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Sample records for acanthamoeba castellanii encystment

  1. Francisella tularensis type A Strains Cause the Rapid Encystment of Acanthamoeba castellanii and Survive in Amoebal Cysts for Three Weeks post Infection

    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.

  2. Acanthamoeba castellanii STAT protein.

    Kicinska, Anna; Leluk, Jacek; Jarmuszkiewicz, Wieslawa

    2014-01-01

    STAT (signal transducers and activators of transcription) proteins are one of the important mediators of phosphotyrosine-regulated signaling in metazoan cells. We described the presence of STAT protein in a unicellular, free-living amoebae with a simple life cycle, Acanthamoeba castellanii. A. castellanii is the only, studied to date, Amoebozoan that does not belong to Mycetozoa but possesses STATs. A sequence of the A. castellanii STAT protein includes domains similar to those of the Dictyostelium STAT proteins: a coiled coil (characteristic for Dictyostelium STAT coiled coil), a STAT DNA-binding domain and a Src-homology domain. The search for protein sequences homologous to A. castellanii STAT revealed 17 additional sequences from lower eukaryotes. Interestingly, all of these sequences come from Amoebozoa organisms that belong to either Mycetozoa (slime molds) or Centramoebida. We showed that there are four separated clades within the slime mold STAT proteins. The A. castellanii STAT protein branches next to a group of STATc proteins from Mycetozoa. We also demonstrate that Amoebozoa form a distinct monophyletic lineage within the STAT protein world that is well separated from the other groups. PMID:25338074

  3. Acanthamoeba castellanii STAT protein.

    Anna Kicinska

    Full Text Available STAT (signal transducers and activators of transcription proteins are one of the important mediators of phosphotyrosine-regulated signaling in metazoan cells. We described the presence of STAT protein in a unicellular, free-living amoebae with a simple life cycle, Acanthamoeba castellanii. A. castellanii is the only, studied to date, Amoebozoan that does not belong to Mycetozoa but possesses STATs. A sequence of the A. castellanii STAT protein includes domains similar to those of the Dictyostelium STAT proteins: a coiled coil (characteristic for Dictyostelium STAT coiled coil, a STAT DNA-binding domain and a Src-homology domain. The search for protein sequences homologous to A. castellanii STAT revealed 17 additional sequences from lower eukaryotes. Interestingly, all of these sequences come from Amoebozoa organisms that belong to either Mycetozoa (slime molds or Centramoebida. We showed that there are four separated clades within the slime mold STAT proteins. The A. castellanii STAT protein branches next to a group of STATc proteins from Mycetozoa. We also demonstrate that Amoebozoa form a distinct monophyletic lineage within the STAT protein world that is well separated from the other groups.

  4. Gene discovery in the Acanthamoeba castellanii genome

    Anderson, Iain J.; Watkins, Russell F.; Samuelson, John; Spencer,David F.; Majoros, William H.; Gray, Michael W.; Loftus, Brendan J.

    2005-08-01

    Acanthamoeba castellanii is a free-living amoeba found in soil, freshwater, and marine environments and an important predator of bacteria. Acanthamoeba castellanii is also an opportunistic pathogen of clinical interest, responsible for several distinct diseases in humans. In order to provide a genomic platform for the study of this ubiquitous and important protist, we generated a sequence survey of approximately 0.5 x coverage of the genome. The data predict that A. castellanii exhibits a greater biosynthetic capacity than the free-living Dictyostelium discoideum and the parasite Entamoeba histolytica, providing an explanation for the ability of A. castellanii to inhabit adversity of environments. Alginate lyase may provide access to bacteria within biofilms by breaking down the biofilm matrix, and polyhydroxybutyrate depolymerase may facilitate utilization of the bacterial storage compound polyhydroxybutyrate as a food source. Enzymes for the synthesis and breakdown of cellulose were identified, and they likely participate in encystation and excystation as in D. discoideum. Trehalose-6-phosphate synthase is present, suggesting that trehalose plays a role in stress adaptation. Detection and response to a number of stress conditions is likely accomplished with a large set of signal transduction histidine kinases and a set of putative receptorserine/threonine kinases similar to those found in E. histolytica. Serine, cysteine and metalloproteases were identified, some of which are likely involved in pathogenicity.

  5. Acanthamoeba encystment: multifactorial effects of buffers, biocides, and demulcents present in contact lens care solutions

    Kovacs CJ

    2015-10-01

    Full Text Available Christopher J Kovacs, Shawn C Lynch, Marjorie J Rah, Kimberly A Millard, Timothy W Morris Bausch & Lomb Incorporated, Rochester, NY, USA Purpose: To determine whether agents which are purportedly capable of inducing encystment of Acanthamoeba can recapitulate the signal when tested in differing formulations. Methods: In accordance with the International Standard ISO 19045, Acanthamoeba castellanii ATCC 50370 trophozoites were cultured in antibiotic-free axenic medium, treated with test solutions, and encystment rates plus viability were measured via bright field and fluorescent microscopy. Test solutions included phosphate-buffered saline (PBS, borate-buffered saline, biguanide- and hydrogen peroxide (H2O2-based biocides, propylene glycol (PG and povidone (POV ophthalmic demulcents, and one-step H2O2-based contact lens disinfection systems. Results: Only PBS solutions with 0.25 ppm polyaminopropyl biguanide (PAPB and increasing concentrations of PG and POV stimulated A. castellanii encystment in a dose-dependent manner, whereas PBS solutions containing 3% H2O2 and increasing concentrations of PG and POV did not stimulate encystment. Borate-buffered saline and PBS/citrate solutions containing PG also did not stimulate encystment. In addition, no encystment was observed after 24 hours, 7 days, or 14 days of exposures of trophozoites to one-step H2O2 contact lens disinfection products or related solutions. Conclusion: The lack of any encystment observed when trophozoites were treated with existing or new one-step H2O2 contact lens care products, as well as when trophozoites were exposed to various related test solutions, confirms that Acanthamoeba encystment is a complex process which depends upon simultaneous contributions of multiple factors including buffers, biocides, and demulcents. Keywords: propylene glycol, contact lens care system, hydrogen peroxide disinfecting solution

  6. Anaerobic respiration: In vitro efficacy of Nitazoxanide against mitochondriate Acanthamoeba castellanii of the T4 genotype.

    Aqeel, Yousuf; Siddiqui, Ruqaiyyah; Farooq, Maria; Khan, Naveed Ahmed

    2015-10-01

    Acanthamoeba is an opportunistic protist pathogen that is responsible for serious human and animal infection. Being one of the most frequently isolated protists from the environment, it is likely that it readily encounters microaerophilic environments. For respiration under anaerobic or low oxygen conditions in several amitochondriate protists, decarboxylation of pyruvate is catalyzed by pyruvate ferredoxin oxidoreductase instead of pyruvate dehydrogenase. In support, Nitazoxanide, an inhibitor of pyruvate ferredoxin oxidoreductase, is effective and non-mutagenic clinically against a range of amitochondriate protists, Giardia intestinalis, Entamoeba histolytica and Trichomonas vaginalis. The overall aim of the present study was to determine in vitro efficacy of Nitazoxanide against Acanthamoeba castellanii. At micromolar concentrations, the findings revealed that Nitazoxanide neither affected A. castellanii growth or viability nor amoeba-mediated host cell monolayer damage in vitro or extracellular proteolytic activities. Similarly, microaerophilic conditions alone had no significant effects. In contrast, microaerophilic conditions together with Nitazoxanide showed amoebicidal effects and inhibited A. castellanii-mediated host cell monolayer damage as well as extracellular proteases. Using encystation assays, it was observed that Nitazoxanide inhibited trophozoite transformation into cysts both under aerophilic and microaerophilic conditions. Furthermore, pre-treatment of cysts with Nitazoxanide inhibited A. castellanii excystation. These findings are important in the identification of potential targets that could be useful against parasite-specific respiration as well as to understand the basic biology of the life cycle of Acanthamoeba. PMID:26297676

  7. Purification and characterization of transcription factor IIIA from Acanthamoeba castellanii

    Polakowski, Nicholas; Paule, Marvin R.

    2002-01-01

    TFIIIA is required to activate RNA polymerase III transcription from 5S RNA genes. Although all known TFIIIA homologs harbor nine zinc fingers that mediate DNA binding, very limited sequence homology is found among these proteins, which reflects unique properties of some TFIIIA homologs. For example, the Acanthamoeba castellanii homolog directly regulates 5S RNA transcription. We have purified and characterized A.castellanii TFIIIA (AcTFIIIA) as a step toward obtaining a clearer understanding...

  8. Azole Antifungal Agents To Treat the Human Pathogens Acanthamoeba castellanii and Acanthamoeba polyphaga through Inhibition of Sterol 14α-Demethylase (CYP51)

    Lamb, David C.; Warrilow, Andrew G. S.; Rolley, Nicola J.; Parker, Josie E.; Nes, W. David; Smith, Stephen N; Kelly, Diane E.; Kelly, Steven L.

    2015-01-01

    In this study, we investigate the amebicidal activities of the pharmaceutical triazole CYP51 inhibitors fluconazole, itraconazole, and voriconazole against Acanthamoeba castellanii and Acanthamoeba polyphaga and assess their potential as therapeutic agents against Acanthamoeba infections in humans. Amebicidal activities of the triazoles were assessed by in vitro minimum inhibition concentration (MIC) determinations using trophozoites of A. castellanii and A. polyphaga. In addition, triazole e...

  9. Temperature-induced membrane-lipid adaptation in Acanthamoeba castellanii.

    Jones, A L; Hann, A C; Harwood, J L; Lloyd, D

    1993-02-15

    A method has been developed for the separation of the major membrane fractions of Acanthamoeba castellanii after growth at different temperatures. The acyl-lipid compositions of individual membrane fractions, microsomal membranes, plasma membrane and mitochondria were analysed after a shift in culture temperature from 30 degrees C to 15 degrees C. The major change in lipid composition observed was an alteration in the relative proportions of oleate and linoleate. This reciprocal change was seen in all the membrane fractions, but occurred most rapidly in the phosphatidylcholine of the microsomal fraction. Thus, there appears to be a rapid induction of delta 12-desaturase activity in A. castellanii after a downward shift in growth temperature. Changes were also seen in the proportions of the n-6 C20 fatty acids, with a decrease in the proportions of icosadienoate and increases of icosatrienoate and arachidonate. However, unlike the alteration in oleate/linoleate ratios, this change was not seen in all the individual lipids of each membrane fraction. PMID:8439295

  10. Artemether Exhibits Amoebicidal Activity against Acanthamoeba castellanii through Inhibition of the Serine Biosynthesis Pathway

    Deng, Yihong; Ran, Wei; Man, Suqin; Li, Xueping; Gao, Hongjian; Tang, Wei; Tachibana, Hiroshi; Cheng, Xunjia

    2015-01-01

    Acanthamoeba sp. parasites are the causative agents of Acanthamoeba keratitis, fatal granulomatous amoebic encephalitis, and cutaneous infections. However, there are currently no effective drugs for these organisms. Here, we evaluated the activity of the antimalarial agent artemether against Acanthamoeba castellanii trophozoites and identified potential targets of this agent through a proteomic approach. Artemether exhibited in vitro amoebicidal activity in a time- and dose-dependent manner a...

  11. Interaction of Escherichia coli K1 and K5 with Acanthamoeba castellanii Trophozoites and Cysts

    Matin, Abdul; Jung, Suk-Yul

    2011-01-01

    The existence of symbiotic relationships between Acanthamoeba and a variety of bacteria is well-documented. However, the ability of Acanthamoeba interacting with host bacterial pathogens has gained particular attention. Here, to understand the interactions of Escherichia coli K1 and E. coli K5 strains with Acanthamoeba castellanii trophozoites and cysts, association assay, invasion assay, survival assay, and the measurement of bacterial numbers from cysts were performed, and nonpathogenic E. ...

  12. Acanthamoeba castellanii induces host cell death via a phosphatidylinositol 3-kinase-dependent mechanism

    Sissons, James; Kim, Kwang Sik; Stins, Monique; Jayasekera, Samantha; Alsam, Selwa; Khan, Naveed Ahmed

    2005-01-01

    Granulomatous amoebic encephalitis due to Acanthamoeba castellanii is a serious human infection with fatal consequences, but it is not clear how the circulating amoebae interact with the blood-brain barrier and transmigrate into the central nervous system. We studied the effects of an Acanthamoeba encephalitis isolate belonging to the T1 genotype on human brain microvascular endothelial cells, which constitute the blood-brain barrier. Using an apoptosis-specific enzyme-linked immunosorbent as...

  13. Acanthamoeba castellanii Induces Host Cell Death via a Phosphatidylinositol 3-Kinase-Dependent Mechanism

    Sissons, James; Kim, Kwang Sik; Stins, Monique; Jayasekera, Samantha; Alsam, Selwa; Khan, Naveed Ahmed

    2005-01-01

    Granulomatous amoebic encephalitis due to Acanthamoeba castellanii is a serious human infection with fatal consequences, but it is not clear how the circulating amoebae interact with the blood-brain barrier and transmigrate into the central nervous system. We studied the effects of an Acanthamoeba encephalitis isolate belonging to the T1 genotype on human brain microvascular endothelial cells, which constitute the blood-brain barrier. Using an apoptosis-specific enzyme-linked immunosorbent assay, we showed that Acanthamoeba induces programmed cell death in brain microvascular endothelial cells. Next, we observed that Acanthamoeba specifically activates phosphatidylinositol 3-kinase. Acanthamoeba-mediated brain endothelial cell death was abolished using LY294002, a phosphatidylinositol 3-kinase inhibitor. These results were further confirmed using brain microvascular endothelial cells expressing dominant negative forms of phosphatidylinositol 3-kinase. This is the first demonstration that Acanthamoeba-mediated brain microvascular endothelial cell death is dependent on phosphatidylinositol 3-kinase. PMID:15845472

  14. Superdiffusion dominates intracellular particle motion in the supercrowded space of pathogenic Acanthamoeba castellanii

    Reverey, J F; Bao, H; Leippe, M; Metzler, R; Selhuber-Unkel, C

    2015-01-01

    Acanthamoebae are free-living protists and human pathogens, whose cellular functions and pathogenicity strongly depend on the transport of intracellular vesicles and granules through the cytosol. Using high-speed live cell imaging in combination with single-particle tracking analysis, we show here that the motion of endogenous intracellular particles in the size range from a few hundred nanometers to several micrometers in Acanthamoeba castellanii is strongly superdiffusive and influenced by cell locomotion, cytoskeletal elements, and myosin II. We demonstrate that cell locomotion significantly contributes to intracellular particle motion, but is clearly not the only origin of superdiffusivity. By analyzing the contribution of microtubules, actin, and myosin II motors we show that myosin II is a major driving force of intracellular motion in A. castellanii. The cytoplasm of A. castellanii is supercrowded with intracellular vesicles and granules, such that significant intracellular motion can only be achieved ...

  15. Effects of pesticides, polychlorinated biphenyls and metals on the growth and reproduction of Acanthamoeba castellanii

    Prescott, L.M.; Kubovec, M.K.; Tryggestad, D.

    1977-07-01

    The effects of pollutants (pesticides, PCB and metals) were studied in the free-living amoeba, Acanthamoeba castellanii. Eight pesticides were used--the insecticides dieldrin, aldrin and sevin, and the herbicides linuron, stam F-34, IPC, atrazine and simazine. It was shown that the sensitivity of A. castellanii to pesticides varied greatly. The population growth was inhibited by linuron, stam F-34, IPC, sevin and atrazine at a level of 10 mg/l. The polychlorinated biphenyl, Arochor 1254, had no significant effect at a concentration of 0.01 mg/l (10 ppb). The studies with metal ions showed that A. castellanii was unaffected by moderately high levels of Cu and Zn, but was sensitive to the presence of Pb and mercuric ions.

  16. The role of Src kinase in the biology and pathogenesis of Acanthamoeba castellanii

    Siddiqui Ruqaiyyah

    2012-06-01

    Full Text Available Abstract Background Acanthamoeba species are the causative agents of fatal granulomatous encephalitis in humans. Haematogenous spread is thought to be a primary step, followed by blood–brain barrier penetration, in the transmission of Acanthmaoeba into the central nervous system, but the associated molecular mechanisms remain unclear. Here, we evaluated the role of Src, a non-receptor protein tyrosine kinase in the biology and pathogenesis of Acanthamoeba. Methods Amoebistatic and amoebicidal assays were performed by incubating amoeba in the presence of Src kinase-selective inhibitor, PP2 (4-amino-5-(4-chlorophenyl-7-(t-butylpyrazolo[3,4-d]pyrimidine and its inactive analog, PP3 (4-amino-7-phenylpyrazolo[3,4-d]pyrimidine. Using this inhibitor, the role of Src kinase in A. castellanii interactions with Escherichia coli was determined. Zymographic assays were performed to study effects of Src kinase on extracellular proteolytic activities of A. castellanii. The human brain microvascular endothelial cells were used to determine the effects of Src kinase on A. castellanii adhesion to and cytotoxicity of host cells. Results Inhibition of Src kinase using a specific inhibitor, PP2 (4-amino-5-(4 chlorophenyl-7-(t-butylpyrazolo [3,4-d] pyrimidine but not its inactive analog, PP3 (4-amino-7-phenylpyrazolo[3,4-d] pyrimidine, had detrimental effects on the growth of A. castellanii (keratitis isolate, belonging to the T4 genotype. Interestingly, inhibition of Src kinase hampered the phagocytic ability of A. castellanii, as measured by the uptake of non-invasive bacteria, but, on the contrary, invasion by pathogenic bacteria was enhanced. Zymographic assays revealed that inhibition of Src kinases reduced extracellular protease activities of A. castellanii. Src kinase inhibition had no significant effect on A. castellanii binding to and cytotoxicity of primary human brain microvascular endothelial cells, which constitute the blood–brain barrier

  17. Cellular Response of the Amoeba Acanthamoeba castellanii to Chlorine, Chlorine Dioxide, and Monochloramine Treatments ▿

    Mogoa, Emerancienne; Bodet, Charles; Morel, Franck; Rodier, Marie-Hélène; Legube, Bernard; Héchard, Yann

    2011-01-01

    Acanthamoeba castellanii is a free-living amoebae commonly found in water systems. Free-living amoebae might be pathogenic but are also known to bear phagocytosis-resistant bacteria, protecting these bacteria from water treatments. The mode of action of these treatments is poorly understood, particularly on amoebae. It is important to examine the action of these treatments on amoebae in order to improve them. The cellular response to chlorine, chlorine dioxide, and monochloramine was tested o...

  18. Azole Antifungal Agents To Treat the Human Pathogens Acanthamoeba castellanii and Acanthamoeba polyphaga through Inhibition of Sterol 14α-Demethylase (CYP51).

    Lamb, David C; Warrilow, Andrew G S; Rolley, Nicola J; Parker, Josie E; Nes, W David; Smith, Stephen N; Kelly, Diane E; Kelly, Steven L

    2015-08-01

    In this study, we investigate the amebicidal activities of the pharmaceutical triazole CYP51 inhibitors fluconazole, itraconazole, and voriconazole against Acanthamoeba castellanii and Acanthamoeba polyphaga and assess their potential as therapeutic agents against Acanthamoeba infections in humans. Amebicidal activities of the triazoles were assessed by in vitro minimum inhibition concentration (MIC) determinations using trophozoites of A. castellanii and A. polyphaga. In addition, triazole effectiveness was assessed by ligand binding studies and inhibition of CYP51 activity of purified A. castellanii CYP51 (AcCYP51) that was heterologously expressed in Escherichia coli. Itraconazole and voriconazole bound tightly to AcCYP51 (dissociation constant [Kd] of 10 and 13 nM), whereas fluconazole bound weakly (Kd of 2,137 nM). Both itraconazole and voriconazole were confirmed to be strong inhibitors of AcCYP51 activity (50% inhibitory concentrations [IC50] of 0.23 and 0.39 μM), whereas inhibition by fluconazole was weak (IC50, 30 μM). However, itraconazole was 8- to 16-fold less effective (MIC, 16 mg/liter) at inhibiting A. polyphaga and A. castellanii cell proliferation than voriconazole (MIC, 1 to 2 mg/liter), while fluconazole did not inhibit Acanthamoeba cell division (MIC, >64 mg/liter) in vitro. Voriconazole was an effective inhibitor of trophozoite proliferation for A. castellanii and A. polyphaga; therefore, it should be evaluated in trials versus itraconazole for controlling Acanthamoeba infections. PMID:26014948

  19. Swedish isolates of Vibrio cholerae enhance their survival when interacted intracellularly with Acanthamoeba castellanii

    Shanan, Salah; Bayoumi, Magdi; Saeed, Amir; Sandström, Gunnar; Abd, Hadi

    2016-01-01

    Vibrio cholerae is a Gram-negative bacterium that occurs naturally in aquatic environment. Only V. cholerae O1 and V. cholerae O139 produce cholera toxin and cause cholera, other serogroups can cause gastroenteritis, open wounds infection, and septicaemia. V. cholerae O1 and V. cholerae O139 grow and survive inside Acanthamoeba castellanii. The aim of this study is to investigate the interactions of the Swedish clinical isolates V. cholerae O3, V. cholerae O4, V. cholerae O5, V. cholerae O11,...

