Scola, Bernard

    2015-03-01

    To detect new potential pathogens in hospital water, we isolated free-living amoebae in water samples taken from three different hospitals in Marseille (France). The samples were inoculated in media containing saline buffer and various bacteria as nutrient sources. The isolated amoebae were identified by gene sequencing. Among the 105 water samples, taken from 19 sites, we isolated 14 amoebae, of which 9 Vermamoeba vermiformis and 5 Acanthamoeba sp. None of the amoebae showed the presence of obligate bacterial endosymbionts. Because V. vermiformis was most commonly isolated, we used an axenic collection strain to isolate amoeba-resistant bacteria from the same sites. The isolated bacterial species included Stenotrophomonas maltophilia and Legionella sp. Legionella taurinensis was isolated for the first time in association with amoebae. A strict intracellular bacterium was isolated, that may represent a new genus among the Chlamydiales. We propose that it be named "Candidatus Rubidus massiliensis". Our study shows that the isolation and identification of new pathogens associated with amoebae, which were previously performed using Acanthamoeba sp., should instead use V. vermiformis because this organism is more commonly associated with humans and is an essential complement of Acanthamoeba sp. co-culture to study the ecology of hospital water supplies. Copyright © 2015 Elsevier Ltd. All rights reserved.

  10. Identification of Naegleria fowleri proteins linked to primary amoebic meningoencephalitis.

    Science.gov (United States)

    Jamerson, Melissa; Schmoyer, Jacqueline A; Park, Jay; Marciano-Cabral, Francine; Cabral, Guy A

    2017-03-01

    Naegleria fowleri (N. fowleri) causes primary amoebic meningoencephalitis, a rapidly fatal disease of the central nervous system. N. fowleri can exist in cyst, flagellate or amoebic forms, depending on environmental conditions. The amoebic form can invade the brain following introduction into the nasal passages. When applied intranasally to a mouse model, cultured N. fowleri amoebae exhibit low virulence. However, upon serial passage in mouse brain, the amoebae acquire a highly virulent state. In the present study, a proteomics approach was applied to the identification of N. fowleri amoeba proteins whose expression was associated with the highly virulent state in mice. Mice were inoculated intranasally with axenically cultured amoebae or with mouse-passaged amoebae. Examination by light and electron microscopy revealed no morphological differences. However, mouse-passaged amoebae were more virulent in mice as indicated by exhibiting a two log10 titre decrease in median infective dose 50 (ID50). Scatter plot analysis of amoebic lysates revealed a subset of proteins, the expression of which was associated with highly virulent amoebae. MS-MS indicated that this subset contained proteins that shared homology with those linked to cytoskeletal rearrangement and the invasion process. Invasion assays were performed in the presence of a select inhibitor to expand on the findings. The collective results suggest that N. fowleri gene products linked to cytoskeletal rearrangement and invasion may be candidate targets in the management of primary amoebic meningoencephalitis.

  11. Modulation of Zinc Homeostasis in Acanthamoeba castellanii as a Possible Antifungal Strategy against Cryptococcus gattii

    Directory of Open Access Journals (Sweden)

    Nicole S. Ribeiro

    2017-08-01

    Full Text Available Cryptococcus gattii is a basidiomycetous yeast that can be found in the environment and is one of the agents of cryptococcosis, a life-threatening disease. During its life cycle, cryptococcal cells take hold inside environmental predators such as amoebae. Despite their evolutionary distance, macrophages and amoebae share conserved similar steps of phagocytosis and microbial killing. To evaluate whether amoebae also share other antifungal strategies developed by macrophages, we investigated nutritional immunity against cryptococcal cells. We focused on zinc homeostasis modulation in Acanthamoeba castellanii infected with C. gattii. The intracellular proliferation rate (IPR in amoebae was determined using C. gattii R265 and mutants for the ZIP1 gene, which displays defects of growth in zinc-limiting conditions. We detected a reduced IPR in cells lacking the ZIP1 gene compared to wild-type strains, suggesting that amoebae produce a low zinc environment to engulfed cells. Furthermore, flow cytometry analysis employing the zinc probe Zinpyr-1 confirmed the reduced concentration of zinc in cryptococcal-infected amoebae. qRT-PCR analysis of zinc transporter-coding genes suggests that zinc export by members of the ZnT family would be involved in the reduced intracellular zinc concentration. These results indicate that amoebae may use nutritional immunity to reduce fungal cell proliferation by reducing zinc availability for the pathogen.

  12. Assessing the Risk of Primary Amoebic Meningoencephalitis from Swimming in the Presence of Environmental Naegleria fowleri

    Science.gov (United States)

    Cabanes, Pierre-André; Wallet, France; Pringuez, Emmanuelle; Pernin, Pierre

    2001-01-01

    Free-living Naegleria fowleri amoebae cause primary amoebic meningoencephalitis (PAM). Because of the apparent conflict between their ubiquity and the rarity of cases observed, we sought to develop a model characterizing the risk of PAM after swimming as a function of the concentration of N. fowleri. The probability of death from PAM as a function of the number of amoebae inhaled is modeled according to results obtained from animals infected with amoeba strains. The calculation of the probability of inhaling one or more amoebae while swimming is based on a double hypothesis: that the distribution of amoebae in the water follows a Poisson distribution and that the mean quantity of water inhaled while swimming is 10 ml. The risk of PAM for a given concentration of amoebae is then obtained by summing the following products: the probability of inhaling n amoebae × the probability of PAM associated with inhaling these n amoebae. We chose the lognormal model to assess the risk of PAM because it yielded the best analysis of the studentized residuals. Nonetheless, the levels of risk thereby obtained cannot be applied to humans without correction, because they are substantially greater than those indicated by available epidemiologic data. The curve was thus adjusted by a factor calculated with the least-squares method. This provides the PAM risk in humans as a function of the N. fowleri concentration in the river. For example, the risk is 8.5 × 10−8 at a concentration of 10 N. fowleri amoebae per liter. PMID:11425704

  13. Intranasal coadministration of Cholera toxin with amoeba lysates modulates the secretion of IgA and IgG antibodies, production of cytokines and expression of pIgR in the nasal cavity of mice in the model of Naegleria fowleri meningoencephalitis.

    Science.gov (United States)

    Carrasco-Yepez, Maricela; Campos-Rodriguez, Rafael; Lopez-Reyes, Israel; Bonilla-Lemus, Patricia; Rodriguez-Cortes, Antonio Yahve; Contis-Montes de Oca, Arturo; Jarillo-Luna, Adriana; Miliar-Garcia, Angel; Rojas-Hernandez, Saul

    2014-11-01

    The nasal mucosa is the first contact with antigens to induce IgA response. The role of this site has rarely been studied. We have shown than intranasal administration with Naegleria fowleri lysates plus Cholera toxin (CT) increased the protection (survival up to 100%) against N. fowleri infection in mice and apparently antibodies IgA and IgG together with polymorphonuclear (PMN) cells avoid the attachment of N. fowleri to apical side of the nasal epithelium. We also observed that nasal immunization resulted in the induction of antigen-specific IgG subclasses (IgG1 and IgG2a) in nasal washes at days 3 and 9 after the challenge and IgA and IgG in the nasal cavity, compared to healthy and infected mice. We found that immunization with both treatments, N. fowleri lysates plus CT or CT alone, increased the expression of the genes for alpha chain, its receptor (pIgR), and it also increased the expression of the corresponding proteins evidenced by the ∼65 and ∼74kDa bands, respectively. Since the production of pIgR, IgA and IgG antibodies, is up-regulated by some factors, we analyzed the expression of genes for IL-10, IL-6, IFN-γ, TNF-α and IL-1β by using RT-PCR of nasal passages. Immunization resulted in an increased expression of IL-10, IL-6, and IFN-γ cytokines. We also aimed to examine the possible influences of immunization and challenge on the production of inflammatory cytokines (TNF-α and IL-1β). We observed that the stimulus of immunization inhibits the production of TNF-α compared to the infected group where the infection without immunization causes an increase in it. Thus, it is possible that the coexistence of selected cytokines produced by our immunization model may provide a highly effective immunological environment for the production of IgA, IgG and pIgR as well as a strong activation of the PMN in mucosal effector tissue such as nasal passages. Copyright © 2014 Elsevier Inc. All rights reserved.

  14. Scrotal abscess, a rare case of extra intestinal amoebiasis.

    Science.gov (United States)

    Prasetyo, R H

    2015-09-01

    The majority of amoeba infection are asymptomatic, but clinically intestinal amoebiasis or extra intestinal amoebiasis may result. Genital amoebiasis is very rare manifestation of extra intestial amoebiasis, but a case of amoebic scrotal abscess, seen in Surabaya. The invasive form of Entamoeba histolytica trophozoite was seen in Giemsa stained aspirate of the abscess. In case of an abscess bacteria are primarly considered, but the case presented here shows that amoeba can be the cause, although very rarely. Thus when bacteriological diagnostics are negative amoeba should be considered, especially in case of brown-reddish colored and foul smelling pus.

  15. Pathogenic and nonpathogenic Acanthamoeba spp. in thermally polluted discharges and surface waters

    Energy Technology Data Exchange (ETDEWEB)

    de Jonckheere, J.F.

    1981-02-01

    During spring and autumn, the total number of amoebae and the number of acanthamoeba species able to grow at 37 degrees C were determined in six thermally polluted factory discharges and the surrounding surface waters. The isolated Acanthamoeba strains were studied for growth in axenic medium, cytopathic effect in Vito cell cultures, and virulence in mice. Although more amoebae were isolated in autumn, the number of Acanthamoeba species was lower than in spring, when the percent of pathogenic strains among the isolates was highest. Higher concentrations of amoebae were found in warm discharges, and more virulent strains occurred in thermal discharges than in surface waters.

  16. Pathogenic and nonpathogenic Acanthamoeba spp. in thermally polluted discharges and surface waters

    International Nuclear Information System (INIS)

    de Jonckheere, J.F.

    1981-01-01

    During spring and autumn, the total number of amoebae and the number of acanthamoeba species able to grow at 37 degrees C were determined in six thermally polluted factory discharges and the surrounding surface waters. The isolated Acanthamoeba strains were studied for growth in axenic medium, cytopathic effect in Vito cell cultures, and virulence in mice. Although more amoebae were isolated in autumn, the number of Acanthamoeba species was lower than in spring, when the percent of pathogenic strains among the isolates was highest. Higher concentrations of amoebae were found in warm discharges, and more virulent strains occurred in thermal discharges than in surface waters

  17. One-Celled Creatures From the Black Lagoon

    Science.gov (United States)

    McClellan, Ray E.

    1974-01-01

    Describes methods of developing and using cultures of microoganisms suitable for use in elementary school science classes. Emphasis is placed upon the characteristics and culturing of amoeba, paramecia, euglena, rotifers, and algae. (JR)

  18. The four new species of Myxomycetes of South-East Maharashtra ...

    African Journals Online (AJOL)

    McRoy

    the true slime-moulds) are organisms which possess an assimilative phase of free living, multinucleate, mobile mass of protoplasm called as the plasmodium like amoebae, and a sporulating phase consisting of a mass of spores typically borne.

  19. Analysis of Rheb in the cellular slime mold Dictyostelium discoideum

    Indian Academy of Sciences (India)

    2014-01-27

    , these free living amoebae come to common collecting points in response ..... slightly slow growers as compared to wild-type but with no obvious alterations in the developmental profile (data not shown). Our results show that ...

  20. Short Communication

    African Journals Online (AJOL)

    with gastrointestinal tract include. Entamoeba histolytica (Amoeba), Giardia lamblia (flagellate) and Balantidia spp. (a ciliate) which cause amoebiasis, giardiasis and balantidiasis, respectively (Prekarski, 1986). The general symptoms include abdominal pain, flatulence and frequent passage of loose stools. Other symptoms.

  1. Phospholipids of Entamoeba invadens

    NARCIS (Netherlands)

    Vliet, H.H.D.M. van; Kamp, J.A.F. op den; Deenen, L.L.M. van

    1975-01-01

    The major phosphoglycerides present in Entamoeba invadens are phosphatidylcholine, phosphatidylethanolamine, phosphatidylserine and phosphatidylinositol. Furthermore, three different sphingolipids could be isolated from the amoeba. In addition to sphingomyelin and a phosphonolipid, ceramide

  2. Ammonia differentially suppresses the cAMP chemotaxis of anterior ...

    Indian Academy of Sciences (India)

    Unknown

    produced ... using an agar substratum dispersed amoeba drop assay. For comparison, we also include ALC assays on a pre- .... tic response (lack of significant ALC movement to the periphery) was analysed by χ 2 analysis (Data Desk). 3. Results.

  3. Disentangling the effects of water chemistry and substratum structure on moss-dwelling unicellular and multicellular micro-organisms in spring-fens

    Czech Academy of Sciences Publication Activity Database

    Hájková, P.; Bojková, J.; Fránková, Markéta; Opravilová, V.; Hájek, M.; Kintrová, K.; Horsák, M.

    2011-01-01

    Roč. 70, suppl.1 (2011), s. 54-64 ISSN 1129-5767 Institutional research plan: CEZ:AV0Z60050516 Keywords : diatoms * rotifers * testate amoebae Subject RIV: EF - Botanics Impact factor: 1.207, year: 2011

  4. Unusual Vermamoeba Vermiformis Strain Isolated from Snow in Mount Teide, Tenerife, Canary Islands, Spain

    OpenAIRE

    Maria Reyes-Batlle; Maryam Niyyati; Carmen M. Martín-Navarro; Atteneri López-Arencibia; Basilio Valladares; Enrique Martínez-Carretero; José E. Piñero; Jacob Lorenzo-Morales

    2015-01-01

    Background: Free-living amoebae (FLA) are protozoa that are widely distributed in the environment mainly in water and soil related habitats. Thermophilic amoebae are among the most common FLA in water bodies, being Vermamoeba vermiformis one of the most common species reported worldwide from these sources. Interestingly, V. vermiformis has often been reported to survive at high temperatures and osmotic pressure worldwide.Materials and Methods: In this study, snow samples were collected from M...

  5. Developmental Cycle and Genome Analysis of ?Rubidus massiliensis,? a New Vermamoeba vermiformis Pathogen

    OpenAIRE

    Bou Khalil, Jacques Y.; Benamar, Samia; Baudoin, Jean-Pierre; Croce, Olivier; Blanc-Tailleur, Caroline; Pagnier, Isabelle; Raoult, Didier; La Scola, Bernard

    2016-01-01

    The study of amoeba-associated Chlamydiae is a dynamic field in which new species are increasingly reported. In the present work, we characterized the developmental cycle and analyzed the genome of a new member of this group associated with Vermamoeba vermiformis, we propose to name Rubidus massiliensis. This bacterium is well adapted to its amoeba host and do not reside inside of inclusion vacuoles after phagocytosis. It has a developmental cycle typical of this family of bacteria, with a tr...

  6. Developmental Cycle and Genome Analysis of Protochlamydia massiliensis sp. nov. a New Species in the Parachlamydiacae Family

    OpenAIRE

    Benamar, Samia; Bou Khalil, Jacques Y.; Blanc-Tailleur, Caroline; Bilen, Melhem; Barrassi, Lina; La Scola, Bernard

    2017-01-01

    Amoeba-associated microorganisms (AAMs) are frequently isolated from water networks. In this paper, we report the isolation and characterization of Protochlamydia massiliensis, an obligate intracellular Gram-negative bacterium belonging to the Parachlamydiaceae family in the Chlamydiales order, from a cooling water tower. This bacterium was isolated on Vermamoeba vermiformis. It has a multiple range of hosts among amoeba and is characterized by a typical replication cycle of Chlamydiae with a...

  7. Ciliates rapidly enhance the frequency of conjugation between Escherichia coli strains through bacterial accumulation in vesicles

    OpenAIRE

    Matsuo, Junji; Oguri, Satoshi; Nakamura, Shinji; Hanawa, Tomoko; Fukumoto, Tatsuya; Hayashi, Yasuhiro; Kawaguchi, Kouhei; Mizutani, Yoshihiko; Yao, Takashi; Akizawa, Kouzi; Suzuki, Haruki; Simizu, Chikara; Matsuno, Kazuhiko; Kamiya, Shigeru; Yamaguchi, Hiroyuki

    2010-01-01

    The mechanism underlying bacterial conjugation through protozoa was investigated. Kanamycin-resistant Escherichia coli SM10λ+ carrying pRT733 with TnphoA was used as donor bacteria and introduced by conjugation into ciprofloxacin-resistant E. coli clinical isolate recipient bacteria. Equal amounts of donor and recipient bacteria were mixed together in the presence or absence of protozoa (ciliates, free-living amoebae, myxamoebae) in Page's amoeba saline for 24 h. Transconjugants were selected...

  8. The Use of Physarum for Testing of Toxicity/Mutagenicity

    Science.gov (United States)

    1984-04-19

    grow logarithmically, not synchronously, but the nuclear DNA content indicates that they are predominantly in haploid G2 -phase (Mohberg and Rusch, 1971...amoebae growing on bacteria can be induced to flagellate by washing and suspending in water or buffer (Aldrich, 1967; Mir and Wright, 1978). When...Colonia strain (Wheals, 1970), however, have a single mating type and can form plasmodia within clones, a haploid amoeba developing into a haploid

  9. Pathogenic Naegleria fowleri and non-pathogenic Naegleria lovaniensis exhibit differential adhesion to, and invasion of, extracellular matrix proteins

    OpenAIRE

    Jamerson, Melissa; da Rocha-Azevedo, Bruno; Cabral, Guy A.; Marciano-Cabral, Francine

    2012-01-01

    Naegleria fowleri and Naegleria lovaniensis are closely related free-living amoebae found in the environment. N. fowleri causes primary amoebic meningoencephalitis (PAM), a rapidly fatal disease of the central nervous system, while N. lovaniensis is non-pathogenic. N. fowleri infection occurs when the amoebae access the nasal passages, attach to the nasal mucosa and its epithelial lining, and migrate to the brain. This process involves interaction with components of the host extracellular mat...

  10. Vegetation history and human impact during the last 300 years recorded in a German peat deposit

    OpenAIRE

    van der Linden, M.; Vickery, E.; Charman, D.J.; Broekens, P.; van Geel, B.

    2008-01-01

    A peat core from the Barschpfuhl kettlehole mire in north-east Germany was analysed for multiproxy indicators (plant macrofossils, pollen/non-pollen microfossils, testate amoebae, colorimetric humification, carbon/nitrogen ratios, bulk density, loss on ignition), to investigate the effects of climate change and human impact on vegetation and peat accumulation during the last c. 300 years. 14C wiggle-match dating was applied for high-precision dating. Testate amoebae assemblages were used to r...

  11. Effects of human impact and climate change during the last 350 years recorded in a Swedish raised bog deposit

    OpenAIRE

    van der Linden, M.; Vickery, E.; Charman, D.J.; van Geel, B.

    2008-01-01

    A peat core from an ombrotrophic mire in central Sweden was analysed for multi-proxy indicators (plant macrofossils, pollen/non-pollen microfossils, testate amoebae, colorimetric humification, carbon/nitrogen ratios, bulk densities, loss on ignition), to evaluate the relative contributions of climate change and human impact on vegetation and peat accumulation during the last c. 350 years. 14C wiggle-match dating was applied for high precision dating. Testate amoebae assemblages were used to r...

  12. The occurrence of Naegleria fowleri in recreational waters in Arizona.

    Science.gov (United States)

    Sifuentes, Laura Y; Choate, Brittany L; Gerba, Charles P; Bright, Kelly R

    2014-09-19

    Naegleria fowleri is a free-living amoeba found in waters in warmer regions that causes primary amoebic meningoencephalitis, a rare but almost universally fatal disease. The goal of this project was to assess the occurrence of N. fowleri and other thermophilic amoebae in 33 recreational surface waters across Arizona to determine if their presence could be correlated with seasonal or other environmental factors. First, 1-L grab samples were collected over two years and analyzed using polymerase chain reaction and amoebae viability. Seasonality was observed, with N. fowleri and thermophilic amoebae (20% and 30%, respectively) being detected more often in the winter and spring combined than in the summer and fall combined (7.9% and 9.5%, respectively). The spring and fall both had an average temperature of 18°C, yet had different occurrence data (18.2% versus 5.9% for N. fowleri, respectively; 27.3% versus 0% for viable amoebae, respectively). These results are in stark contrast to previous studies in which N. fowleri has been found almost exclusively during warmer months. Over the two-year study, N. fowleri was detected in six and thermophilic amoebae in eight of the 33 recreational water bodies. Five of these were lakes near Phoenix that tested positive for N. fowleri and thermophilic amoebae over multiple seasons. These lakes differed significantly (P ≤ 0.05) from the other 28 surface waters, with a lower average temperature in the spring, a higher temperature in the fall, a higher pH and turbidity in the summer, and a lower electro-conductivity in the spring. They also had lower Escherichia coli and heterotrophic bacteria levels during colder months. Future N. fowleri monitoring in Arizona should focus on these five lakes to further elucidate the factors that contribute to the low occurrence of this amoeba in the summer or which might explain why these lakes appear to be reservoirs for the organism.

  13. Adaptation of Francisella tularensis to the Mammalian Environment Is Governed by Cues Which Can Be Mimicked In Vitro▿ †

    OpenAIRE

    Hazlett, Karsten R. O.; Caldon, Seth D.; McArthur, Debbie G.; Cirillo, Kerry A.; Kirimanjeswara, Girish S.; Magguilli, Micheal L.; Malik, Meenakshi; Shah, Aaloki; Broderick, Scott; Golovliov, Igor; Metzger, Dennis W.; Rajan, Krishna; Sellati, Timothy J.; Loegering, Daniel J.

    2008-01-01

    The intracellular bacterium Francisella tularensis survives in mammals, arthropods, and freshwater amoeba. It was previously established that the conventional media used for in vitro propagation of this microbe do not yield bacteria that mimic those harvested from infected mammals; whether these in vitro-cultivated bacteria resemble arthropod- or amoeba-adapted Francisella is unknown. As a foundation for our goal of identifying F. tularensis outer membrane proteins which are expressed during ...

  14. Possible environmental sources of Acanthamoeba spp in contact lens wearers.

    OpenAIRE

    Seal, D; Stapleton, F; Dart, J

    1992-01-01

    The water supply and dust samples from the home environment (bathrooms and kitchens) of 50 wearers of contact lenses (CLs) were cultured for the presence of free-living amoebae. CL cases, solutions, and water taps were cultured for bacteria, which amoebae require for growth. Acanthamoeba spp were isolated from water drawn from six bathroom cold water taps (tank supplied), five in the presence of limescale, and from one kitchen cold water tap (mains supplied). There was an association between ...