  20. Acanthamoeba castellanii of the T4 genotype is a potential environmental host for Enterobacter aerogenes and Aeromonas hydrophila

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

    2013-01-01

    Background 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. Methods Here, we used a keratitis isolate of A. castellanii of the T4 genotyp...

  1. Swedish isolates of Vibrio cholerae enhance their survival when interacted intracellularly with Acanthamoeba castellanii

    Salah Shanan

    2016-04-01

    Full Text Available Vibrio cholerae is a Gram-negative bacterium that occurs naturally in aquatic environment. Only V. cholerae O1 and V. cholerae O139 produce cholera toxin and cause cholera, other serogroups can cause gastroenteritis, open wounds infection, and septicaemia. V. cholerae O1 and V. cholerae O139 grow and survive inside Acanthamoeba castellanii. The aim of this study is to investigate the interactions of the Swedish clinical isolates V. cholerae O3, V. cholerae O4, V. cholerae O5, V. cholerae O11, and V. cholerae O160 with A. castellanii. The interaction between A. castellanii and V. cholerae strains was studied by means of amoeba cell counts, viable counts of the bacteria in the absence or presence of amoebae, and of the intracellularly growing bacteria, visualised by electron microscopy. These results show that all V. cholerae can grow and survive outside and inside the amoebae, disclosing that V. cholerae O3, V. cholerae O4, V. cholerae O5, V. cholerae O11, and V. cholerae O160 all can be considered as facultative intracellular bacteria.

  2. Caspase-like proteins: Acanthamoeba castellanii metacaspase and Dictyostelium discoideum paracaspase, what are their functions?

    Entsar Saheb; Wendy Trzyna; John Bush

    2014-12-01

    Caspases are cysteine proteases that are important regulators of programmed cell death in animals. Two novel relatives to members of the caspase families metacaspases and paracaspase have been discovered. Metacaspase type-1 was identified in Acanthamoeba castellanii, an opportunistic protozoan parasite that causes severe diseases in humans. Paracaspase was found in the non-pathogenic protozoan Dictyostelium discoideum. Since their discovery in Acanthamoeba and Dictyostelium, metacaspases and paracaspases have remained poorly characterized. At present we do not have sufficient data about the molecular function of these caspase-like proteins or their role, if any, in programmed cell death. How these caspase proteins function at the molecular level is an important area of study that will provide insight into their potential for treatment therapies against Acanthamoeba infection and other similar parasitic protozoan. Additionally, finding the molecular functions of these caspase-like proteins will provide information concerning their role in more complex organisms.The aim of this article was to review recent discoveries about metacaspases and paracaspases as regulators of apoptotic and non-apoptotic processes.

  3. Rapid induction of microsomal delta 12(omega 6)-desaturase activity in chilled Acanthamoeba castellanii.

    Jones, A L; Lloyd, D; Harwood, J L

    1993-11-15

    The activity of microsomal delta 12-desaturase in Acanthamoeba castellanii was increased after growing cultures were chilled from the optimal growth temperature (30 degrees C) to 15 degrees C. This increase was detectable in microsomes isolated from organisms subjected to only 10 min chilling. The mechanism of induction was investigated. The increase in activity on chilling was greatly reduced when protein synthesis was blocked before the temperature shift. Thus the major mechanism for the induction of delta 12-desaturase is increased protein synthesis. delta 12-Desaturase activity was higher when assayed at 20 degrees C than when assayed at 30 degrees C, but these changes were not due to the increased solubility of O2 at 20 degrees C. The major substrate of delta 12-desaturase was found to be 1-acyl-2-oleoyl phosphatidylcholine. PMID:8250841

  4. Oxygen induces fatty acid (n-6)-desaturation independently of temperature in Acanthamoeba castellanii.

    Thomas, K; Rutter, A; Suller, M; Harwood, J; Lloyd, D

    1998-03-20

    Induction of a microsomal oleate delta12 (n-6) desaturase which is mainly responsible for an increase in membrane lipid unsaturation at low temperature has been observed in the free-living amoeba Acanthamoeba castellanii. In this study we show that the enzyme can also be regulated by oxygen independently of temperature in batch cultures grown to O2-limitation. Raising the oxygen concentration from below the lower limit of detection (< 0.1 microM) to approximately air-saturation (230 microM), whilst maintaining the growth temperature constant (30 degrees C), increased lipid unsaturation and elevated n-6-desaturase activity 2.3-fold. Addition of the protein synthesis inhibitor, anisomycin, showed that increased desaturase activity was due to new protein synthesis rather than activation of pre-existing enzyme. These observations are important for future studies of the mechanism of temperature adaptation in poikilotherms. PMID:9541030

  5. Molecular Characterization and Sequencing of a Gene Encoding Mannose Binding Protein in an Iranian Isolate of Acanthamoeba castellanii as a Major Agent of Acanthamoeba Keratitis

    SH Farnia

    2008-07-01

    Full Text Available Background: Acanthamoeba castellanii is the important cause of amoebic keratitis in Iran. The key molecule in pathogenesis of Acanthamoeba keratitis is Mannose Binding Protein (MBP led to adhesion of amoeba to corneal epithelium. Subsequent to adhesion other cytopathic effects occur. The goal of this study was to identify the molecular characterization of a gene encoding MBP in an Iranian isolate of A.castellanii in order to pave the way for further investigations such as new therapeutic advances or immunization. Methods: A.castellanii was cultured on non nutrient agar. Extraction of DNA was performed by phenol-chloroform method. After designing a pair of primer for the gene encoding MBP, PCR analysis was performed. Finally, the PCR product has been sequenced and the result submitted to the gene data banks. Results: An MBP gene of 1081 nucleotides was sequenced. This fragment contained three introns and encodes a protein with 194 amino acids. Homology search by Blast program showed a significant homology with the MBP gene in gene data banks (96%. Besides, the identity of amino acids with the other MBPs in gene data banks was about 86%. Conclusion: We isolated and sequenced a gene fragment encoding MBP in an Iranian isolate of A.castellanii. Molecular characterization of this important gene is the first step in pursuing researches such as developing better therapeutic agents, immunization of population at risk or even developing a diagnostic tool by PCR techniques.

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

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

    2012-01-01

    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....... 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...

  7. Tracking amino acid's uptake into the protozoan Acanthamoeba castellanii by stable-isotope labelling and Raman spectral imaging

    Naemat, Abida; Elsheikha, Hany M.; Notingher, Ioan

    2016-04-01

    The capacity of pathogens to acquire nutrients from their host cells is one of the most fundamental aspects of infection biology. Hence, measuring the patterns of nutrients' uptake by pathogens is essential for understanding the interactions of pathogens with eukaryotic host cells. In this study, we optimized a technique that allows fast and non-destructive measurement of the amino acid Phenylalanine (Phe) acquired by the trophozoite stage of the protozoan Acanthamoeba castellanii (A. castellanii) as they engage with individual human retinal pigment epithelial cells (ARPE-19). ARPE-19 host cells were pre-saturated with Deuterated Phe (L-Phe(D8)) to replace the native substrate Phe (L-Phe). The uptake of L-Phe(D8) by A. castellanii trophozoites was measured by Raman microspectroscopy. This approach allowed us to characterize the uptake patterns of this essential amino acid into A. castellanii trophozoites at a single cell level. At 24 hours post infection (PI) A. castellanii trophozoites are capable of salvaging L-Phe(D8) from host cells. The uptake pattern was time-dependent during the first 24 hours of infection and complete substitution with L-Phe(D8) in all parasites was detected at 48 hours PI. On the other hand, isolated A. castellanii trachyzoites (grown without host cells) did not show significant uptake for L-Phe(D8) from the media; only achieved an uptake ratio of 16-18% of L-Phe(D8) from the culture medium after 24 hours. These findings demonstrate the potential of combining Raman microspectroscopy and stable isotope labelling approaches to elucidate the role of metabolism in mediating A. castellanii interaction with host cells.

  8. Oxygen induction of a novel fatty acid n-6 desaturase in the soil protozoon, Acanthamoeba castellanii.

    Rutter, Andrew J; Thomas, Katie L; Herbert, Derek; Henderson, R James; Lloyd, David; Harwood, John L

    2002-11-15

    Induction of fatty acid desaturation is very important for the temperature adaptation of poikilotherms. However, in oxygen-limited late-exponential-phase Acanthamoeba castellanii cultures, oxygen alone was able to induce increased activity of a fatty acid desaturase that converts oleate into linoleate and which has been implicated in the temperature adaptation of this organism. Experiments with Delta(10)-nonadecenoate showed that the enzyme is an n -6 desaturase rather than a Delta(12)-desaturase. It also used preferentially 1-acyl-2-oleoyl-phosphatidylcholine as substrate and NAD(P)H as electron donor. The involvement of cytochrome b (5) as an intermediate electron carrier was shown by difference spectra measurements and anti-(cytochrome b (5)) antibody experiments. Of the three protein components of the desaturase complex, oxygen only increased the activity of the terminal (cyanide-sensitive) protein during n -6 desaturase induction. The induction of this terminal protein paralleled well the increase in overall oleate n -6 desaturation. The ability of oxygen to induce oleate desaturase independently of temperature in this lower eukaryotic animal model is of novel intrinsic interest, as well as being important for the design of future experiments to determine the molecular mechanism of temperature adaptation in poikilotherms. PMID:12153399

  9. In Vitro Efficacies of Clinically Available Drugs against Growth and Viability of an Acanthamoeba castellanii Keratitis Isolate Belonging to the T4 Genotype

    Baig, Abdul Mannan; Iqbal, Junaid; Khan, Naveed Ahmed

    2013-01-01

    The effects of clinically available drugs targeting muscarinic cholinergic, adrenergic, dopaminergic, and serotonergic receptors; intracellular calcium levels and/or the function of calcium-dependent biochemical pathways; ion channels; and cellular pumps were tested against a keratitis isolate of Acanthamoeba castellanii belonging to the T4 genotype. In vitro growth inhibition (amoebistatic) assays were performed by incubating A. castellanii with various concentrations of drugs in the growth ...

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

    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. PMID:26074248

  11. The role of Src kinase in the biology and pathogenesis of Acanthamoeba castellanii

    Siddiqui Ruqaiyyah; Iqbal Junaid; Maugueret Marie-josée; Khan Naveed

    2012-01-01

    Abstract Background Acanthamoeba species are the causative agents of fatal granulomatous encephalitis in humans. Haematogenous spread is thought to be a primary step, followed by blood–brain barrier penetration, in the transmission of Acanthmaoeba into the central nervous system, but the associated molecular mechanisms remain unclear. Here, we evaluated the role of Src, a non-receptor protein tyrosine kinase in the biology and pathogenesis of Acanthamoeba. Methods Amoebistatic and amoebicidal...

  12. A bifunctional Delta12,Delta15-desaturase from Acanthamoeba castellanii directs the synthesis of highly unusual n-1 series unsaturated fatty acids.

    Sayanova, Olga; Haslam, Richard; Guschina, Irina; Lloyd, David; Christie, William W; Harwood, John L; Napier, Johnathan A

    2006-12-01

    The free-living soil protozoon Acanthamoeba castellanii synthesizes a range of polyunsaturated fatty acids, the balance of which can be altered by environmental changes. We have isolated and functionally characterized in yeast a microsomal desaturase from A. castellanii, which catalyzes the sequential conversion of C(16) and C(18) Delta9-monounsaturated fatty acids to di- and tri-unsaturated forms. In the case of C(16) substrates, this bifunctional A. castellanii Delta12,Delta15-desaturase generated a highly unusual fatty acid, hexadecatrienoic acid (16:3Delta(9,12,15)(n-1)). The identification of a desaturase, which can catalyze the insertion of a double bond between the terminal two carbons of a fatty acid represents a new addition to desaturase functionality and plasticity. We have also co-expressed in yeast the A. castellanii bifunctional Delta12,Delta15-desaturase with a microsomal Delta6-desaturase, resulting in the synthesis of the highly unsaturated C(16) fatty acid hexadecatetraenoic acid (16:4Delta(6,9,12,15)(n-1)), previously only reported in marine microorganisms. Our work therefore demonstrates the feasibility of the heterologous synthesis of polyunsaturated fatty acids of the n-1 series. The presence of a bifunctional Delta12,Delta15-desaturase in A. castellanii is also considered with reference to the evolution of desaturases and the lineage of this protist. PMID:16950768

  13. Effect of environmental stress factors on the uptake and survival of Campylobacter jejuni in Acanthamoeba castellanii

    Bui, Thanh Xuan; Qvortrup, Klaus; Wolff, Anders;

    2012-01-01

    Background: Campylobacter jejuni is a major cause of bacterial food-borne illness in Europe and North America. The mechanisms allowing survival in the environment and transmission to new hosts are not well understood. Environmental free-living protozoa may facilitate both processes. Pre-exposure to...... heat, starvation, oxidative or osmotic stresses encountered in the environment may affect the subsequent interaction of C. jejuni with free-living protozoa. To test this hypothesis, we examined the impact of environmental stress on expression of virulence-associated genes (ciaB, dnaJ, and htrA) of C...... could be recovered at this time point. Overall, pre-exposure of C. jejuni to environmental stresses did not promote intracellular survival in A. castellanii. Conclusions: Together, these findings suggest that the stress response in C. jejuni and its interaction with A. castellanii are complex and...

  14. Lack of Outer Membrane Protein A Enhances the Release of Outer Membrane Vesicles and Survival of Vibrio cholerae and Suppresses Viability of Acanthamoeba castellanii

    Soni Priya Valeru

    2014-01-01

    Full Text Available Vibrio cholerae, the causative agent of the diarrhoeal disease cholera, survives in aquatic environments. The bacterium has developed a survival strategy to grow and survive inside Acanthamoeba castellanii. It has been shown that V. cholerae expresses outer membrane proteins as virulence factors playing a role in the adherence to interacted host cells. This study examined the role of outer membrane protein A (OmpA and outer membrane vesicles (OMVs in survival of V. cholerae alone and during its interaction with A. castellanii. The results showed that an OmpA mutant of V. cholerae survived longer than wild-type V. cholerae when cultivated alone. Cocultivation with A. castellanii enhanced the survival of both bacterial strains and OmpA protein exhibited no effect on attachment, engulfment, and survival inside the amoebae. However, cocultivation of the OmpA mutant of V. cholerae decreased the viability of A. castellanii and this bacterial strain released more OMVs than wild-type V. cholerae. Surprisingly, treatment of amoeba cells with OMVs isolated from the OmpA mutant significantly decreased viable counts of the amoeba cells. In conclusion, the results might highlight a regulating rule for OmpA in survival of V. cholerae and OMVs as a potent virulence factor for this bacterium towards eukaryotes in the environment.

  15. Acanthamoeba and Fusarium interactions: A possible problem in keratitis.

    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. PMID:26851515

  16. Use of In Vitro Assays To Determine Effects of Human Serum on Biological Characteristics of Acanthamoeba castellanii

    Sissons, James; Alsam, Selwa; Stins, Monique; Rivas, Antonio Ortega; Morales, Jacob Lorenzo; Faull, Jane; Khan, Naveed Ahmed

    2006-01-01

    Normal human serum inhibits Acanthamoeba (encephalitis isolate) binding to and cytotoxicity of human brain microvascular endothelial cells, which constitute the blood-brain barrier. Zymographic assays revealed that serum inhibits extracellular protease activities of acanthamoebae. But it is most likely that inhibition of specific properties of acanthamoebae is a consequence of the initial amoebicidal-amoebistatic effects induced by serum. For example, serum exhibited amoebicidal effects; i.e., up to 50% of the exposed trophozoites were killed. The residual subpopulation, although viable, remained static over longer incubations. Interestingly, serum enhanced the phagocytic ability of acanthamoebae, as measured by bacterial uptake. Overall, our results demonstrate that human serum has inhibitory effects on Acanthamoeba growth and viability, protease secretions, and binding to and subsequent cytotoxicity for brain microvascular endothelial cells. Conversely, Acanthamoeba phagocytosis was stimulated by serum. PMID:16825391

  17. Use of In Vitro Assays To Determine Effects of Human Serum on Biological Characteristics of Acanthamoeba castellanii

    Sissons, James; Alsam, Selwa; Stins, Monique; Rivas, Antonio Ortega; Morales, Jacob Lorenzo; Faull, Jane; Khan, Naveed Ahmed

    2006-01-01

    Normal human serum inhibits Acanthamoeba (encephalitis isolate) binding to and cytotoxicity of human brain microvascular endothelial cells, which constitute the blood-brain barrier. Zymographic assays revealed that serum inhibits extracellular protease activities of acanthamoebae. But it is most likely that inhibition of specific properties of acanthamoebae is a consequence of the initial amoebicidal-amoebistatic effects induced by serum. For example, serum exhibited amoebicidal effects; i.e....

  18. A ribosomal protein gene cluster is encoded in the mitochondrial DNA of Dictyostelium discoideum: UGA termination codons and similarity of gene order to Acanthamoeba castellanii.

    Iwamoto, M; Pi, M; Kurihara, M; Morio, T; Tanaka, Y

    1998-04-01

    We sequenced a region of about 14.5 kb downstream from the ribosomal protein L11 gene (rpl11) in the mitochondrial DNA (54+/-2 kb) of the cellular slime mold Dictyostelium discoideum. Sequence analysis revealed that eleven ribosomal protein genes and six open reading frames (ORFs) formed a cluster arranged in the order: rpl11-orf189-rps12-rps7-rpl2-rps19-+ ++orf425-orf1740-rpl16-rpl14-orf188- rps14-rps8-rpl6-rps13-orf127-orf796. This order was very similar to that of homologous genes in Acanthamoeba castellanii mitochondrial DNA. The N-terminal region of ORF425 and the C-terminal region of ORF1740 had partial similarities to the S3 ribosomal protein of other organisms. The termination codons of rpl16 and orf188 were UGA, which has not hitherto been found in genes encoded in D. discoideum mitochondrial DNA. PMID:9560439

  19. Localization of myosin IC and myosin II in Acanthamoeba castellanii by indirect immunofluorescence and immunogold electron microscopy

    1990-01-01

    Polyclonal antisera have been raised against purified Acanthamoeba myosin II and to a synthetic 26 amino acid peptide that corresponds in sequence to the phosphorylation site of Acanthamoeba myosin IC. These antisera are specific for their respective antigens as determined by immunoblotting after SDS-PAGE of total cell lysates. By using the antisera, localization studies were performed by indirect immunofluorescence and by immunogold electron microscopy. Myosin II occurred in the cell cytopla...

  20. Establishment and validation of whole-cell based fluorescence assays to identify anti-mycobacterial compounds using the Acanthamoeba castellanii-Mycobacterium marinum host-pathogen system.

    Sébastien Kicka

    Full Text Available Tuberculosis is considered to be one of the world's deadliest disease with 2 million deaths each year. The need for new antitubercular drugs is further exacerbated by the emergence of drug-resistance strains. Despite multiple recent efforts, the majority of the hits discovered by traditional target-based screening showed low efficiency in vivo. Therefore, there is heightened demand for whole-cell based approaches directly using host-pathogen systems. The phenotypic host-pathogen assay described here is based on the monitoring of GFP-expressing Mycobacterium marinum during infection of the amoeba Acanthamoeba castellanii. The assay showed straight-forward medium-throughput scalability, robustness and ease of manipulation, demonstrating its qualities as an efficient compound screening system. Validation with a series of known antitubercular compounds highlighted the advantages of the assay in comparison to previously published macrophage-Mycobacterium tuberculosis-based screening systems. Combination with secondary growth assays based on either GFP-expressing D. discoideum or M. marinum allowed us to further fine-tune compound characterization by distinguishing and quantifying growth inhibition, cytotoxic properties and antibiotic activities of the compounds. The simple and relatively low cost system described here is most suitable to detect anti-infective compounds, whether they present antibiotic activities or not, in which case they might exert anti-virulence or host defense boosting activities, both of which are largely overlooked by classical screening approaches.

  1. Photochemotherapeutic strategies against Acanthamoeba keratitis

    Siddiqui, Ruqaiyyah; Khan, Naveed Ahmed

    2012-01-01

    Here, we determined the potential of photochemotherapy, namely the application of photodynamic compounds followed by exposure to a suitable source of UV-visible radiation against corneal pathogen, Acanthamoeba. Organometallic macromolecule, tin porphyrin [Sn(IV)porphyrin] was synthesized and purity confirmed using nuclear magnetic resonance spectroscopy. The Sn(IV)porphyrin was tested against a keratitis isolate of Acanthamoeba castellanii belonging to the T4 genotype using growth and viabili...

  2. Interactions of Neuropathogenic Escherichia coli K1 (RS218) and Its Derivatives Lacking Genomic Islands with Phagocytic Acanthamoeba castellanii and Nonphagocytic Brain Endothelial Cells

    Yousuf, Farzana Abubakar; Yousuf, Zuhair; Iqbal, Junaid; Siddiqui, Ruqaiyyah; Khan, Hafsa; Khan, Naveed Ahmed

    2014-01-01

    Here we determined the role of various genomic islands in E. coli K1 interactions with phagocytic A. castellanii and nonphagocytic brain microvascular endothelial cells. The findings revealed that the genomic islands deletion mutants of RS218 related to toxins (peptide toxin, α-hemolysin), adhesins (P fimbriae, F17-like fimbriae, nonfimbrial adhesins, Hek, and hemagglutinin), protein secretion system (T1SS for hemolysin), invasins (IbeA, CNF1), metabolism (D-serine catabolism, dihydroxyacetone, glycerol, and glyoxylate metabolism) showed reduced interactions with both A. castellanii and brain microvascular endothelial cells. Interestingly, the deletion of RS218-derived genomic island 21 containing adhesins (P fimbriae, F17-like fimbriae, nonfimbrial adhesins, Hek, and hemagglutinin), protein secretion system (T1SS for hemolysin), invasins (CNF1), metabolism (D-serine catabolism) abolished E. coli K1-mediated HBMEC cytotoxicity in a CNF1-independent manner. Therefore, the characterization of these genomic islands should reveal mechanisms of evolutionary gain for E. coli K1 pathogenicity. PMID:24818136

  3. Interactions of Neuropathogenic Escherichia coli K1 (RS218 and Its Derivatives Lacking Genomic Islands with Phagocytic Acanthamoeba castellanii and Nonphagocytic Brain Endothelial Cells

    Farzana Abubakar Yousuf

    2014-01-01

    Full Text Available Here we determined the role of various genomic islands in E. coli K1 interactions with phagocytic A. castellanii and nonphagocytic brain microvascular endothelial cells. The findings revealed that the genomic islands deletion mutants of RS218 related to toxins (peptide toxin, α-hemolysin, adhesins (P fimbriae, F17-like fimbriae, nonfimbrial adhesins, Hek, and hemagglutinin, protein secretion system (T1SS for hemolysin, invasins (IbeA, CNF1, metabolism (D-serine catabolism, dihydroxyacetone, glycerol, and glyoxylate metabolism showed reduced interactions with both A. castellanii and brain microvascular endothelial cells. Interestingly, the deletion of RS218-derived genomic island 21 containing adhesins (P fimbriae, F17-like fimbriae, nonfimbrial adhesins, Hek, and hemagglutinin, protein secretion system (T1SS for hemolysin, invasins (CNF1, metabolism (D-serine catabolism abolished E. coli K1-mediated HBMEC cytotoxicity in a CNF1-independent manner. Therefore, the characterization of these genomic islands should reveal mechanisms of evolutionary gain for E. coli K1 pathogenicity.

  4. SJL mice infected with Acanthamoeba castellanii develop central nervous system autoimmunity through the generation of cross-reactive T cells for myelin antigens

    Massilamany, Chandirasegaran; Marciano-Cabral, Francine; Rocha-Azevedo, Bruno da;

    2014-01-01

    SJL mice reminiscent of the diseases induced with their corresponding cognate peptides. We now demonstrate that mice infected with ACA also show the generation of cross-reactive T cells, predominantly for PLP 139-151, as evaluated by T cell proliferation and IAs/dextramer staining. We verified that......, suggesting that ACA infection can trigger CNS autoimmunity in the presence of preexisting repertoire of autoreactive T cells. Taken together, the data provide novel insights into the pathogenesis of Acanthamoeba infections, and the potential role of infectious agents with mimicry epitopes to self-antigens in...

  5. Biophysical Studies of Processes Involved in Acanthamoeba Infections : Contact Lens Contamination, Adhesion, and Intracellular Dynamics

    Reverey, Julia Franziska

    2014-01-01

    Acanthamoeba castellanii can cause a severe and painful, acanthamoeba keratitis. In this disease, the acanthamoebae reach the eye via contaminated contact lenses. In the eye, they invade cornea cells and kill them. The first step in this process is the formation of a close contact between the acanthamoeba and the target-cell. This contact is mainly mediated by mannose. Subsequently, cytolytic factors move to the contact site and are released leading to the death of the target-cell. The single...

  6. Pathogenic strains of Acanthamoeba are recognized by TLR4 and initiated inflammatory responses in the cornea.

    Hassan Alizadeh

    Full Text Available Free-living amoebae of the Acanthamoeba species are the causative agent of Acanthamoeba keratitis (AK, a sight-threatening corneal infection that causes severe pain and a characteristic ring-shaped corneal infiltrate. Innate immune responses play an important role in resistance against AK. The aim of this study is to determine if Toll-like receptors (TLRs on corneal epithelial cells are activated by Acanthamoeba, leading to initiation of inflammatory responses in the cornea. Human corneal epithelial (HCE cells constitutively expressed TLR1, TLR2, TLR3, TLR4, and TLR9 mRNA, and A. castellanii upregulated TLR4 transcription. Expression of TLR1, TLR2, TLR3, and TLR9 was unchanged when HCE cells were exposed to A. castellanii. IL-8 mRNA expression was upregulated in HCE cells exposed to A. castellanii. A. castellanii and lipopolysaccharide (LPS induced significant IL-8 production by HCE cells as measured by ELISA. The percentage of total cells positive for TLR4 was higher in A. castellanii stimulated HCE cells compared to unstimulated HCE cells. A. castellanii induced upregulation of IL-8 in TLR4 expressing human embryonic kidney (HEK-293 cells, but not TLR3 expressing HEK-293 cells. TLR4 neutralizing antibody inhibited A. castellanii-induced IL-8 by HCE and HEK-293 cells. Clinical strains but not soil strains of Acanthamoeba activated TLR4 expression in Chinese hamster corneas in vivo and in vitro. Clinical isolates but not soil isolates of Acanthamoeba induced significant (P< 0.05 CXCL2 production in Chinese hamster corneas 3 and 7 days after infection, which coincided with increased inflammatory cells in the corneas. Results suggest that pathogenic species of Acanthamoeba activate TLR4 and induce production of CXCL2 in the Chinese hamster model of AK. TLR4 may be a potential target in the development of novel treatment strategies in Acanthamoeba and other microbial infections that activate TLR4 in corneal cells.

  7. Cysteine Protease Inhibitor (AcStefin) Is Required for Complete Cyst Formation of Acanthamoeba

    Lee, Jung-Yub; Song, Su-Min; Moon, Eun-Kyung; Lee, Yu-Ran; Jha, Bijay Kumar; Danne, Dinzouna-Boutamba Sylvatrie; Cha, Hee-Jae; Yu, Hak Sun; Kong, Hyun-Hee; Chung, Dong-Il; Hong, Yeonchul

    2013-01-01

    The encystation of Acanthamoeba leads to the formation of resilient cysts from vegetative trophozoites. This process is essential for parasite survival under unfavorable conditions, such as those associated with starvation, low temperatures, and biocides. Furthermore, cysteine proteases have been implicated in the massive turnover of intracellular components required for encystation. Thus, strict modulation of the activities of cysteine proteases is required to protect Acanthamoeba from intra...

  8. Acanthamoeba species in Swimming Pools of Cairo, Egypt.

    Ahmad Al-Herrawy

    2014-06-01

    Full Text Available The free-living amoebae Acanthamoeba spp. have been recognized as etiologic agents of amoebic encephalitis, keratitis, otitis, lung lesions and other skin infections mainly in immuno-compromised individuals. The purpose of this study is to detect the presence of Acanthamoeba in swimming pools in Egypt using a polymerase chain reaction (PCR method.Water samples were collected from 10 different swimming pools in Cairo, Egypt. Samples were cultured on non-nutrient agar for the detection of Acanthamoeba isolates that were confirmed by PCR amplification using genus specific primers. The molecularly confirmed Acanthamoeba isolates were morphologically identified to the species level.Members of genus Acanthamoeba were detected in 49.2% of the examined swimming-pool water samples. Morphologically, six Acanthamoeba species were isolated from the examined swimming pool water namely A. polyphaga, A.castellanii, A. rhysodes, A. mauritaniensis, A. royreba and A. triangularis. All the identified species of Acanthamoeba were molecularly confirmed to be related to the genus Acanthamoeba.The isolated species of Acanthamoeba could provoke variable degrees of infections to the swimmers. The culture method is cheaper and easier than PCR techniques that are faster for the detection of free-living amoebae.

  9. Acanthamoeba meningoencephalitis

    S R Chandra

    2014-01-01

    Full Text Available Report of a case of young immunocompetent male adult with autopsy proven acanthamoeba meningoencephalitis. The patient presented with a protracted febrile illness of 3 months duration with features of meningoencephalitis, this was followed by rapid deterioration while on anti tuberculous therapy and steroids and ended fatally. His magnetic resonance imaging showed features of hemorrhagic meningoencephalitis and magnetic resonance spectroscopy showed choline peak. Autopsy revealed necrotizing meningoencephalitis and intraocular colonization due to acanthamoeba.

  10. F. novicida-Infected A. castellanii Does Not Enhance Bacterial Virulence in Mice

    Ozanic, Mateja; Gobin, Ivana; Brezovec, Martin; Marecic, Valentina; Trobonjaca, Zlatko; Abu Kwaik, Yousef; Santic, Marina

    2016-01-01

    Francisella tularensis is a facultative intracellular bacterium that causes tularemia in humans and animals. Epidemiology of tularemia worldwide is often associated with water-borne transmission, which includes mosquitoes and amoebae as the potential host reservoirs of the bacteria in water environment. In vitro studies showed intracellular replication of F. tularensis within Acanthamoeba castellanii and Hartmanella vermiformis cells. While infection of amoeba by Legionella pneumophila has been shown to enhance infectivity of L. pneumophila the role of F. tularensis-infected protozoa in the pathogenesis of tularemia is not known. We used 6 h coculture of A. castellanii and F. novicida for investigation of the effect of inhaled amoeba on the pathogenesis of tularemia on in vivo model. Balb/c mice were infected intratracheally with F. novicida or with F. novicida-infected A. castellanii. Surprisingly, infection with F. novicida-infected A. castellanii did not lead to bronchopneumonia in Balb/c mice, and Francisella did not disseminate into the liver and spleen. Upon inhalation, F. novicida infects a variety of host cells, though neutrophils are the predominant cells early during infection in the lung infiltrates of pulmonary tularemia. The numbers of neutrophils in the lungs of Balb/c mice were significantly lower in the infection of mice with F. novicida-infected A. castellanii in comparison to group of mice infected only with F. novicida. These results demonstrate that following inoculation of mice with F. novicida-infected A. castellanii, mice did not develop tularemia. PMID:27242974

  11. Metabolic profiling of the protozoan parasite Entamoeba invadens revealed activation of unpredicted pathway during encystation.

    Ghulam Jeelani

    Full Text Available Encystation, which is cellular differentiation from the motile, proliferative, labile trophozoite form to the dormant, resistant cyst form, is a crucial process found in parasitic and free-living protozoa such as Entamoeba, Giardia, Acanthamoeba, and Balamuthia. Since encystation is an essential process to deal with the adverse external environmental changes during the life cycle, and often integral to the transmission of the diseases, biochemical understanding of the process potentially provides useful measures against the infections caused by this group of protozoa. In this study, we investigated metabolic and transcriptomic changes that occur during encystation in Entamoeba invadens, the reptilian sibling of mammal-infecting E. histolytica, using capillary electrophoresis-tandem mass spectrometry-based metabolite profiling and DNA microarray-based expression profiling. As the encystation progressed, the levels of majority of metabolites involved in glycolysis and nucleotides drastically decreased, indicating energy generation is ceased. Furthermore, the flux of glycolysis was redirected toward chitin wall biosynthesis. We found remarkable temporal increases in biogenic amines such as isoamylamine, isobutylamine, and cadaverine, during the early period of encystation, when the trophozoites form large multicellular aggregates (precyst. We also found remarkable induction of γ-aminobutyric acid (GABA during encystation. This study has unveiled for the first time the dynamics of the transcriptional and metabolic regulatory networks during encystation, and should help in better understanding of the process in pathogenic eukaryotes, and further development of measures controlling infections they cause.

  12. Survival of taylorellae in the environmental amoeba Acanthamoeba castellanii

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

    2014-01-01

    Background 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...

  13. Failure of chemotherapy in the first reported cases of Acanthamoeba keratitis in Pakistan

    Siddiqui, Ruqaiyyah; Chaudhry, Tanveer; Lakhundi, Sahreena; Ahmad, Khabir; Khan, Naveed Ahmed

    2014-01-01

    Acanthamoeba keratitis is a painful and progressive infection of the cornea that can result in loss of vision. Here, for the first time in Pakistan, we report two cases of Acanthamoeba keratitis. The first patient was a 37-year-old female who presented with severe itching, redness, pain, along with loss of vision. The patient was a regular soft contact lens wearer. The second patient was a 25-year-old female who had been using soft contact lenses for the past two years. She presented with a burning sensation and extreme pain, along with loss of vision. Both patients were treated for a possible microbial keratitis with topical moxifloxacin hydrochloride drops, vancomycin drops, propamidine isethionate ointment, amphotericin B drops, and amikacin drops. However, the response was inadequate and both patients were referred for corneal transplant. Acanthamoeba castellanii was isolated by placing contact lenses and contact lens cases on non-nutrient agar plates containing a lawn of non-invasive Escherichia coli K-12 HB101 bacteria. The polymerase chain reaction (PCR) using genus-specific probes confirmed the identity of Acanthamoeba spp., whereas the morphological characteristics of trophozoites and cysts were suggestive of A. castellanii in both cases. With growing use of contact lenses for vision correction/cosmetic use coupled with sub-standard lens care in this region and the possibility of non-contact lens-associated Acanthamoeba keratitis, a need for increased awareness of this sight-threatening infection is discussed further. PMID:24548160

  14. Effect of microwave treatment to acanthamoeba: a possibility of contact lens storage case sterilization?

    Microbially contaminated contact lens storage cases are considered to be a predisposing risk factor for Acanthamoeba keratitis. What kind of effect does microwave irradiation have on contact lens cases contaminated with Acanthamoeba cysts and trophozoites? Different types of contact lens cases were contaminated with trophozoites and cysts of three different Acanthamoeba species (A. hatchetti, A. castellanii, A. comandoni) and exposed to microwave irradiation for 3, 5 and 8 minutes respectively. One of the three test series was run with dehydrated cysts. After an irradiation period of 3 minutes under humid conditions cysts and trophozoites of all three strains were completely destroyed. Dehydrated cysts of A. hatchetti and A. castellanii were not killed by irradiation. There were no negative effects of irradiation on the contact lens cases themselves. Acanthamoeba cysts and trophozoites are effectively killed by microwave irradiation on condition that irradiation takes place in humid conditions. This can be easily achieved by filling the contact lens storage cases with tap water. We recommend microwave irradiation as a cheap and save method for the sterilization of contact lens cases in order to avoid a possible Acanthamoeba infection of the eye. (author)

  15. Regulation of carbohydrate metabolism during Giardia encystment

    Vliegenthart, J.F.G.; Jarroll, E.L.; Macechko, P.T.; Steimle, P.A.; Bulik, D.; Karr, C.D.; Keulen, Harry van; Paget, P.A.

    2001-01-01

    Giardia intestinalis trophozoites encyst when they are exposed to bile. During encystment, events related to the inducible synthesis of a novel N-acetyl-d-galactosamine (GalNAc) homopolymer, occur. Within the first 6 h of encystment, mRNA for glucosamine 6-P isomerase (GPI), the first inducible enzy

  16. Cellulose degradation: a therapeutic strategy in the improved treatment of Acanthamoeba infections

    Lakhundi, Sahreena; Siddiqui, Ruqaiyyah; Khan, Naveed Ahmed

    2015-01-01

    Acanthamoeba is an opportunistic free-living amoeba that can cause blinding keratitis and fatal brain infection. Early diagnosis, followed by aggressive treatment is a pre-requisite in the successful treatment but even then the prognosis remains poor. A major drawback during the course of treatment is the ability of the amoeba to enclose itself within a shell (a process known as encystment), making it resistant to chemotherapeutic agents. As the cyst wall is partly made of cellulose, thus cel...

  17. Inactivation of Acanthamoeba spp. and Other Ocular Pathogens by Application of Cold Atmospheric Gas Plasma.

    Heaselgrave, Wayne; Shama, Gilbert; Andrew, Peter W; Kong, Michael G

    2016-05-15

    Currently there are estimated to be approximately 3.7 million contact lens wearers in the United Kingdom and 39.2 million in North America. Contact lens wear is a major risk factor for developing an infection of the cornea known as keratitis due to poor lens hygiene practices. While there is an international standard for testing disinfection methods against bacteria and fungi (ISO 14729), no such guidelines exist for the protozoan Acanthamoeba, which causes a potentially blinding keratitis most commonly seen in contact lens wearers, and as a result, many commercially available disinfecting solutions show incomplete disinfection after 6 and 24 h of exposure. Challenge test assays based on international standard ISO 14729 were used to determine the antimicrobial activity of cold atmospheric gas plasma (CAP) against Pseudomonas aeruginosa, Candida albicans, and trophozoites and cysts of Acanthamoeba polyphaga and Acanthamoeba castellanii P. aeruginosa and C. albicans were completely inactivated in 0.5 min and 2 min, respectively, and trophozoites of A. polyphaga and A. castellanii were completely inactivated in 1 min and 2 min, respectively. Furthermore, for the highly resistant cyst stage of both species, complete inactivation was achieved after 4 min of exposure to CAP. This study demonstrates that the CAP technology is highly effective against bacterial, fungal, and protozoan pathogens. The further development of this technology has enormous potential, as this approach is able to deliver the complete inactivation of ocular pathogens in minutes, in contrast to commercial multipurpose disinfecting solutions that require a minimum of 6 h. PMID:26994079

  18. Biology and pathogenesis of Acanthamoeba

    Siddiqui Ruqaiyyah; Khan Naveed

    2012-01-01

    Abstract Acanthamoeba is a free-living protist pathogen, capable of causing a blinding keratitis and fatal granulomatous encephalitis. The factors that contribute to Acanthamoeba infections include parasite biology, genetic diversity, environmental spread and host susceptibility, and are highlighted together with potential therapeutic and preventative measures. The use of Acanthamoeba in the study of cellular differentiation mechanisms, motility and phagocytosis, bacterial pathogenesis and ev...