  15. Allovahlkampfia spelaea Causing Keratitis in Humans.

    Science.gov (United States)

    Tolba, Mohammed Essa Marghany; Huseein, Enas Abdelhameed Mahmoud; Farrag, Haiam Mohamed Mahmoud; Mohamed, Hanan El Deek; Kobayashi, Seiki; Suzuki, Jun; Ali, Tarek Ahmed Mohamed; Sugano, Sumio

    2016-07-01

    Free-living amoebae are present worldwide. They can survive in different environment causing human diseases in some instances. Acanthamoeba sp. is known for causing sight-threatening keratitis in humans. Free-living amoeba keratitis is more common in developing countries. Amoebae of family Vahlkampfiidae are rarely reported to cause such affections. A new genus, Allovahlkampfia spelaea was recently identified from caves with no data about pathogenicity in humans. We tried to identify the causative free-living amoeba in a case of keratitis in an Egyptian patient using morphological and molecular techniques. Pathogenic amoebae were culture using monoxenic culture system. Identification through morphological features and 18S ribosomal RNA subunit DNA amplification and sequencing was done. Pathogenicity to laboratory rabbits and ability to produce keratitis were assessed experimentally. Allovahlkampfia spelaea was identified as a cause of human keratitis. Whole sequence of 18S ribosomal subunit DNA was sequenced and assembled. The Egyptian strain was closely related to SK1 strain isolated in Slovenia. The ability to induce keratitis was confirmed using animal model. This the first time to report Allovahlkampfia spelaea as a human pathogen. Combining both molecular and morphological identification is critical to correctly diagnose amoebae causing keratitis in humans. Use of different pairs of primers and sequencing amplified DNA is needed to prevent misdiagnosis.

  16. Possible environmental sources of Acanthamoeba spp in contact lens wearers.

    Science.gov (United States)

    Seal, D; Stapleton, F; Dart, J

    1992-01-01

    The water supply and dust samples from the home environment (bathrooms and kitchens) of 50 wearers of contact lenses (CLs) were cultured for the presence of free-living amoebae. CL cases, solutions, and water taps were cultured for bacteria, which amoebae require for growth. Acanthamoeba spp were isolated from water drawn from six bathroom cold water taps (tank supplied), five in the presence of limescale, and from one kitchen cold water tap (mains supplied). There was an association between the presence of limescale in water and direct culture for free-living amoebae, suggesting that scale provides a favourable microenvironment for amoebae. Acanthamoebae were also found in dust from around one washbasin. Nineteen of 50 CL cases, 12/122 CL care rinsing solutions, and 59/100 cold water taps yielded Gram negative bacteria which could be ingested by amoebae. It is concluded from this study that CLs should not be washed in first-drawn tank-fed cold water, especially if limescale is present, and that soft CLs should be rinsed in manufactured single-use, sterile solutions. Rigid CL and CL cases should only be washed with boiled tap water (preferably hot), or single-use sterile solutions, and stored dry to prevent multiplication of amoebae and Gram negative bacteria. PMID:1627513

  17. Receptor-mediated uptake of Legionella pneumophila by Acanthamoeba castellanii and Naegleria lovaniensis.

    Science.gov (United States)

    Declerck, P; Behets, J; De Keersmaecker, B; Ollevier, F

    2007-12-01

    Investigation of the attachment and uptake of Legionella pneumophila by Acanthamoeba castellanii and Naegleria lovaniensis, as these are two critical steps in the subsequent bacterial survival in both amoeba hosts. Initially, the mode of Legionella uptake was examined using inhibitors of microfilament-dependent and receptor-mediated uptake phagocytosis. Secondly, the minimum saccharide structure to interfere with L. pneumophila uptake was determined by means of selected saccharides. Bacterial attachment and uptake by each of the amoeba species occurred through a receptor-mediated endocytosis, which required de novo synthesis of host proteins. Legionella pneumophila showed a high affinity to the alpha1-3D-mannobiose domain of the mannose-binding receptor located on A. castellanii. In contrast, L. pneumophila bacteria had a high affinity for the GalNAcbeta1-4Gal domain of the N-acetyl-D-galactosamine receptor of N. lovaniensis. Our data pointed to a remarkable adaptation of L. pneumophila to invade different amoeba hosts, as the uptake by both amoeba species is mediated by two different receptor families. The fact that L. pneumophila is taken up by two different amoeba species using different receptor families adds further complexity to the host-parasite interaction process, as 14 amoeba species are known to be appropriate Legionella hosts.

  18. The interplay between Entamoeba and enteropathogenic bacteria modulates epithelial cell damage.

    Directory of Open Access Journals (Sweden)

    José Manuel Galván-Moroyoqui

    Full Text Available BACKGROUND: Mixed intestinal infections with Entamoeba histolytica, Entamoeba dispar and bacteria with exacerbated manifestations of disease are common in regions where amoebiasis is endemic. However, amoeba-bacteria interactions remain largely unexamined. METHODOLOGY: Trophozoites of E. histolytica and E. dispar were co-cultured with enteropathogenic bacteria strains Escherichia coli (EPEC, Shigella dysenteriae and a commensal Escherichia coli. Amoebae that phagocytosed bacteria were tested for a cytopathic effect on epithelial cell monolayers. Cysteine proteinase activity, adhesion and cell surface concentration of Gal/GalNAc lectin were analyzed in amoebae showing increased virulence. Structural and functional changes and induction of IL-8 expression were determined in epithelial cells before and after exposure to bacteria. Chemotaxis of amoebae and neutrophils to human IL-8 and conditioned culture media from epithelial cells exposed to bacteria was quantified. PRINCIPAL FINDINGS: E. histolytica digested phagocytosed bacteria, although S. dysenteriae retained 70% viability after ingestion. Phagocytosis of pathogenic bacteria augmented the cytopathic effect of E. histolytica and increased expression of Gal/GalNAc lectin on the amoebic surface and increased cysteine proteinase activity. E. dispar remained avirulent. Adhesion of amoebae and damage to cells exposed to bacteria were increased. Additional increases were observed if amoebae had phagocytosed bacteria. Co-culture of epithelial cells with enteropathogenic bacteria disrupted monolayer permeability and induced expression of IL-8. Media from these co-cultures and human recombinant IL-8 were similarly chemotactic for neutrophils and E. histolytica. CONCLUSIONS: Epithelial monolayers exposed to enteropathogenic bacteria become more susceptible to E. histolytica damage. At the same time, phagocytosis of pathogenic bacteria by amoebae further increased epithelial cell damage. SIGNIFICANCE

  19. Impact of non-Legionella bacteria on the uptake and intracellular replication of Legionella pneumophila in Acanthamoeba castellanii and Naegleria lovaniensis.

    Science.gov (United States)

    Declerck, P; Behets, J; Delaedt, Y; Margineanu, A; Lammertyn, E; Ollevier, F

    2005-11-01

    In aquatic environments, Legionella pneumophila survives, in association with other bacteria, within biofilms by multiplying in free-living amoebae. The precise mechanisms underlying several aspects of the uptake and intracellular replication of L. pneumophila in amoebae, especially in the presence of other bacteria, remain unknown. In the present study, we examined the competitive effect of selected non-Legionella bacteria (Escherichia coli, Aeromonas hydrophila, Flavobacterium breve, and Pseudomonas aeruginosa) on the uptake of L. pneumophila serogroup 1 by the amoebae Acanthamoeba castellanii and Naegleria lovaniensis. We also investigated their possible influence on the intracellular replication of L. pneumophila in both amoeba species. Our results showed that the non-Legionella bacteria did not compete with L. pneumophila for uptake, suggesting that the amoeba hosts took in L. pneumophila through a specific and presumably highly efficient uptake mechanism. Living and heat-inactivated P. aeruginosa best supported the replication of L. pneumophila in N. lovaniensis and A. castellanii, respectively, whereas for both amoeba species, E. coli yielded the lowest number of replicated L. pneumophila. Furthermore, microscopic examination showed that 100% of the A. castellanii and only 2% of the N. lovaniensis population were infected with L. pneumophila at the end of the experiment. This study clearly shows the influence of some non-Legionella bacteria on the intracellular replication of L. pneumophila in A. castellanii and N. lovaniensis. It also demonstrates the different abilities of the two tested amoeba species to serve as a proper host for the replication and distribution of the human pathogen in man-made aquatic environments such as cooling towers, shower heads, and air conditioning systems with potential serious consequences for human health.

  20. Growth dynamic of Naegleria fowleri in a microbial freshwater biofilm.

    Science.gov (United States)

    Goudot, Sébastien; Herbelin, Pascaline; Mathieu, Laurence; Soreau, Sylvie; Banas, Sandrine; Jorand, Frédéric

    2012-09-01

    The presence of pathogenic free-living amoebae (FLA) such as Naegleria fowleri in freshwater environments is a potential public health risk. Although its occurrence in various water sources has been well reported, its presence and associated factors in biofilm remain unknown. In this study, the density of N. fowleri in biofilms spontaneously growing on glass slides fed by raw freshwater were followed at 32 °C and 42 °C for 45 days. The biofilms were collected with their substrata and characterized for their structure, numbered for their bacterial density, thermophilic free-living amoebae, and pathogenic N. fowleri. The cell density of N. fowleri within the biofilms was significantly affected both by the temperature and the nutrient level (bacteria/amoeba ratio). At 32 °C, the density remained constantly low (1-10 N. fowleri/cm(2)) indicating that the amoebae were in a survival state, whereas at 42 °C the density reached 30-900 N. fowleri/cm(2) indicating an active growth phase. The nutrient level, as well, strongly affected the apparent specific growth rate (μ) of N. fowleri in the range of 0.03-0.23 h(-1). At 42 °C a hyperbolic relationship was found between μ and the bacteria/amoeba ratio. A ratio of 10(6) to 10(7) bacteria/amoeba was needed to approach the apparent μ(max) value (0.23 h(-1)). Data analysis also showed that a threshold for the nutrient level of close to 10(4) bacteria/amoeba is needed to detect the growth of N. fowleri in freshwater biofilm. This study emphasizes the important role of the temperature and bacteria as prey to promote not only the growth of N. fowleri, but also its survival. Copyright © 2012 Elsevier Ltd. All rights reserved.

  1. Interactions between Human Norovirus Surrogates and Acanthamoeba spp.

    Science.gov (United States)

    Hsueh, Tun-Yun

    2015-01-01

    Human noroviruses (HuNoVs) are the most common cause of food-borne disease outbreaks, as well as virus-related waterborne disease outbreaks in the United States. Here, we hypothesize that common free-living amoebae (FLA)—ubiquitous in the environment, known to interact with pathogens, and frequently isolated from water and fresh produce—could potentially act as reservoirs of HuNoV and facilitate the environmental transmission of HuNoVs. To investigate FLA as reservoirs for HuNoV, the interactions between two Acanthamoeba species, A. castellanii and A. polyphaga, as well as two HuNoV surrogates, murine norovirus type 1 (MNV-1) and feline calicivirus (FCV), were evaluated. The results showed that after 1 h of amoeba-virus incubation at 25°C, 490 and 337 PFU of MNV-1/ml were recovered from A. castellanii and A. polyphaga, respectively, while only few or no FCVs were detected. In addition, prolonged interaction of MNV-1 with amoebae was investigated for a period of 8 days, and MNV-1 was demonstrated to remain stable at around 200 PFU/ml from day 2 to day 8 after virus inoculation in A. castellanii. Moreover, after a complete amoeba life cycle (i.e., encystment and excystment), infectious viruses could still be detected. To determine the location of virus associated with amoebae, immunofluorescence experiments were performed and showed MNV-1 transitioning from the amoeba surface to inside the amoeba over a 24-h period. These results are significant to the understanding of how HuNoVs may interact with other microorganisms in the environment in order to aid in its persistence and survival, as well as potential transmission in water and to vulnerable food products such as fresh produce. PMID:25841006

  2. Yersinia pestis Resists Predation by Acanthamoeba castellanii and Exhibits Prolonged Intracellular Survival.

    Science.gov (United States)

    Benavides-Montaño, Javier A; Vadyvaloo, Viveka

    2017-07-01

    Plague is a flea-borne rodent-associated zoonotic disease caused by Yersinia pestis The disease is characterized by epizootics with high rodent mortalities, punctuated by interepizootic periods when the bacterium persists in an unknown reservoir. This study investigates the interaction between Y. pestis and the ubiquitous soil free-living amoeba (FLA) Acanthamoeba castellanii to assess if the bacterium can survive within soil amoebae and whether intracellular mechanisms are conserved between infection of mammalian macrophages and soil amoebae. The results demonstrate that during coculture with amoebae, representative Y. pestis strains of epidemic biovars Medievalis, Orientalis, and Antiqua are phagocytized and able to survive within amoebae for at least 5 days. Key Y. pestis determinants of the intracellular interaction of Y. pestis and phagocytic macrophages, PhoP and the type three secretion system (T3SS), were then tested for their roles in the Y. pestis -amoeba interaction. Consistent with a requirement for the PhoP transcriptional activator in the intracellular survival of Y. pestis in macrophages, a PhoP mutant is unable to survive when cocultured with amoebae. Additionally, induction of the T3SS blocks phagocytic uptake of Y. pestis by amoebae, similar to that which occurs during macrophage infection. Electron microscopy revealed that in A. castellanii , Y. pestis resides intact within spacious vacuoles which were characterized using lysosomal trackers as being separated from the lysosomal compartment. This evidence for prolonged survival and subversion of intracellular digestion of Y. pestis within FLA suggests that protozoa may serve as a protective soil reservoir for Y. pestis IMPORTANCE Yersinia pestis is a reemerging flea-borne zoonotic disease. Sylvatic plague cycles are characterized by an epizootic period during which the disease spreads rapidly, causing high rodent mortality, and an interepizootic period when the bacterium quiescently persists in an

  3. Vernalophrys algivore gen. nov., sp. nov. (Rhizaria: Cercozoa: Vampyrellida), a New Algal Predator Isolated from Outdoor Mass Culture of Scenedesmus dimorphus

    Science.gov (United States)

    Patterson, David J.; Li, Yunguang; Hu, Zixuan; Sommerfeld, Milton; Chen, Yongsheng

    2015-01-01

    Microbial contamination is the main cause of loss of biomass yield in microalgal cultures, especially under outdoor environmental conditions. Little is known about the identities of microbial contaminants in outdoor mass algal cultures. In this study, a new genus and species of vampyrellid amoeba, Vernalophrys algivore, is described from cultures of Scenedesmus dimorphus in open raceway ponds and outdoor flat-panel photobioreactors. This vampyrellid amoeba was a significant grazer of Scenedesmus and was frequently associated with a very rapid decline in algal numbers. We report on the morphology, subcellular structure, feeding behavior, molecular phylogeny, and life cycle. The new amoeba resembles Leptophrys in the shape of trophozoites and pseudopodia and in the mechanism of feeding (mainly by engulfment). It possesses two distinctive regions in helix E10_1 (nucleotides 117 to 119, CAA) and E23_1 (nucleotides 522 and 523, AG) of the 18S rRNA gene. It did not form a monophyletic group with Leptophrys in molecular phylogenetic trees. We establish a new genus, Vernalophrys, with the type species Vernalophrys algivore. The occurrence, impact of the amoeba on mass culture of S. dimorphus, and means to reduce vampyrellid amoeba contamination in Scenedesmus cultures are addressed. The information obtained from this study will be useful for developing an early warning system and control measures for preventing or treating this contaminant in microalgal mass cultures. PMID:25819973

  4. Isolation of an Acanthamoeba strain with intracellular Burkholderia pickettii infection.

    Science.gov (United States)

    Michel, R; Hauröder, B

    1997-04-01

    Burkholderia pickettii is a facultative pathogen that has been isolated from patient sources and environmental sources including respiratory therapy solutions, deionized water and aqueous disinfectants. The organism has been associated with septicemia and respiratory tract infections. In our investigation, Burkholderia pickettii (biovar 2) was for the first time isolated from Acanthamoeba sp. (group II), a free living amoeba species recovered from the wet area of a physiotherapy unit. Pathogenic strains of acanthamoebae may cause amoebic-encephalitis (AE) and keratitis. Light and electron microscopic examinations showed that in a first step, the bacterial were phagocytized by the amoebae. In contrast to Enterobacter cloacae and Escherichia coli that were used as food organisms and digested within food vacuoles, Burkholderia pickettii caused the amoebae to develop large vacuoles filled with completely intact and motile bacteria. 3-5 days after infection, Pseudomonas pickettii had multiplied within the enlarging parasitophorous vacuoles. Ultrastructural changes in the host cells occurred and the amoebae finally underwent rupture or lysis. In cocultivation assays we could not only reinfect the original host amoeba but Acanthamoeba strains from other habitats could successfully be infected with Burkholderia pickettii as well.

  5. Infection of Acanthamoeba castellanii by Chlamydia pneumoniae.

    Science.gov (United States)

    Essig, A; Heinemann, M; Simnacher, U; Marre, R

    1997-04-01

    Chlamydia pneumoniae is an intracellular respiratory pathogen, which, similar to Legionella, might have developed mechanisms to escape the intracellular bactericidal activity of both human host cells and amoeba. We therefore investigated the intracellular growth and survival of C. pneumoniae in Acanthamoeba castellanii by using cell culture, immunofluorescence microscopy, and electron microscopy. A castellanii was incubated with purified elementary bodies of C. pneumoniae TW 183 at a concentration of 10(6) inclusion-forming units (IFU)/ml to give a ratio of approximately 1 IFU of C. pneumoniae per amoeba. Quantitative determination of chlamydial growth within A. castellanii revealed viable and infective C. pneumoniae in the range of 10(4) to 10(5) IFU/ml between days 7 and 14 postinfection. Immunofluorescence analysis and transmission electron microscopy with subsequent immunogold staining confirmed evidence of infection of the amoebae by C. Pneumoniae and additionally revealed that C. pneumoniae entered the typical growth cycle. Our results show that amoebae allow the survival of C. pneumoniae, suggesting that amoebae may serve as an additional reservoir for Chlamydia or Chlamydia-related organisms.

  6. Dictyostelium discoideum as a novel host system to study the interaction between phagocytes and yeasts

    Directory of Open Access Journals (Sweden)

    Barbara Koller

    2016-10-01

    Full Text Available The social amoeba Dictyostelium discoideum is a well-established model organism to study the interaction between bacteria and phagocytes. In contrast, research using D. discoideum as a host model for fungi is rare. We describe a comprehensive study, which uses D. discoideum as a host model system to investigate the interaction with apathogenic (Saccharomyces cerevisiae and pathogenic (Candida sp. yeast. We show that Dictyostelium can be co-cultivated with yeasts on solid media, offering a convenient test to study the interaction between fungi and phagocytes. We demonstrate that a number of D. discoideum mutants increase (atg1-, kil1-, kil2- or decrease (atg6- the ability of the amoebae to predate yeast cells. On the yeast side, growth characteristics, reduced phagocytosis rate, as well as known virulence factors of C. albicans (EFG1, CPH1, HGC1, ICL1 contribute to the resistance of yeast cells against predation by the amoebae. Investigating haploid C. albicans strains, we suggest using the amoebae plate test for screening purposes after random mutagenesis. Finally, we discuss the potential of our adapted amoebae plate test to use D. discoideum for risk assessment of yeast strains.

  7. Protozoa Drive the Dynamics of Culturable Biocontrol Bacterial Communities.

    Science.gov (United States)

    Müller, Maren Stella; Scheu, Stefan; Jousset, Alexandre

    2013-01-01

    Some soil bacteria protect plants against soil-borne diseases by producing toxic secondary metabolites. Such beneficial biocontrol bacteria can be used in agricultural systems as alternative to agrochemicals. The broad spectrum toxins responsible for plant protection also inhibit predation by protozoa and nematodes, the main consumers of bacteria in soil. Therefore, predation pressure may favour biocontrol bacteria and contribute to plant health. We analyzed the effect of Acanthamoeba castellanii on semi-natural soil bacterial communities in a microcosm experiment. We determined the frequency of culturable bacteria carrying genes responsible for the production of the antifungal compounds 2,4-diacetylphloroglucinol (DAPG), pyrrolnitrin (PRN) and hydrogen cyanide (HCN) in presence and absence of A. castellanii. We then measured if amoebae affected soil suppressiveness in a bioassay with sugar beet seedlings confronted to the fungal pathogen Rhizoctonia solani. Amoebae increased the frequency of both DAPG and HCN positive bacteria in later plant growth phases (2 and 3 weeks), as well as the average number of biocontrol genes per bacterium. The abundance of DAPG positive bacteria correlated with disease suppression, suggesting that their promotion by amoebae may enhance soil health. However, the net effect of amoebae on soil suppressiveness was neutral to slightly negative, possibly because amoebae slow down the establishment of biocontrol bacteria on the recently emerged seedlings used in the assay. The results indicate that microfaunal predators foster biocontrol bacterial communities. Understanding interactions between biocontrol bacteria and their predators may thus help developing environmentally friendly management practices of agricultural systems.

  8. Survival of enterohemorrhagic Escherichia coli in the presence of Acanthamoeba castellanii and its dependence on Pho regulon

    Science.gov (United States)

    Chekabab, Samuel Mohammed; Daigle, France; Charette, Steve J; Dozois, Charles M; Harel, Josée

    2012-01-01

    Enterohemorrhagic Escherichia coli (EHEC) are involved in outbreaks of food-borne illness and transmitted to humans through bovine products or water contaminated by cattle feces. Microbial interaction is one of the strategies used by pathogenic bacteria to survive in the environment. Among protozoa, the free-living amoebae are known to host and protect several water-borne pathogens. In this study, the interaction between EHEC and the predacious protozoa Acanthamoeba castellanii was investigated. Using monoculture and cocultures, growth of both organisms was estimated for 3 weeks by total and viable cell counts. The numbers of EHEC were significantly higher when cultured with amoebae than without, and less EHEC shifted into a viable but nonculturable state in the presence of amoebae. Using several mutants, we observed that the Pho regulon is required for EHEC growth when cocultured with amoebae. In contrast, the Shiga toxins (Stx) were not involved in this association phenotype. Cocultures monitored by electron microscopy revealed a loss of the regular rod shape of EHEC and the secretion of multilamellar vesicles by the amoebae, which did not contain bacteria. As the interaction between A. castellanii and EHEC appears beneficial for bacterial growth, this supports a potential role for protozoa in promoting the persistence of EHEC in the environment. PMID:23233434

  9. Interaction of Aspergillus fumigatus conidia with Acanthamoeba castellanii parallels macrophage-fungus interactions.

    Science.gov (United States)

    Van Waeyenberghe, Lieven; Baré, Julie; Pasmans, Frank; Claeys, Myriam; Bert, Wim; Haesebrouck, Freddy; Houf, Kurt; Martel, An

    2013-12-01

    Aspergillus fumigatus and free-living amoebae are common inhabitants of soil. Mechanisms of A. fumigatus to circumvent the amoeba's digestion may facilitate overcoming the vertebrate macrophage defence mechanisms. We performed co-culture experiments using A. fumigatus conidia and the amoeba Acanthamoeba castellanii. Approximately 25% of the amoebae ingested A. fumigatus conidia after 1 h of contact. During intra-amoebal passage, part of the ingested conidia was able to escape the food vacuole and to germinate inside the cytoplasm of A. castellanii. Fungal release into the extra-protozoan environment by exocytosis of conidia or by germination was observed with light and transmission electron microscopy. These processes resulted in structural changes in A. castellanii, leading to amoebal permeabilization without cell lysis. In conclusion, A. castellanii internalizes A. fumigatus conidia, resulting in fungal intracellular germination and subsequent amoebal death. As such, this interaction highly resembles that of A. fumigatus with mammalian and avian macrophages. This suggests that A. fumigatus virulence mechanisms to evade macrophage killing may be acquired by co-evolutionary interactions among A. fumigatus and environmental amoebae. © 2013 John Wiley & Sons Ltd and Society for Applied Microbiology.