  19. Photochemotherapeutic Strategy against Acanthamoeba Infections

    Aqeel, Yousuf; Siddiqui, Ruqaiyyah; Anwar, Ayaz; Shah, Muhammad Raza; Khoja, Shahrukh; Khan, Naveed Ahmed

    2015-01-01

    Acanthamoeba is a protist pathogen that can cause serious human infections, including blinding keratitis and a granulomatous amoebic encephalitis that almost always results in death. The current treatment for these infections includes a mixture of drugs, and even then, a recurrence can occur. Photochemotherapy has shown promise in the treatment of Acanthamoeba infections; however, the selective targeting of pathogenic Acanthamoeba has remained a major concern. The mannose-binding protein is a...

  20. Biology and pathogenesis of Acanthamoeba

    Siddiqui Ruqaiyyah

    2012-01-01

    Full Text Available Abstract Acanthamoeba is a free-living protist pathogen, capable of causing a blinding keratitis and fatal granulomatous encephalitis. The factors that contribute to Acanthamoeba infections include parasite biology, genetic diversity, environmental spread and host susceptibility, and are highlighted together with potential therapeutic and preventative measures. The use of Acanthamoeba in the study of cellular differentiation mechanisms, motility and phagocytosis, bacterial pathogenesis and evolutionary processes makes it an attractive model organism. There is a significant emphasis on Acanthamoeba as a Trojan horse of other microbes including viral, bacterial, protists and yeast pathogens.

  1. The proteome landscape of Giardia lamblia encystation.

    Carmen Faso

    Full Text Available Giardia lamblia is an intestinal protozoan parasite required to survive in the environment in order to be transmitted to a new host. To ensure parasite survival, flagellated trophozoites colonizing the small intestine differentiate into non-motile environmentally-resistant cysts which are then shed in the environment. This cell differentiation process called encystation is characterized by significant morphological remodeling which includes secretion of large amounts of cyst wall material. Although much is known about the transcriptional regulation of encystation and the synthesis and trafficking of cyst wall material, the investigation of global changes in protein content and abundance during G. lamblia encystation is still unaddressed. In this study, we report on the quantitative analysis of the G. lamblia proteome during encystation using tandem mass spectrometry. Quantification of more than 1000 proteins revealed major changes in protein abundance in early, mid and late encystation, notably in constitutive secretory protein trafficking. Early stages of encystation were marked by a striking decrease of endoplasmic reticulum-targeted variant-specific surface proteins and significant increases in cytoskeleton regulatory components, NEK protein kinases and proteins involved in protein folding and glycolysis. This was in stark contrast to cells in the later stages of encystation which presented a surprisingly similar proteome composition to non-encysting trophozoites. Altogether these data constitute the first quantitative atlas of the Giardia proteome covering the whole process of encystation and point towards an important role for post-transcriptional control of gene expression in Giardia differentiation. Furthermore, our data provide a valuable resource for the community-based annotation effort of the G. lamblia genome, where almost 70% of all predicted gene models remains "hypothetical".

  2. The proteome landscape of Giardia lamblia encystation.

    Faso, Carmen; Bischof, Sylvain; Hehl, Adrian B

    2013-01-01

    Giardia lamblia is an intestinal protozoan parasite required to survive in the environment in order to be transmitted to a new host. To ensure parasite survival, flagellated trophozoites colonizing the small intestine differentiate into non-motile environmentally-resistant cysts which are then shed in the environment. This cell differentiation process called encystation is characterized by significant morphological remodeling which includes secretion of large amounts of cyst wall material. Although much is known about the transcriptional regulation of encystation and the synthesis and trafficking of cyst wall material, the investigation of global changes in protein content and abundance during G. lamblia encystation is still unaddressed. In this study, we report on the quantitative analysis of the G. lamblia proteome during encystation using tandem mass spectrometry. Quantification of more than 1000 proteins revealed major changes in protein abundance in early, mid and late encystation, notably in constitutive secretory protein trafficking. Early stages of encystation were marked by a striking decrease of endoplasmic reticulum-targeted variant-specific surface proteins and significant increases in cytoskeleton regulatory components, NEK protein kinases and proteins involved in protein folding and glycolysis. This was in stark contrast to cells in the later stages of encystation which presented a surprisingly similar proteome composition to non-encysting trophozoites. Altogether these data constitute the first quantitative atlas of the Giardia proteome covering the whole process of encystation and point towards an important role for post-transcriptional control of gene expression in Giardia differentiation. Furthermore, our data provide a valuable resource for the community-based annotation effort of the G. lamblia genome, where almost 70% of all predicted gene models remains "hypothetical". PMID:24391747

  3. Molecular Phylogeny of Acanthamoeba

    Kong, Hyun Hee

    2009-01-01

    After morphological grouping of Acanthamoeba by Pussard and Pons, phylogeny of the genus has been always a big topic to the researchers. Because of the variability of morphological characteristics, unchangeable and stable characters have been investigated for phylogenic criteria. Isoenzyme and mitochondrial DNA RFLP (Mt DNA RFLP) analyses revealed different patterns among strains assigned to a same species. Therefore, these characteristics would be considered as tools for strain discriminatio...

  4. Evaluation of Ozone Application in Dental Unit Water Lines Con¬taminated with Pathogenic Acanthamoeba

    Wafaa HIKAL

    2015-10-01

    Full Text Available Background: In this study morphological and molecular characterization of Acan­thamoeba strains, isolated from dental unit waterlines (DUWLs were surveyed and the levels of disinfection achievable in vitro by the application of ozone disinfect­ant to DUWLs were evaluate.Methods: Water samples were collected from air-water syringes, cup fillers and tap water before and at the end of the working day. They were cultured on non-nutrient agar (NNA plates. Species identification was carried out with a PCR assay based on sequence analysis of the 18S rRNA gene. The cellular response to ozone was tested on Acanthamoeba cyst with different doses at different contact time in vitro twice.Results: Prevalence rates for Acanthamoeba contamination were 100, 100 and 72% for air-water syringes, cup fillers and tap water, respectively. The morphological analysis revealed the presence of A. castellanii, A. griffin, A. hatchitti and A. lenticulata. Phylogenetic analysis of the sequences showed the four strains to be closely related to a sequence type (T3, T4, T5 and T11. Acanthamoeba cells were stained with try­pan blue, which revealed killed of Acanthamoeba instantaneously after 10 minutes in ozonized water. There was no growth of Acanthamoeba occurred after ozone treat­ment in water bottles for 5 minutes with a flow rate of 500 mg/hour.Conclusion: Ozone can play an important role in controlling the problem of contami­nation of DUWLs as a potent disinfectant.

  5. Analysis of Giardin expression during encystation of Giardia lamblia

    The present study analyzed giardin transcription in trophozoites and cysts during encystation of Giardia lamblia. Encystment was induced using standard methods, and the number of trophozoites and cysts were counted at various time-points during encystation. At all time points, RNA from both stages...

  6. Acanthamoeba feature a unique backpacking strategy to trap and feed on Listeria monocytogenes and other motile bacteria.

    Doyscher, Dominik; Fieseler, Lars; Dons, Lone; Loessner, Martin J; Schuppler, Markus

    2013-02-01

    Despite its prominent role as an intracellular human pathogen, Listeria monocytogenes normally features a saprophytic lifestyle, and shares many environmental habitats with predatory protozoa. Earlier studies claimed that Acanthamoeba may act as environmental reservoirs for L. monocytogenes, whereas others failed to confirm this hypothesis. Our findings support the latter and provide clear evidence that L. monocytogenes is unable to persist in Acanthamoeba castellanii and A. polyphaga. Instead, external Listeria cells are rapidly immobilized on the surface of Acanthamoeba trophozoites, forming large aggregates of densely packed bacteria that we termed backpacks. While the assembly of backpacks is dependent on bacterial motility, flagellation alone is not sufficient. Electron micrographs showed that the aggregates are held together by filaments of likely amoebal origin. Time-lapse microscopy revealed that shortly after the bacteria are collected, the amoeba can change direction of movement, phagocytose the backpack and continue to repeat the process. The phenomenon was also observed with avirulent L. monocytogenes mutants, non-pathogenic Listeria, and other motile bacteria, indicating that formation of backpacks is not specific for L. monocytogenes, and independent of bacterial pathogenicity or virulence. Hence, backpacking appears to represent a unique and highly effective strategy of Acanthamoeba to trap and feed on motile bacteria. PMID:22925311

  7. Coordinated Changes in Gene Expression Throughout Encystation of Giardia intestinalis.

    Einarsson, Elin; Troell, Karin; Hoeppner, Marc P; Grabherr, Manfred; Ribacke, Ulf; Svärd, Staffan G

    2016-03-01

    Differentiation into infectious cysts through the process of encystation is crucial for transmission and survival of the intestinal protozoan parasite Giardia intestinalis. Hitherto the majority of studies have focused on the early events, leaving late encystation poorly defined. In order to further study encystation, focusing on the later events, we developed a new encystation protocol that generates a higher yield of mature cysts compared to standard methods. Transcriptome changes during the entire differentiation from trophozoites to cysts were thereafter studied using RNA sequencing (RNA-seq). A high level of periodicity was observed for up- and down-regulated genes, both at the level of the entire transcriptome and putative regulators. This suggests the trajectory of differentiation to be coordinated through developmentally linked gene regulatory activities. Our study identifies a core of 13 genes that are consistently up-regulated during initial encystation. Of these, two constitute previously uncharacterized proteins that we were able to localize to a new type of encystation-specific vesicles. Interestingly, the largest transcriptional changes were seen in the late phase of encystation with the majority of the highly up-regulated genes encoding hypothetical proteins. Several of these were epitope-tagged and localized to further characterize these previously unknown genetic components of encystation and possibly excystation. Finally, we also detected a switch of variant specific surface proteins (VSPs) in the late phase of encystation. This occurred at the same time as nuclear division and DNA replication, suggesting a potential link between the processes. PMID:27015092

  8. The pathogenesis of Acanthamoeba infections: current status and future implications

    Khan, Naveed Ahmed

    2004-01-01

    Acanthamoeba are opportunistic protozoan parasites that can cause painful, visionthreatening keratitis. However the pathogenesis and pathophysiology of Acanthamoeba keratitis remain incompletely understood. Most cases of Acanthamoeba keratitis develop as a result of poor hygiene in contact lens care but it is unclear how amoebae transmigrate from the environment into the cornea leading to inflammation, photophobia and blindness. Acanthamoeba keratitis has become increasingly...

  9. Ceratite bilateral por Acanthamoeba: relato de caso Bilateral Acanthamoeba keratitis: case report

    Wilson Nahmatallah Obeid

    2003-12-01

    Full Text Available A ceratite por Acanthamoeba é uma infecção ocular grave que, apesar dos recentes progressos no diagnóstico e tratamento, ainda provoca prolongada morbidade e perda da acuidade visual. Relatamos um caso de ceratite bilateral por Acanthamoeba em usuário de lentes de contato, que é o primeiro caso descrito na literatura brasileira.Acanthamoeba keratitis is a severe ocular infection which even with recent progress in diagnosis and treatment still causes long morbidity and loss of visual acuity. We describe a case of bilateral Acanthamoeba keratitis in a user of contact lenses, which is the first case discussed in Brazilian literature.

  10. The immunological aspects of Acanthamoeba infections

    Naveed A.  Khan

    2005-01-01

    Full Text Available Acanthamoeba is a protozoan pathogen that is responsible for two recognized diseases; i a blinding keratitis, most commonly associated with contact lens wear, and ii a rare but fatal granulomatous encephalitis that usually is limited to immunocompromised patients. The fact that Acanthamoeba infections are generally limited to avascular cornea or immunocompromised patients suggests that the normal immune responses may be sufficient to control and/or eradicate these pathogens. Here we describe our current understanding of the immune responses to Acanthamoeba infections.

  11. Chitin synthetase in encysting Giardia lamblia and Entamoeba invadens

    Das, S.; Gillin, F.D.

    1987-05-01

    Giardia lamblia (Gl) and Entamoeba invadens (Ei) are protozoan parasites with two morphologic stages in their life cycles. Motile trophozoites colonize the intestine of humans and reptiles respectively. Water resistant cysts, which can survive outside the host, transmit infection. In vitro cyst formation of Ei from trophozoites has been reported, and the authors have recently induced in vitro encystation of Gl. Although the cyst walls of both parasites contain chitin, it synthesis by encysting trophozoites has not been reported. The authors now show that encystation conditions greatly increase chitin synthetase (CS) specific activity (incorporation of /sup 3/H GlcNAc from UDP-GlcNAc into TCA-or alcohol-precipitable material). Extracts of encysting Gl incorporated 3.6 nmol/mg protein in 5 hr compared to < 0.005 in controls. Extracts of encysting Fi incorporated 4.8 n mol/mg protein, compared to 1.7 in the control. CS activity of both parasites requires preformed chitin. The Gl enzyme requires a reducing agent, is inhibited by digitonin and the CS inhibitors, polyoxin D and Nikkomycin, but not by tunicamycin. The product is digested by chitinase. Ei enzyme does not require a reducing agent and is stimulated by 1 mg/ml digitonin, but inhibited by higher concentrations. These studies demonstrate CS enzymes which may play important roles in encystation of Gl and Ei.

  12. Chitin synthetase in encysting Giardia lamblia and Entamoeba invadens

    Giardia lamblia (Gl) and Entamoeba invadens (Ei) are protozoan parasites with two morphologic stages in their life cycles. Motile trophozoites colonize the intestine of humans and reptiles respectively. Water resistant cysts, which can survive outside the host, transmit infection. In vitro cyst formation of Ei from trophozoites has been reported, and the authors have recently induced in vitro encystation of Gl. Although the cyst walls of both parasites contain chitin, it synthesis by encysting trophozoites has not been reported. The authors now show that encystation conditions greatly increase chitin synthetase (CS) specific activity (incorporation of 3H GlcNAc from UDP-GlcNAc into TCA-or alcohol-precipitable material). Extracts of encysting Gl incorporated 3.6 nmol/mg protein in 5 hr compared to < 0.005 in controls. Extracts of encysting Fi incorporated 4.8 n mol/mg protein, compared to 1.7 in the control. CS activity of both parasites requires preformed chitin. The Gl enzyme requires a reducing agent, is inhibited by digitonin and the CS inhibitors, polyoxin D and Nikkomycin, but not by tunicamycin. The product is digested by chitinase. Ei enzyme does not require a reducing agent and is stimulated by 1 mg/ml digitonin, but inhibited by higher concentrations. These studies demonstrate CS enzymes which may play important roles in encystation of Gl and Ei

  13. Development of a new oxygen consumption rate assay in cultures of Acanthamoeba (Protozoa: Lobosea) and its application to evaluate viability and amoebicidal activity in vitro.

    Heredero-Bermejo, I; Criado-Fornelio, A; Soliveri, J; Díaz-Martín, J A; Matilla-Fuentes, J; Sánchez-Arias, J A; Copa-Patiño, J L; Pérez-Serrano, J

    2015-08-01

    A new fluorometric method has been developed for measuring the oxygen consumption rate (OCR) of Acanthamoeba cultures in microplates and for screening molecules with amoebicidal activity against this microorganism. The use of a biofunctional matrix (containing an oxygen-sensitive fluorogenic probe) attached to the microplate wells allowed continuous measurement of OCR in the medium, hence assessment of amoebic growth. The new OCR method applied to cell viability yielded a linear relationship and monitoring was much quicker than with indirect viability assays previously used. In addition, two drugs were tested in a cytotoxicity assay monitored by the new OCR viability test. With this procedure, the standard amoebicidal drug chlorhexidine digluconate showed an IC50 of 3.53 + 1.3 mg/l against Acanthamoeba polyphaga and 3.19 + 1.2 mg/l against Acanthamoeba castellanii, whereas a cationic dendrimer [G1Si(NMe3+)4] showed an IC50 of 6.42 + 1.3 mg/l against A. polyphaga. These data agree with previous studies conducted in our laboratory. Therefore, the new OCR method has proven powerful and quick for amoebicidal drug screening and is likely to be applied in biochemical studies concerning protozoa respiration and metabolism. PMID:25956947

  14. Acanthamoeba induces cell-cycle arrest in host cells

    Sissons, J.; Alsam, S.; Jayasekera, S.; Kim, K S; Stins, M; Khan, Naveed Ahmed

    2004-01-01

    Acanthamoeba can cause fatal granulomatous amoebic encephalitis (GAE) and eye keratitis. However, the pathogenesis and pathophysiology of these emerging diseases remain unclear. In this study, the effects of Acanthamoeba on the host cell cycle using human brain microvascular endothelial cells (HBMEC) and human corneal epithelial cells (HCEC) were determined. Two isolates of Acanthamoeba belonging to the T1 genotype (GAE isolate) and T4 genotype (keratitis isolate) were used, which showed seve...

  15. An update on Acanthamoeba keratitis: diagnosis, pathogenesis and treatment

    Lorenzo-Morales Jacob; Khan Naveed A.; Walochnik Julia

    2015-01-01

    Free-living amoebae of the genus Acanthamoeba are causal agents of a severe sight-threatening infection of the cornea known as Acanthamoeba keratitis. Moreover, the number of reported cases worldwide is increasing year after year, mostly in contact lens wearers, although cases have also been reported in non-contact lens wearers. Interestingly, Acanthamoeba keratitis has remained significant, despite our advances in antimicrobial chemotherapy and supportive care. In part, this is due to an inc...

  16. Is there evidence of sexual reproduction (meiosis) in Acanthamoeba?

    Khan, Naveed A.; Siddiqui, Ruqaiyyah

    2015-01-01

    Evolution of independently breeding species into males and females (gametes) has remained a puzzle. Given the significant advantages of sexual reproduction over asexual reproduction as a long-term species survival strategy; here, we pose the question whether there is some form of meiosis in Acanthamoeba species, which represents our ancient lineage. The recently available Acanthamoeba genome revealed several genes implicated in meiosis in sexual eukaryotes such as Spo11, Mre11, Rad50, Rad51, Rad52, Mnd1, Dmc1, Msh, and Mlh, suggesting that Acanthamoeba is capable of some form of meiosis, inferring the presence of sexual reproduction in Acanthamoeba, and that meiosis evolved early in eukaryotic evolution. PMID:25800982

  17. Heat Shock Protein 90 regulates encystation in Entamoeba

    Meetali eSingh

    2015-10-01

    Full Text Available Enteric protozoan Entamoeba histolytica is a major cause of debilitating diarrheal infection worldwide with high morbidity and mortality. Even though the clinical burden of this parasite is very high, this infection is categorized as a neglected disease. Parasite is transmitted through feco-oral route and exhibit two distinct stages namely – trophozoites and cysts. Mechanism and regulation of encystation is not clearly understood. Previous studies have established the role of Heat shock protein 90 (Hsp90 in regulating stage transition in various protozoan parasites like Giardia, Plasmodium, Leishmania and Toxoplasma. Our study for the first time reports that Hsp90 plays a crucial role in life cycle of Entamoeba as well. We identify Hsp90 to be a negative regulator of encystation in Entamoeba. We also show that Hsp90 inhibition interferes with the process of phagocytosis in Entamoeba. Overall, we show that Hsp90 plays an important role in virulence and transmission of Entamoeba.

  18. Encystation commitment in Giardia duodenalis: a long and winding road

    Argüello-Garciá R.

    2009-12-01

    Full Text Available Cholesterol and bile salts are relevant modulators of Giardia encystation. Although several molecules within signaling cascades have been identified, and changes in their expression observed during giardial encystation, their underlying interactions leading to expression of cyst wall markers (CWPs and precursors of the GalNAc homopolymer are not well defined. Recent experimental data and the completion of the Giardia Genome Project Database (GiardiaDB allow us now to consider the role of bile salts as “natural stimuli” and the potential involvement of a Raf/MEK/ERK pathway mediating cholesterol-regulated expression of cyst-specific genes. These new findings may provide promising targets for diagnostics, drug design and prophylactic intervention against giardiasis.

  19. Prevalence of encysted Toxoplasma gondii in raptors from Alabama.

    Lindsay, D S; Smith, P C; Hoerr, F J; Blagburn, B L

    1993-12-01

    Little is known about the prevalence of encysted Toxoplasma gondii in wild birds. We examined the hearts and breast muscles from 101 raptors for encysted T. gondii. All of the raptors had been submitted for necropsy to the State Veterinary Diagnostic Laboratory, Auburn, Alabama. Tissues were digested in acid-pepsin solution and inoculated into groups of 3-5 laboratory mice. Toxoplasma gondii was isolated from 27 of 101 (26.7%) raptors: 8 of 12 (66.7%) red-shouldered hawks (Buteo lineatus), 13 of 27 (41.1%) red-tailed hawks (Buteo jamaicensis), 1 of 4 (25%) Cooper's hawks (Accipiter cooperi), 1 of 5 (20%) great horned owls (Bubo virginianus), 4 of 15 (26.7%) barred owls (Strix varia), and 1 of 3 (33.3%) kestrels (Falco sparverius). Toxoplasma gondii was not isolated from 3 broad-winged hawks (Buteo platypterus), 3 sharp-shinned hawks (Accipiter striatus), 6 barn owls (Tyto alba), 9 screech owls (Asio otus), a Mississippi kite (Ictinia misisippiensis), 2 golden eagles (Aquila chrysaetos), a bald eagle (Haliaeetus leucocephalus), 4 ospreys (Pandion haliaetus), 4 turkey vultures (Cathartes aura), or 2 black vultures (Coragyps atratus). No significant difference (P > 0.05) in prevalence was detected based on sex using chi-square analysis. Chi-square analysis of the data demonstrated that adult raptors had encysted stages of T. gondii significantly (P < 0.05) more often than did immature raptors. PMID:8277379

  20. Current Status of Acanthamoeba in Iran: A Narrative Review Article.

    Maryam Niyyati

    2015-06-01

    Full Text Available Free-living amoebae belonging to the genus Acanthamoeba have an environmental distribution. Amoebic keratitis due to these protozoan parasites continue to rise in Iran and worldwide. In Iran, there are various researches regarding both morphological and molecular identification of Acanthamoeba spp. in environmental and clinical samples. However, there is no thorough review about Acanthamoeba genotypes and their distribution in environmental sources such as water, dust and biofilm in Iran. Besides, according to increasing cases of Amoebic keratitis in the region awareness regarding the pathogenic potential of these sight-threatening amoebae is of utmost importance.We conducted a thorough review based on the database sources such as MEDLINE, PubMed and Google scholar. No restrictions were placed on study date, study design or language of publication. We searched all valuable and relevant information considering the occurrence of the Acanthamoeba in both environmental and clinical samples.According to our thorough review Acanthamoeba belonging to T4 genotype is the most prevalent type strain in environmental and clinical samples in several regions in Iran and worldwide, however, there are reports regarding Acanthamoeba belonging to other genotypes such as T2, T3, T5, T6 and T11 and the mentioned point could leads us to more researches with the goal of presenting the real genotype dominance of Acanthamoeba and related disease in the country.Overall, the present review will focus on present status of genotypes of Acanthamoeba in Iran during recent years.

  1. An update on Acanthamoeba keratitis: diagnosis, pathogenesis and treatment

    Lorenzo-Morales Jacob

    2015-01-01

    Full Text Available Free-living amoebae of the genus Acanthamoeba are causal agents of a severe sight-threatening infection of the cornea known as Acanthamoeba keratitis. Moreover, the number of reported cases worldwide is increasing year after year, mostly in contact lens wearers, although cases have also been reported in non-contact lens wearers. Interestingly, Acanthamoeba keratitis has remained significant, despite our advances in antimicrobial chemotherapy and supportive care. In part, this is due to an incomplete understanding of the pathogenesis and pathophysiology of the disease, diagnostic delays and problems associated with chemotherapeutic interventions. In view of the devastating nature of this disease, here we present our current understanding of Acanthamoeba keratitis and molecular mechanisms associated with the disease, as well as virulence traits of Acanthamoeba that may be potential targets for improved diagnosis, therapeutic interventions and/or for the development of preventative measures. Novel molecular approaches such as proteomics, RNAi and a consensus in the diagnostic approaches for a suspected case of Acanthamoeba keratitis are proposed and reviewed based on data which have been compiled after years of working on this amoebic organism using many different techniques and listening to many experts in this field at conferences, workshops and international meetings. Altogether, this review may serve as the milestone for developing an effective solution for the prevention, control and treatment of Acanthamoeba infections.

  2. An update on Acanthamoeba keratitis: diagnosis, pathogenesis and treatment

    Lorenzo-Morales, Jacob; Khan, Naveed A.; Walochnik, Julia

    2015-01-01

    Free-living amoebae of the genus Acanthamoeba are causal agents of a severe sight-threatening infection of the cornea known as Acanthamoeba keratitis. Moreover, the number of reported cases worldwide is increasing year after year, mostly in contact lens wearers, although cases have also been reported in non-contact lens wearers. Interestingly, Acanthamoeba keratitis has remained significant, despite our advances in antimicrobial chemotherapy and supportive care. In part, this is due to an incomplete understanding of the pathogenesis and pathophysiology of the disease, diagnostic delays and problems associated with chemotherapeutic interventions. In view of the devastating nature of this disease, here we present our current understanding of Acanthamoeba keratitis and molecular mechanisms associated with the disease, as well as virulence traits of Acanthamoeba that may be potential targets for improved diagnosis, therapeutic interventions and/or for the development of preventative measures. Novel molecular approaches such as proteomics, RNAi and a consensus in the diagnostic approaches for a suspected case of Acanthamoeba keratitis are proposed and reviewed based on data which have been compiled after years of working on this amoebic organism using many different techniques and listening to many experts in this field at conferences, workshops and international meetings. Altogether, this review may serve as the milestone for developing an effective solution for the prevention, control and treatment of Acanthamoeba infections. PMID:25687209

  3. Acanthamoeba: epidimiology, pathogenicity and evaluation of effectiveness of recent drugs

    To study the epidimiology of Acanthamoeba and to evaluate the effectiveness of some recent drugs against parasite. The study was carried out from March to May 2005 at the ophthalmic clinic of King Fahad Hospital, Saudi Arabia. Samples of rinsing solutions and saline of contact lens, tap water, Swimming pool water and Soil from Hufof city Saudi Arabia were tested. Mice were used to infect them via intranasal inoculation from isolated culture strain for confirming pathogenicity. Rokitamycin, polymixin B, suramin and chloropromazin were used to study their effects on Acanthamoeba growth in vitro. Acanthamoeba were detected in 20% of solution of contact lens, 20% of tap water, 50% of swimming pool samples and 40% of soil samples. All animals died or were sacrificed and had Acanthamoeba isolated from their organs. Higher percentage of growth inhibition of Acanthamoeba cultured was shown by chloropromazine and rokitamycin after 21 days (100%), while Polymyin B and Suramin showed 83% and 64% inhibition respectively. Acanthamoeba isolated in significant percent of environmental sources. Pathogenicity of organism was confirmed in mice. Contact lens wearers should be aware of the risks associated with Acanthamoeba. Rokitamycin and chlorpromazine showed good inhibition in vitro. (author)

  4. Diversity and Seasonal Impact of Acanthamoeba Species in a Subtropical Rivershed

    Po-Min Kao

    2013-01-01

    Full Text Available This study evaluated the presence of Acanthamoeba species in the Puzih River watershed, which features typical subtropical monsoon climate and is located just above the Tropic of Cancer in Taiwan. The relationship between the seasonal and geographical distributions of Acanthamoeba species in this rivershed was also investigated. Acanthamoeba species were detected in water samples using the amoebal enrichment culture method and confirmed by PCR. A total of 136 water samples were included in this study, 16 (11.7% of which contained Acanthamoeba species. Samples with the highest percentage of Acanthamoeba (32.4% were obtained during the summer season, mainly from upstream areas. The identified species in the four seasons included Acanthamoeba palestinensis (T2, Acanthamoeba sp. IS2/T4 (T4, Acanthamoeba lenticulata (T5, Acanthamoeba hatchetti (T11, Acanthamoeba healyi (T12, and Acanthamoeba jacobsi (T15. The most frequently identified Acanthamoeba genotype was T4 (68.7%. Acanthamoeba genotype T4 is responsible for Acanthamoeba keratitis and should be considered for associated human health risk potential in the rivershed.

  5. Acanthamoeba encephalitis: A Case Report and Review of Therapy

    A Zamora

    2014-01-01

    Full Text Available Background: Acanthamoeba is a rare cause of encephalitis yet is associated with high mortality. Treatment protocols vary greatly and generally include combination therapy across a wide spectrum of antiinfective classes. Case Description: A 63-year-old male who underwent renal transplantation presented 6 months after transplantation with depressed level of consciousness. Imaging of the head with computerized tomography showed an enhancing lesion suspicious for brain abscess. Biopsy of the lesion showed Acanthamoeba cysts. The patient was treated with sulfadiazine, fluconazole, flucytosine, azithromycin, and miltefosine but without success. We review recently published cases of Acanthamoeba encephalitis with an emphasis on treatment protocols and outcomes. Conclusion: Free-living protozoans such as Acanthamoeba are ubiquitous in the environment and should be suspected in immunosuppressed persons who present with central nervous system findings and brain abscess. Biopsy is critical to establish the etiology so that appropriate combination therapy can be deployed.

  6. Is there evidence of sexual reproduction (meiosis) in Acanthamoeba?

    Khan, Naveed A.; Siddiqui, Ruqaiyyah

    2015-01-01

    Evolution of independently breeding species into males and females (gametes) has remained a puzzle. Given the significant advantages of sexual reproduction over asexual reproduction as a long-term species survival strategy; here, we pose the question whether there is some form of meiosis in Acanthamoeba species, which represents our ancient lineage. The recently available Acanthamoeba genome revealed several genes implicated in meiosis in sexual eukaryotes such as Spo11, Mre...

  7. The immunological aspects of Acanthamoeba infections

    Naveed A.  Khan

    2005-01-01

    Acanthamoeba is a protozoan pathogen that is responsible for two recognized diseases; i) a blinding keratitis, most commonly associated with contact lens wear, and ii) a rare but fatal granulomatous encephalitis that usually is limited to immunocompromised patients. The fact that Acanthamoeba infections are generally limited to avascular cornea or immunocompromised patients suggests that the normal immune responses may be sufficient to control and/or eradicate these pathogens. Here we describ...

  8. Proteases as Markers for Differentiation of Pathogenic and Nonpathogenic Species of Acanthamoeba

    Khan, Naveed A.; Jarroll, Edward L.; Panjwani, Noorjahan; Cao, Zhiyi; Paget, Timothy A.

    2000-01-01

    Acanthamoeba keratitis is a vision-threatening infection caused by pathogenic species of the genus Acanthamoeba. Although not all Acanthamoeba spp. can cause keratitis, it is important to differentiate pathogenic species and isolates from nonpathogens. Since extracellular proteases may play a role in ocular pathology, we used colorimetric, cytopathic, and zymographic assays to assess extracellular protease activity in pathogenic and nonpathogenic Acanthamoeba. Colorimetric assays, using azo-linked protein as a substrate, showed extracellular protease activity in Acanthamoeba-conditioned medium and differentiated pathogenic and nonpathogenic Acanthamoeba. Monolayers of immortalized corneal epithelial cells in four-well plates were used for cytopathic effect (CPE) assays. Pathogenic Acanthamoeba isolates exhibited marked CPE on immortalized corneal epithelial cells, while nonpathogenic isolates did not exhibit CPE. Protease zymography was performed with Acanthamoeba-conditioned medium as well as with Acanthamoeba- plus epithelial-cell-conditioned medium. The zymographic protease assays showed various banding patterns for different strains of Acanthamoeba. In pathogenic Acanthamoeba isolates, all protease bands were inhibited by phenylmethylsulfonyl fluoride (PMSF), suggesting serine type proteases, while in nonpathogenic strains only partial inhibition was observed by using PMSF. The pathogenic Acanthamoeba strains grown under typical laboratory conditions without epithelial cells exhibited one overexpressed protease band of 107 kDa in common; this protease was not observed in nonpathogenic Acanthamoeba strains. The 107-kDa protease exhibited activity over a pH range of 5 to 9.5. PMID:10921939

  9. Azasterols impair Giardia lamblia proliferation and induces encystation

    The effects of sterol biosynthesis inhibitors on growth and fine structure of Giardia lamblia P1 strain cultures were analyzed. Azasterols demonstrated high efficacy in killing cells. The IC50 values for 22,26-azasterol and 24(R,S),25-epiminolanosterol were 7 μM and 170 nM, respectively. Morphological analysis showed that azasterols induced changes in G. lamblia ultrastructure. The most significant alterations were: (a) considerable increase of the size of the peripheral vesicles, which are part of the parasite endosomal-lysosomal system; (b) appearance of autophagosomal structures; and (c) induction of differentiation, followed by an abnormal enlargement of encystation secretory vesicles. We propose that azasterols are effective chemotherapeutic drugs against Giardia lamblia in vitro and may have another target in cells besides sterol biosynthesis

  10. Proteases as Markers for Differentiation of Pathogenic and Nonpathogenic Species of Acanthamoeba

    Khan, Naveed A.; Jarroll, Edward L.; Panjwani, Noorjahan; Cao, Zhiyi; Paget, Timothy A.

    2000-01-01

    Acanthamoeba keratitis is a vision-threatening infection caused by pathogenic species of the genus Acanthamoeba. Although not all Acanthamoeba spp. can cause keratitis, it is important to differentiate pathogenic species and isolates from nonpathogens. Since extracellular proteases may play a role in ocular pathology, we used colorimetric, cytopathic, and zymographic assays to assess extracellular protease activity in pathogenic and nonpathogenic Acanthamoeba. Colorimetric assays, using azo-l...

  11. Identification of the Major Cysteine Protease of Giardia and Its Role in Encystation*S⃞

    DuBois, Kelly N.; Abodeely, Marla; Sakanari, Judy; Craik, Charles S.; Lee, Malinda; McKerrow, James H.; Sajid, Mohammed

    2008-01-01

    Giardia lamblia is a protozoan parasite and the earliest branching clade of eukaryota. The Giardia life cycle alternates between an asexually replicating vegetative form and an infectious cyst form. Encystation and excystation are crucial processes for the survival and transmission of Giardia. Cysteine proteases in Giardia have been implicated in proteolytic processing events that enable the continuance of the life cycle throughout encystation and excystation. Using qu...

  12. A case of radial keratoneuritis in non-Acanthamoeba keratitis

    Mutoh T

    2012-09-01

    Full Text Available Tetsuya Mutoh, Yukihiro Matsumoto, Makoto ChikudaDepartment of Ophthalmology, Dokkyo Medical University Koshigaya Hospital, Saitama, JapanAbstract: A case of non-Acanthamoeba keratitis with radial keratoneuritis, which is thought to be pathognomonic for Acanthamoeba keratitis, is reported. A healthy 32-year-old woman with a history of frequent replacement of her contact lenses due to wear was examined at Dokkyo Medical University Koshigaya Hospital (Saitama, Japan and found to have a slight corneal opacity that was accompanied by radial keratoneuritis. Based on both the patient’s clinical findings and past history, the presence of Acanthamoeba keratitis was highly suspected. However, direct light microscopy of corneal scrapings stained by the Parker ink–potassium hydroxide method only found Acanthamoeba-type material in the specimen collected at her initial visit. In all other specimens collected from the patient, no Acanthamoeba was found either when using the same method or when performing cultures of the surgical debridement of the corneal lesion. In addition, topical antifungal eye drops, systemic antifungal drugs, and surgical debridement were also not effective in this case. Since a precise diagnosis could not be made, the patient was treated with topical 0.1% betamethasone sodium, which ultimately resulted in a dramatic improvement of her corneal inflammation. At 23 days after initiation of topical administration of 0.1% betamethasone sodium, visual acuity was 20/250, with a slight corneal opacity noted at the original site of infection. The outcome of the current case suggests that radial keratoneuritis is not always pathognomonic for Acanthamoeba keratitis.Keywords: radial keratoneuritis, non-Acanthamoeba keratitis, topical corticosteroid

  13. IDENTIFICATION OF Pseudomonas spp. AS AMOEBA-RESISTANT MICROORGANISMS IN ISOLATES OF Acanthamoeba

    Vinicius José Maschio

    2015-02-01

    Full Text Available Acanthamoeba is a “Trojan horse” of the microbial world. The aim of this study was to identify the presence of Pseudomonas as an amoeba-resistant microorganism in 12 isolates of Acanthamoeba. All isolates showed the genus Pseudomonas spp. as amoeba-resistant microorganisms. Thus, one can see that the Acanthamoeba isolates studied are hosts of Pseudomonas.

  14. Actin, RhoA, and Rab11 Participation during Encystment in Entamoeba invadens

    M. Herrera-Martínez

    2013-01-01

    Full Text Available In the genus Entamoeba, actin reorganization is necessary for cyst differentiation; however, its role is still unknown. The aim of this work was to investigate the role of actin and encystation-related proteins during Entamoeba invadens encystation. Studied proteins were actin, RhoA, a small GTPase involved through its effectors in the rearrangement of the actin cytoskeleton; Rab11, a protein involved in the transport of encystation vesicles; and enolase, as an encystment vesicles marker. Results showed a high level of polymerized actin accompanied by increased levels of RhoA-GTP during cell rounding and loss of vacuoles. Cytochalasin D, an actin polymerization inhibitor, and Y27632, an inhibitor of RhoA activity, reduced encystment in 80%. These inhibitors also blocked cell rounding, disposal of vacuoles, and the proper formation of the cysts wall. At later times, F-actin and Rab11 colocalized with enolase, suggesting that Rab11 could participate in the transport of the cyst wall components through the F-actin cytoskeleton. These results suggest that actin cytoskeleton rearrangement is playing a decisive role in determining cell morphology changes and helping with the transport of cell wall components to the cell surface during encystment of E. invadens.

  15. The Transcriptional Response to Encystation Stimuli in Giardia lamblia Is Restricted to a Small Set of Genes ▿†

    Morf, Laura; Spycher, Cornelia; Rehrauer, Hubert; Fournier, Catharine Aquino; Morrison, Hilary G.; Hehl, Adrian B.

    2010-01-01

    The protozoan parasite Giardia lamblia undergoes stage differentiation in the small intestine of the host to an environmentally resistant and infectious cyst. Encystation involves the secretion of an extracellular matrix comprised of cyst wall proteins (CWPs) and a β(1-3)-GalNAc homopolymer. Upon the induction of encystation, genes coding for CWPs are switched on, and mRNAs coding for a Myb transcription factor and enzymes involved in cyst wall glycan synthesis are upregulated. Encystation in vitro is triggered by several protocols, which call for changes in bile concentrations or availability of lipids, and elevated pH. However, the conditions for induction are not standardized and we predicted significant protocol-specific side effects. This makes reliable identification of encystation factors difficult. Here, we exploited the possibility of inducing encystation with two different protocols, which we show to be equally effective, for a comparative mRNA profile analysis. The standard encystation protocol induced a bipartite transcriptional response with surprisingly minor involvement of stress genes. A comparative analysis revealed a core set of only 18 encystation genes and showed that a majority of genes was indeed upregulated as a side effect of inducing conditions. We also established a Myb binding sequence as a signature motif in encystation promoters, suggesting coordinated regulation of these factors. PMID:20693303

  16. The transcriptional response to encystation stimuli in Giardia lamblia is restricted to a small set of genes.

    Morf, Laura; Spycher, Cornelia; Rehrauer, Hubert; Fournier, Catharine Aquino; Morrison, Hilary G; Hehl, Adrian B

    2010-10-01

    The protozoan parasite Giardia lamblia undergoes stage differentiation in the small intestine of the host to an environmentally resistant and infectious cyst. Encystation involves the secretion of an extracellular matrix comprised of cyst wall proteins (CWPs) and a β(1-3)-GalNAc homopolymer. Upon the induction of encystation, genes coding for CWPs are switched on, and mRNAs coding for a Myb transcription factor and enzymes involved in cyst wall glycan synthesis are upregulated. Encystation in vitro is triggered by several protocols, which call for changes in bile concentrations or availability of lipids, and elevated pH. However, the conditions for induction are not standardized and we predicted significant protocol-specific side effects. This makes reliable identification of encystation factors difficult. Here, we exploited the possibility of inducing encystation with two different protocols, which we show to be equally effective, for a comparative mRNA profile analysis. The standard encystation protocol induced a bipartite transcriptional response with surprisingly minor involvement of stress genes. A comparative analysis revealed a core set of only 18 encystation genes and showed that a majority of genes was indeed upregulated as a side effect of inducing conditions. We also established a Myb binding sequence as a signature motif in encystation promoters, suggesting coordinated regulation of these factors. PMID:20693303

  17. In vitro Effect of Monosaccharides on the Virulence of Acanthamoeba Isolated from Patients with Amoebic Keratitis

    Y. Maroofi

    2007-04-01

    Full Text Available Introduction & Objective: Acanthamoeba is free-living amoeba that is found in soil, water, air as well as in human pharynx. Acanthamoeba is causative agent of granulomatose amoebic encephalitis (GAE in immunosuppressed and AIDS individuals and amoebic keratitis in people who use the lens. Pathogenic species of Acanthamoeba have protein receptors named mannose binding protein (MBP. Acanthamoeba via MBP adhere to the glycoproteins included mannose. Acanthamoeba adhesion to the target cells induces a protease secretion is called mannose inducing protein-133 (MIP-133. Exogense mannose can inhibit the adherence of Acanthamoeba; also, it can increase the cytopathatic effect (CPE through increase the secretion of MIP-133. In the present work, the effect of monosaccharides on the virulance of Acanthamoeba isolated from patient with amoebic keratitis, in HeLa cell culture was investigated.Materials & Methods: The isolates were cultured in HeLa cell culture, then 100, 50, 10, 1 and 0.1 mM of galactose, glucose and mannose were added to plates. Plates were observed with invert microscope in 8, 16, 32, 48, and 72 hours after culture.Results: Data implicated that mannose (100 mM showed the highest effect on increasing cytopathy of Acanthamoeba in HeLa cell culture. Meanwhile, galactose (100 mM increased the virulence of Acanthamoeba in the cell culture after 32 hours.Conclusion: Adding mannose and galactose to HeLa cell culture contain Acanthamoeba can increase the virulence of the parasite significantly.