  10. Protozoa Drive the Dynamics of Culturable Biocontrol Bacterial Communities.

    Directory of Open Access Journals (Sweden)

    Maren Stella Müller

    Full Text Available Some soil bacteria protect plants against soil-borne diseases by producing toxic secondary metabolites. Such beneficial biocontrol bacteria can be used in agricultural systems as alternative to agrochemicals. The broad spectrum toxins responsible for plant protection also inhibit predation by protozoa and nematodes, the main consumers of bacteria in soil. Therefore, predation pressure may favour biocontrol bacteria and contribute to plant health. We analyzed the effect of Acanthamoeba castellanii on semi-natural soil bacterial communities in a microcosm experiment. We determined the frequency of culturable bacteria carrying genes responsible for the production of the antifungal compounds 2,4-diacetylphloroglucinol (DAPG, pyrrolnitrin (PRN and hydrogen cyanide (HCN in presence and absence of A. castellanii. We then measured if amoebae affected soil suppressiveness in a bioassay with sugar beet seedlings confronted to the fungal pathogen Rhizoctonia solani. Amoebae increased the frequency of both DAPG and HCN positive bacteria in later plant growth phases (2 and 3 weeks, as well as the average number of biocontrol genes per bacterium. The abundance of DAPG positive bacteria correlated with disease suppression, suggesting that their promotion by amoebae may enhance soil health. However, the net effect of amoebae on soil suppressiveness was neutral to slightly negative, possibly because amoebae slow down the establishment of biocontrol bacteria on the recently emerged seedlings used in the assay. The results indicate that microfaunal predators foster biocontrol bacterial communities. Understanding interactions between biocontrol bacteria and their predators may thus help developing environmentally friendly management practices of agricultural systems.

  11. Legionella pneumophila decreases velocity of Acanthamoeba castellanii.

    Science.gov (United States)

    Mengue, Luce; Richard, Freddie-Jeanne; Caubet, Yves; Rolland, Steven; Héchard, Yann; Samba-Louaka, Ascel

    2017-12-01

    Acanthamoeba castellanii is a free-living amoeba commonly found in aquatic environment. It feeds on bacteria even if some bacteria resist amoebal digestion. Thus, A. castellanii is described as a Trojan horse able to harbor pathogenic bacteria. L. pneumophila is one of the amoeba-resisting bacteria able to avoid host degradation by phagocytosis and to multiply inside the amoeba. When infecting its host, L. pneumophila injects hundreds of effectors via a type IV secretion system that change physiology of the amoeba to its profit. In this study, we assess mobility of A. castellanii upon infection with L. pneumophila. Electron-microscopy analysis of amoebae revealed a reduction of acanthopodia on cells infected with L. pneumophila. Analysis of velocity showed that migration of A. castellanii infected with L. pneumophila was significantly impaired compare to uninfected cells. Taken together, infection with L. pneumophila could prevent formation of cytoplasmic extensions such as acanthopodia with consequences on the shape, adherence and mobility of A. castellanii. Copyright © 2017 Elsevier Inc. All rights reserved.

  12. Species-specific immunity induced by infection with Entamoeba histolytica and Entamoeba moshkovskii in mice.

    Directory of Open Access Journals (Sweden)

    Chikako Shimokawa

    Full Text Available Entamoeba histolytica, the parasitic amoeba responsible for amoebiasis, causes approximately 100,000 deaths every year. There is currently no vaccine against this parasite. We have previously shown that intracecal inoculation of E. histolytica trophozoites leads to chronic and non-healing cecitis in mice. Entamoeba moshkovskii, a closely related amoeba, also causes diarrhea and other intestinal disorders in this model. Here, we investigated the effect of infection followed by drug-cure of these species on the induction of immunity against homologous or heterologous species challenge. Mice were infected with E. histolytica or E. moshkovskii and treated with metronidazole 14 days later. Re-challenge with E. histolytica or E. moshkovskii was conducted seven or 28 days following confirmation of the clearance of amoebae, and the degree of protection compared to non-exposed control mice was evaluated. We show that primary infection with these amoebae induces a species-specific immune response which protects against challenge with the homologous, but not a heterologous species. These findings pave the way, therefore, for the identification of novel amoebae antigens that may become the targets of vaccines and provide a useful platform to investigate host protective immunity to Entamoeba infections.

  13. Species-specific immunity induced by infection with Entamoeba histolytica and Entamoeba moshkovskii in mice.

    Science.gov (United States)

    Shimokawa, Chikako; Culleton, Richard; Imai, Takashi; Suzue, Kazutomo; Hirai, Makoto; Taniguchi, Tomoyo; Kobayashi, Seiki; Hisaeda, Hajime; Hamano, Shinjiro

    2013-01-01

    Entamoeba histolytica, the parasitic amoeba responsible for amoebiasis, causes approximately 100,000 deaths every year. There is currently no vaccine against this parasite. We have previously shown that intracecal inoculation of E. histolytica trophozoites leads to chronic and non-healing cecitis in mice. Entamoeba moshkovskii, a closely related amoeba, also causes diarrhea and other intestinal disorders in this model. Here, we investigated the effect of infection followed by drug-cure of these species on the induction of immunity against homologous or heterologous species challenge. Mice were infected with E. histolytica or E. moshkovskii and treated with metronidazole 14 days later. Re-challenge with E. histolytica or E. moshkovskii was conducted seven or 28 days following confirmation of the clearance of amoebae, and the degree of protection compared to non-exposed control mice was evaluated. We show that primary infection with these amoebae induces a species-specific immune response which protects against challenge with the homologous, but not a heterologous species. These findings pave the way, therefore, for the identification of novel amoebae antigens that may become the targets of vaccines and provide a useful platform to investigate host protective immunity to Entamoeba infections.

  14. Identification of Conserved ABC Importers Necessary for Intracellular Survival of Legionella pneumophila in Multiple Hosts

    Directory of Open Access Journals (Sweden)

    Amrita Lama

    2017-11-01

    Full Text Available It is established that the human pathogen Legionella pneumophila becomes significantly augmented for infection of macrophages after intracellular growth in amoebae when compared to like-strains cultivated in laboratory media. Based on this observation, we reasoned that the most critical virulence determinants of L.p. are expressed by responding to stimuli generated by the protozoan host specifically; a process we term “protozoan-priming.” We sought to identify L.p. virulence factors that were required for replication in amoebae in order to highlight the genes necessary for production of the most infectious form of the bacterium. Using a transposon mutagenesis screen, we successfully identified 12 insertions that produced bacteria severely attenuated for growth in amoebae, while retaining a functional Dot/Icm type IVb secretion system. Seven of these insertion mutants were found dispensable for growth in macrophages, revealing attractive therapeutic targets that reside upstream of the pathogen-human interface. Two candidates identified, lpg0730 and lpg0122 were required for survival and replication in amoebae and macrophage host cells. Both genes are conserved among numerous important human pathogenic bacteria that can persist or replicate in amoebae. Each gene encodes a component of an ATP binding cassette (ABC transport complex of unknown function. We demonstrate the lpg0730 ortholog in Francisella tularensis subsp. novicida to be essential for colonization of both protozoan and mammalian host cells, highlighting conserved survival mechanisms employed by bacteria that utilize protozoa as an environmental reservoir for replication.

  15. Arcella peruviana sp. nov. (Amoebozoa: Arcellinida, Arcellidae), a new species from a tropical peatland in Amazonia.

    Science.gov (United States)

    Reczuga, Monika K; Swindles, Graeme T; Grewling, Łukasz; Lamentowicz, Mariusz

    2015-10-01

    There has only been one study on the ecology of testate amoebae from Amazonian peatlands, despite Amazonia being a biodiversity hotspot of global importance. During analysis of litter samples from Aucayacu peatland, western (Peruvian) Amazonia, we discovered a testate amoeba with a distinct morphology unlike any other species reported previously. We describe a new species, Arcella peruviana, based on its distinct morphology, compare it to morphologically similar species and provide information about its ecology. This new species is characterised by a distinct cruciform aperture (diameter ranges between 12 and 17μm) which is slightly invaginated. The test is small (height 43-57μm) and polygonal in cross-section. Our discovery suggests the existence of an unknown diversity of testate amoebae in Amazonia. The absence of the new Arcella species in more intensively-sampled regions supports the view that protists have restricted distributions. Copyright © 2015 Elsevier GmbH. All rights reserved.

  16. Meningoencephalitis due to the amoeboflagellate Naegleria fowleri in ruminants in Algeria

    Directory of Open Access Journals (Sweden)

    Benterki Mohamed Seghir

    2016-01-01

    Full Text Available Primary amoebic meningoencephalitis (PAM is a fatal infection in most cases, caused by the amoeba flagellate Naegleria fowleri. This report describes the first cases of PAM in Algeria, in a cow and a ewe from Batna, north-eastern Algeria. The death of both ruminants occurred a week after the first clinical manifestations. The cerebrospinal fluid, after staining with May-Grünwald-Giemsa, showed the presence of amoebae cells. Histological sections revealed numerous amoebae in all parts of the brain. The presence of N. fowleri was confirmed using a species-specific real-time PCR in histological tissue sections. The two PAM cases were reported during the hot season, and the source of infection is very likely the water where the cattle came to drink. Particular attention should be focused on this type of infection in aquatic environments when the temperature is high and preventive measures must be taken to avoid the proliferation of N. fowleri.

  17. The use of dimethyl sulfoxide in contact lens disinfectants is a potential preventative strategy against contracting Acanthamoeba keratitis.

    Science.gov (United States)

    Siddiqui, Ruqaiyyah; Aqeel, Yousuf; Khan, Naveed Ahmed

    2016-10-01

    Acanthamoeba castellanii is the causative agent of blinding keratitis. Though reported in non-contact lens wearers, it is most frequently associated with improper use of contact lens. For contact lens wearers, amoebae attachment to the lens is a critical first step, followed by amoebae binding to the corneal epithelial cells during extended lens wear. Acanthamoeba attachment to surfaces (biological or inert) and migration is an active process and occurs during the trophozoite stage. Thus retaining amoebae in the cyst stage (dormant form) offers an added preventative measure in impeding parasite traversal from the contact lens onto the cornea. Here, we showed that as low as 3% DMSO, abolished A. castellanii excystation. Based on the findings, it is proposed that DMSO should be included in the contact lens disinfectants as an added preventative strategy against contracting Acanthamoeba keratitis. Copyright © 2016 British Contact Lens Association. Published by Elsevier Ltd. All rights reserved.

  18. Arcobacter butzleri survives within trophozoite of Acanthamoeba castellanii.

    Science.gov (United States)

    Villanueva, María P; Medina, Gustavo; Fernández, Heriberto

    2016-01-01

    The survival of three Arcobacter butzleri strains inside Acanthamoeba castellanii was assessed using axenic cultures of A. castellanii that were inoculated with the tested strains and incubated at 26°C under aerobic conditions for 240h. The behavior of bacteria in contact with amoebae was monitored using phase contrast microscopy. The bacterial survival rate within amoebae was assessed through counting colony forming units, using the gentamicin protection assay. All A. butzleri strains were able to survive during 240h within the amoebae, thus suggesting that (i) A. butzleri resists the amoebic digestion processes at least for the analyzed time; (ii) that A. castellanii could serve as an environmental reservoir for this bacterium, probably acting as a transmission vehicle for A. butzleri. Copyright © 2016 Asociación Argentina de Microbiología. Publicado por Elsevier España, S.L.U. All rights reserved.

  19. Acanthamoeba culbertsoni isolated from a clinical case with intraocular dissemination: Structure and in vitro analysis of the interaction with hamster cornea and MDCK epithelial cell monolayers.

    Science.gov (United States)

    González-Robles, Arturo; Omaña-Molina, Maritza; Salazar-Villatoro, Lizbeth; Flores-Maldonado, Catalina; Lorenzo-Morales, Jacob; Reyes-Batlle, María; Arnalich-Montiel, Francisco; Martínez-Palomo, Adolfo

    2017-12-01

    Acanthamoeba culbertsoni trophozoites, previously isolated from a human keratitis case with severe intraocular damage, were maintained in axenic culture. Co-incubation of amoebae with MDCK cell monolayers demonstrated an apparent preference of the amoebae to introduce themselves between the cells. The trophozoites appeared to cross the cell monolayer through the tight junctions, which resulted in decreased trans-epithelial resistance (TER) measurements. Unexpectedly, after co-incubation of amoebae with hamster corneas, we observed that the trophozoites were able to cross the different cell layers and reach the corneal stroma after only 12 h of interaction, in contrast to other Acanthamoeba species. These observations suggest that this A. culbertsoni isolate is particularly pathogenic. Further research with diverse methodologies needs to be performed to explain the unique behavior of this Acanthamoeba strain. Copyright © 2017 Elsevier Inc. All rights reserved.

  20. Necrotizing Meningoencephalitis in a Captive Black and White Ruffed Lemur (Varecia variegata variegata) Caused by Acanthamoeba T4 Genotype.

    Science.gov (United States)

    Gaide, N; Pelandakis, M; Robveille, C; Albaric, O; Jouvion, G; Souchon, M; Risler, A; Abadie, J

    2015-11-01

    A mature male, black and white ruffed lemur (Varecia variegata variegata) died in a zoological garden after a 4-day history of lethargy and non-responsive convulsions. Necropsy and histopathological examinations revealed acute necrotizing and haemorrhagic meningoencephalitis with intralesional amoebas confirmed by immunohistochemistry. Acanthamoeba T4 genotype was identified as the causative agent of the brain lesion, based on amplification and sequencing of 18S ribosomal RNA genes. The presence of free-living amoebas in water and mud from the lemur's environment was investigated by morphological and molecular analyses. The two predominant genera, representing 80% of isolated amoebas, were Naegleria spp. and Acanthamoeba spp. All Acanthamoeba isolates belonged to the T4 genotype. To the author's knowledge, this is the first report of a meningoencephalitis due to Acanthamoeba T4 genotype in Lemuridae with concurrent analysis of pathological tissues and environment. Copyright © 2015 Elsevier Ltd. All rights reserved.

  1. Role of bacterial surface structures on the interaction of Klebsiella pneumoniae with phagocytes.

    Directory of Open Access Journals (Sweden)

    Catalina March

    Full Text Available Phagocytosis is a key process of the immune system. The human pathogen Klebsiella pneumoniae is a well known example of a pathogen highly resistant to phagocytosis. A wealth of evidence demonstrates that the capsule polysaccharide (CPS plays a crucial role in resistance to phagocytosis. The amoeba Dictyostelium discoideum shares with mammalian macrophages the ability to phagocytose and kill bacteria. The fact that K. pneumoniae is ubiquitous in nature and, therefore, should avoid predation by amoebae, poses the question whether K. pneumoniae employs similar means to counteract amoebae and mammalian phagocytes. Here we developed an assay to evaluate K. pneumoniae-D. discoideum interaction. The richness of the growth medium affected the threshold at which the cps mutant was permissive for Dictyostelium and only at lower nutrient concentrations the cps mutant was susceptible to predation by amoebae. Given the critical role of bacterial surface elements on host-pathogen interactions, we explored the possible contribution of the lipopolysaccharide (LPS and outer membrane proteins (OMPs to combat phagoyctosis by D. discoideum. We uncover that, in addition to the CPS, the LPS O-polysaccharide and the first core sugar participate in Klebsiella resistance to predation by D. discoideum. K. pneumoniae LPS lipid A decorations are also necessary to avoid predation by amoebae although PagP-dependent palmitoylation plays a more important role than the lipid A modification with aminoarabinose. Mutants lacking OMPs OmpA or OmpK36 were also permissive for D. discoideium growth. Except the LPS O-polysaccharide mutants, all mutants were more susceptible to phagocytosis by mouse alveolar macrophages. Finally, we found a correlation between virulence, using the pneumonia mouse model, and resistance to phagocytosis. Altogether, this work reveals novel K. pneumoniae determinants involved in resistance to phagocytosis and supports the notion that Dictyostelium amoebae

  2. Polarization and charge transfer in the hydration of chloride ions

    International Nuclear Information System (INIS)

    Zhao Zhen; Rogers, David M.; Beck, Thomas L.

    2010-01-01

    A theoretical study of the structural and electronic properties of the chloride ion and water molecules in the first hydration shell is presented. The calculations are performed on an ensemble of configurations obtained from molecular dynamics simulations of a single chloride ion in bulk water. The simulations utilize the polarizable AMOEBA force field for trajectory generation and MP2-level calculations are performed to examine the electronic structure properties of the ions and surrounding waters in the external field of more distant waters. The ChelpG method is employed to explore the effective charges and dipoles on the chloride ions and first-shell waters. The quantum theory of atoms in molecules (QTAIM) is further utilized to examine charge transfer from the anion to surrounding water molecules. The clusters extracted from the AMOEBA simulations exhibit high probabilities of anisotropic solvation for chloride ions in bulk water. From the QTAIM analysis, 0.2 elementary charges are transferred from the ion to the first-shell water molecules. The default AMOEBA model overestimates the average dipole moment magnitude of the ion compared to the quantum mechanical value. The average magnitude of the dipole moment of the water molecules in the first shell treated at the MP2-level, with the more distant waters handled with an AMOEBA effective charge model, is 2.67 D. This value is close to the AMOEBA result for first-shell waters (2.72 D) and is slightly reduced from the bulk AMOEBA value (2.78 D). The magnitude of the dipole moment of the water molecules in the first solvation shell is most strongly affected by the local water-water interactions and hydrogen bonds with the second solvation shell, rather than by interactions with the ion.

  3. Mechanisms associated with phagocytosis of Arcobacter butzleri by Acanthamoeba castellanii.

    Science.gov (United States)

    Medina, Gustavo; Flores-Martin, Sandra; Fonseca, Belchiolina; Otth, Carola; Fernandez, Heriberto

    2014-05-01

    Acanthamoeba castellanii is a free-living amoeba widely found in environmental matrices such as soil and water. Arcobacter butzleri is an emerging potential zoonotic pathogen that can be isolated from environmental water sources, where they can establish endosymbiotic relationships with amoebas. The aim of this study was to describe the implication of mannose-binding proteins and membrane-associated receptors of glucose and galactose present in the amoebic membrane, during the attachment of Arcobacter butzleri by blocking with different saccharides. Another objective was to describe the signaling pathways involved in phagocytosis of these bacteria using specific inhibitors and analyze the implication of phagolysosome formation on the survival of Arcobacter butzleri inside the amoeba. We infer that the attachment of Arcobacter butzleri to the amoeba is a process which involves the participation of mannose-binding proteins and membrane-associated receptors of glucose and galactose present in the amoeba. We also demonstrated an active role of protozoan actin polymerization in the phagocytosis of Arcobacter butzleri and a critical involvement of PI3K and RhoA pathways. Further, we demonstrated that the tyrosine kinase-induced actin polymerization signal is essential in Acanthamoeba-mediated bacterial uptake. Through phagolysosomal formation analysis, we conclude that the survival of Arcobacter butzleri inside the amoeba could be related with the ability to remain inside vacuoles not fused with lysosomes, or with the ability to retard the fusion between these structures. All these results help the understanding of the bacterial uptake mechanisms used by Acanthamoeba castellanii and contribute to evidence of the survival mechanisms of Arcobacter butzleri.

  4. Survival of the fattest

    DEFF Research Database (Denmark)

    Morgani, Sophie M; Brickman, Joshua M

    2013-01-01

    Experiments on the social amoeba Dictyostelium discoideum show that the origins of lineage bias in this system lie in the nutritional history of individual cells. Clues to the molecular basis for this process suggest similar forces may be at work in early mammalian development.......Experiments on the social amoeba Dictyostelium discoideum show that the origins of lineage bias in this system lie in the nutritional history of individual cells. Clues to the molecular basis for this process suggest similar forces may be at work in early mammalian development....

  5. Ribosomal RNA genes challenge the monophyly of the Hyalospheniidae (Amoebozoa: Arcellinida)

    DEFF Research Database (Denmark)

    Lara, Enrique; Heger, Thierry J; Ekelund, Flemming

    2008-01-01

    causes confusion in the phylogeny, taxonomy and the debate on cosmopolitanism of free-living protists. Here we present a SSU rRNA-based phylogeny of the Hyalospheniidae including the most common species. Similar to the filose testate amoebae of the order Euglyphida the most basal clades have a terminal......To date only five partial and two complete SSU rRNA gene sequences are available for the lobose testate amoebae (Arcellinida). Consequently, the phylogenetic relationships among taxa and the definition of species are still largely dependant on morphological characters of uncertain value, which...

  6. Acanthamoeba castellanii of the T4 genotype is a potential environmental host for Enterobacter aerogenes and Aeromonas hydrophila.

    Science.gov (United States)

    Yousuf, Farzana Abubakar; Siddiqui, Ruqaiyyah; Khan, Naveed Ahmed

    2013-06-07

    Acanthamoeba can interact with a wide range of microorganisms such as viruses, algae, yeasts, protists and bacteria including Legionella pneumophila, Pseudomonas aeruginosa, Vibrio cholerae, Helicobacter pylori, Listeria monocytogenes, Mycobacterium spp., and Escherichia coli. In this capacity, Acanthamoeba has been suggested as a vector in the transmission of bacterial pathogens to the susceptible hosts. Here, we used a keratitis isolate of A. castellanii of the T4 genotype and studied its interactions with two bacterial genera which have not been tested before, Enterobacter aerogenes, and Aeromonas hydrophila, as well as E. coli. Assays were performed to determine bacterial association with and invasion of A. castellanii. Additionally, bacterial survival intracellular of A. castellanii trophozoites as well as cysts was determined. All three bacterial isolates tested, associated, invaded, and survived inside A. castellanii trophozoites as well as A. castellanii cysts. However, E. aerogenes and E. coli exhibited significantly reduced association with and invasion of A. castellanii as compared with A. hydrophila (P castellanii trophozoites, while amoeba remained intact; however A. hydrophila exhibited higher survival inside amoebae (14.54 ± 3.3 bacteria:amoeba ratio) compared with E. aerogenes (3.96 ± 0.7 bacteria:amoeba ratio) and E. coli (5.85 ± 1.1 bacteria:amoeba ratio). A. hydrophila, E. coli, and E. aerogenes remained viable during the encystment process and exhibited higher levels of recovery from mature cysts (14.13 ± 0.89 A. hydrophila:amoeba ratio, 10.13 ± 1.17 E. aerogenes:amoeba ratio, and 11.95 ± 0.7 E. coli:amoeba ratio). A. hydrophila and E. aerogenes also joined the ranks of other bacteria that could benefit from A. castellanii. Because cysts can be airborne, these findings suggest that Acanthamoeba is a potential vector in the transmission of A. hydrophila and E. aerogenes to susceptible hosts.