  18. A retrospective study of nine cases of Acanthamoeba keratitis

    Tetsuya Mutoh

    2010-10-01

    Full Text Available Tetsuya Mutoh, Isao Ishikawa, Yukihiro Matsumoto, Makoto ChikudaDokkyo Medical University Koshigaya Hospital, Saitama, JapanPurpose: To evaluate the clinical features of Acanthamoeba keratitis in nine patients diagnosed at Dokkyo Medical University Koshigaya Hospital, Saitama, Japan.Methods: In nine eyes of nine patients, Acanthamoeba keratitis was diagnosed by direct light microscopy of corneal scrapings stained by the Parker ink-potassium hydroxide method between September 2006 and September 2009. Their clinical features and course were studied retrospectively. Antifungal eye drops, systemic antifungal therapy, and surgical debridement of the corneal lesions were performed in all patients.Results: At presentation, the clinical stage was initial in six cases, transient in one case, and complete in two cases. The patients were all contact lens wearers who had washed their lens storage cases with tap water. After treatment, final visual acuity was improved in six cases, unchanged in one case, and worse in two cases. The patient with the worst final vision (hand motion had rheumatoid arthritis and was taking oral prednisolone, which led to corneal perforation and prevented adequate debridement from being done.Conclusion: Acanthamoeba keratitis is closely related to wearing contact lenses and washing the lens storage case with tap water. Although final visual acuity improved after treatment in most patients, insufficient surgical debridement resulted in a poor visual prognosis.Keywords: surgical debridement, Acanthamoeba keratitis, contact lens wearers

  19. Salmonella pneumonia complicated with encysted empyema in an immunocompromised youth: Case report and literature Review.

    Saeed, Nermin Kamal

    2016-01-01

    In this case report we described a Bahraini male patient of twenty years of age, a smoker and diagnosed with stage IV B Hodgkin lymphoma. He presented with fever, nonproductive cough, upper back pain and shortness of breath due to right upper lobe pneumonia with right encysted pleural effusion. Salmonella enterica serotype Enteritidis was isolated from the sputum. He was successfully treated with 2 weeks of ceftriaxone followed by 2 weeks of oral cefixime. This was the first case of encysted empyema caused by Salmonella enterica serotype Enteritidis reported in the Kingdom of Bahrain. The different aspects of pulmonary Salmonella infections were discussed and the literature was reviewed. PMID:27131011

  20. Identification and properties of proteases from an Acanthamoeba isolate capable of producing granulomatous encephalitis

    Jarroll Edward L; Lightfoot Mary; Goldsworthy Graham; Alsam Selwa; Sissons James; Khan Naveed

    2006-01-01

    Abstract Background Granulomatous amoebic encephalitis due to Acanthamoeba is often a fatal human disease. However, the pathogenesis and pathophysiology of Acanthamoeba encephalitis remain unclear. In this study, the role of extracellular Acanthamoeba proteases in central nervous system pathogenesis and pathophysiology was examined. Results Using an encephalitis isolate belonging to T1 genotype, we observed two major proteases with approximate molecular weights of 150 KD and 130 KD on SDS-PAG...

  1. Isolation of Acanthamoeba-Specific Antibodies from a Bacteriophage Display Library

    Khan, Naveed A.; Greenman, John; Topping, Katherine P.; Victoria C. Hough; Temple, Graham S.; Paget, Timothy A.

    2000-01-01

    Acanthamoeba causes opportunistic eye infections in humans, which can lead to severe keratitis and may ultimately result in blindness. Current methods for identifying this organism rely on culture and microscopy. In this paper, we describe the isolation of antibody fragments that can be used for the unequivocal identification of Acanthamoeba. A bacteriophage antibody display library was used to isolate antibody fragments that bind specifically to Acanthamoeba. Individual clones were studied b...

  2. Acanthamoeba spp. in Contact Lenses from Healthy Individuals from Madrid, Spain

    Gomes, Thiago dos Santos; Magnet, Angela; Izquierdo, Fernando; Vaccaro, Lucianna; Redondo, Fernando; Bueno, Sara; Sánchez, Maria Luisa; Angulo, Santiago; Fenoy, Soledad; Hurtado, Carolina; del Aguila, Carmen

    2016-01-01

    Purpose Acanthamoeba keratitis (AK) is a painful and potentially blinding corneal infection caused by Acanthamoeba spp. In Madrid, environmental studies have demonstrated a high presence of these free-living amoebae in tap water. Since most of AK cases occur in contact lenses (CL) wearers with inadequate hygiene habits, the presence of Acanthamoeba in discarded CL has been studied and compared with other common etiological agents of keratitis, such as Pseudomonas aeruginosa and Staphylococcus...

  3. A Case of Non-Contact Lens related Acanthamoeba keratitis in Malaysia

    Mohamed Kamel, A. G.; Faridah, H.; Yusof, S; Norazah, A.; Nakisah, M. A.

    2005-01-01

    Acanthamoeba is a ubiquitous free-living amoeba and is responsible for an uncommon yet increasingly diagnosed keratitis in humans. Acanthamoeba keratitis is perhaps the most challenging ocular infection to manage successfully and it can result in permanently impaired vision or blindness. Although contact lens use is the principal risk factor, about 10% of cases occur following trauma and exposure to contaminated soil or water. Cases of Acanthamoeba keratitis involving contact lens wearers hav...

  4. Isolation of Acanthamoeba-Specific Antibodies from a Bacteriophage Display Library

    Khan, Naveed A.; Greenman, John; Topping, Katherine P.; Hough, Victoria C.; Temple, Graham S.; Paget, Timothy A.

    2000-01-01

    Acanthamoeba causes opportunistic eye infections in humans, which can lead to severe keratitis and may ultimately result in blindness. Current methods for identifying this organism rely on culture and microscopy. In this paper, we describe the isolation of antibody fragments that can be used for the unequivocal identification of Acanthamoeba. A bacteriophage antibody display library was used to isolate antibody fragments that bind specifically to Acanthamoeba. Individual clones were studied by enzyme-linked immunosorbent assay, flow cytometry, and immunofluorescence. Four antibody clones that specifically bind to Acanthamoeba spp. were identified. PMID:10835006

  5. Acanthamoeba protease activity promotes allergic airway inflammation via protease-activated receptor 2.

    Mi Kyung Park

    Full Text Available Acanthamoeba is a free-living amoeba commonly present in the environment and often found in human airway cavities. Acanthamoeba possesses strong proteases that can elicit allergic airway inflammation. To our knowledge, the aeroallergenicity of Acanthamoeba has not been reported. We repeatedly inoculated mice with Acanthamoeba trophozoites or excretory-secretory (ES proteins intra-nasally and evaluated symptoms and airway immune responses. Acanthamoeba trophozoites or ES proteins elicited immune responses in mice that resembled allergic airway inflammation. ES proteins had strong protease activity and activated the expression of several chemokine genes (CCL11, CCL17, CCL22, TSLP, and IL-25 in mouse lung epithelial cells. The serine protease inhibitor phenyl-methane-sulfonyl fluoride (PMSF inhibited ES protein activity. ES proteins also stimulated dendritic cells and enhanced the differentiation of naive T cells into IL-4-secreting T cells. After repeated inoculation of the protease-activated receptor 2 knockout mouse with ES proteins, airway inflammation and Th2 immune responses were markedly reduced, but not to basal levels. Furthermore, asthma patients had higher Acanthamoeba-specific IgE titers than healthy controls and we found Acanthamoeba specific antigen from house dust in typical living room. Our findings suggest that Acanthamoeba elicits allergic airway symptoms in mice via a protease allergen. In addition, it is possible that Acanthamoeba may be one of the triggers human airway allergic disease.

  6. Effect of Antimicrobial Compounds on Balamuthia mandrillaris Encystment and Human Brain Microvascular Endothelial Cell Cytopathogenicity▿

    Siddiqui, Ruqaiyyah; Matin, Abdul; Warhurst, David; Stins, Monique; Khan, Naveed Ahmed

    2007-01-01

    Cycloheximide, ketoconazole, or preexposure of organisms to cytochalasin D prevented Balamuthia mandrillaris-associated cytopathogenicity in human brain microvascular endothelial cells, which constitute the blood-brain barrier. In an assay for inhibition of cyst production, these three agents prevented the production of cysts, suggesting that the biosynthesis of proteins and ergosterol and the polymerization of actin are important in cytopathogenicity and encystment.

  7. Taken to the limit--Is desiccation stress causing precocious encystment of trematode parasites in snails?

    O'Dwyer, Katie; Poulin, Robert

    2015-12-01

    When hosts experience environmental stress, the quantity and quality of resources they provide for parasites may be diminished, and host longevity may be decreased. Under stress, parasites may adopt alternative strategies to avoid fitness reductions. Trematode parasites typically have complex life cycles, involving asexual reproduction in a gastropod first intermediate host. A rare phenomenon, briefly mentioned in the literature, and termed 'precocious encystment' involves the next stage in the parasites' life cycle (metacercarial cyst) forming within the preceding stage (redia), while still inside the snail. In the trematode Parorchis sp. NZ using rocky shore snails exposed to long periods outside water, we hypothesised that this might be an adaptive strategy against desiccation, preventing parasite emergence from the snail. To test this, we first investigated the effect of prolonged desiccation on the survival of two species of high intertidal snails. Secondly, we measured the reproductive output (cercarial production) of the parasite under wet and dry conditions. Finally, we quantified the influence of desiccation stress on the occurrence of precocious encystment. Snail mortality was higher under dry conditions, indicating stress, and it was somewhat exacerbated for infected snails. Parasite reproductive output differed between wet and dry conditions, with parasites of snails kept in dry conditions producing more cercariae when placed in water. Little variation was observed in the occurrence of precocious encystment, although some subtle patterns emerged. Given the stresses associated with living in high intertidal environments, we discuss precocious encystment as a possible stress response in this trematode parasite. PMID:26344863

  8. ENCYSTATION AND EXPRESSION OF CYST ANTIGENS BY 'GIARDIA LAMBLIA' IN VITRO

    The cyst form of Giardia lamblia is responsible for transmission of giardiasis, a major waterborne intestinal disease. These studies demonstrate for the first time expression of cyst antigens and encystation of G. lamblia in vitro by both morphologic and immunologic criteria. The...

  9. Viability of Acanthamoeba after exposure to a multipurpose disinfecting contact lens solution and two hydrogen peroxide systems

    Hiti, K; Walochnik, J.; Haller-Schober, E M; Faschinger, C; Aspöck, H.

    2002-01-01

    Background/aim: Contact lens cases contaminated with Acanthamoeba are a major risk factor for an infection of the eye. In this study the anti-Acanthamoeba activity of three different contact lens storage solutions was tested.

  10. The generation gap: Proteome changes and strain variation during encystation in Giardia duodenalis.

    Emery, Samantha J; Pascovi, Dana; Lacey, Ernest; Haynes, Paul A

    2015-05-01

    The prevalence of Giardia duodenalis in humans is partly owed to its direct and simple life cycle, as well as the formation of the environmentally resistant and infective cysts. Proteomic and transcriptomic studies have previously analysed the encystation process using the well-characterised laboratory genomic strain, WB C6. This study presents the first quantitative study of encystation using pathogenically relevant and alternative assemblage A strains: the human-derived BRIS/82/HEPU/106 (H-106)and avian-derived BRIS/95/HEPU/2041 (B-2041). We utilised tandem MS/MS with a label-free quantitative approach to compare cysts and trophozoite life stages for strain variation, as well as confirm universal encystation markers of assemblage A. A total of 1061 non-redundant proteins were identified from both strains, including trophozoite- and cyst-specific proteomes and life-stage differentially expressed proteins. Additionally, 24 proteins previously classified in the literature as encystation-specific were confirmed as strain-independent markers of encystation. Functional cluster analysis of differentially expressed proteins saw significant overlap between strains, including protein trafficking and localisation in cysts, NEK kinase function, and carbohydrate metabolism in trophozoites. Two significant points of strain specific adaptations in cysts were also identified. B-2041 possessed major up-regulation of the ankyrin repeat protein 21.1 family compared to H-106. Furthermore, cysts of B-2041 retained near-complete VSP variant diversity between cysts and trophozoites, while H-106 lost 45% of its VSP variant diversity between life cycle stages, a constriction previously observed in studies of WB C6. This is the first report of strain variation in the cyst stage in G. duodenalis, and highlights cyst variation and its impacts on reinfection and life cycle success. PMID:26045354

  11. Uptake and Replication of Salmonella enterica in Acanthamoeba rhysodes

    Tezcan-Merdol, Dilek; Ljungström, Marianne; Winiecka-Krusnell, Jadwiga; Linder, Ewert; Engstrand, Lars; Rhen, Mikael

    2004-01-01

    The ability of salmonellae to become internalized and to survive and replicate in amoebae was evaluated by using three separate serovars of Salmonella enterica and five different isolates of axenic Acanthamoeba spp. In gentamicin protection assays, Salmonella enterica serovar Dublin was internalized more efficiently than Salmonella enterica serovar Enteritidis or Salmonella enterica serovar Typhimurium in all of the amoeba isolates tested. The bacteria appeared to be most efficiently internal...

  12. Acanthamoeba on Sabouraud's agar from a patient with keratitis

    Baradkar, Vasant; Samal, Badhuli; Mali, Swapna A; Kulkarni, Ketaki; Shastri, Jayanthi

    2011-01-01

    A 25-year-old transgender patient came with complaints of watery discharge, red eye and photophobia in the left eye since 2 days. The patient had a history of wearing colored contact lenses since 4 years and cleaning the lens with tap water. Culture of lenses on Mac Conkey and blood agar yielded Klebsiella pneumoniae and Pseudomonas aeruginosa. Sabouroud's agar showed yeast cells and double-walled cysts of Acanthamoeba species. On further incubation of Sabouroud's agar, the cysts transformed ...

  13. Optimized methods for Legionella pneumophila release from its Acanthamoeba hosts

    Dietersdorfer, Elisabeth; Cervero-Aragó, Sílvia; Sommer, Regina; Kirschner, Alexander K.; Walochnik, Julia

    2016-01-01

    Background Free-living amoebae (FLA) and particularly acanthamoebae serve as vehicles and hosts for Legionella pneumophila, among other pathogenic microorganisms. Within the amoebae, L. pneumophila activates a complex regulatory pathway that enables the bacteria to resist amoebal digestion and to replicate. Moreover, the amoebae provide the bacteria protection against harsh environmental conditions and disinfectants commonly used in engineered water systems. To study this ecological relations...

  14. The Isolation and Detection of Acanthamoeba Keratitis in Rural Water Sources of Arak, Iran

    Mosayebi, M. (PhD

    2014-05-01

    Full Text Available Background and Objective: Acanthamoeba species are free-living protozoa that can be isolated from all environments. They can bring about different diseases in healthy individuals and immune suppressed patients, for example, Granulomatous Amoebic Encephalitis (GAE, Acanthamoeba Keratitis (AK, Cutaneous and Nasopharyngeal infections. The aim of this study was to evaluate the Acanthamoeba prevalence in rural water sources of Markazi province. Material and Methods: In this cross sectional study, 54 water samples were collected from 36 villages of Markus province. First, the Samples were filtered by filter paper (watchman 42. Next, the filtered paper were placed in page saline solution and centrifuged. Then, the obtained sediment was cultured on non-nutrient agar (NNA plates overlaid with heat-killed Escherichia coli. After that, the provided smear (after 4 - 7 days stained with Geimsa. Results: The samples were positive (33; 61.11% and negative (21; 38.89% for Acanthamoeba cyst. The best result for isolation of Acanthamoeba cysts was obtained after shaking of filter paper. Conclusion: A high percentage of rural water sources have been contaminated with Acanthamoeba, which can be the major factor in causing human infections. Therefore, some effective methods are required to prevent from water sources contamination. Keywords: Acanthamoeba; Acanthamoeba Keratitis; Markazi Province; Rural; Water Sources

  15. Rab11 and actin cytoskeleton participate in Giardia lamblia encystation, guiding the specific vesicles to the cyst wall.

    Araceli Castillo-Romero

    Full Text Available BACKGROUND: Giardia passes through two stages during its life cycle, the trophozoite and the cyst. Cyst formation involves the synthesis of cyst wall proteins (CWPs and the transport of CWPs into encystation-specific vesicles (ESVs. Active vesicular trafficking is essential for encystation, but the molecular machinery driving vesicular trafficking remains unknown. The Rab proteins are involved in the targeting of vesicles to several intracellular compartments through their association with cytoskeletal motor proteins. METHODOLOGY AND PRINCIPAL FINDINGS: In this study, we found a relationship between Rab11 and the actin cytoskeleton in CWP1 transport. Confocal microscopy showed Rab11 was distributed throughout the entire trophozoite, while in cysts it was translocated to the periphery of the cell, where it colocalized with ESVs and microfilaments. Encystation was also accompanied by changes in rab11 mRNA expression. To evaluate the role of microfilaments in encystation, the cells were treated with latrunculin A. Scanning electron microscopy showed this treatment resulted in morphological damages to encysted parasites. The intensity of fluorescence-labeled Rab11 and CWP1 in ESVs and cyst walls was reduced, and rab11 and cwp1 mRNA levels were down-regulated. Furthermore, knocking down Rab11 with a hammerhead ribozyme resulted in an up to 80% down-regulation of rab11 mRNA. Although this knockdown did not appear lethal for trophozoites and did not affect cwp1 expression during the encystation, confocal images showed CWP1 was redistributed throughout the cytosol. CONCLUSIONS AND SIGNIFICANCE: Our results indicate that Rab11 participates in the early and late encystation stages by regulating CWP1 localization and the actin-mediated transport of ESVs towards the periphery. In addition, alterations in the dynamics of actin affected rab11 and cwp1 expression. Our results provide new information about the molecules involved in Giardia encystation and

  16. Influence of cadmium exposure on the incidence of first intermediate host encystment by Echinoparyphium recurvatum cercariae in Lymnaea peregra

    Morley, Neil; Crane, M.; Lewis, J W

    2006-01-01

    The effect of cadmium exposure of the snail first intermediate host Lymnaea peregra on the incidence of encystment of Echinoparyphium recurvatum (Digenea: Echinostomatidae) cercariae without emergence from the snail was investigated. Exposure to 100mg l21 Cd for 72 h caused a significant increase in the incidence of first host encystment when compared to controls. In addition, autometallographic staining of E. recurvatum daughter rediae and developing cercariae showed that there was metal acc...

  17. Encysted hydrocele of the cord presenting as a groin swelling in an adult: a case report

    Bibekananda Mahapatra

    2015-09-01

    Full Text Available The hydrocele of the cord presents as a translucent swelling in the inguinal canal or scrotum, separately above the testis. It is more commonly seen in childhood and is rare in adults. It comprises of two varieties based on their communication with the peritoneal cavity. The encysted hydrocele of the cord does not communicate with the peritoneal cavity above or the tunica vaginalis below. The other variety known as the funicular hydrocele communicates with the peritoneal cavity above but does not communicate with the tunica vaginalis below. This case report presents the case of a 22 year old young male with an inguinal swelling. Clinical differential diagnoses were of inguinal lymphadenitis, encysted hydrocele of the cord and irreducible hernia. The aim of this report is to highlight the role of ultrasonogram in early diagnosis and surgical excision as mainstay of treatment. [Int J Res Med Sci 2015; 3(9.000: 2475-2477

  18. Effect of Antimicrobial Compounds on Balamuthia mandrillaris Encystment and Human Brain Microvascular Endothelial Cell Cytopathogenicity▿

    Siddiqui, Ruqaiyyah; Matin, Abdul; Warhurst, David; Stins, Monique; Khan, Naveed Ahmed

    2007-01-01

    Cycloheximide, ketoconazole, or preexposure of organisms to cytochalasin D prevented Balamuthia mandrillaris-associated cytopathogenicity in human brain microvascular endothelial cells, which constitute the blood-brain barrier. In an assay for inhibition of cyst production, these three agents prevented the production of cysts, suggesting that the biosynthesis of proteins and ergosterol and the polymerization of actin are important in cytopathogenicity and encystment. PMID:17875991

  19. Toxicity of cadmium and zinc to encystment and in vitro excystment of Parorchis acanthus (Digenea: Philophthalmidae)

    Morley, N.J.; Crane, M.; Lewis, J W

    2001-01-01

    The toxicity of cadmium, zinc and cadmium}zinc mixtures at concentrations ranging from 1000 to 50000 lg}l were investigated against cercariae and metacercariae of Parorchis acanthus obtained from the dog whelk Nucella lapillus. Cercarial encystment at concentrations of 25000 lg}l or higher was signi®cantly impaired by all test metals; however, at lower concentrations only zinc demonstrated toxicity. Mixtures of cadmium and zinc had a synergistic effect compared with single metal toxicity but ...

  20. Rac Regulates Giardia lamblia Encystation by Coordinating Cyst Wall Protein Trafficking and Secretion

    Krtková, Jana; Thomas, Elizabeth B.; Alas, Germain C. M.; Schraner, Elisabeth M.; Behjatnia, Habib R.; Hehl, Adrian B.

    2016-01-01

    ABSTRACT Encystation of the common intestinal parasite Giardia lamblia involves the production, trafficking, and secretion of cyst wall material (CWM). However, the molecular mechanism responsible for the regulation of these sequential processes remains elusive. Here, we examined the role of GlRac, Giardia’s sole Rho family GTPase, in the regulation of endomembrane organization and cyst wall protein (CWP) trafficking. Localization studies indicated that GlRac is associated with the endoplasmic reticulum (ER) and the Golgi apparatus-like encystation-specific vesicles (ESVs). Constitutive GlRac signaling increased levels of the ER marker PDI2, induced ER swelling, reduced overall CWP1 production, and promoted the early maturation of ESVs. Quantitative analysis of cells expressing constitutively active hemagglutinin (HA)-tagged GlRac (HA-RacCA) revealed fewer but larger ESVs than control cells. Consistent with the phenotype of premature maturation of ESVs in HA-RacCA-expressing cells, constitutive GlRac signaling resulted in increased CWP1 secretion and, conversely, morpholino depletion of GlRac blocked CWP1 secretion. Wild-type cells unexpectedly secreted large quantities of CWP1 into the medium, and free CWP1 was used cooperatively during cyst formation. These results, in part, could account for the previously reported observation that G. lamblia encysts more efficiently at high cell densities. These studies of GlRac show that it regulates encystation at several levels, and our findings support its coordinating role as a regulator of CWP trafficking and secretion. The central role of GlRac in regulating membrane trafficking and the cytoskeleton, both of which are essential to Giardia parasitism, further suggests its potential as a novel target for drug development to treat giardiasis. PMID:27555307

  1. Encysted hydrocele of the cord presenting as a groin swelling in an adult: a case report

    Bibekananda Mahapatra; Anuradha Panchal; Abhishek Mahadik; Qurratulain Chougle

    2015-01-01

    The hydrocele of the cord presents as a translucent swelling in the inguinal canal or scrotum, separately above the testis. It is more commonly seen in childhood and is rare in adults. It comprises of two varieties based on their communication with the peritoneal cavity. The encysted hydrocele of the cord does not communicate with the peritoneal cavity above or the tunica vaginalis below. The other variety known as the funicular hydrocele communicates with the peritoneal cavity above but does...

  2. A small heat shock/α-crystallin protein from encysted Artemia embryos suppresses tubulin denaturation

    Day, Rossalyn M.; Gupta, Jagdish S.; MacRae, Thomas H.

    2003-01-01

    Small heat shock/α-crystallin proteins function as molecular chaperones, protecting other proteins from irreversible denaturation by an energy-independent process. The brine shrimp, Artemia franciscana, produces a small heat shock/α-crystallin protein termed p26, found in embryos undergoing encystment, diapause, and metabolic arrest. These embryos withstand long-term anoxia and other stresses normally expected to cause death, a property likely dependent on molecular chaperone activity. The as...

  3. Genotypic, physiological, and biochemical characterization of potentially pathogenic Acanthamoeba isolated from the environment in Cairo, Egypt.

    Tawfeek, Gihan Mostafa; Bishara, Sawsan Abdel-Hamid; Sarhan, Rania Mohammad; ElShabrawi Taher, Eman; ElSaady Khayyal, Amira

    2016-05-01

    Acanthamoebae are the most common opportunistic amphizoic protozoa that cause life-threatening granulomatous amoebic encephalitis in immunocompromised individuals and sight-threatening amoebic keratitis (AK) in contact lens wearers. The present work aimed to determine the presence of Acanthamoeba isolates in different environmental sources: water, soil, and dust in Cairo, Egypt and to characterize the pathogenic potential of the isolated Acanthamoeba using physiological and biochemical assays as well as determination of the genotypes in an attempt to correlate pathogenicity with certain genotypes. The study included the collection of 22 corneal scrapings from patients complaining of symptoms and signs indicative of acanthamoeba keratitis (AK) and 75 environmental samples followed by cultivation on non-nutrient agar plates preseeded with E. coli. Positive samples for Acanthamoeba were subjected to osmo- and thermo-tolerance assays and zymography analysis. Potentially pathogenic isolates were subjected to PCR amplification using genus-specific primer pair. Isolates were classified at the genotype level based on the sequence analysis of Acanthamoeba 18S rRNA gene (diagnostic fragment 3). The total detection rate for Acanthamoeba in environmental samples was 33.3 %, 31.4 % in water, 40 % in soil, and 20 % in dust samples. Three and two Acanthamoeba isolates from water and soil sources, respectively, had the potential for pathogenicity as they exhibited full range of pathogenic traits. Other 12 isolates were designated as weak potential pathogens. Only ten of the environmental isolates were positive in PCR and were classified by genotype analysis into T4 genotype (70 %), T3 (10 %) and T5 (20 %). Potential pathogens belonged to genotypes T4 (from water) and T5 (from soil) while weak potential pathogens belonged to genotypes T3 (from water) and T4 (from water and soil). Additionally, T7 genotype was isolated from keratitis patients. There is a considerable

  4. Isolation and genotyping of Acanthamoeba spp. from recreational soil of parks in Tehran, Iran

    M Niyyati

    2013-11-01

    Full Text Available Background & aim: Acanthamoeba is a genus of free-living amoebae found in environmental sources. These amphizoic amoebae can lead to severe human disease such as encephalitis and keratitis. Acanthamoeba transmits to humans through contact with soil and dust from scratching the skin. The aim of the present study was to identify the genotypes of Acanthamoeba in parks of the city of Tehran using molecular and morphological - based methods. Methods: In this study, 52 samples of soil were collected from 17 parks in Tehran. Samples were then filtered and cultured on 1.5% non-nutrient agar. DNA extraction and PCR amplification was performed using genus specific primers. Sequencing analysis and BLAST search were done for genotype identification. Results: Out of 52 soil samples, 14 strain (26.9% were positive for Acanthamoeba amoebae by microscopic observation. Out Of 14 positive isolates, 9 (17.3% were positive for Acanthamoeba using genus specific primer pairs. Of 14 strains, 9 were sequenced successfully. Genotype identification was revealed that all strains were belonged to T4 type. T4 genotypes among strains are human pathogens. Conclusions: Identification of pathogenic Acanthamoeba belonging to T4 genotype in recreational parks could be of utmost importance. Results of this study show that soil contamination, particularly in parks where children play and assemble is a sanitary risk for them. Key words: Acanthamoeba, Genotypes, Soil, Park

  5. A Case of Non-Contact Lens related Acanthamoeba keratitis in Malaysia

    Mohamed Kamel, A. G.

    2005-01-01

    Full Text Available Acanthamoeba is a ubiquitous free-living amoeba and is responsible for an uncommon yet increasingly diagnosed keratitis in humans. Acanthamoeba keratitis is perhaps the most challenging ocular infection to manage successfully and it can result in permanently impaired vision or blindness. Although contact lens use is the principal risk factor, about 10% of cases occur following trauma and exposure to contaminated soil or water. Cases of Acanthamoeba keratitis involving contact lens wearers have previously been reported in Malaysia but this is the first time, a non-contact lens relatedAcanthamoeba keratitis is reported. The case involved a 28 year old Indonesian male construction worker who had a trauma of the right eye. While working his eye was struck by some sand and dust particles and he quickly washed his eye with water from an open tank at the construction site. He then experienced pain, redness, glaring and blurring of vision of the right eye. The diagnosis was missed at the initial presentation but subsequent culture of the corneal scraping demonstrated Acanthamoeba as the aetiological agent. The history, clinical findings, diagnosis and treatment of non-contact lens related Acanthamoeba keratitis are briefly discussed in this communication. We hope to create awareness especially among the medical and paramedical staff about the existence of this infection in the country and fully support the consideration of Acanthamoeba keratitis as part of the differential diagnosis of most cases of presumed microbial keratitis.

  6. Evaluation of immunoperoxidase staining technique in the diagnosis of Acanthamoeba keratitis

    Sharma Savitri

    2001-01-01

    Full Text Available Purpose: We describe a simple procedure of Immunoperoxidase (IP technique, using indigenously raised antibody, to screen corneal scrapings for Acanthamoeba cysts and trophozoites. This study sought to determine the utility of this test in the diagnosis of Acanthamoeba keratitis. Methods: A high titre polyclonal antibody against a local clinical isolate (axenic of Acanthamoeba species (trophozoite lysate antigen was raised in rabbits and used for standardization of IP technique for corneal scrapings. Twenty two smears of corneal scrapings, collected from patients showing Acanthamoeba cysts in corneal scrapings stained with calcofluorwhite (pool-1 and patients showing no cysts in similar scrapings (pool-2, were coded and stained by IP technique by a masked technician. All 22 patients had also been tested for bacteria, fungus, and Acanthamoeba in their corneal scrapings by smears and cultures. IP stained smears were examined for organisms including cysts and trophozoites of Acanthamoeba and background staining by two observers masked to the results of other smears and cultures. The validity of the IP test in detection of Acanthamoeba cysts and trophozoites was measured by sensitivity, specificity, positive predictive value and negative predictive value in comparison (McNemar test for paired comparison with calcofluor white staining and culture. Results: Based on the readings of observer 1 and compared to calcofluor white staining, the IP test had a sensitivity of 100%, a specificity of 94%, positive predictive value of 80% and negative predictive value of 100%. When compared to culture, the values were 83%, 100%, 100% and 94% respectively. Trophozoites missed in calcofluor white stained smears, were detected in 2 out of 6 cases of culture-positive Acanthamoeba keratitis. The Kappa coefficient of interobserver agreement was determined as fair (30.4%. Conclusion: The immunoperoxidase technique is a simple and useful test in the diagnosis of

  7. Acanthamoeba spp. in Contact Lenses from Healthy Individuals from Madrid, Spain

    Gomes, Thiago dos Santos; Magnet, Angela; Izquierdo, Fernando; Vaccaro, Lucianna; Redondo, Fernando; Bueno, Sara; Sánchez, Maria Luisa; Angulo, Santiago; Fenoy, Soledad; Hurtado, Carolina; del Aguila, Carmen

    2016-01-01

    Purpose Acanthamoeba keratitis (AK) is a painful and potentially blinding corneal infection caused by Acanthamoeba spp. In Madrid, environmental studies have demonstrated a high presence of these free-living amoebae in tap water. Since most of AK cases occur in contact lenses (CL) wearers with inadequate hygiene habits, the presence of Acanthamoeba in discarded CL has been studied and compared with other common etiological agents of keratitis, such as Pseudomonas aeruginosa and Staphylococcus aureus. Methods One hundred and seventy-seven healthy individuals from Madrid contributed their discarded CL and answered a questionnaire on hygiene habits. DNA was extracted from the CL solution and analyzed by real-time PCR for Acanthamoeba, Pseudomonas aeruginosa and Staphylococcus aureus. These CL and their solutions were also cultured on non-nutrient agar to isolate Acanthamoeba. Results Among the 177 samples, Acanthamoeba DNA was detected in 87 (49.2%), P. aeruginosa DNA in 14 (7.9%) and S. aureus DNA in 19 (10.7%). Cultivable amoebae, however, were observed in only one sample (0.6%). This isolate was genotyped as T4. The habits reported by this CL owner included some recognized risk factors for AK, but in this study only the practice of “not cleaning the CL case” presented some statistical significant association with Acanthamoeba DNA presence. Detection of the investigated bacterial DNA did not demonstrate statistical significant association with the studied practices, but the presence of P. aeruginosa revealed a possible inhibition of Acanthamoeba in these samples. Conclusions The PCR results suggest a high presence of Acanthamoeba spp. in healthy CL wearers from Madrid, but we can assume that CL solutions are properly disinfecting the CL since only 1.1% of the positive PCR samples correspond to viable amoebae and, after four years, only one participant reported stronger ocular problems. Nevertheless, more studies are necessary to corroborate this hypothesis. PMID

  8. Acanthamoeba keratitis in a non-contact lens wearer with human immunodeficiency virus

    Hansen, Birgitte Rønde; Kronborg, Gitte

    2003-01-01

    Acanthamoeba keratitis is potentially blinding and often associated with contact lens wearing. A human immunodeficiency virus (HIV)-positive patient, a non-contact lens wearer, presented with keratitis. She experienced a protracted course of disease, characterized by exacerbations and remissions......, and was treated with various topical antibiotics and steroids. 13 months after symptom onset the eye was removed owing to serious scarring of cornea and unbearable pain. Microbiological and histopathological examination of the cornea showed Acanthamoeba. In non-contact lens wearers suffering from...... Acanthamoeba keratitis the diagnosis is delayed, pathognomonic features are often not seen and visual outcome is usually poor. There is no known relation between HIV infection and Acanthamoeba keratitis....

  9. Proteomic approach to reveal the proteins associated with encystment of the ciliate Euplotes encysticus.

    Jiwu Chen

    Full Text Available In order to identify and reveal the proteins related to encystment of the ciliate Euplotes encysticus, we analyzed variation in the abundance of the proteins isolated from the resting cyst comparing with proteins in the vegetative cell. 2-D electrophoresis, MALDI-TOF MS techniques and Bioinformatics were used for proteome separation, quantification and identification. The comparative proteomics studies revealed 26 proteins with changes on the expression in the resting cysts, including 12 specific proteins and 14 differential proteins. 12 specific proteins and 10 out of the 14 differential proteins were selected and identified by MALDI-TOF MS. The identified specific proteins with known functions included type II cytoskeletal 1, keratin, Nop16 domain containing protein, protein arginine n-methyltransferase, epsilon-trimethyllysine hydroxylase and calpain-like protein. The identified differential proteins with known functions included Lysozyme C, keratinocyte growth factor, lysozyme homolog AT-2, formate acetyltransferase, alpha S1 casein and cold-shock protein. We discussed the functions of these proteins as well as their contribution in the process of encystment. These identified proteins covered a wide range of molecular functions, including gene regulation, RNA regulation, proteins degradation and oxidation resistance, stress response, material transport and cytoskeleton organization. Therefore, differential expression of these proteins was essential for cell morphological and physiological changes during encystment. This suggested that the peculiar proteins and differential proteins might play important roles in the process of the vegetative cells transforming into the resting cysts. These observations may be novel findings that bring new insights into the detailed mechanisms of dormancy.

  10. Differentiation of Naegleria fowleri from Acanthamoeba species by using monoclonal antibodies and flow cytometry.

    Flores, B M; Garcia, C A; Stamm, W E; Torian, B E

    1990-01-01

    Monoclonal antibodies to Naegleria fowleri and Acanthamoeba polyphaga were analyzed by enzyme-linked immunosorbent assay, indirect immunofluorescence microscopy, and fluorescence flow cytometry to assess specificity and cross-reactivity with axenically cultured N. fowleri and Acanthamoeba spp. Four monoclonal antibodies to N. fowleri were specific for N. fowleri and had no reactivity to A. polyphaga. Similarly, four monoclonal antibodies to A. polyphaga did not react with N. fowleri. Two of t...

  11. Distribution of Acanthamoeba Genotypes Isolated from Recreational and Therapeutic Geothermal Water Sources in Southwestern Iran

    Niyyati, Maryam; Saberi, Reza; Latifi, Alireza; LASJERDI, Zohreh

    2016-01-01

    A comprehensive survey was conducted along 10 km of geothermal rivers in southwestern Iran. A total of 40 water samples were tested for the presence of Acanthamoeba spp., and genotypes were determined by targeting the diagnostic fragment 3 region of the 18S rRNA gene. The pathogenic potential of all positive isolates was also identified using tolerance ability test. High occurrences of Acanthamoeba (50%) were detected in the sampling areas. Based on sequencing analysis, isolates belonging to ...

  12. Detection of Acanthamoeba and Toxoplasma in River Water Samples by Molecular Methods in Iran.

    Mohammad Reza Mahmoudi

    2015-06-01

    Full Text Available Free-living amoebae such as Acanthamoeba species may act as carriers of Cryptosporidium and Toxoplasma oocysts, thus, may play an important role in the water-borne transmission of these parasites. In the present study, a loop mediated isothermal amplification (LAMP method for detection of Toxoplasma and a PCR assay were developed for investigation of Acanthamoeba in environmental water samples.A total of 34 samples were collected from the surface water in Guilan Province. Water samples were filtrated with membrane filters and followed by DNA extraction. PCR and LAMP methods used for detection of the protozoan parasites Acanthamoeba and Toxoplasma respectively.Totally 30 and 2 of 34 samples were positive for Acanthamoeba and Toxoplasma oocysts respectively. Two samples were positive for both investigated parasites.The investigated water supplies, are contaminated by Toxoplasma and Acanthamoeba (oocystes. Acanthamoeba may play an important role in water-borne transmission of Toxoplasma in the study area. For the first time in Iran, protocol of LAMP method was used effectively for the detection of Toxoplasma in surface water samples in Iran.

  13. Presence of Acanthamoeba spp.in water purification plants in southern England

    Shanmuganathan V; Khan NA

    2009-01-01

    Objective:To identify the prevalence of Acanthamoeba in drinking water treatment plants during the course of the purification processes.Methods:Samples were taken from two drinking water purification plants and moni-tored for the presence of Acanthamoeba in order to estimate the removal capacity of treatment methods em-ployed.Water samples were collected at each step in the purification,during the one year survey,and ana-lysed for the presence of Acanthamoeba spp.by plating on bacterial-seeded plates.Results:The results showed that amoebae were present in surface raw waters in 100 % of the samples tested.Acanthamoeba spp.were iso-lated from 71 % and 57 % of the water samples collected from post flat-bottom clarifier 1 and post-sedimenta-tion plant respectively.Considering the outflow drinking waters,the removal capacity was 100 % in both puri-fication plants monitored.The occurrence of Acanthamoeba was not associated with seasonality.Conclusion:These findings confirm that water purification plants employing methods of flocculation,sedimentation,and fil-tration in combination with activated charcoal filtration,ozonisation and chlorination exhibited sufficient Acan-thamoeba removal capacity and the presence of amoebae in the tap water may be due to older plumbing,water storage tanks,tap water hygiene,and /or environmental settings.