  7. A Bio-Inspired Method for the Constrained Shortest Path Problem

    Directory of Open Access Journals (Sweden)

    Hongping Wang

    2014-01-01

    optimization, crew scheduling, network routing and so on. It is an open issue since it is a NP-hard problem. In this paper, we propose an innovative method which is based on the internal mechanism of the adaptive amoeba algorithm. The proposed method is divided into two parts. In the first part, we employ the original amoeba algorithm to solve the shortest path problem in directed networks. In the second part, we combine the Physarum algorithm with a bio-inspired rule to deal with the CSP. Finally, by comparing the results with other method using an examples in DCLC problem, we demonstrate the accuracy of the proposed method.

  8. Gamma-irradiated bacterial preparation having anti-tumor activity

    International Nuclear Information System (INIS)

    Vass, A.A.; Tyndall, R.L.; Terzaghi-Howe, P.

    1999-01-01

    This application describes a bacterial preparation from Pseudomonas species isolated number s ign15 ATCC 55638 that has been exposed to gamma radiation exhibits cytotoxicity that is specific for neoplastic carcinoma cells. A method for obtaining a bacterial preparation having antitumor activity consists of suspending a bacterial isolate in media and exposing the suspension to gamma radiation. A bacterial preparation of an aged culture of an amoeba-associated bacteria exhibits anti-reverse transcriptase activity. A method for obtaining a bacterial preparation having anti-reverse transcriptase activity from an amoeba-associated bacterial isolate grown to stationary phase is disclosed

  9. An individual-level selection model for the apparent altruism ...

    Indian Academy of Sciences (India)

    Further, it must be capable of beingmodified to cover social behaviour in CSMs with an extracellular stalk, as well as in sorocarpic amoebae whose stalk cellsare viable. With regard to D. ... From this perspective, all cells behave in a manner that is 'selfish' rather than 'altruistic', albeit withdifferent expectations of success.

  10. To what extent do food preferences explain the trophic position of heterotrophic and mixotrophic microbial consumers in a Sphagnum peatland?

    Science.gov (United States)

    Jassey, Vincent E J; Meyer, Caroline; Dupuy, Christine; Bernard, Nadine; Mitchell, Edward A D; Toussaint, Marie-Laure; Metian, Marc; Chatelain, Auriel P; Gilbert, Daniel

    2013-10-01

    Although microorganisms are the primary drivers of biogeochemical cycles, the structure and functioning of microbial food webs are poorly studied. This is the case in Sphagnum peatlands, where microbial communities play a key role in the global carbon cycle. Here, we explored the structure of the microbial food web from a Sphagnum peatland by analyzing (1) the density and biomass of different microbial functional groups, (2) the natural stable isotope (δ(13)C and δ(15)N) signatures of key microbial consumers (testate amoebae), and (3) the digestive vacuole contents of Hyalosphenia papilio, the dominant testate amoeba species in our system. Our results showed that the feeding type of testate amoeba species (bacterivory, algivory, or both) translates into their trophic position as assessed by isotopic signatures. Our study further demonstrates, for H. papilio, the energetic benefits of mixotrophy when the density of its preferential prey is low. Overall, our results show that testate amoebae occupy different trophic levels within the microbial food web, depending on their feeding behavior, the density of their food resources, and their metabolism (i.e., mixotrophy vs. heterotrophy). Combined analyses of predation, community structure, and stable isotopes now allow the structure of microbial food webs to be more completely described, which should lead to improved models of microbial community function.

  11. Autonomous and non-autonomous traits mediate social cooperation

    Indian Academy of Sciences (India)

    2011-07-08

    Jul 8, 2011 ... In the trishanku (triA−) mutant of the social amoeba Dictyostelium discoideum, aggregates are smaller than usual and the spore mass is located mid-way up the ... Department of Molecular Reproduction, Development and Genetics, Indian Institute of Science, Bangalore 560 012, India; Division of Basic and ...

  12. The evolution of a distributed operating system

    NARCIS (Netherlands)

    van Renesse, Robbert; Tanenbaum, Andrew S.; Mullender, Sape J.; Schröder-Preikschat, Wolfgang; Zimmer, Wolfgang

    AMOEBA is a research project to build a true distributed operating system using the object model. Under the COST11-ter MANDIS project this work was extended to cover wide-area networks. Besides describing the system, this paper discusses the successive versions in the implementation of its model,

  13. Using Sparse Capabilities in a Distributed Operating System

    NARCIS (Netherlands)

    Tanenbaum, Andrew S.; Mullender, Sape J.; van Renesse, Robbert

    Most distributed operating systems constructed to date have lacked a unifying mechanism for naming and protection. In this paper we discuss a system, Amoeba, that uses capabilities for naming and protecting objects. In contrast to traditional, centralized operating systems, in which capabilities are

  14. Characterization of Entamoeba histolytica-induced ...

    Indian Academy of Sciences (India)

    However, it requires intact cells, as purified lectin failed to induce dephosphorylation in Jurkat cells. The nonpathogenic, but morphologically identical amoeba, Entamoeba moshkovskii also did not induce dephosphorylation in target cells. Treatment of Jurkat cells with phosphotyrosine phosphatase inhibitors has shown that ...

  15. Formation, succession and landscape history of Central-European summit raised bogs: A multiproxy study from the Hrubý Jeseník Mountains

    Czech Academy of Sciences Publication Activity Database

    Dudová, Lydie; Hájková, Petra; Buchtová, H.; Opravilová, V.

    2013-01-01

    Roč. 23, č. 2 (2013), s. 230-242 ISSN 0959-6836 R&D Projects: GA ČR GA206/08/0389 Institutional support: RVO:67985939 Keywords : bog succession * Larix * testate amoebae Subject RIV: EF - Botanics Impact factor: 3.794, year: 2013

  16. Půdní mikrofauna, krytenky (Protozoa: Testacea) a půdní hlístice (Nematoda), vybraných lokalit CHKO a BR Křivoklátsko

    Czech Academy of Sciences Publication Activity Database

    Háněl, Ladislav

    2011-01-01

    Roč. 31, - (2011), s. 209-214 ISSN 0231-5807 R&D Projects: GA ČR(CZ) GA206/93/0276; GA ČR(IO) GA526/06/1348 Institutional research plan: CEZ:AV0Z60660521 Keywords : Křivoklátsko Protected Landscape Area * deciduous forest * testate amoebae Subject RIV: EH - Ecology, Behaviour

  17. Diffusion-Assisted Aggregation and Synchronization in Dictyostelium discoideum

    Science.gov (United States)

    Nagano, Seido

    1998-05-01

    In biological pattern formation, chemotaxis and cell adhesion are essential. However, we lack quantitative data and a theory to understand their coordination. The cellular dynamics theory presented can clarify how Dictyostelium discoideum amoebae use diffusible cyclic adenosine 3',5'-monophosphate, and coordinate chemotaxis and cell adhesion during aggregation.

  18. Characterization of a human-pathogenic Acanthamoeba griffini isolated from a contact lens-wearing keratitis patient in Spain.

    Science.gov (United States)

    Heredero-Bermejo, I; Criado-Fornelio, A; De Fuentes, I; Soliveri, J; Copa-Patiño, J L; Pérez-Serrano, J

    2015-02-01

    Amoebae were isolated from contact lenses of a symptomatic lens wearer in Spain. Protozoa were characterized by studying their morphology, biology, protease activity and the 18S rRNA gene sequence. Morphology of the organism was observed by light microscopy, scanning electron microscopy and transmission electron microscopy. Its structure corresponded to an amphizoic amoeba. The protozoa grew well at 37 °C and poorly at lower temperatures. In addition, it was capable of lysing mammalian cells in vitro. A major 56 kDa proteolytic enzyme was observed in amoeba crude extracts by gelatin-sodium dodecyl sulphate-polyacrylamide gel electrophoresis. Most proteolytic enzymes in protozoa extracts showed significant activity over a wide range of pH (3-9) and temperature (8-45 °C) values. The assays on inhibition of protease activity indicated strongly that enzymes detected in amoeba extracts corresponded to serine proteases and, to a lesser extent, cysteine proteases. The use of proteinase inhibitors on a tissue culture model proved that the proteinase activity is critical for developing focal lesions in HeLa cell monolayers. Finally, partial sequencing of the 18S ribosomal RNA gene and phylogenetic analyses indicated that the isolate is closely related to Acanthamoeba griffini H37 from the UK (T3 genotype).

  19. Relationship between Legionella pneumophila and Acanthamoeba polyphaga: physiological status and susceptibility to chemical inactivation.

    Science.gov (United States)

    Barker, J; Brown, M R; Collier, P J; Farrell, I; Gilbert, P

    1992-01-01

    Survival studies were conducted on Legionella pneumophila cells that had been grown intracellularly in Acanthamoeba polyphaga and then exposed to polyhexamethylene biguanide (PHMB), benzisothiazolone (BIT), and 5-chloro-N-methylisothiazolone (CMIT). Susceptibilities were also determined for L. pneumophila grown under iron-sufficient and iron-depleted conditions. BIT was relatively ineffective against cells grown under iron depletion; in contrast, iron-depleted conditions increased the susceptibilities of cells to PHMB and CMIT. The activities of all three biocides were greatly reduced against L. pneumophila grown in amoebae. PHMB (1 x MIC) gave 99.99% reductions in viability for cultures grown in broth within 6 h and no detectable survivors at 24 h but only 90 and 99.9% killing at 6 h and 24 h, respectively, for cells grown in amoebae. The antimicrobial properties of the three biocides against A. polyphaga were also determined. The majority of amoebae recovered from BIT treatment, but few, if any, survived CMIT treatment or exposure to PHMB. This study not only shows the profound effect that intra-amoebal growth has on the physiological status and antimicrobial susceptibility of L. pneumophila but also reveals PHMB to be a potential biocide for effective water treatment. In this respect, PHMB has significant activity, below its recommended use concentrations, against both the host amoeba and L. pneumophila. PMID:1514790

  20. Growth of Legionella pneumophila in Acanthamoeba castellanii enhances invasion.

    Science.gov (United States)

    Cirillo, J D; Falkow, S; Tompkins, L S

    1994-01-01

    Legionella pneumophila is considered to be a facultative intracellular parasite. Therefore, the ability of these bacteria to enter, i.e., invade, eukaryotic cells is expected to be a key pathogenic determinant. We compared the invasive ability of bacteria grown under standard laboratory conditions with that of bacteria grown in Acanthamoeba castellanii, one of the protozoan species that serves as a natural host for L. pneumophila in the environment. Amoeba-grown L. pneumophila cells were found to be at least 100-fold more invasive for epithelial cells and 10-fold more invasive for macrophages and A. castellanii than were L. pneumophila cells grown on agar. Comparison of agar- and amoeba-grown L. pneumophila cells by light and electron microscopy demonstrated dramatic differences in the morphology and structure of the bacteria. Analyses of protein expression in the two strains of bacteria suggest that these phenotypic differences may be due to the expression of new proteins in amoeba-grown L. pneumophila cells. In addition, the amoeba-grown bacteria were found to enter macrophages via coiling phagocytosis at a higher frequency than agar-grown bacteria did. Replication of L. pneumophila in protozoans present in domestic water supplies may be necessary to produce bacteria that are competent to enter mammalian cells and produce human disease. Images PMID:8039895

  1. Clustering algorithm evaluation and the development of a replacement for procedure 1. [for crop inventories

    Science.gov (United States)

    Lennington, R. K.; Johnson, J. K.

    1979-01-01

    An efficient procedure which clusters data using a completely unsupervised clustering algorithm and then uses labeled pixels to label the resulting clusters or perform a stratified estimate using the clusters as strata is developed. Three clustering algorithms, CLASSY, AMOEBA, and ISOCLS, are compared for efficiency. Three stratified estimation schemes and three labeling schemes are also considered and compared.

  2. The pathogenic amoeboflagellate Naegleria fowleri: environmental isolations, competitors, ecologic interactions, and the flagellate-empty habitat hypothesis.

    Science.gov (United States)

    Griffin, J L

    1983-05-01

    From several surveys of environmental sites, the virulent human pathogen, Naegleria fowleri, was isolated from a pond in Georgia, a sewage treatment plant in Missouri, and from the Potomac and Anacostia rivers near and in Washington, D.C. Widely scattered, sparse populations seemed only a potential threat to human health at the time of sampling. The data support an estimate that the sites sampled contain 10,000 typical, low temperature, bactivorous amoebae for each heat tolerant amoeba able to grow at 45 degrees C. Heat tolerant competitors were much more common than N. fowleri. Naegleria lovaniensis, which is heat tolerant but nonpathogenic, was isolated from and downstream from an open air thermal pollution temperature gradient. Hot piles of composting sewage sludge yielded no amoeboflagellates, many heat tolerant (45-49 degrees C) amoebae, and one thermophilic (52 degrees C) Acanthamoeba. Features of the methods used include two-stage incubation to increase isolation of sparse organisms and distinction of N. fowleri from almost all other amoebae on agar plates. The flagellate-empty habitat hypothesis postulates a general model in which human intervention and/or natural events remove usual competitors and the ability to transform to a motile flagellate confers an advantage in recolonizing.

  3. Susceptibility of pathogenic and nonpathogenic Naegleria spp. to complement-mediated lysis.

    Science.gov (United States)

    Whiteman, L Y; Marciano-Cabral, F

    1987-10-01

    The susceptibility of four species of Naegleria amoebae to complement-mediated lysis was determined. The amoebicidal activity of normal human serum (NHS) and normal guinea pig serum (NGPS) for Naegleria amoebae was measured by an in vitro cytotoxicity assay. Release of radioactivity from amoebae labeled with [3H]uridine and visual observation with a compound microscope were used as indices of lysis. Highly pathogenic mouse-passaged N. fowleri was less susceptible to the lytic effects of NHS and NGPS than the weakly pathogenic, axenically grown N. fowleri or N. australiensis and the nonpathogenic amoebae N. gruberi and N. lovaniensis. However, both pathogenic and nonpathogenic Naegleria spp. depleted complement as assessed by total hemolytic activity. Treatment of serum with EDTA, heat (56 degrees C, 30 min), cobra venom factor, or antibody to C3 or C9 complement components decreased the amoebicidal activity of NHS. The presence of specific agglutinating antibody to N. fowleri enhanced the amoebicidal activity of NGPS for N. fowleri.

  4. Unusual cause of paraplegia in a child of 5 years

    African Journals Online (AJOL)

    2014-07-04

    Jul 4, 2014 ... There were contractures of both ankles and he had no bladder or bowel control. Blood count and chemistry were normal. A serology for amoeba and cysticercosis .... neural foramina into the muscle planes, as in our case. The invasive nature of the disease in the spine makes total removal and therefore ...

  5. Genetic Determinism vs. Phenotypic Plasticity in Protist Morphology

    Czech Academy of Sciences Publication Activity Database

    Mulot, M.; Marcisz, K.; Grandgirard, L.; Lara, E.; Kosakyan, Anush; Robroek, B. J. M.; Lamentowicz, M.; Payne, R. J.; Mitchell, E.A.D.

    2017-01-01

    Roč. 64, č. 6 (2017), s. 729-739 ISSN 1066-5234 Institutional support: RVO:60077344 Keywords : Body size * protozoa * soil moisture * testate amoebae * water table depth Subject RIV: EB - Genetics ; Molecular Biology OBOR OECD: Genetics and heredity (medical genetics to be 3) Impact factor: 2.692, year: 2016

  6. Creative research in the chemical industry – Four decades in ...

    Indian Academy of Sciences (India)

    Unknown

    2006-07-05

    Jul 5, 2006 ... loids,2,3 followed by practical training for 4 months at the Ciba laboratories in Basel. Earlier I had done my PhD with Prof T R Govindachari at Presidency ... rameters for effects on the cardiovascular, central nervous and endocrine systems and in infectious diseases due to bacteria, amoeba and helminths.

  7. Vermistella arctica n. sp. Nominates the Genus Vermistella as a Candidate for Taxon with Bipolar Distribution

    Czech Academy of Sciences Publication Activity Database

    Tyml, Tomáš; Kostka, Martin; Ditrich, O.; Dyková, I.

    2016-01-01

    Roč. 63, č. 2 (2016), s. 210-219 ISSN 1066-5234 R&D Projects: GA ČR(CZ) GBP505/12/G112 Institutional support: RVO:60077344 Keywords : actin * biogeography * bipolar distribution * free-living amoebae * phylogeny * polar regions * SSU Subject RIV: EG - Zoology Impact factor: 2.692, year: 2016

  8. The Life History of Flabellula baltica Smirnov (Gymnamoebae, Rhizopoda): Adaptations to a Spatially and Temporally Heterogeneous Environment

    DEFF Research Database (Denmark)

    Fenchel, Tom

    2010-01-01

    The polymorphic life history of the marine naked amoeba Flabellula baltica was studied. It can be interpreted in terms of adaptations to an environment that is patchy in time and space and it represents trade-off between longevity during starvation and the ability to initiate multiplication soon...

  9. Life under the Microscope: Children's Ideas about Microbes

    Science.gov (United States)

    Allen, Michael; Bridle, Georgina; Briten, Elizabeth

    2015-01-01

    Microbes (by definition) are tiny living things that are only visible through a microscope and include bacteria, viruses, fungi, and protoctists (mainly single-celled life forms such as amoebae and algae). Although people are familiar with the effects of microbes, such as infectious disease and food spoilage, because of their lack of visibility,…

  10. Late Holocene human impact and climate change recorded in a North Swedish peat deposit

    NARCIS (Netherlands)

    van der Linden, M.; Barke, J.; Vickery, E.; Charman, D.J.; van Geel, B.

    2008-01-01

    A peat core from a mire with poor fen lawns and ombrotrophic hummock strings in Northern Sweden was examined (plant macrofossils, pollen/non-pollen microfossils, testate amoebae, colorimetric humification, carbon/nitrogen ratios, bulk densities, loss on ignition) to investigate the effects of

  11. Journal of Biosciences | Indian Academy of Sciences

    Indian Academy of Sciences (India)

    It has also been shown that although this organism contains sequence homologs of genes which are known to control the cell cycle of most eukaryotes, these genes may be structurally altered and their equivalent function yet to be demonstrated in amoeba. The available information suggests that surveillance mechanisms ...

  12. An individual-level selection model for the apparent altruism ...

    Indian Academy of Sciences (India)

    Amotz Zahavi

    2018-02-16

    Feb 16, 2018 ... Bacterial transformation:distribution, shared mechanisms and divergent control. Nat. Rev. Microbiol. 12 181–196. Kang S, Tice AK, Spiegel FW, Silberman JD, Pánek T, Cˇ epicka I,. Kostka M, Kosakyan A, Alcântara DMC, Roger AJ, et al. 2017. Between a pod and a hard test: the deep evolution of amoebae.

  13. Regulation of TORC2 complex in Dictyostelium discoideum

    NARCIS (Netherlands)

    Khanna, Ankita

    2016-01-01

    Dictyostelium is an amoeba that lives in the soil where it feeds on bacteria. During scarcity of food, Dictyostelium cells undergo a highly regulated developmental process in which the cells aggregate by chemotaxing towards pulsatile emission of extracellular cAMP from a signaling center; the cells

  14. Expression of matrix metalloproteinases in Naegleria fowleri and their role in invasion of the central nervous system.

    Science.gov (United States)

    Lam, Charlton; Jamerson, Melissa; Cabral, Guy; Carlesso, Ana Maris; Marciano-Cabral, Francine

    2017-10-01

    Naegleria fowleri is a free-living amoeba found in freshwater lakes and ponds and is the causative agent of primary amoebic meningoencephalitis (PAM), a rapidly fatal disease of the central nervous system (CNS). PAM occurs when amoebae attach to the nasal epithelium and invade the CNS, a process that involves binding to, and degradation of, extracellular matrix (ECM) components. This degradation is mediated by matrix metalloproteinases (MMPs), enzymes that have been described in other pathogenic protozoa, and that have been linked to their increased motility and invasive capability. These enzymes also are upregulated in tumorigenic cells and have been implicated in metastasis of certain tumours. In the present study, in vitro experiments linked MMPs functionally to the degradation of the ECM. Gelatin zymography demonstrated enzyme activity in N. fowleri whole cell lysates, conditioned media and media collected from invasion assays. Western immunoblotting indicated the presence of the metalloproteinases MMP-2 (gelatinase A), MMP-9 (gelatinase B) and MMP-14 [membrane type-1 matrix metalloproteinase (MT1-MMP)]. Highly virulent mouse-passaged amoebae expressed higher levels of MMPs than weakly virulent axenically grown amoebae. The functional relevance of MMPs in media was indicated through the use of the MMP inhibitor, 1,10-phenanthroline. The collective in vitro results suggest that MMPs play a critical role in vivo in invasion of the CNS and that these enzymes may be amenable targets for limiting PAM.

  15. Biocidal efficacy of monochloramine against planktonic and biofilm-associated Naegleria fowleri cells.

    Science.gov (United States)

    Goudot, S; Herbelin, P; Mathieu, L; Soreau, S; Banas, S; Jorand, F P A

    2014-04-01

    Free-living amoebae (FLA) in aqueous systems are a problem for water network managers and health authorities because some are pathogenic, such as Naegleria fowleri, and they have also been reported to operate as reservoirs and vectors of several pathogenic bacteria. Therefore, to better control the occurrence of such amoebae, we evaluate the efficacy of monochloramine against planktonic forms (trophozoites and cysts) and also biofilm-associated cells of N. fowleri as FLA are often associated with biofilms. From a freshwater biofilm growing in a pilot reactor and inoculated with N. fowleri, we obtained Ct values ranging from 4 to 17 mg Cl2 min l(-1) at 25°C and pH 8·2 on both planktonic and biofilm associated cells. In addition, the inactivation pattern of biofilm associated was intermediate between those of trophozoïtes and cysts. The monochloramine efficiency varies with the life stage of N. fowleri (trophozoïte, cyst, and biofilm-associated). The sensitivity to disinfectant of amoeba, that is, trophozoïtes and cysts, in the biofilm life stage is as high as that of their planktonic cyst form. This study gives Ct values for cysts and biofilm-associated N. fowleri. This may impact on water treatment strategies against amoebae and should be considered when controlling N. fowleri in man-made water systems such as cooling towers or hot water systems. © 2014 The Society for Applied Microbiology.

  16. Naegleria fowleri: contact-dependent secretion of electrondense granules (EDG).

    Science.gov (United States)

    Chávez-Munguía, Bibiana; Villatoro, Lizbeth Salazar; Omaña-Molina, Maritza; Rodríguez-Monroy, Marco Aurelio; Segovia-Gamboa, Norma; Martínez-Palomo, Adolfo

    2014-07-01

    The free living amoeba Naegleria fowleri is pathogenic to humans but also to other mammalians. These amoebae may invade the nasal mucosa and migrate into the brain causing cerebral hemorrhagic necrosis, a rapidly fatal infection. Knowledge of the cytolytic mechanism involved in the destruction of brain tissues by Naegleria trophozoites is limited. In other amoebic species, such as Entamoeba histolytica, we have previously reported the possible lytic role of small cytoplasmic components endowed with proteolytic activities, known as electrondense granules (EDG). Using transmission electron microscopy we now report that EDG, seldom found in long term cultured N. fowleri, are present in abundance in trophozoites recovered from experimental mice brain lesions. Numerous EDG were also observed in amoebae incubated with collagen substrates or cultured epithelial cells. SDS-PAGE assays of concentrated supernatants of these trophozoites, containing EDG, revealed proteolytic activities. These results suggest that EDG may have a clear role in the cytopathic mechanisms of this pathogenic amoeba. Copyright © 2014 Elsevier Inc. All rights reserved.

  17. Occurrence of Thermotolerant Hartmannella Vermiformis and Naegleria Spp. in Hot Springs of Ardebil Province, Northwest Iran

    Directory of Open Access Journals (Sweden)

    E Nazemalhosseini Mojarad

    2012-06-01

    Full Text Available Background: Geothermal waters could be suitable niches for thermophilic free living amoebae including Naegleria and Hartmannella. Ardebil Province, northwest Iran is popular for having many hot springs for recreational and health purposes activity. The present research is the first molecular based investigation regarding the presence of Naegleria and Hartmannella in the hot springs of Ardebil Province in Iran.Methods: Overall, 30 water samples were taken from waters of thermal hot springs in Ardebil Province, Iran during 2010-2011. All collected samples were transferred to Dept. of Parasitology and Mycology, Shahid Beheshti University of Medical Sciences, Tehran, Iran. Cultivation of concentrated water samples was performed using culture-enrichment method. Cloning of the target amoebae was obtained and morphological and molecular analysis was done using page key combined with two sets of primers, respectively. Sequence analysis and homology search was used for strains identification.Results: Of 30 water samples, 8 (26.7% were positive for thermotolerant Vahlkampfiids and Hartman­nella based on morphological characteristics of vegetative form and double walled cysts. Cloning of the target amoebae were done successfully. Sequencing of the positive isolates revealed that the strains belonged to Naegleria (N. carteri and N. spp and H. vermiformis.Conclusion: The result highlights a need for improved filtration and disinfection and periodic monitoring of recreational thermal waters in order to prevent disease related to free- living amoebae. This is the first comprehensive molecular study of thermophilic Naegleria and Hartmannella in hot springs of Iran.

  18. Occurrence of Thermotolerant Hartmannella vermiformis and Naegleria Spp. in Hot Springs of Ardebil Province, Northwest Iran.

    Science.gov (United States)

    Solgi, R; Niyyati, M; Haghighi, A; Mojarad, E Nazemalhosseini

    2012-01-01

    Geothermal waters could be suitable niches for thermophilic free living amoebae including Naegleria and Hartmannella. Ardebil Province, northwest Iran is popular for having many hot springs for recreational and health purposes activity. The present research is the first molecular based investigation regarding the presence of Naegleria and Hartmannella in the hot springs of Ardebil Province in Iran. Overall, 30 water samples were taken from waters of thermal hot springs in Ardebil Province, Iran during 2010-2011. All collected samples were transferred to Dept. of Parasitology and Mycology, Shahid Beheshti University of Medical Sciences, Tehran, Iran. Cultivation of concentrated water samples was performed using culture-enrichment method. Cloning of the target amoebae was obtained and morphological and molecular analysis was done using page key combined with two sets of primers, respectively. Sequence analysis and homology search was used for strains identification. Of 30 water samples, 8 (26.7%) were positive for thermotolerant Vahlkampfiids and Hartmannella based on morphological characteristics of vegetative form and double walled cysts. Cloning of the target amoebae were done successfully. Sequencing of the positive isolates revealed that the strains belonged to Naegleria (N. carteri and N. spp) and H. vermiformis. The result highlights a need for improved filtration and disinfection and periodic monitoring of recreational thermal waters in order to prevent disease related to free- living amoebae. This is the first comprehensive molecular study of thermophilic Naegleria and Hartmannella in hot springs of Iran.

  19. Journal of Biosciences | Indian Academy of Sciences

    Indian Academy of Sciences (India)

    Entamoeba histolytica and Giardia lamblia are microaerophilic protists, which have long been considered models of ancient pre-mitochondriate eukaryotes. As transitional eukaryotes, amoebae and giardia appeared to lack organelles of higher eukaryotes and to depend upon energy metabolism appropriate for anaerobic ...

  20. Meeting on the Microbiology of Soils, Autumn 2001: Estimation of protozoan diversity in soil

    DEFF Research Database (Denmark)

    Ekelund, Flemming

    2002-01-01

    Different methods of estimating protozoan diversity in soil are discussed in this paper, with the major emphasis on heterotrophic flagellates. Although many species of ciliates and testate amoebae seem to be unique to the soil environment, the communities of heterotrophic flagellates and naked am...

  1. Microfaunal primary succession on the volcanic island Surtsey

    DEFF Research Database (Denmark)

    Frederiksen, Helle B.; Kraglund, H. O.; Ekelund, F.

    2001-01-01

    and nematodes were absent and only amoebae and heterotrophic flagellates were found. Most of the protozoan populations we examined were unable to survive salinity levels corresponding to seawater. We therefore conclude that many of soil protozoa populations on Surtsey arrived to the island as airborne cysts...

  2. Differentiation of Entamoeba histolytica, Entamoeba dispar and ...

    African Journals Online (AJOL)

    2016-06-28

    Jun 28, 2016 ... The protozoan parasite Entamoeba histolytica is estimated to infect 50 million people and causes ... protozoan and its presence in clinical specimens does not justify treatment.[8] It has actually been ..... food handlers with amoeba infection initially diagnosed by microscopy. Parasite 2008;15:65-68. 8.

  3. Eukaryotic checkpoints are absent in the cell division cycle of ...

    Indian Academy of Sciences (India)

    It has also been shown that although this organism contains sequence homologs of genes which are known to control the cell cycle of most eukaryotes, these genes may be structurally altered and their equivalent function yet to be demonstrated in amoeba. The available information suggests that surveillance mechanisms ...

  4. Gastrodiscoides hominis infection in a Nigerian- case report

    African Journals Online (AJOL)

    No cyst or trophozoite of amoeba was found in her stool. Malaria parasite was not detected in her weekly blood film throughout the period of follow up. Heamatological investigations showed haematocrit. (PCV) of 22% (Normal value is 36-44%), red blood cell. (RBC) morphology showed microcytosis and hypochro- mia.

  5. Galactose/N-acetylgalactosamine lectin: the coordinator of host cell ...

    Indian Academy of Sciences (India)

    The Gal/GalNAc lectin binds galactose and N-acetylgalactosamine allowing the adherence of amoebas to host cells. Involvement of the lectin in the pathogenesis of E. histolytica infection will be reviewed in this paper. The lectin has been shown to have very specific and substantial effects on adherence, cytotoxicity, and ...

  6. Biology Today symposium | Mid Year Meetings | Events | Indian ...

    Indian Academy of Sciences (India)

    11.15, D. P. KASBEKAR, Centre for Cellular and Molecular Biology, Hyderabad Neurospora abhors a transposon. 11.45, IMRAN SIDDIQI, Centre for Cellular and Molecular Biology, Hyderabad Meiotic chromosome organization. 12.15, VIDYANAND NANJUNDIAH, Indian Institute of Science, Bengaluru Social amoebae.

  7. Arachidonic acid is a chemoattractant for Dictyostelium discoideum ...

    Indian Academy of Sciences (India)

    Cyclic AMP (cAMP) is a natural chemoattractant of the social amoeba Dictyostelium discoideum. It is detected by cell surface cAMP receptors. Besides a signalling cascade involving phosphatidylinositol 3,4,5-trisphosphate (PIP3), Ca2+ signalling has been shown to have a major role in chemotaxis. Previously, we have ...

  8. Comparing the Dictyostelium and Entamoeba genomes reveals an ancient split in the Conosa lineage.

    Directory of Open Access Journals (Sweden)

    Jie Song

    2005-12-01

    Full Text Available The Amoebozoa are a sister clade to the fungi and the animals, but are poorly sampled for completely sequenced genomes. The social amoeba Dictyostelium discoideum and amitochondriate pathogen Entamoeba histolytica are the first Amoebozoa with genomes completely sequenced. Both organisms are classified under the Conosa subphylum. To identify Amoebozoa-specific genomic elements, we compared these two genomes to each other and to other eukaryotic genomes. An expanded phylogenetic tree built from the complete predicted proteomes of 23 eukaryotes places the two amoebae in the same lineage, although the divergence is estimated to be greater than that between animals and fungi, and probably happened shortly after the Amoebozoa split from the opisthokont lineage. Most of the 1,500 orthologous gene families shared between the two amoebae are also shared with plant, animal, and fungal genomes. We found that only 42 gene families are distinct to the amoeba lineage; among these are a large number of proteins that contain repeats of the FNIP domain, and a putative transcription factor essential for proper cell type differentiation in D. discoideum. These Amoebozoa-specific genes may be useful in the design of novel diagnostics and therapies for amoebal pathologies.

  9. Fish-isolated strains of Hartmannella vermiformis Page, 1967: morphology, phylogeny and molecular diagnosis of the species in tissue lesions

    Czech Academy of Sciences Publication Activity Database

    Dyková, Iva; Pindová, Z.; Fiala, Ivan; Dvořáková, Helena; Macháčková, Blanka

    2005-01-01

    Roč. 52, č. 4 (2005), s. 295-303 ISSN 0015-5683 R&D Projects: GA AV ČR IAA6022202 Institutional research plan: CEZ:AV0Z60220518 Keywords : amphizoic amoeba e * Hartmannella vermiformis * morphology Subject RIV: EA - Cell Biology Impact factor: 1.138, year: 2005

  10. Prevalence and antimicrobial profiles of Salmonella serovars from ...

    African Journals Online (AJOL)

    ADEYEYE

    2014-01-21

    Jan 21, 2014 ... nematodes, amoebas and Salmonella. Journal of the Association of Official. Analytical Chemists 67(3): 613–615. Sirichote P, Bangtrakulnonth A, Tianmanee K,. Unahalekhaka A, Oulai A, Chittaphithakchai. S, Kheorwrod W & Hendriksen RS (2010). Serotypes and antimicrobial resistance of. Salmonella ...

  11. Pan African Medical Journal - Vol 26, No 1 (2017)

    African Journals Online (AJOL)

    Free-living amoebae isolated in the Central African Republic: epidemiological and molecular aspects · EMAIL FREE FULL TEXT EMAIL FREE FULL TEXT DOWNLOAD FULL TEXT DOWNLOAD FULL TEXT. Alain Farra, Claudine Bekondi, Vianney Tricou, Jean Robert Mbecko, Antoine Talarmin, 1-10.

  12. Use Integrated Management of Childhood Illnesses guidelines: a ...

    African Journals Online (AJOL)

    etiologic agent is parasitic mainly amoeba and giardia. Objective: The objective of this study was to assess the use of IMCI guidelines in children presenting with diarrhea at Ghindae Hospital. Methods: The study was a prospective in which all children below the age of 5 years, who presented primarily with all forms of ...

  13. An Acrasin-Like Attractant from Yeast Extract Specific for Dictyostelium lacteum

    NARCIS (Netherlands)

    Mato, José M.; Haastert, Peter J.M. van; Krens, Frans A.; Konijn, Theo M.

    1977-01-01

    The transition of the unicellular to the multicellular stage in Dictyostelium lacteum is not mediated by cyclic AMP. The attractant for aggregative amoebae of this cellular slime mold species was isolated from yeast extract and purified more than 1000-fold without a significant loss of activity.

  14. [Detection and differentiation of Entamoeba histolytica and Entamoeba dispar by polymerase chain reaction in a community in Zulia State, Venezuela].

    Science.gov (United States)

    Rivero, Zulbey; Bracho, Angela; Calchi, Marinella; Díaz, Iris; Acurero, Ellen; Maldonado, Adriana; Chourio, Glenis; Arráiz, Nailet; Corzo, Gilbert

    2009-01-01

    Differential identification of Entamoeba histolytica and Entamoeba dispar is essential for both appropriate patient treatment and epidemiological purposes. To determine the prevalence of these amoeba infections in Santa Rosa de Agua (Maracaibo, Zulia State, Venezuela), a PCR assay using specific primers for each species was standardized and applied. 204 stool samples were analyzed through direct microscopic examination with SSF (0.85%) and lugol, formol-ether concentration, and PCR. Under direct microscopy, 42 individuals (20.58%) presented the E. histolytica/E. dispar complex. Meanwhile PCR showed 47 positive cases for these amoebas: 22 E. histolytica (10.78%), 16 E. dispar (7.84%), and 9 (4.41%) mixed infections. There was no significant difference in the presence of E. histolytica and/or E. dispar according to either gender or age. There were no cases of these amoebas in children under 2 years of age. Observed frequency of E. histolytica (31/204) shows the endemic nature of amoeba infection in this community.

  15. Amoebapore is an important virulence factor of Entamoeba histolytica

    Indian Academy of Sciences (India)

    Unknown

    point the role of amoebapore in this process. For this purpose we used E. coli cells expressing GFP protein. Incubation of bacteria with amoeba results in fast (1 h) phagocytosis of numerous (> 100) bacteria which, as seen by confocal fluorescence microscope (figure 8), are rapidly taken up into vacuoles of the trophozoite.

  16. Quality assessment of drinking water from different sources in Lafia ...

    African Journals Online (AJOL)

    Yeast (Candida spp), filamentous fungi, the protozoa (Amoeba spp and Paramecium spp), algae, particularly Spirogyra spp were detected in hand dug wells, the three streams and River Amba. The detection of these organisms in dinking water is an indication of the unsuitability of the water sources. Protection of water ...

  17. The Use of Extended Wear Contact Lenses in the Aviation Environment: An Army-wide Study.

    Science.gov (United States)

    1992-09-01

    intervention techniques. Acanthamoeba infection Corneal infection associated with contact lens wear also has been linked to organisms of the amoebae...genus Acanthamoeba . While 83 percent of the Acanthamoeba -infected individuals in a recent retrospective clinical study were contact lens wearers, there... Acanthamoeba keratitis are: history of corneal trauma, exposure to contaminated water, and contact lens wear (Stehr-Green, Bailey, and Visvesvara, 1989

  18. Acanthamoeba and Fusarium interactions: A possible problem in keratitis.

    Science.gov (United States)

    Nunes, Thais Esther Teixeira; Brazil, Nathalya Tesch; Fuentefria, Alexandre Meneghello; Rott, Marilise Brittes

    2016-05-01

    The incidence of Acanthamoeba and Fusarium species has increased in contact lens-related infectious keratitis. They share several environments and cases of co-infection have been reported. The interaction between the amoebae and other microorganisms may result in significant changes for both, like increased virulence in mammalian hosts. In this study, we evaluated the interaction of three Acanthamoeba castellanii strains with Fusarium conidia and the possible implications on keratitis. F. conidia were internalized by A. castellanii strains and were able to germinate inside the amoebae. The co-culture with the live amoebae, as well as the amoebal culture supernatant and lysate, increased the fungal growth significantly. Moreover, live F. solani and its culture supernatant enhanced the survival of amoebae, but in a different way in each amoebal strain. The encystment of the A. castellanii strain re-isolated from rat lung was increased by the fungus. These results show that A. castellanii and F. solani interaction may have an important influence on survival of both, and specially indicate a possible effect on virulence characteristics of these microorganisms. These data suggest that the A. castellanii-F. solani interaction may cause severe impacts on keratitis. Copyright © 2016 Elsevier B.V. All rights reserved.

  19. Experimental study on pathogenic potential of six Acanthamoeba strains isolated from fish

    Czech Academy of Sciences Publication Activity Database

    Veverková, Marie; Dyková, Iva; Pecková, Hana

    2002-01-01

    Roč. 49, č. 3 (2002), s. 243-245 ISSN 0015-5683 R&D Projects: GA AV ČR KSK6005114 Institutional research plan: CEZ:AV0Z6022909 Keywords : amoeba * fish * pathology Subject RIV: EA - Cell Biology Impact factor: 0.515, year: 2002

  20. Francisella tularensis type A Strains Cause the Rapid Encystment of Acanthamoeba castellanii and Survive in Amoebal Cysts for Three Weeks post Infection

    Energy Technology Data Exchange (ETDEWEB)

    El-Etr, S H; Margolis, J; Monack, D; Robison, R; Cohen, M; Moore, E; Rasley, A

    2009-07-28

    Francisella tularensis, the causative agent of the zoonotic disease tularemia, has recently gained increased attention due to the emergence of tularemia in geographical areas where the disease has been previously unknown, and the organism's potential as a bioterrorism agent. Although F. tularensis has an extremely broad host range, the bacterial reservoir in nature has not been conclusively identified. In this study, the ability of virulent F. tularensis strains to survive and replicate in the amoeba Acanthamoeba castellanii was explored. We observe that A. castellanii trophozoites rapidly encyst in response to F. tularensis infection and that this rapid encystment phenotype (REP) is caused by factor(s) secreted by amoebae and/or F. tularensis into the co-culture media. Further, our results indicate that in contrast to LVS, virulent strains of F. tularensis can survive in A. castellanii cysts for at least 3 weeks post infection and that induction of rapid amoeba encystment is essential for survival. In addition, our data indicate that pathogenic F. tularensis strains block lysosomal fusion in A. castellanii. Taken together, these data suggest that the interactions between F. tularensis strains and amoeba may play a role in the environmental persistence of F. tularensis.

  1. Strategies of the protozoan parasite Entamoeba histolytica to evade ...

    Indian Academy of Sciences (India)

    Unknown

    cally in the detoxification of endogenous reactive oxygen species, might be an important enzyme to protect the amoebae against oxidative attack of activated host phago- cytic (Bruchhaus et al 1993, 1997; Flores et al 1993). 4. Resistance of E. histolytica to RNI. Thus, if E. histolytica has the enzymatic potential to resist ROI ...

  2. Meeting on the Microbiology of Soils, Autumn 2001: Estimation of protozoan diversity in soil

    DEFF Research Database (Denmark)

    Ekelund, Flemming

    2002-01-01

    amoebae are probably best considered as restricted versions of their aquatic counterparts. Soil protozoa are difficult to observe directly, hence culture techniques are usually used. These techniques enable us to explore certain functional aspects, but their major drawback is that some soil protozoa...

  3. Catecholamines in Entamoebae: recent (re) discoveries

    Indian Academy of Sciences (India)

    Keywords. Acanthamoeba; catecholamines; encystation; Entamoeba; epinephrine; Hartmannella; histamine; serotonin. Abstract. Free-living and enteric amoebae have similar two-stage life cycles, and both organisms depend on being able to monitor environmental conditions to determine whether to continue multiplying as ...

  4. Determinants that govern the recognition and uptake of Escherichia coli O157 : H7 by Acanthamoeba castellanii.

    Science.gov (United States)

    Arnold, Jason W; Spacht, Drew; Koudelka, Gerald B

    2016-10-01

    Predation by phagocytic predators is a major source of bacterial mortality. The first steps in protozoan predation are recognition and consumption of their bacterial prey. However, the precise mechanisms governing prey recognition and phagocytosis by protists, and the identities of the molecular and cellular factors involved in these processes are, as yet, ill-characterized. Here, we show that that the ability of the phagocytic bacterivorous amoebae, Acanthamoeba castellanii, to recognize and internalize Escherichia coli, a bacterial prey, varies with LPS structure and composition. The presence of an O-antigen carbohydrate is not required for uptake of E. coli by A. castellanii. However, O1-antigen types, not O157 O-antigen types, inhibit recognition and uptake of bacteria by amoeba. This finding implies that O-antigen may function as an antipredator defence molecule. Recognition and uptake of E. coli by A. castellanii is mediated by the interaction of mannose-binding protein located on amoebae's surface with LPS carbohydrate. Phagocytic mammalian cells also use mannose-binding lectins to recognize and/or mediate phagocytosis of E. coli. Nonetheless, A. castellanii's mannose binding protein apparently displays no sequence similarity with any known metazoan mannose binding protein. Hence, the similarity in bacterial recognition mechanisms of amoebae and mammalian phagocytes may be a result of convergent evolution. © 2016 John Wiley & Sons Ltd.

  5. Acanthamoeba and Dictyostelium as Cellular Models for Legionella Infection

    Directory of Open Access Journals (Sweden)

    A. Leoni Swart

    2018-03-01

    Full Text Available Environmental bacteria of the genus Legionella naturally parasitize free-living amoebae. Upon inhalation of bacteria-laden aerosols, the opportunistic pathogens grow intracellularly in alveolar macrophages and can cause a life-threatening pneumonia termed Legionnaires' disease. Intracellular replication in amoebae and macrophages takes place in a unique membrane-bound compartment, the Legionella-containing vacuole (LCV. LCV formation requires the bacterial Icm/Dot type IV secretion system, which translocates literally hundreds of “effector” proteins into host cells, where they modulate crucial cellular processes for the pathogen's benefit. The mechanism of LCV formation appears to be evolutionarily conserved, and therefore, amoebae are not only ecologically significant niches for Legionella spp., but also useful cellular models for eukaryotic phagocytes. In particular, Acanthamoeba castellanii and Dictyostelium discoideum emerged over the last years as versatile and powerful models. Using genetic, biochemical and cell biological approaches, molecular interactions between amoebae and Legionella pneumophila have recently been investigated in detail with a focus on the role of phosphoinositide lipids, small and large GTPases, autophagy components and the retromer complex, as well as on bacterial effectors targeting these host factors.

  6. Strategies of the protozoan parasite Entamoeba histolytica to evade ...

    Indian Academy of Sciences (India)

    Unknown

    and the subsequent migration of the amoeba to the liver where abscesses are formed (Espinosa-Cantellano and. Martinez-Paloma 2000; Stanley 2001). ..... activity of bacterial peroxiredoxins; Nature (London) 407. 211–215. Cario E and Podolsky D K 2000 Differential alteration in intes- tinal epithelial cell expression of ...

  7. Genomics approaches in the understanding of Entamoeba ...

    African Journals Online (AJOL)

    STORAGESEVER

    2009-04-20

    Apr 20, 2009 ... transfer of bacterial genes into the E. histolytica genome was observed, and evidence that these ... The parasite genome also encodes a large number of novel receptor kinases and contains expan- ... studies were performed using RNA isolated from amoeba that have chemotaxed towards TNF (close to the ...

  8. Microbial- and isothiocyanate-mediated control of Phytophthora and Pythium species

    Science.gov (United States)

    M.F. Cohen; E. Yamamoto; E. Condeso; B.L. Anacker; N. Rank; M. Mazzola

    2008-01-01

    Plant pathogens of the oomycete lineage share common susceptibilities to many biotic and abiotic stresses. We are investigating the potential of antagonistic bacteria, isothiocyanates, and mycophagous amoebae to control diseases caused by Phytophthora spp., including the etiologic agent of sudden oak death, Phytophthora ramorum (...

  9. Autonomous and non-autonomous traits mediate social cooperation ...

    Indian Academy of Sciences (India)

    2011-07-08

    Jul 8, 2011 ... In the trishanku (triA−) mutant of the social amoeba Dictyostelium discoideum, aggregates are smaller than usual and the spore mass is located mid-way up the stalk, not at the apex. We have monitored aggregate territory size, spore allocation and fruiting body morphology in chimaeric groups of ...

  10. Analysis of Rheb in the cellular slime mold Dictyostelium discoideum

    Indian Academy of Sciences (India)

    2014-01-27

    Jan 27, 2014 ... of TOR in regulating various aspects of cellular function. ... It is a protist growing and dividing mitotically as long as food is abundant. On starvation, these free living amoebae come to common collecting points in response to the chemoattractant ..... indicates that artificial Rheb expression does not have.

  11. The revised classification of eukaryotes

    Czech Academy of Sciences Publication Activity Database

    Adl, S.; Simpson, A. G. B.; Lane, C. E.; Lukeš, Julius; Bass, D.; Bowser, S. S.; Brown, M W.; Burki, F.; Dunthorn, M.; Hampl, V.; Heiss, A.; Hoppenrath, M.; Lara, E.; Gall, L. L.; Lynn, D. H.; McManus, H.; Mitchell, E. A. D.; Mozley-Stanridge, S. E.; Parfrey, L. W.; Pawlowski, J.; Rueckert, S.; Shadwick, L.; Schoch, C.L.; Smirnov, A.; Spiegel, F. W.

    2012-01-01

    Roč. 59, č. 5 (2012), s. 429-514 ISSN 1066-5234 Institutional support: RVO:60077344 Keywords : Algae * amoebae * biodiversity * ciliates * flagellates * fungi * parasites * protozoa * systematics * taxonomy Subject RIV: EB - Genetics ; Molecular Biology Impact factor: 2.162, year: 2012 http://onlinelibrary.wiley.com/doi/10.1111/j.1550-7408.2012.00644.x/pdf

  12. Cell fate choice and social evolution in Dictyostelium discoideum ...

    Indian Academy of Sciences (India)

    Cell fate choice and social evolution in. Dictyostelium discoideum: Interplay of morphogens and heterogeneities. Attempts to understand the development of the social amoeba Dictyostelium discoideum keep throwing up surprises and drive home the point that here too, as in any biological situation, no explanation can.

  13. Litter quality as driving factor for plant nutrition via grazing of protozoa on soil microorganisms.

    Science.gov (United States)

    Koller, Robert; Robin, Christophe; Bonkowski, Michael; Ruess, Liliane; Scheu, Stefan

    2013-08-01

    Plant residues provide a major source of nitrogen (N) for plant growth. Litter N mineralization varies with litter carbon-to-nitrogen (C-to-N) ratio and presence of bacterial-feeding fauna. We assessed the effect of amoebae, major bacterial feeders in soil, on mineralization of litter of low (high quality) and high C-to-N ratio (low quality) and evaluated consequences for plant growth. We used stable isotopes to determine plant N uptake from litter and plant C partitioning. Stable isotope probing of phospholipid fatty acids was used to follow incorporation of plant C into microorganisms. Amoebae increased plant N uptake independent of litter quality and thereby the biomass of shoots and roots by 33% and 66%, respectively. Plant allocation of total (13)C to roots in low (42%) exceeded that of high-quality litter treatments (26%). Amoebae increased plant allocation of (13)C to roots by 37%. Microbial community structure and incorporation of (13)C into PLFAs varied significantly with litter quality and in the low-quality litter treatment also with the presence of amoebae. Overall, the results suggest that in particular at low nutrient conditions, root-derived C fosters the mobilization of bacterial N by protozoa, thereby increasing plant growth when microorganisms and plants compete for nutrients. © 2013 Federation of European Microbiological Societies. Published by John Wiley & Sons Ltd. All rights reserved.

  14. Decoding Non-Coding DNA: Trash or Treasure?

    Indian Academy of Sciences (India)

    organisms such as frogs and amoebae have much more DNA per cell compared to mammals. (2) In addition, the G value paradox states that the total number of protein-coding genes is also not proportional to biological complexity and even alternative splic- ing (the process by which the cell can combine different exons to.

  15. What Entamoeba histolytica and Giardia lamblia tell us about the ...

    Indian Academy of Sciences (India)

    Unknown

    Entamoeba histolytica and Giardia lamblia are microaerophilic protists, which have long been considered models of ancient pre-mitochondriate eukaryotes. As transitional eukaryotes, amoebae and giardia appeared to lack organelles of higher eukaryotes and to depend upon energy metabolism appropriate for anaerobic ...

  16. Lethal action of thermal neutron to living cells

    International Nuclear Information System (INIS)

    Kawai, Kenichi; Akahoshi, Mitsuhiko

    1975-01-01

    Using Amoeba Proteus (DNA content was 5x10 -10 g/cell), action mechanism of thermal neutron was described by using death of cells as an index. Using mononuclear type (M-type) and polynuclear type (P-type) of Amoeba, investigation was made as to whether the number of nucleus contained in cell was related to death of cell by radiation. As compared to gamma-ray, marked difference between sensitivity of both types of Amoeba was not recognized. It was suggested that nucleus was important as a place of death of cell by thermal neutron. In order to demonstrate that reaction of 31 P(n, γ) 32 Pwas a main cause of lethality, the following experiment was carried out. 1) In order to investigate that radical reaction was not a main cause of lethal action by thermal neutron, increase and decrease of radical reaction were observed by adding AET, Cysteine and BudR. As to thermal neutron action, effect of its chemical factors did not appeared. 2) In order to demonstrate that nuclear reaction of constituent atom of living body played an important part in death of cell, increase and decrease of nuclear reaction within cell were observed by taking Na 2 , B 4 and O 7 into Amoeba. It was suggested that nuclear reaction of cell constituent atom itself was important in death of cell. 3) Potentiation effect of gamma-ray and thermal neutron was investigated and its mechanism was discussed. (K. Serizawa)

  17. Amoebic meningoencephalitis: 16 fatalities.

    Science.gov (United States)

    Cerva, L; Novăk, K

    1968-04-05

    The parasitologic examination of pathologic brain tissue from 16 cases of acute plurulent meningoencephalitis occurring between 1962 and 1965 in northern Bohemia disclosed massive infection of the central nervous system by amoebas of the limax type. The common source was an indoor swimming pool.

  18. Potentially pathogenic Acanthamoeba genotype T4 isolated from dental units and emergency combination showers.

    Science.gov (United States)

    Castro-Artavia, Esteban; Retana-Moreira, Lissette; Lorenzo-Morales, Jacob; Abrahams-Sandí, Elizabeth

    2017-12-01

    Acanthamoeba is the genus of free-living amoebae that is most frequently isolated in nature. To date, 20 Acanthamoeba genotypes have been described. Genotype T4 is responsible for approximately 90% of encephalitis and keratitis cases. Due to the ubiquitous presence of amoebae, isolation from environmental sources is not uncommon; to determine the clinical importance of an isolation, it is necessary to have evidence of the pathogenic potential of amoebae. The aim of this study was to physiologically characterise 8 Acanthamoeba T4 isolates obtained from dental units and emergency combination showers and to determine their pathogenic potential by employing different laboratory techniques. Eight axenic cultures of Acanthamoeba genotype T4 were used in pathogenic potential assays. Osmotolerance, thermotolerance, determination and characterisation of extracellular proteases and evaluation of cytopathic effects in MDCK cells were performed. All of the isolates were osmotolerant, thermotolerant and had serine proteases from 44-122 kDa. Two isolates had cytopathic effects on the MDCK cell monolayer. The presence of Acanthamoeba T4 with pathogenic potential in areas such as those tested in this study reaffirms the need for adequate cleaning and maintenance protocols to reduce the possibility of infection with free-living amoebae.

  19. Protacanthamoeba bohemica n. sp., isolated from the liver of tench, Tinca tinca (Linnaeus, 1758)

    Czech Academy of Sciences Publication Activity Database

    Dyková, Iva; Veverková-Fialová, M.; Fiala, Ivan; Dvořáková, Helena

    2005-01-01

    Roč. 44, č. 4 (2005), s. 369-376 ISSN 0065-1583 R&D Projects: GA AV ČR IAA6022202 Institutional research plan: CEZ:AV0Z60220518 Keywords : amphizoic amoebae * Protacanthamoeba bohemica * morphology Subject RIV: EA - Cell Biology Impact factor: 0.987, year: 2005

  20. Photodynamic inactivation of Acanthamoeba polyphaga with curcuminoids: an in vitro study

    Science.gov (United States)

    Corrêa, Thaila Q.; Geralde, Mariana C.; Carvalho, Mariana T.; Bagnato, Vanderlei S.; Kurachi, Cristina; de Souza, Clovis W. O.

    2016-03-01

    Acanthamoeba polyphaga are free-living amoebae that can be considered potentially pathogenic organisms by cause serious human infections, including keratitis and granulomatous amoebic encephalitis that usually results in death. Photodynamic inactivation (PDI) has been used for the biological control of microorganisms and can be promise in the control of Acanthamoeba infections. This study evaluated the in vitro effectiveness of PDI in A. polyphaga using curcuminoids salt as photosensitizer (PS) besides observing morphological changes caused by this PS in this organism, in confocal microscopy. A. polyphaga trophozoites were grown at 37°C in PYG medium for 48 to 72 hours. After, the trophozoites were incubated with PS solution during one hour and the samples were irradiated using light-emitting diodes at 460 nm at light doses 30 and 50 J/cm2. The results revealed reduction of 27.7%, 61.4% and 82.5% at 30 J/cm2 and 75.2%, 85.0% and 95.9% at 50 J/cm2, respectively, at curcuminoid salt concentrations of 500, 1000 and 1500 μg/mL. Through fluorescence images, it was possible to visualize the curcuminoid salt's uptake by the trophozoites. The PS showed toxicity to amoebae, in the dark, but the irradiation in PDI contributed to amoebae death effect. These data suggest that PDI may be an application of therapeutic intervention against Acanthamoeba infections, since it was effective in the inactivation of these amoebae.

  1. Isolation of pathogenic Naegleria from artificially heated waters

    Energy Technology Data Exchange (ETDEWEB)

    Tyndall, R L; Willaert, E; Stevens, A R; Coutant, C C

    1977-01-01

    Investigations were undertaken to determine whether heated waters facilitate the proliferation of free-living amoeba that cause primary amoebic meningoencephalitis. Water samples were taken close to the discharges of power plants situated on lakes or rivers in Florida and Texas and from cooling towers in Tennessee. The water temperatures ranged from 29 to 42/sup 0/C. Water samples were also taken from several lakes in Florida and Texas without associated power plants. The water temperatures of these ranged from 30/sup 0/ to 34/sup 0/C. Twenty-five-250-ml samples were filtered through membranes. Samples taken from the control lakes and cooling towers showed no growth of pathogenic amoeba, whereas growth was obtained from 2 of the 8 lakes and rivers in Florida and from 1 of the 7 man-made lakes in Texas that were artificially heated. The amoebae were identified as belonging to the genus Naegleria from their trophozoite and cyst structure, ability to grow at 45/sup 0/C, to transform into flagellates, and to produce primary amebic meningoencephalitis (PAME) in mice after intranasal instillation. Their identification as N. fowleri was confirmed by indirect immunofluorescent analysis with antiserum produced against N. fowleri. These findings indicate that artificial heating of waters may facilitate the growth of pathogenic free living amoeba.

  2. Structure and function of a unique pore-forming protein from a pathogenic acanthamoeba

    NARCIS (Netherlands)

    Michalek, M.; Sönnichsen, F.D.; Wechselberger, R.W.; Wienk, H.L.J.; Leippe, M.; et al., [No Value

    2013-01-01

    Human pathogens often produce soluble protein toxins that generate pores inside membranes, resulting in the death of target cells and tissue damage. In pathogenic amoebae, this has been exemplified with amoebapores of the enteric protozoan parasite Entamoeba histolytica. Here we characterize

  3. Relationship between Legionella pneumophila and Acanthamoeba polyphaga: Physiological status and susceptibility to chemical inactivation

    Energy Technology Data Exchange (ETDEWEB)

    Barker, J.; Farrell, I. (Aston Univ., Aston Triangle, Birmingham (United Kingdom)); Brown, M.R.W.; Collier, P.J.; Gilbert, P. (Univ. of Manchester (United Kingdom))

    1992-08-01

    Survival studies were conducted on Legionella pneumophila cells that had been grown intracellulary in Acanthamoeba polyphaga and then exposed to polyhexamethylene biguanide (PHMB), benzisothiazolone (BIT), and 5-chloro-N-methylisothiazolone (CMIT). Susceptibilities were also determined for L. pneumophila grown under iron-sufficient and iron-depleted conditions. BIT was relatively ineffective against cells to PHMB and CMIT. The activities of all three biocides were greatly reduced against L. pneumophila grown in amoebae. PHMB (1 [times] MIC) gave 99.99% reductions in viability for cultures grown in broth within 6 h and no detectable survivors at 24 h but only 90 and 99.9% killing at 6 h and 24 h, respectively, for cells grown in amoebae. The antimicrobial properties of the three biocides against A. polyphaga were also determined. The majority of amoebae recovered from BIT treatment, but few, if any, survived CMIT treatment or exposure of PHMB. This study not only shows the profound effect that intra-amoebal growth has on the physiological status and antimicrobial susceptibility of L. pneumophila but also reveals PHMB to be a potential biocide for effective water treatment. In this respect, PHMB has significant activity, below its recommended use concentrations, against both the host amoeba and L. pneumophila.

  4. Evaluation of the effectiveness of tea tree oil in treatment of Acanthamoeba infection.

    Science.gov (United States)

    Hadaś, Edward; Derda, Monika; Cholewiński, Marcin

    2017-03-01

    Eye diseases caused by amoebae from the genus Acanthamoeba are usually chronic and severe, and their treatment is prolonged and not very effective. The difficulties associated with therapy have led to attempts at finding alternative treatment methods. Particularly popular is searching for cures among drugs made of plants. However, no substances with total efficacy in treating Acanthamoeba keratitis have been identified.Results of our semi in vivo studies of tea tree oil simulating eyeball infection demonstrated 100% effectiveness in the case of both trophozoites and cysts of amoebae from the genus Acanthamoeba. The action of tea tree oil indicates that this is the first substance with a potential ability to quickly and effectively remove the amoebae from the eye. Tea tree oil has the ability to penetrate tissues, which allows it to destroy amoebae in both the shallow and deep layers of the cornea. The present research into the use of tea tree oil in the therapy of Acanthamoeba infection is the first study of this type in parasitology. It offers tremendous potential for effective treatment of Acanthamoeba keratitis and other diseases caused by these protozoa.

  5. Potentially pathogenic Acanthamoeba genotype T4 isolated from dental units and emergency combination showers

    Directory of Open Access Journals (Sweden)

    Esteban Castro-Artavia

    Full Text Available BACKGROUND Acanthamoeba is the genus of free-living amoebae that is most frequently isolated in nature. To date, 20 Acanthamoeba genotypes have been described. Genotype T4 is responsible for approximately 90% of encephalitis and keratitis cases. Due to the ubiquitous presence of amoebae, isolation from environmental sources is not uncommon; to determine the clinical importance of an isolation, it is necessary to have evidence of the pathogenic potential of amoebae. OBJECTIVE The aim of this study was to physiologically characterise 8 Acanthamoeba T4 isolates obtained from dental units and emergency combination showers and to determine their pathogenic potential by employing different laboratory techniques. METHODS Eight axenic cultures of Acanthamoeba genotype T4 were used in pathogenic potential assays. Osmotolerance, thermotolerance, determination and characterisation of extracellular proteases and evaluation of cytopathic effects in MDCK cells were performed. FINDINGS All of the isolates were osmotolerant, thermotolerant and had serine proteases from 44-122 kDa. Two isolates had cytopathic effects on the MDCK cell monolayer. MAIN CONCLUSION The presence of Acanthamoeba T4 with pathogenic potential in areas such as those tested in this study reaffirms the need for adequate cleaning and maintenance protocols to reduce the possibility of infection with free-living amoebae.

  6. Microbial diversities (16S and 18S rDNA gene pyrosequencing) and environmental pathogens within drinking water biofilms grown on the common premise plumbing materials unplasticized polyvinylchloride and copper

    Science.gov (United States)

    Drinking water (DW) biofilm communities influence the survival of opportunistic pathogens, e.g. Legionella pneumophila, via parasitization of free-living amoebae such as Acanthamoebae. Yet knowledge about the microbial composition of DW biofilms developed on common in-premise pl...

  7. Journal of Biosciences | Indian Academy of Sciences

    Indian Academy of Sciences (India)

    Keywords. Acanthamoeba; catecholamines; encystation; Entamoeba; epinephrine; Hartmannella; histamine; serotonin. Abstract. Free-living and enteric amoebae have similar two-stage life cycles, and both organisms depend on being able to monitor environmental conditions to determine whether to continue multiplying as ...

  8. Journal of Biosciences | Indian Academy of Sciences

    Indian Academy of Sciences (India)

    Entamoeba histolytica killing of host cells is contact dependent and mediated by a Gal/GalNAc lectin. Upon contact with amoeba a rapid and extensive dephosphorylation of tyrosine phosphorylated host cell proteins is observed. This effect is mediated by the Gal/GalNAc lectin. However, it requires intact cells, as purified ...

  9. Stenamoeba polymorpha, a New Species Isolated from Domesticated Horse Equus ferus caballus.

    Science.gov (United States)

    Peglar, Michael T; Nerad, Thomas A; Anderson, O Roger

    2016-11-01

    A new species of lobosean amoeba, Stenamoeba polymorpha n. sp., was isolated from the diarrheic stool of a domesticated horse in Great Falls Virginia, U.S. It shares characteristics with the five other described Stenamoeba species. However, electron microscopy revealed S. polymorpha has a substantially thickened cell surface lamina. Under light microscopy, the amoebae had a dynamic polymorphic appearance because hyaloplasm readily formed and resorbed subpseudopodia from any peripheral region of the cell. While in locomotion, the amoebae produced subpseudopodia that led and alternated the direction of movement with an apparent zigzag path. Sometimes, stationary amoebae had a vertical cell posture that was substantially taller than wide or long. The new species description is also supported by small subunit ribosomal ribonucleic acid gene analyses using phylogenies with both broad and narrow taxon sampling, pairwise nucleotide comparisons, and in-silico secondary structure predictions. The latter suggested Stenamoeba spp. discriminatory motifs in variable region eight. Stenamoeba polymorpha n. sp. is the first of its genus to be recovered from mammal feces, the first to yield a potential group one intron and the second described from North America. Here, the phylogenies suggest Stenamoeba limacina is this new species' nearest known relative. © 2016 The Author(s) Journal of Eukaryotic Microbiology © 2016 International Society of Protistologists.

  10. Effects of human impact and climate change during the last 350 years recorded in a Swedish raised bog deposit

    NARCIS (Netherlands)

    van der Linden, M.; Vickery, E.; Charman, D.J.; van Geel, B.

    2008-01-01

    A peat core from an ombrotrophic mire in central Sweden was analysed for multi-proxy indicators (plant macrofossils, pollen/non-pollen microfossils, testate amoebae, colorimetric humification, carbon/nitrogen ratios, bulk densities, loss on ignition), to evaluate the relative contributions of

  11. United polarizable multipole water model for molecular mechanics simulation

    Energy Technology Data Exchange (ETDEWEB)

    Qi, Rui; Wang, Qiantao; Ren, Pengyu, E-mail: pren@mail.utexas.edu [Department of Biomedical Engineering, The University of Texas at Austin, Austin, Texas 78712 (United States); Wang, Lee-Ping; Pande, Vijay S. [Department of Chemistry, Stanford University, Stanford, California 94305 (United States)

    2015-07-07

    We report the development of a united AMOEBA (uAMOEBA) polarizable water model, which is computationally 3–5 times more efficient than the three-site AMOEBA03 model in molecular dynamics simulations while providing comparable accuracy for gas-phase and liquid properties. In this coarse-grained polarizable water model, both electrostatic (permanent and induced) and van der Waals representations have been reduced to a single site located at the oxygen atom. The permanent charge distribution is described via the molecular dipole and quadrupole moments and the many-body polarization via an isotropic molecular polarizability, all located at the oxygen center. Similarly, a single van der Waals interaction site is used for each water molecule. Hydrogen atoms are retained only for the purpose of defining local frames for the molecular multipole moments and intramolecular vibrational modes. The parameters have been derived based on a combination of ab initio quantum mechanical and experimental data set containing gas-phase cluster structures and energies, and liquid thermodynamic properties. For validation, additional properties including dimer interaction energy, liquid structures, self-diffusion coefficient, and shear viscosity have been evaluated. The results demonstrate good transferability from the gas to the liquid phase over a wide range of temperatures, and from nonpolar to polar environments, due to the presence of molecular polarizability. The water coordination, hydrogen-bonding structure, and dynamic properties given by uAMOEBA are similar to those derived from the all-atom AMOEBA03 model and experiments. Thus, the current model is an accurate and efficient alternative for modeling water.

  12. Acanthamoeba (T4) trophozoites cross the MDCK epithelium without cell damage but increase paracellular permeability and transepithelial resistance by modifying tight junction composition.

    Science.gov (United States)

    Flores-Maldonado, Catalina; González-Robles, Arturo; Salazar-Villatoro, Lizbeth; Omaña-Molina, Maritza; Gallardo, Juan Manuel; González-Lázaro, Mónica; Hernández-Ramírez, Verónica Ivonne; Talamás-Rohana, Patricia; Lorenzo-Morales, Jacob; Martínez-Palomo, Adolfo

    2017-12-01

    Free-living amoebae of the genus Acanthamoeba are protozoa ubiquitously found in nature. Some species of the genus are potentially pathogenic for humans provoking keratitis in healthy individuals, often in contact lens wearers and opportunistic infections such as pneumonitis, fatal granulomatous encephalitis and skin infections, particularly in immunocompromised individuals. The pathogenic mechanisms of these amoebae are poorly understood, however it had been suggested that contact dependent mechanisms are important during invasion, regardless of the epithelia type, since amoebae penetrate epithelia separating tight junction (TJ). This study was undertaken to determine whether Acanthamoeba sp. (T4) damages the barrier function of the TJ in MDCK epithelial monolayers. Actin cytoskeleton staining and electron microscopy analyses were performed; paracellular permeability and TJ sealing were evaluated by apicobasolateral diffusion of ruthenium red and transepithelial resistance (TER) measurements; immunofluorescence and Western blot assays were performed to locate and estimate expression of TJ protein claudins 2 (Cldn2) and 4 (Cldn4). The results show that Acanthamoeba sp. crosses the MDCK monolayer without altering the actin cytoskeleton or the morphology of the cells. When trophozoites or conditioned medium interact with the monolayer, paracellular diffusion of ruthenium red increases. After 6 h, the amoebae, but not their conditioned medium, increase the TER, and Cldn2 is removed from the TJ, and its overall content in the cells diminishes, while Cldn4 is targeted to the TJ without changing its expression level. In conclusion Acanthamoeba (T4) crosses MDCK monolayer without damaging the cells, increasing permeability and TER through Cldn2 degradation, and redirecting Cldn4 to TJ. These results strongly suggest that contact-dependent mechanisms are relevant during amoebae invasion. Copyright © 2017 Elsevier Inc. All rights reserved.

  13. Development of a modified gentamicin protection assay to investigate the interaction between Campylobacter jejuni and Acanthamoeba castellanii ATCC 30010.

    Science.gov (United States)

    Dirks, Brian P; Quinlan, Jennifer J

    2014-05-01

    Campylobacter jejuni is one of the leading causes of diarrheal illness worldwide. It is persistent in the environment and on poultry despite its microaerophilic nature and sensitivity to dessication and pH. Studies have demonstrated that C. jejuni co-incubated with Acanthamoeba spp. may be protected from harmful environmental factors. Research in this area, however has included a range of different methodologies for co-incubation, recovery of bacteria and amoebae, and verification of internalization. In this study a modified gentamicin protection assay (mGPA) was developed with a standardized co-incubation procedure. The mGPA addresses limitations of the traditional GPA by providing quantification of the rate of internalization, or lack of internalization, of C. jejuni by Acanthamoeba castellanii. The mGPA described here utilizes tubes instead of cell culture plates allowing for determination of exact numbers of A. castellanii and C. jejuni to be co-incubated prior to addition to tubes. Additionally, the mGPA allows for the incorporation of C. jejuni-only controls to determine the fate of C. jejuni throughout the assay in the absence of A. castellanii. Using the mGPA it was determined that on average 1.6×10(5) C. jejuni (or 0.006% of initial 1×10(9) inoculum) survive the assay in the absence of A. castellanii. Additionally, results obtained with the mGPA demonstrated that while co-incubation with amoebae sometimes (56% of co-incubations) provided a protective effect for C. jejuni, in other cases it did not provide any protective effect (39% of co-incubations), and in at least one case there was a statistically significant higher recovery of C. jejuni in controls when compared to C. jejuni co-incubated with amoebae. The modified gentamicin protection assay described here allows better quantification of the rate and incidence of internalization of bacteria by amoebae. Use of the standardized mGPA developed here with varying environmental parameters and/or strains

  14. Stenotrophomonas maltophilia and Vermamoeba vermiformis relationships: bacterial multiplication and protection in amoebal-derived structures.

    Science.gov (United States)

    Cateau, Estelle; Maisonneuve, Elodie; Peguilhan, Samuel; Quellard, Nathalie; Hechard, Yann; Rodier, Marie-Helene

    2014-12-01

    Stenotrophomonas maltophilia, a bacteria involved in healthcare-associated infections, can be found in hospital water systems. Other microorganisms, such as Free Living amoebae (FLA), are also at times recovered in the same environment. Amongst these protozoa, many authors have reported the presence of Vermamoeba vermiformis. We show here that this amoeba enhances S. maltophilia growth and harbors the bacteria in amoebal-derived structures after 28 days in harsh conditions. These results highlight the fact that particular attention should be paid to the presence of FLA in hospital water systems, because of their potential implication in survival and growth of pathogenic bacterial species. Copyright © 2014 Institut Pasteur. Published by Elsevier Masson SAS. All rights reserved.

  15. Developmental Cycle and Genome Analysis ofProtochlamydia massiliensissp. nov. a New Species in theParachlamydiacaeFamily.

    Science.gov (United States)

    Benamar, Samia; Bou Khalil, Jacques Y; Blanc-Tailleur, Caroline; Bilen, Melhem; Barrassi, Lina; La Scola, Bernard

    2017-01-01

    Amoeba-associated microorganisms (AAMs) are frequently isolated from water networks. In this paper, we report the isolation and characterization of Protochlamydia massiliensis , an obligate intracellular Gram-negative bacterium belonging to the Parachlamydiaceae family in the Chlamydiales order, from a cooling water tower. This bacterium was isolated on Vermamoeba vermiformis . It has a multiple range of hosts among amoeba and is characterized by a typical replication cycle of Chlamydiae with a particularity, recently shown in some chlamydia, which is the absence of inclusion vacuoles in the V. vermiformis host, adding by this a new member of Chlamydiae undergoing developmental cycle changes in the newly adapted host V. vermiformis . Draft genome sequencing revealed a chromosome of 2.86 Mb consisting of four contigs and a plasmid of 92 Kb.

  16. Protochlamydia phocaeensis sp. nov., a new Chlamydiales species with host dependent replication cycle.

    Science.gov (United States)

    Bou Khalil, Jacques Y; Benamar, Samia; Di Pinto, Fabrizio; Blanc-Tailleur, Caroline; Raoult, Didier; La Scola, Bernard

    2017-06-01

    Chlamydiae are pathogenic and symbiotic bacteria, which form an important part of amoeba-associated microorganisms. In this paper, we report the isolation, developmental cycle and genome analysis of Protochlamydia phocaeensis sp. nov., an obligate intracellular parasite with a large host spectrum, able to infect Acanthamoeba castellanii, Acanthamoeba polyphaga, and Vermamoeba vermiformis. The genome size is 3,424,182 bp with a GC content of 42%. This bacterium displayed a particular developmental cycle depending on the infected host. The P. phocaeensis showed typical inclusion vacuoles in A. castellanii, while these were absent in V. vermiformis. Since "Chlamydiae-amoebae" interactions are supposed to depend on the chlamydial species, our findings speculate that variations in the developmental cycle of certain Chlamydiae are also host dependent. Copyright © 2017 Institut Pasteur. Published by Elsevier Masson SAS. All rights reserved.

  17. Description of Neoparamoeba longipodia n. sp. and a new strain of Neoparamoeba aestuarina (Page, 1970) (Amoebozoa, Dactylopodida) from deep-sea habitats.

    Science.gov (United States)

    Volkova, Ekaterina; Kudryavtsev, Alexander

    2017-10-01

    Two strains of lobose amoebae have been isolated from the deep-sea bottom sediments of the Sea of Japan (3.6km deep) and Western Atlantic Ocean (5.1km deep). Amoebae of both strains have a dactylopodial mophotype, intracellular kinetoplastid symbiont (Perkinsela-like organism) and have no microscales on the cell surface. The morphology and molecular data of the Sea of Japan strain allow us to unambiguously identify it as Neoparamoeba aestuarina (Page, 1970). At the same time, the Atlantic strain is described as a new species Neoparamoeba longipodia as it differs from other species of the genus Neoparamoeba in morphology and gene sequence data. The data presented expand the range of known habitats for the genus Neoparamoeba and permit further analysis of the phylogenetic relationships within this clade with the expanded set of molecular data. Copyright © 2017 Elsevier GmbH. All rights reserved.

  18. The design of a real-time distributed system

    International Nuclear Information System (INIS)

    Hertzberger, L.O.; Tuynman, F.; Mullender, S.J.; Poletiek, G.; Vermeulen, J.C.; Renesse, R. van; Tanenbaum, A.S.

    1986-01-01

    In modern physics experiments an increasing number and variety of programmable processors is used. As a consequence, a software environment is needed that provides an integrated approach to development, testing and use of real-time distributed software. This contribution is based on work being done in the AMOEBA Distributed Operating System Project and the FADOS Real-Time Distributed Operating System Project. A short description of both systems is presented as an example of how basic real-time operating system services can be organized. AMOEBA is the result of fundamental research in the field of distributed operating systems, while FADOS has been designed for applications as encountered in experimental high-energy physics. (Auth.)

  19. Systemic acanthamoebiasis associated with canine distemper in dogs in the semiarid region of Paraíba, Brazil

    Directory of Open Access Journals (Sweden)

    Maria T.S. Frade

    2015-02-01

    Full Text Available Infections by free-living amoebae can cause systemic disease in animals and humans. We describe the epidemiological, clinical and pathological aspects of disseminated acanthamoebiasis associated with canine distemper in three dogs of the semiarid region of Paraíba, Northeastern Brazil. Affected dogs developed progressive neurological and respiratory signs that progressed to death within in two to 20 days. Gross lesions were irregular and with yellow-reddish nodules randomly distributed in the lungs, heart, kidneys, spleen, lymph nodes, adrenals, and intestine. One dog had foci of malacia in the parietal cortex and another one in nucleus of brain basis. Histologically, pyogranulomas with areas of necrosis and hemorrhage in all organs affected were observed, associated with myriads of intralesional amoebic trophozoites. All three cases were concomitant canine distemper, that possibly triggered immunosuppression in the dogs. The diagnosis was performed through microscopic findings of infection by free-living amoebae and confirmed Acanthamoeba sp. by immunohistochemistry

  20. Acanthamoeba pearcei n. sp. (Protozoa: Amoebida) from sewage contaminated sediments.

    Science.gov (United States)

    Nerad, T A; Sawyer, T K; Lewis, E J; McLaughlin, S M

    1995-01-01

    Seabottom sediments from a discontinued Philadelphia-Camden 40-Mile ocean sewage disposal site were cultured for cyst-forming free-living amoebae. Barge delivered wastes were discharged at the site from 1973 until 1980 when the site was closed. One station at the southeast margin of the site was sampled at a depth of approximately 50 m, twice in 1978 and once in 1982, 1983 and 1984. Sediment from the 1978 collection yielded Acanthamoeba polyphaga, Vahlkampfia sp., and an unknown amoeba with stellate endocysts similar to those of A. astronyxis. Trophozoites and cysts of the isolate were typical of those described for the genus Acanthamoeba. Biochemical tests employing enzyme electrophoresis and morphological studies on live and stained specimens showed that the isolate was distinct from other well-described species within the family Acanthamoebidae Sawyer & Griffin, 1975.

  1. Effect of bacteria on survival and growth of Acanthamoeba castellanii.

    Science.gov (United States)

    Wang, X; Ahearn, D G

    1997-04-01

    The growth and survival of Acanthamoeba castellanii in the presence of Gram-negative bacteria such as Pseudomonas aeruginosa, Escherichia coli, Serratia marcescens, and Stenotrophomonas maltophilia varied with the densities and species of bacteria. All species of bacteria suspended in a buffered saline at densities of 10(5) to 10(6)/ml supported the growth and survival of 10(6)/ml trophozoites of Acanthamoeba castellanii in a buffered saline solution. At densities of bacteria to amoebae of 100:1 or greater, growth and survival of A. castellanii were suppressed, particularly by P. aeruginosa. In an enrichment medium, the rapid growth of most co-inoculated bacteria inhibited the growth and survival of the amoeba.

  2. Migration patterns of pathogenic and nonpathogenic Naegleria spp.

    Science.gov (United States)

    Thong, Y H; Ferrante, A

    1986-01-01

    Four species of Naegleria were tested for their ability to migrate under agarose. Pathogenic N. fowleri strains exhibited rapid locomotion at 37 degrees C. Environmental isolates of N. fowleri moved faster than clinical isolates which had been kept in axenic culture for longer periods, and this result was confirmed by using the 84-2205-7 strain kept in axenic culture for 1 or 5 months. Nonpathogenic N. gruberi strains migrated actively at 28 degrees C but not at 37 degrees C; moreover, even at 28 degrees C, active amoebae constituted only a small proportion of the whole. The temperature-tolerant, nonpathogenic species N. lovaniensis moved more slowly than N. fowleri at 37 degrees C. In contrast, N. australiensis, which is temperature tolerant as well as pathogenic for mice, migrated at a rate comparable to that of N. fowleri. There appears to be a direct correlation between the locomotive ability of free-living amoebae and their pathogenic potential.

  3. Balamuthia mandrillaris: Further morphological observations of trophozoites by light, scanning and transmission electron microscopy.

    Science.gov (United States)

    González-Robles, Arturo; Lares-Villa, Fernando; Lares-Jiménez, Luis Fernando; Omaña-Molina, Maritza; Salazar-Villatoro, Lizbeth; Martínez-Palomo, Adolfo

    2015-10-01

    Additional morphological features of Balamuthia mandrillaris observed by light and electron microscopy are reported. Trophozoites were extremely pleomorphic: their cell shapes ranged from rounded to elongated and sometimes they appeared exceptionally stretched out and branched. By transmission electron microscopy it was possible to observe two different cytoplasmic areas, the ectoplasm and the endoplasm and often sections of rough endoplasmic reticulum were found in the transition zone. The cytoplasm was very fibrogranular and most of the organelles typically found in eukaryotic cells were observed. A particular finding was the presence of numerous mitochondria with a different structure from those of other free-living amoebae. The observations reported here may reinforce the morphological knowledge of this amoeba and provide a background for further analyses. Copyright © 2015 Elsevier Inc. All rights reserved.

  4. Isolation and molecular characterization of Acanthamoeba and Balamuthia mandrillaris from combination shower units in Costa Rica.

    Science.gov (United States)

    Retana-Moreira, Lissette; Abrahams-Sandí, Elizabeth; Cabello-Vílchez, Alfonso Martín; Reyes-Batlle, María; Valladares, Basilio; Martínez-Carretero, Enrique; Piñero, José E; Lorenzo-Morales, Jacob

    2014-11-01

    Free living amoebae (FLA) are ubiquitous protozoa, which may behave as parasites under certain conditions. Four genera are recognized as causal agents of infections in humans and animals: Naegleria, Sappinia, Acanthamoeba and Balamuthia. This work determines the presence of FLA in combination shower units and employs molecular biology for the characterization of isolates. The morphological analysis and partial sequencing of the 18S rDNA gene revealed the presence of Acanthamoeba genotype T4 in 30% of the units sampled. In addition to Acanthamoeba cysts, trophozoites with morphological characteristics similar to Balamuthia were identified. PCR assay using the mitochondrial 16S rRNA gene as a target confirmed the identification of the amoeba as Balamuthia mandrillaris. Up to date, this is the first report of the isolation of B. mandrillaris in Central America and the fifth report worldwide.

  5. Effects of industrial chemicals and radioactive materials in biological systems

    International Nuclear Information System (INIS)

    Gangopadhyay, A.; Chatterjee, S.

    1987-01-01

    Much has been written on the effects of radiation and toxic chemicals on biological systems. In this communication general considerations regarding these topics will be discussed very briefly; the major emphasis will be focused on the effects of chemicals, namely ethyl methane sulfonate (EMS) on Amoeba, Advantages to the use of amoeba for studying the effects of radiation and chemicals include the following: large mononucleate unicellular organisms having a long generation time; opportunity to study cellular organelles and biochemical and genetic alterations in a single cell system; and a long cell cycle, the stages of which can be synchronized without resorting to chemical treatment or temperature shock and thereby readily permitting study at defined stages of the cell's life cycle. This, in turn, is discussed in light of current disposal methods for this type of waste and how it might be safely disposed of

  6. New Entamoeba group in howler monkeys (Alouatta spp.) associated with parasites of reptiles.

    Science.gov (United States)

    Villanueva-García, Claudia; Gordillo-Chávez, Elías José; Baños-Ojeda, Carlos; Rendón-Franco, Emilio; Muñoz-García, Claudia Irais; Carrero, Julio César; Córdoba-Aguilar, Alex; Maravilla, Pablo; Galian, José; Martínez-Hernández, Fernando; Villalobos, Guiehdani

    2017-08-01

    Our knowledge of the parasite species present in wildlife hosts is incomplete. Protozoans such as amoebae of the genus Entamoeba infect a large variety of vertebrate species, including NHPs. However, traditionally, their identification has been accomplished through microscopic evaluation; therefore, amoeba species have not always been identified correctly. We searched for Entamoeba spp. using a fragment of the small subunit rDNA in free-ranging howler monkeys (Alouatta palliata and A. pigra) from southeast Mexico. One hundred fifty five samples were collected, with 46 from A. palliata and 109 from A. pigra and 8 of the total samples were positive. We detected a new clade of Entamoeba, which was separated from other described species but closer to E. insolita, as well as an unnamed sequence typically found in iguana species with low shared identity values (reptiles.

  7. Isolation of the etiological agent of primary amoebic meningoencephalitis from artificially heated waters

    Energy Technology Data Exchange (ETDEWEB)

    Stevens, A.R.; Tyndall, R.L.; Coutant, C.C.; Willaert, E.

    1977-12-01

    To determine whether artificial heating of water by power plant discharges facilitates proliferation of the pathogenic free-living amoebae that cause primary amoebic meningoencephalitis, water samples (250 ml) were taken from discharges within 3,000 feet (ca. 914.4 m) of power plants and were processed for amoeba culture. Pathogenic Naegleria fowleri grew out of water samples from two of five lakes and rivers in Florida and from one of eight man-made lakes in Texas. Pathogenic N. fowleri did not grow from water samples taken from cooling towers and control lakes, the latter of which had no associated power plants. The identification of N. fowleri was confirmed by pathogenicity in mice and by indirect immunofluorescence analyses, by using a specific antiserum.

  8. Concentration of Naegleria fowleri in natural waters used for recreational purposes in Sonora, Mexico (November 2007-October 2008).

    Science.gov (United States)

    Lares-Villa, Fernando; Hernández-Peña, Claudia

    2010-09-01

    A survey was designed to know the concentration of Naegleria fowleri in recreational areas in Hornos, Sonora, during a year. Samples were taken monthly at La Isleta and Las Palmas and the total amoeba counts were obtained by the most probable number method (MPN). The identification of N. fowleri was made by PCR. The maximum concentration of total thermophilic amoebae was 9175 MPN/L for La Isleta and 3477 MPN/L for Las Palmas. Thermophilic Naegleria were present mainly during summer and fall. October's concentrations were up to 201 MPN/L, at both places. The maximum concentrations of N. fowleri were 201 MPN/L and 18 MPN/L for La Isleta and Las Palmas respectively, and were isolated from August to October. The presence of N. fowleri in these particular natural bodies of water reinforces the need for adaptation of preventive measures to avoid cases of primary amoebic meningoencephalitis. Copyright 2010 Elsevier Inc. All rights reserved.

  9. Primary amoebic meningoencephalitis: first reported case from Rohtak, North India

    Directory of Open Access Journals (Sweden)

    Naveen Gupta

    Full Text Available A fatal case of primary amoebic encephalitis (PAM in a 20 year old boy, a proven case of acute leukemic leukemia (ALL type L2, in remission is described. No history of swimming could be elicited. The clinical presentation, the isolation of the amoeba from the cerebrospinal fluid (CSF, the poor response to amphotericin B, and the ultimately fatal outcome are all consistent with the diagnosis of PAM. On the basis of its ability to grow at temperature 42ºC and 45ºC, morphology of trophozoite, and the presence of flagellate forms in CSF, the amoeba was identified as Naegleria fowleri. Other drugs used in combination with amphotericin B are tetracycline, rifampicin, and miconazole. A possibility of PAM should always be considered in all cases of acute purulent meningoencephalitis in which no bacteria or fungus are found.

  10. Developmental Cycle and Genome Analysis of Protochlamydia massiliensis sp. nov. a New Species in the Parachlamydiacae Family

    Directory of Open Access Journals (Sweden)

    Samia Benamar

    2017-08-01

    Full Text Available Amoeba-associated microorganisms (AAMs are frequently isolated from water networks. In this paper, we report the isolation and characterization of Protochlamydia massiliensis, an obligate intracellular Gram-negative bacterium belonging to the Parachlamydiaceae family in the Chlamydiales order, from a cooling water tower. This bacterium was isolated on Vermamoeba vermiformis. It has a multiple range of hosts among amoeba and is characterized by a typical replication cycle of Chlamydiae with a particularity, recently shown in some chlamydia, which is the absence of inclusion vacuoles in the V. vermiformis host, adding by this a new member of Chlamydiae undergoing developmental cycle changes in the newly adapted host V. vermiformis. Draft genome sequencing revealed a chromosome of 2.86 Mb consisting of four contigs and a plasmid of 92 Kb.

  11. Acanthamoeba castellanii is not be an adequate model to study human adenovirus interactions with macrophagic cells.

    Science.gov (United States)

    Maisonneuve, Elodie; Cateau, Estelle; Leveque, Nicolas; Kaaki, Sihem; Beby-Defaux, Agnès; Rodier, Marie-Hélène

    2017-01-01

    Free living amoebae (FLA) including Acanthamoeba castellanii, are protozoa that feed on different microorganisms including viruses. These microorganisms show remarkable similarities with macrophages in cellular structures, physiology or ability to phagocyte preys, and some authors have therefore wondered whether Acanthamoeba and macrophages are evolutionary related. It has been considered that this amoeba may be an in vitro model to investigate relationships between pathogens and macrophagic cells. So, we intended in this study to compare the interactions between a human adenovirus strain and A. castellanii or THP-1 macrophagic cells. The results of molecular and microscopy techniques following co-cultures experiments have shown that the presence of the adenovirus decreased the viability of macrophages, while it has no effect on amoebic viability. On another hand, the viral replication occurred only in macrophages. These results showed that this amoebal model is not relevant to explore the relationships between adenoviruses and macrophages in in vitro experiments.

  12. Acanthamoeba castellanii is not be an adequate model to study human adenovirus interactions with macrophagic cells.

    Directory of Open Access Journals (Sweden)

    Elodie Maisonneuve

    Full Text Available Free living amoebae (FLA including Acanthamoeba castellanii, are protozoa that feed on different microorganisms including viruses. These microorganisms show remarkable similarities with macrophages in cellular structures, physiology or ability to phagocyte preys, and some authors have therefore wondered whether Acanthamoeba and macrophages are evolutionary related. It has been considered that this amoeba may be an in vitro model to investigate relationships between pathogens and macrophagic cells. So, we intended in this study to compare the interactions between a human adenovirus strain and A. castellanii or THP-1 macrophagic cells. The results of molecular and microscopy techniques following co-cultures experiments have shown that the presence of the adenovirus decreased the viability of macrophages, while it has no effect on amoebic viability. On another hand, the viral replication occurred only in macrophages. These results showed that this amoebal model is not relevant to explore the relationships between adenoviruses and macrophages in in vitro experiments.

  13. Effects of urban particulate deposition on microbial communities living in bryophytes: an experimental study.

    Science.gov (United States)

    Meyer, C; Bernard, N; Moskura, M; Toussaint, M L; Denayer, F; Gilbert, D

    2010-10-01

    Our previous in situ study showed that bryophyte-microorganism complexes were affected by particulate atmospheric pollution. Here, the effect of urban particulate wet deposits on microbial communities living in bryophytes was studied under controlled conditions. An urban particulate solution was prepared with particles extracted from analyzer' filters and nebulized on bryophytes in treatments differing in frequency and quantity. The bryophytes did not accumulate metallic trace elements, which were present in very weak concentrations. However, in treated microcosms the total microbial biomass and the biomasses of cyanobacteria, active testate amoebae and fungi significantly decreased in response to the deposition of particles. These results confirm that microbial communities living in terrestrial bryophytes could be more sensitive indicators of atmospheric pollution than bryophytes. Moreover, they suggest that unicellular predators--such as testate amoebae--could be especially useful microbial indicators, since they seem to be both directly and indirectly affected by pollution. Copyright © 2010 Elsevier Inc. All rights reserved.

  14. Chew on this: Amoebic trogocytosis and host cell killing by Entamoeba histolytica

    Science.gov (United States)

    Ralston, Katherine S.

    2015-01-01

    Entamoeba histolytica was named “histolytica” (histo-: tissue; lytic-: dissolving) for its ability to destroy host tissues. Direct killing of host cells by the amoebae is likely to be the driving factor that underlies tissue destruction, but the mechanism was unclear. We recently showed that after attaching to host cells, amoebae bite off and ingest distinct host cell fragments, and that this contributes to cell killing. Here we review this process, termed “amoebic trogocytosis” (trogo-: nibble), and how this process interplays with phagocytosis, or whole cell ingestion, in this organism. “Nibbling” processes have been described in other microbes and in multicellular organisms. The discovery of amoebic trogocytosis in E. histolytica may also shed light on an evolutionarily conserved process for intercellular exchange. PMID:26070402

  15. Primary Amoebic Meningoencephalitis in an Iranian Infant

    Directory of Open Access Journals (Sweden)

    Zahra Movahedi

    2012-01-01

    Full Text Available Introduction. Naegleria fowleri, a free living amoeba, can cause devastating and deadly diseases in humans. This is the first report of primary amoebic meningoencephalitis from Iran. Case report. A five-month-old male infant presented with the history of fever and eye gaze for three days, after beginning of bacterial meningitis, a plain and contrast CT revealed communicated hydrocephalus. In the repeat of CSF analysis on microscopic examination of wet preparation of CSF, Naegleria Fowleri was seen. Then, Amphotericin B and Rifampin were started. On followup, two months later, the patient was totally asymptomatic. Conclusion. Though occurrence of PAM is rare, this unusual disease has grave prognosis, so infection with free living amoebas must be considered in differential diagnosis of pediatric patients of purulent meningitis without evidence of bacteria on Gram’s stain and imaging findings, nonspecific brain edema on CT or hydrocephalus even without history of contact.

  16. Quantitation by flow microfluorometry of total cellular DNA in Acanthamoeba

    Energy Technology Data Exchange (ETDEWEB)

    Coulson, P.B.; Tyndall, R.

    1978-01-01

    The DNA content of five speciea of Acanthamoeba was determined by flow microfluorometry. Acanthamoeba castellanii (AC-30), acanthamoeba polyphaga (APG and P-23), acanthamoeba rhysodes, acanthamoeba culbertsoni (A-1), and acanthamoeba royreba were grown in a casitone based medium 24 to 48 hr. The trophozoites were harvested, fixed in 70% ethanol (acidified), pretreated with RNase, stained with propidium diiodide, and evaluated for DNA-bound fluorescence. All species tested had DNA values between 2.0 to 5.0 pg/cell. These results placed DNA/cell values of Acanthamoeba slightly lower than DNA/cell values of other eucaryotic cells and much lower than Amoeba proteus values. These results indicate that FMF may be a useful adjunct in distinguishing Acanthamoeba cells from either eucaryotic cells or some other amoeba. However, differences in DNA/cell between species of Acanthamoeba are small and would not be useful in identification of species.

  17. Detection of glycoproteins in the Acanthamoeba plasma membrane

    Energy Technology Data Exchange (ETDEWEB)

    Paatero, G.I.L. (Abo Akademi (Finland)); Gahmberg, C.G. (Univ. of Helsinki (Finland))

    1988-11-01

    In the present study the authors have shown that glycoproteins are present in the plasma membrane of Acanthamoeba castellanii by utilizing different radioactive labeling techniques. Plasma membrane proteins in the amoeba were iodinated by {sup 125}I-lactoperoxidase labeling and the solubilized radiolabeled glycoproteins were separated by lectin-Sepharose affinity chromatography followed by polyacrylamide gel electrophoresis. The periodate/NaB{sup 3}H{sub 4} and galactose oxidase/NaB{sup 3}H{sub 4} labeling techniques were used for labeling of surface carbohydrates in the amoeba. Several surface-labeled glycoproteins were observed in addition to a diffusely labeled region with M{sub r} of 55,000-75,000 seen on electrophoresis, which could represent glycolipids. The presence of glycoproteins in the plasma membrane of Acanthamoeba castellanii was confirmed by metabolic labeling with ({sup 35}S)methionine followed by lectin-Sepharose affinity chromatography and polyacrylamide gel electrophoresis.

  18. Detection of glycoproteins in the Acanthamoeba plasma membrane

    International Nuclear Information System (INIS)

    Paatero, G.I.L.; Gahmberg, C.G.

    1988-01-01

    In the present study the authors have shown that glycoproteins are present in the plasma membrane of Acanthamoeba castellanii by utilizing different radioactive labeling techniques. Plasma membrane proteins in the amoeba were iodinated by 125 I-lactoperoxidase labeling and the solubilized radiolabeled glycoproteins were separated by lectin-Sepharose affinity chromatography followed by polyacrylamide gel electrophoresis. The periodate/NaB 3 H 4 and galactose oxidase/NaB 3 H 4 labeling techniques were used for labeling of surface carbohydrates in the amoeba. Several surface-labeled glycoproteins were observed in addition to a diffusely labeled region with M r of 55,000-75,000 seen on electrophoresis, which could represent glycolipids. The presence of glycoproteins in the plasma membrane of Acanthamoeba castellanii was confirmed by metabolic labeling with [ 35 S]methionine followed by lectin-Sepharose affinity chromatography and polyacrylamide gel electrophoresis

  19. Comparison of formalin-ethyl ether sedimentation, formalin-ethyl acetate sedimentation, and zinc sulfate flotation techniques for detection of intestinal parasites.

    OpenAIRE

    Truant, A L; Elliott, S H; Kelly, M T; Smith, J H

    1981-01-01

    Formalin-ethyl ether sedimentation, Formalin-ethyl acetate sedimentation, and zinc sulfate flotation techniques were compared using over 250 clinical parasitology specimens. Fifty positive specimens were identified, and a variety of parasites, including amoebae, flagellates, cestodes, nematodes, and trematodes, were encountered. The Formalin-ether and Formalin-ethyl acetate sedimentation procedures gave identical results for the detection of cysts, ova, and larvae, and these methods offered a...

  20. Involvement of Sib Proteins in the Regulation of Cellular Adhesion in Dictyostelium discoideum▿ †

    OpenAIRE

    Cornillon, Sophie; Froquet, Romain; Cosson, Pierre

    2008-01-01

    Molecular mechanisms ensuring cellular adhesion have been studied in detail in Dictyostelium amoebae, but little is known about the regulation of cellular adhesion in these cells. Here, we show that cellular adhesion is regulated in Dictyostelium, notably by the concentration of a cellular secreted factor accumulating in the medium. This constitutes a quorum-sensing mechanism allowing coordinated regulation of cellular adhesion in a Dictyostelium population. In order to understand the mechani...

  1. Cooling systems

    International Nuclear Information System (INIS)

    Coutant, C.C.

    1978-01-01

    Progress on the thermal effects project is reported with regard to physiology and distribution of Corbicula; power plant effects studies on burrowing mayfly populations; comparative thermal responses of largemouth bass from northern and southern populations; temperature selection by striped bass in Cherokee Reservoir; fish population studies; and predictive thermoregulation by fishes. Progress is also reported on the following; cause and ecological ramifications of threadfin shad impingement; entrainment project; aquaculture project; pathogenic amoeba project; and cooling tower drift project

  2. First Report of Vermamoeba vermiformis in the Island of El Hierro, Canary Islands, Spain

    OpenAIRE

    María Reyes-Batlle; Maryam Niyyati; Carolina Wagner; Carmen M. Martín-Navarro; Atteneri López-Arencibia; Ines Sifaoui; Basilio Valladares; Enrique Martínez-Carretero; José E. Piñero; Jacob Lorenzo-Morales

    2016-01-01

    Background: Free-living amoebae (FLA) are group of protozoa distributed worldwide in many habitats mainly water and soil related sources. Some members of FLA are able to act as opportunistic pathogens and are environmental carriers of other pathogenic agents such as bacteria and viruses. Vermamoeba vermiformis is a highly abundant FLA species in water bodies and has recently gained environmental importance as it acts as a vehicle of many pathogenic bacteria such as Legionella pneumophila.Case...

  3. New Brazilian giant viruses isolation from environmental samples using a panel of protozoa.

    OpenAIRE

    Fábio Pio Dornas; Fábio Pio Dornas; Jacques Yaacoub Bou Khalil; Isabelle ePagnier; Didier eRaoult; Jonatas eAbrahao; Bernard eLa Scola

    2015-01-01

    The Megavirales are a newly described order capable of infecting different types of eukaryotic hosts. For the most part, the natural host is unknown. Several methods have been used to detect these viruses, with large discrepancies between molecular methods and co-cultures. To isolate giant viruses, we propose the use of different species of amoeba as a cellular support. The aim of this work was to isolate new Brazilian giant viruses by comparing the protozoa Acanthamoeba castellanii, A. polyp...

  4. Effect of temperature and colonization of Legionella pneumophila and Vermamoeba vermiformis on bacterial community composition of copper drinking water biofilms

    OpenAIRE

    Buse, Helen Y.; Ji, Pan; Gomez?Alvarez, Vicente; Pruden, Amy; Edwards, Marc A.; Ashbolt, Nicholas J.

    2017-01-01

    Summary It is unclear how the water?based pathogen, Legionella pneumophila (Lp), and associated free?living amoeba (FLA) hosts change or are changed by the microbial composition of drinking water (DW) biofilm communities. Thus, this study characterized the bacterial community structure over a 7?month period within mature (>?600?day?old) copper DW biofilms in reactors simulating premise plumbing and assessed the impact of temperature and introduction of Lp and its FLA host, Vermamoeba vermifor...

  5. Molecular detection of Acanthamoeba spp., Naegleria fowleri and Vermamoeba (Hartmannella) vermiformis as vectors for Legionella spp. in untreated and solar pasteurized harvested rainwater

    OpenAIRE

    Dobrowsky, Penelope H.; Khan, Sehaam; Cloete, Thomas E.; Khan, Wesaal

    2016-01-01

    Background Legionella spp. employ multiple strategies to adapt to stressful environments including the proliferation in protective biofilms and the ability to form associations with free-living amoeba (FLA). The aim of the current study was to identify Legionella spp., Acanthamoeba spp., Vermamoeba (Hartmannella) vermiformis and Naegleria fowleri that persist in a harvested rainwater and solar pasteurization treatment system. Methods Pasteurized (45??C, 65??C, 68??C, 74??C, 84??C and 93??C) a...

  6. The first isolation of Cochliopodium gulosum Schaeffer, 1926 (Lobosea, Himatismenida) since its initial description. I. light-microscopical investigation

    OpenAIRE

    Kudryavtsev, Alexander

    1999-01-01

    A marine lobose amoeba possessing characteristic features of the genus Cochliopodium (Hertwig and Lesser, 1874) Kudryavtsev, 1999 was found in the upper layer of sand at the beach of Keret Island, Kandalaksha Bay, the White Sea. Light-microscopical identity of this isolate and Cochliopodium gulosum Schaeffer, 1926 has been shown. This species has never been isolated and studied since its initial description. Its validity and generic position are confirmed by the results of light-microscopical...

  7. Etude de mécanismes de l'endocytose chez Dictyostelium discoideum. Caractérisation des effets inhibiteurs de la caféine et de la cycloheximide. Analyse du pH endosomal.

    OpenAIRE

    Gonzalez-Hernandez, Carlos

    1991-01-01

    Endocytosis pathways in Dictyostelium discoideum amoeba were investigated by the study of endocytic compartment acidification. The mechanisms of inhibition by caffeine and cycloheximide, two drugs that are not perturbing cellular energetic metabolism, were also explored.Fluorescein isothiocyanate labeled dextran (FITC dextran) was used as a fluid phase marker to quantify pinocytosis kinetics. Fluid internalization is more active in nutritive medium as compared to plain buffer. In both cases, ...

  8. Electrotaxis: Cell Directional Movement in Electric Fields.

    Science.gov (United States)

    Sroka, Jolanta; Zimolag, Eliza; Lasota, Slawomir; Korohoda, Wlodzimierz; Madeja, Zbigniew

    2018-01-01

    Electrotaxis plays an important role during embryogenesis, inflammation, wound healing, and tumour metastasis. However, the mechanisms at play during electrotaxis are still poorly understood. Therefore intensive studies on signaling pathways involved in this phenomenon should be carried out. In this chapter, we described an experimental system for studying electrotaxis of Amoeba proteus, mouse embryonic fibroblasts (MEF), Walker carcinosarcoma cells WC256, and bone marrow adherent cells (BMAC).

  9. Use of monoclonal antibodies to distinguish pathogenic Naegleria fowleri (cysts, trophozoites, or flagellate forms) from other Naegleria species.

    OpenAIRE

    Sparagano, O; Drouet, E; Brebant, R; Manet, E; Denoyel, G A; Pernin, P

    1993-01-01

    Monoclonal antibodies (MAbs) reactive to the pathogenic amoeba Naegleria fowleri were analyzed by enzyme-linked immunosorbent assay (ELISA), indirect immunofluorescence assay, Western blotting (immunoblotting), and radioimmunoprecipitation assay (RIPA). Two MAbs (3A4 and 5D12) showed reactivity by ELISA with all N. fowleri strains tested and no reactivity with the five other Naegleria species, N. lovaniensis, N. gruberi, N. australiensis, N. jadini, and N. andersoni. These MAbs reacted with t...

  10. Nuclease activity of Legionella pneumophila Cas2 promotes intracellular infection of amoebal host cells.

    Science.gov (United States)

    Gunderson, Felizza F; Mallama, Celeste A; Fairbairn, Stephanie G; Cianciotto, Nicholas P

    2015-03-01

    Legionella pneumophila, the primary agent of Legionnaires' disease, flourishes in both natural and man-made environments by growing in a wide variety of aquatic amoebae. Recently, we determined that the Cas2 protein of L. pneumophila promotes intracellular infection of Acanthamoeba castellanii and Hartmannella vermiformis, the two amoebae most commonly linked to cases of disease. The Cas2 family of proteins is best known for its role in the bacterial and archeal clustered regularly interspaced short palindromic repeat (CRISPR)-CRISPR-associated protein (Cas) system that constitutes a form of adaptive immunity against phage and plasmid. However, the infection event mediated by L. pneumophila Cas2 appeared to be distinct from this function, because cas2 mutants exhibited infectivity defects in the absence of added phage or plasmid and since mutants lacking the CRISPR array or any one of the other cas genes were not impaired in infection ability. We now report that the Cas2 protein of L. pneumophila has both RNase and DNase activities, with the RNase activity being more pronounced. By characterizing a catalytically deficient version of Cas2, we determined that nuclease activity is critical for promoting infection of amoebae. Also, introduction of Cas2, but not its catalytic mutant form, into a strain of L. pneumophila that naturally lacks a CRISPR-Cas locus caused that strain to be 40- to 80-fold more infective for amoebae, unequivocally demonstrating that Cas2 facilitates the infection process independently of any other component encoded within the CRISPR-Cas locus. Finally, a cas2 mutant was impaired for infection of Willaertia magna but not Naegleria lovaniensis, suggesting that Cas2 promotes infection of most but not all amoebal hosts. Copyright © 2015, American Society for Microbiology. All Rights Reserved.

  11. Aquatic organisms as amber inclusions and examples from a modern swamp forest

    OpenAIRE

    Schmidt, Alexander R.; Dilcher, David L.

    2007-01-01

    To find aquatic organisms in tree resin may seem to be highly unlikely, but the fossil record provides numerous amber-preserved limnetic arthropods (e.g., water beetles, water striders, and crustaceans) and microorganisms (e.g., bacteria, algae, ciliates, testate amoebae, and rotifers). Here we explain the frequently discussed process of embedding aquatic organisms in tree resin based on field studies in a Florida swamp forest. Different aquatic arthropods and all major groups of limnetic mic...

  12. Occurrence and Control of Legionella in Recycled Water Systems

    OpenAIRE

    Jjemba, Patrick K.; Johnson, William; Bukhari, Zia; LeChevallier, Mark W.

    2015-01-01

    Legionella pneumophila is on the United States Environmental Protection Agency (USEPA) Candidate Contaminant list (CCL) as an important pathogen. It is commonly encountered in recycled water and is typically associated with amoeba, notably Naegleria fowleri (also on the CCL) and Acanthamoeba sp. No legionellosis outbreak has been linked to recycled water and it is important for the industry to proactively keep things that way. A review was conducted examine the occurrence of Legionella and it...

  13. Compilation of 1986 Annual Reports of the Navy ELF (Extremely Low Frequency) Communications System Ecological Monitoring Program. Volume 2. TABS D-G.

    Science.gov (United States)

    1987-07-01

    diploid organisms. For this reason, the analytical and statistical techniques developed for isoenzyme analyses of higher organisms is used in the present...determined by varying the number of bacteria for amoeba growth and then following growth rate and maximum yield. Culture chambers, containing...polyphaga show differences between isolates consistent with a diploid organism. Nei’s method for measuring genetic difference (Net, 1972) Is used In

  14. Variation in the SSUrDNA of the Genus Protostelium Leads to a New Phylogenetic Understanding of the Genus and of the Species Concept for Protostelium mycophaga (Protosteliida, Amoebozoa).

    Science.gov (United States)

    Shadwick, John D L; Silberman, Jeffery D; Spiegel, Frederick W

    2017-10-16

    Members of the genus Protostelium (including P. mycophaga, P. nocturnum, and P. okumukumu) are protosteloid amoebae commonly found in terrestrial habitats on dead plant matter. They, along with the closely allied nominal genus Planoprotostelium, containing the single species Pl. aurantium, all have an amoeboid trophic stage with acutely pointed subpseudopodia and orange lipid droplets in the granuloplasm. These amoebae form stalked fruiting bodies topped with a single, usually deciduous spore. The species are identified based on their fruiting body morphologies except for Pl. aurantium which looks similar to P. mycophaga in fruiting morphology, but has amoebae that can make flagella in liquid medium. We built phylogenetic trees using nuclear small subunit ribosomal DNA sequences of 35 isolates from the genera Protostelium and Planoprotostelium and found that (1) the nonflagellated P. nocturnum and P. okumukumu branch basally in the genus Protostelium, (2) the flagellate, Pl. aurantium falls within the genus Protostelium in a monophyletic clade with the nominal variety, P. mycophaga var. crassipes, (3) the cultures initially identified as Protostelium mycophaga can be divided into at least three morphologically recognizable taxa, P. aurantium n. comb., P. apiculatum n. sp., and P. m. rodmani n. subsp., as well as a paraphyletic assemblage that includes the remainder of the P. mycophaga morphotype. These findings have implications for understanding the ecology, evolution, and diversity of these amoeboid organisms and for using these amoebae as models for other amoeboid groups. © 2017 The Author(s) Journal of Eukaryotic Microbiology © 2017 International Society of Protistologists.

  15. Didymium-like myxogastrids (class Mycetozoa) as endocommensals of sea urchins (Sphaerechinus granularis)

    Czech Academy of Sciences Publication Activity Database

    Dyková, Iva; Lom, Jiří; Dvořáková, Helena; Pecková, Hana; Fiala, Ivan

    2007-01-01

    Roč. 54, č. 1 (2007), s. 1-12 ISSN 0015-5683 R&D Projects: GA ČR GA206/05/2384; GA MŠk LC522 Institutional research plan: CEZ:AV0Z60220518 Keywords : Didymium-like myxogastrids * Myxogastrea * sea-urchin amoeba strains * SSU rDNA * flagellar apparatus * phylogeny Subject RIV: EA - Cell Biology Impact factor: 1.000, year: 2007

  16. Naegleria fowleri in cooling circuits of industrial and power plants in North Moravia

    Energy Technology Data Exchange (ETDEWEB)

    Cerva, L.; Simanov, L.

    1983-01-01

    Both nonvirulent (15 strains) and virulent (1 strain) variants of Naegleria fowleri were isolated from the cooling circuits in seven of ten examined plants in North Moravia. No amoebae were found in waters with a high salinity (electric conductivity more than 614 microseconds) and with a low content of phosphates and oxygen. Other results of the physical, chemical and bacteriological examinations showed no direct bearing on the occurrence of N. fowleri.

  17. Solving inverse problems for biological models using the collage method for differential equations.

    Science.gov (United States)

    Capasso, V; Kunze, H E; La Torre, D; Vrscay, E R

    2013-07-01

    In the first part of this paper we show how inverse problems for differential equations can be solved using the so-called collage method. Inverse problems can be solved by minimizing the collage distance in an appropriate metric space. We then provide several numerical examples in mathematical biology. We consider applications of this approach to the following areas: population dynamics, mRNA and protein concentration, bacteria and amoeba cells interaction, tumor growth.

  18. Coupling of cytoskeleton functions for fibroblast locomotion

    DEFF Research Database (Denmark)

    Couchman, J R; Lenn, M; Rees, D A

    1985-01-01

    Using a chick cell phenotype specialised for locomotion with morphometric measurements made possible by modern instrumentation technology, we have reinvestigated motile functions in fibroblast locomotion. Quantitative analysis of rapid fluctuations in cell form and organelle distribution during l...... function of microtubules to direct the flow towards multiple foci on the leading edge, and so determine cell polarity. Such a mechanism of locomotion for fibroblasts has many features consistent with evidence for other cell types, especially amoebae and leukocytes....

  19. Solvation structure and dynamics of Ni{sup 2+}(aq) from a polarizable force field

    Energy Technology Data Exchange (ETDEWEB)

    Mareš, Jiří, E-mail: jiri.mares@oulu.fi; Vaara, Juha

    2014-10-31

    Highlights: • We parameterize the Ni{sup 2+} ion within the AMOEBA polarizable forcefield. • Besides vdW parameters, we fit also polarizability, Thole damping and charge. • We use an empirical adjustment to account for the transition into condensed phase. • Very good structural and dynamical properties of Ni{sup 2+}(aq) are demonstrated. - Abstract: An aqueous solution of Ni{sup 2+} has often been used as a prototypic transition-metal system for experimental and theoretical studies in nuclear and electron-spin magnetic resonance (NMR and ESR). Molecular dynamics (MD) simulation of Ni{sup 2+}(aq) has been a part of many of these studies. As a transition metal complex, its MD simulation is particularly difficult using common force fields. In this work, we parameterize the Ni{sup 2+} ion for a simulation of the aqueous solution within the modern polarizable force field AMOEBA. We show that a successful parameterization is possible for this specific case when releasing the physical interpretation of the electrostatic and polarization parameters of the force field. In doing so, particularly the Thole damping parameter and also the ion charge and polarizability were used as fitting parameters. The resulting parameterizations give in a MD simulation good structural and dynamical properties of the [Ni(H{sub 2}O){sub 6}]{sup 2+} complex, along with the expected excellent performance of AMOEBA for the water solvent. The presented parameterization is appropriate for high-accuracy simulations of both structural and dynamic properties of Ni{sup 2+}(aq). This work documents possible approaches of parameterization of a transition metal within the AMOEBA force field.

  20. Characterization of a Legionella micdadei mip mutant

    DEFF Research Database (Denmark)

    O'Connell, W A; Bangsborg, Jette Marie; Cianciotto, N P

    1995-01-01

    The pathogenesis of Legionella micdadei is dependent upon its ability to infect alveolar phagocytes. To better understand the basis of intracellular infection by this organism, we examined the importance of its Mip surface protein. In Legionella pneumophila, Mip promotes infection of both human m...... into the phagocyte. Similarly, the mutant was less able to parasitize Hartmannella amoebae. Taken together, these data argue that Mip specifically potentiates intracellular growth by L. micdadei....