  14. Isolation of Acanthamoeba isolates belonging to T2, T3, T4 and T7 genotypes from environmental samples in Ankara, Turkey

    Kiliç, A.; Tanyuksel, M.; Sissons, J.; Jayasekera, S.; Khan, Naveed Ahmed

    2004-01-01

    Acanthamoeba keratitis is a blinding infection that is becoming increasingly important in human health. Early diagnosis is a prerequisite for successful treatment and requires identification of Acanthamoeba at the genotypic level. The genus Acanthamoeba consists of both pathogenic and non-pathogenic species and has been recently classified into 13 different genotypes, T1-T12 and T14. More importantly, 95% of Acanthamoeba isolates that produce keratitis belong to T4 genotypes. In this study, w...

  15. [Use of the presence of cellulose in cellular wall of Acanthamoeba cysts for diagnostic purposes].

    Derda, Monika; Hadaś, Edward

    2009-01-01

    Species identification within the genus Acanthamoeba is based predominantly on morphological and biochemical features. It is labor-intensive and requires cloning and axenization. We described a novel immunocytochemical method for the identification of Acanthamoeba spp. based on selective binding of Clostridium cellulovorans cellulase to protozoan cyst wall cellulose. Free-living amoebae isolated from different water sources by filtration and subsequent cultivation on non-nutrient agar were assigned to genera Acanthamoeba, Naegleria or Hartmannella using morphological taxonomic criteria. Tissues samples from experimentally infected mice were fixed in formalin and for sectioning embedded in paraffin or snap frozen. The Cellulose-Binding Domain of C. cellulovorans cellulase (CBD) obtained as a recombinant protein, were coupled to the fluorescent dye using Alexa Fluor350, 488, 568 - Protein Labelling Kit or labelled with the biotin using EZ-Link Sulfo-NHS-Biotin. All coupling procedures were performed according to the methods provided by manufacturers. For staining with CBD conjugate, slides containing cysts collected from the agar plates or tissue sections were immersed with PBS and incubated with CBD for 30 min at room temperature, washed 3 times with PBS. For staining with CBD-biotin slides containing cysts were incubated with biotinylated CBD for 30 min at room temperature. Subsequent washings in changes of PBS were followed by the incubation with Strept ABComplex/HRP, for 30 min at room temperature, than 3,3 diaminobenzidine tetrahydrochloride was added for 15 min. Slides were rinsed with water, dried and examined in the light microscope. We showed that cellulose could be easily detected by immunofluorescence using conjugated CBD in the inner cyst wall of Acanthamoeba spp. The reference strains of Acanthamoeba spp. and all Acanthamoeba strains isolated from water and from tissues of infected animals gave positive reaction. CBD prepared as a biotynylated protein

  16. Acanthamoeba Keratitis Presenting as Dendritic Keratitis in a Soft Contact Lens Wearer

    Edward Yip Yeung

    2002-03-01

    Full Text Available Acanthamoeba keratitis is a rare cause of corneal infection in Taiwan, which can resultin devastating visual outcomes. A 37-year-old woman, who wore soft contact lenses, sufferedfrom severe pain in her left eye. Biomicroscopy revealed dendritic keratitis, radialkeratoneuritis, and fine keratic precipitates on her cornea. Culture, using non-nutrient agarplate seeded with Escherichia coli, resulted in heavy growth of Acanthamoeba. The inpatienttreatment, including topical neomycin-polymyxin B and metronidazole (0.5% eyedrops,oral ketoconazole, and then oral prednisolone, successfully controlled the cornealinfection. The best-corrected visual acuity was 0.9 without any evidence of recurrence ofinfection after 21 months of follow up. Acanthamoeba keratitis can present as dendritic keratitis,which mimics herpes simplex infection, thus, delays appropriate treatment. Early diagnosisand judicious treatment are essential for restoring the vision and avoiding the subsequentneed of penetrating keratoplasty.

  17. Validation of Real-Time PCR for Laboratory Diagnosis of Acanthamoeba Keratitis▿

    Thompson, Paul P.; Kowalski, Regis P.; Shanks, Robert M. Q.; Gordon, Y. Jerold

    2008-01-01

    Confirmation of Acanthamoeba keratitis by laboratory diagnosis is the first step in the treatment of this vision-threatening disease. Two real-time PCR TaqMan protocols (the Rivière and Qvarnstrom assays) were developed for the detection of genus-specific Acanthamoeba DNA but lacked clinical validation. We have adapted these assays for the Cepheid SmartCycler II system (i) by determining their real-time PCR limits of detection and amplification efficiencies, (ii) by determining their ability ...

  18. Failure of chemotherapy in the first reported cases of Acanthamoeba keratitis in Pakistan

    Siddiqui, Ruqaiyyah; Chaudhry, Tanveer; Lakhundi, Sahreena; Ahmad, Khabir; Khan, Naveed Ahmed

    2014-01-01

    Acanthamoeba keratitis is a painful and progressive infection of the cornea that can result in loss of vision. Here, for the first time in Pakistan, we report two cases of Acanthamoeba keratitis. The first patient was a 37-year-old female who presented with severe itching, redness, pain, along with loss of vision. The patient was a regular soft contact lens wearer. The second patient was a 25-year-old female who had been using soft contact lenses for the past two years. She presented with a b...

  19. Eukaryotic cell encystation and cancer cell dormancy: is a greater devil veiled in the details of a lesser evil?

    Baig, Abdul Mannan; Khan, Naveed Ahmed; Abbas, Farhat

    2015-01-01

    Cancer cell dormancy is the main cause of cancer recurrence and failure of therapy as dormant cells evade not only the anticancer drugs but also the host immune system. These dormant cells veil themselves from detection by imaging and/or using biomarkers, which imposes an additional problem in targeting such cells. A similar form of hibernation process known as encystation is studied in detail for pathogenic unicellular eukaryotic microorganisms. By examination using microarray gene expressio...

  20. Acanthamoeba genotype T4 from the UK and Iran and isolation of the T2 genotype from clinical isolates

    A.H. Maghsood; Sissons, J.; Rezaian, M.; Nolder, D; Warhurst, D.; Khan, Naveed Ahmed

    2005-01-01

    The majority of the keratitis-causing Acanthamoeba isolates are genotype T4. In an attempt to determine whether predominance of T4 isolates in Acanthamoeba keratitis is due to greater virulence or greater prevalence of this genotype, Acanthamoeba genotypes were determined for 13 keratitis isolates and 12 environmental isolates from Iran. Among 13 clinical isolates, eight (61.5 %) belonged to T4, two (15.3 %) belonged to T3 and three (23 %) belonged to the T2 genotype. In contrast, the majorit...

  1. Eukaryotic cell encystation and cancer cell dormancy: is a greater devil veiled in the details of a lesser evil?

    Baig, Abdul Mannan; Khan, Naveed Ahmed; Abbas, Farhat

    2015-01-01

    Cancer cell dormancy is the main cause of cancer recurrence and failure of therapy as dormant cells evade not only the anticancer drugs but also the host immune system. These dormant cells veil themselves from detection by imaging and/or using biomarkers, which imposes an additional problem in targeting such cells. A similar form of hibernation process known as encystation is studied in detail for pathogenic unicellular eukaryotic microorganisms. By examination using microarray gene expression profiles, immunocytochemistry tools, and siRNAs during the process of encystation, understanding the covert features of cancer cell dormancy as proposed could be possible. This knowledge can be extended to dormant cancer cells to uncover the mechanisms that underlie this ghost, yet dangerous state of human cancers. We propose a strategy to induce dormancy and exit this state by application of knowledge gained from the encystation induction and retrieval processes in pathogenic eukaryotic microorganisms. Given that early detection and characterization of dormant malignant tumor cells is important as a general strategy to monitor and prevent the development of overt metastatic disease, this homology may enable the design of therapies that could either awake the dormant cell from dormancy to make it available for therapies or prolong such a phase to make cancer appear as a chronic disease. PMID:25859414

  2. Isolation and identification of Acanthamoeba spp. from thermal swimming pools and spas in Southern Brazil.

    Fabres, Laura Fuhrich; Rosa Dos Santos, Sayonara Peixoto; Benitez, Lisianne Brittes; Rott, Marilise Brittes

    2016-06-01

    Free-living amoebae (FLA) are widely distributed in soil and water. A few number of them are implicated in human disease: Acanthamoeba spp., Naegleria fowleri, Balamuthia mandrillaris and Sappinia diploidea. Species of Acanthamoeba can cause keratitis and brain infections. In this study, 72 water samples were taken from both hot tubs and thermal swimming pools in the city of Porto Alegre, RS, Brazil, to determine the presence of Acanthamoeba in the water as well as perform the phenotypic and genotypic characterization of the isolates. The identification of the isolates was based on the cysts morphology and PCR amplification using genus-specific oligonucleotides. When the isolates were submitted to PCR reaction only 8 were confirmed as belonging to the genus Acanthamoeba. The sequences analysis when compared to the sequences in the GenBank, showed genotype distribution in group T3 (12,5%), T5 (12,5%), T4 (25%) and T15 (50%). The results of this study confirmed the presence of potentially pathogenic isolates of free living amoebae in hot swimming pool and spas which can present risks to human health. PMID:27078644

  3. Molecular Identification and Sequencing of Mannose Binding Protein (MBP Gene of Acanthamoeba palestinensis

    M Rezaeian

    2010-02-01

    Full Text Available "nBackground: Acanthamoeba keratitis develops by pathogenic Acanthamoeba such as A. pal­es­tinen­sis. Indeed this species is one of the known causative agents of amoebic keratitis in Iran. Mannose Binding Protein (MBP is the main pathogenicity factors for developing this sight threatening disease. We aimed to characterize MBP gene in pathogenic Acanthamoeba isolates such as A. palestinensis."nMethods: This experimental research was performed in the School of Public Health, Tehran University of Medical Sciences, Tehran, Iran during 2007-2008.  A. palestinensis was grown on 2% non-nutrient agar overlaid with Escherichia coli. DNA extraction was performed using phenol-chloroform method. PCR reaction and amplification were done using specific primer pairs of MBP. The amplified fragment were purified and sequenced. Finally, the obtained fragment was deposited in the gene data bank."nResults: A 900 bp PCR-product was recovered after PCR reaction. Sequence analysis of the purified PCR product revealed a gene with 943 nucleotides. Homology analysis of the ob­tained sequence showed 81% similarity with the available MBP gene in the gene data bank. The fragment was deposited in the gene data bank under accession number EU678895"nConclusion: MBP is known as the most important factor in Acanthamoeba pathogenesis cas­cade. Therefore, characterization of this gene can aid in developing better therapeutic agents and even immunization of high-risk people.

  4. Free-living Amoebae (FLA: morphological and molecular identification of Acanthamoeba in dental unit water

    Trabelsi H.

    2010-03-01

    Full Text Available The aim of our study was to detect free-living Amoebae (FLA by morphological methods and to identify Acanthamoeba spp. by PCR in the dental unit water lines (DUWL. Materials and methods: it was a prospective study dealing with 196 water samples collected from DUWL (94 samples taken in the early morning before materials flush and patient consultations and 102 samples taken after consultations. At the same time, 39 samples from tap water were realized. Results: 135 (69 % samples were positives by the morphological study with morphotypical diversity. The predominant morphotype was the monopodial (39.2 %. 18 strains of Acanthamoeba spp. were detected in DUW (13.3 % and three strains in tap water (10 %. The amplification of 18S rDNA gene of these strains of Acanthamoeba spp. was positive for all samples. Conclusion: the FLA and Acanthamoeba were isolated both in tap water and in dental unit. The amoeba pathogenicity has not been demonstrated after oral or dental contamination; but the presence of intracellular and pathogenic bacteria in the amoeba could be a source of microbiological risks for patients in case of deep dental care or immunodepression. The improvement of this dental unit was necessary by putting a filter of 0.2 microns porosity before the arrival of the water in hand-pieces allowing the limitation of FLA passage.

  5. Acanthamoeba keratitis in a non-contact lens wearer with human immunodeficiency virus

    Hansen, Birgitte Rønde; Kronborg, Gitte

    2003-01-01

    , and was treated with various topical antibiotics and steroids. 13 months after symptom onset the eye was removed owing to serious scarring of cornea and unbearable pain. Microbiological and histopathological examination of the cornea showed Acanthamoeba. In non-contact lens wearers suffering from...

  6. Fatal Granulomatous Amoebic Encephalitis Caused by Acanthamoeba in a Patient With Kidney Transplant: A Case Report.

    Salameh, Ahmad; Bello, Nancy; Becker, Jennifer; Zangeneh, Tirdad

    2015-09-01

    Granulomatous amoebic encephalitis (GAE) due to Acanthamoeba is almost a uniformly fatal infection in immune-compromised hosts despite multidrug combination therapy. We report a case of GAE in a female who received a deceased donor kidney graft. She was treated with a combination of miltefosine, pentamidine, sulfadiazine, fluconazole, flucytosine, and azithromycin. PMID:26280011

  7. Comparative efficacy of moxidectin and ivermectin against hypobiotic and encysted cyathostomes and other equine parasites.

    Xiao, L; Herd, R P; Majewski, G A

    1994-05-01

    Efficacies of moxidectin and ivermectin were compared in four groups of eight ponies with natural parasite infections: placebo (Control), oral moxidectin gel at 0.3 mg kg-1 of body weight (Mox 0.3), oral moxidectin gel at 0.4 mg kg-1 of body weight (Mox 0.4), and oral ivermectin paste at 0.2 mg kg-1 of body weight (Ivermectin). Fecal samples were taken 0 and 2 weeks after treatment. Animals were necropsied and worms were collected 2 weeks after treatment. Moxidectin and ivermectin showed similar efficacy (99%) against adult cyathostomes, Strongylus spp., Triodontophorus spp. and Habronema muscae. Both drugs were also more than 98% effective against luminal cyathostome and Oxyuris equi fourth stage larvae (L4). Neither drug was effective (0-10.1%) against hypobiotic early third stage cyathostome larvae (EL3). Moxidectin was moderately effective (62.6-79.1%) in removing encysted cyathostome late third stage larvae (LL3) and L4, whereas ivermectin was ineffective (0%) against these stages. By contrast, ivermectin was 95.4% effective against Gasterophilus spp. third instar stage, whereas moxidectin was only 0-20.4% effective. PMID:8091622

  8. Habitat diversity and adaptation to environmental stress in encysted embryos of the crustacean Artemia

    Joshua A Tanguay; Reno C Reyes; James S Clegg

    2004-12-01

    Encysted embryos (cysts) of the brine shrimp, Artemia, provide excellent opportunities for the study of biochemical and biophysical adaptation to extremes of environmental stress in animals. Among other virtues, this organism is found in a wide variety of hypersaline habitats, ranging from deserts, to tropics, to mountains. One adaptation implicated in the ecological success of Artemia is p26, a small heat shock protein that previous evidence indicates plays the role of a molecular chaperone in these embryos. We add to that evidence here. We summarize recently published work on thermal tolerance and stress protein levels in embryos from the San Francisco Bay (SFB) of California inoculated into experimental ponds in southern Vietnam where water temperatures are much higher. New results on the relative contents of three stress proteins (hsp70, artemin and p26) will be presented along with data on cysts of A. tibetiana collected from the high plateau of Tibet about 4.5 km above sea level. Unpublished results on the stress protein artemin are discussed briefly in the context of this paper, and its potential role as an RNA chaperone. Interestingly, we show that the substantial tolerance of A. franciscana embryos to ultraviolet (UV) light does not seem to result from intracellular biochemistry but, rather, from their surrounding thick shell, a biophysical adaptation of considerable importance since these embryos receive heavy doses of UV in nature.

  9. Infection in a rat model reactivates attenuated virulence after long-term axenic culture of Acanthamoeba spp

    Carolina De Marco Verissimo

    2013-11-01

    Full Text Available Prolonged culturing of many microorganisms leads to the loss of virulence and a reduction of their infective capacity. However, little is known about the changes in the pathogenic strains of Acanthamoeba after long culture periods. Our study evaluated the effect of prolonged culturing on the invasiveness of different isolates of Acanthamoeba in an in vivo rat model. ATCC strains of Acanthamoeba, isolates from the environment and clinical cases were evaluated. The in vivo model was effective in establishing the infection and differentiating the pathogenicity of the isolates and re-isolates. The amoebae cultured in the laboratory for long periods were less virulent than those that were recently isolated, confirming the importance of passing Acanthamoeba strains in animal models.

  10. Acanthamoeba genotypes T2, T4, and T11 in soil sources from El Hierro island, Canary Islands, Spain.

    Reyes-Batlle, María; Zamora-Herrera, Jonadab; Vargas-Mesa, Alejandro; Valerón-Tejera, Marco Antonio; Wagner, Carolina; Martín-Navarro, Carmen Ma; López-Arencibia, Atteneri; Sifaoui, Ines; Martínez-Carretero, Enrique; Valladares, Basilio; Piñero, José E; Lorenzo-Morales, Jacob

    2016-08-01

    The genus Acanthamoeba includes pathogenic strains which are causative agents of keratitis and encephalitis that often may end fatal in humans and other animals. In the present study, forty soil samples were collected in the island of El Hierro, Canary Islands, Spain, and checked for the presence of Acanthamoeba. Samples were cultivated onto 2 % non-nutrient agar plates seeded with a layer of heat killed Escherichia coli. Amplification by PCR and sequencing of the DF3 region of the 18S rDNA of Acanthamoeba was carried out in order to confirm morphological identification of the amoebae. Furthermore, Acanthamoeba spp. was isolated from 47.5 % of soil samples. Moreover, genotypes T2, T4, and T11 were identified in these samples. To the best of our knowledge, this is the first study to establish genotypes T2, T4, and T11 in soil sources from El Hierro island. PMID:27075307

  11. Acanthamoeba spp. in domestic tap water in houses of contact lens wearers in the metropolitan area of Mexico City.

    Bonilla-Lemus, Patricia; Ramírez-Bautista, Gerardo A; Zamora-Muñoz, Claudia; Ibarra-Montes, María Del Rocío; Ramírez-Flores, Elizabeth; Hernández-Martínez, María Dolores

    2010-09-01

    A survey was carried out in the metropolitan area of Mexico City to determine the presence of Acanthamoeba in the tap water of houses of contact lens wearers. Water samples were taken from the mains water entry, bathroom sinks and storage containers (roof tanks, cisterns) of 27 houses; and from the solution contained in the contact lens cases. Samples were filtered and cultured onto NNE medium. The isolates were identified based on their morphological features and pathogenicity. Total and fecal coliforms, water temperature, pH, dissolved oxygen and residual free-chlorine were measured by standard methods. Forty five isolates of Acanthamoeba from 200 water samples were obtained. The highest number of amoebae was isolated from cisterns and roof tanks. Most Acanthamoeba isolates were non-pathogenic, however, their presence in tap water is a potential hazard since some species can cause Acanthamoeba keratitis and granulomatous amoebic encephalitis. PMID:19995560

  12. Hypoxia attenuates inflammatory mediators production induced by Acanthamoeba via Toll-like receptor 4 signaling in human corneal epithelial cells

    Pan, Hong [Department of Ophthalmology, Qilu Hospital, Shandong University, 107, Wenhua Xi Road, Jinan 250012 (China); The Key Laboratory of Cardiovascular Remodeling and Function Research, Chinese Ministry of Education and Chinese Ministry of Public Health, Qilu Hospital, Shandong University, 107, Wenhua Xi Road, Jinan 250012 (China); Wu, Xinyi, E-mail: xywu8868@163.com [Department of Ophthalmology, Qilu Hospital, Shandong University, 107, Wenhua Xi Road, Jinan 250012 (China)

    2012-04-13

    Highlights: Black-Right-Pointing-Pointer Hypoxia attenuates Acanthamoeba-induced the production of IL-8 and IFN-{beta}. Black-Right-Pointing-Pointer Hypoxia inhibits TLR4 expression in a time-dependent manner in HCECs. Black-Right-Pointing-Pointer Hypoxia inhibits Acanthamoeba-induced the activation of NF-{kappa}B and ERK1/2 in HCECs. Black-Right-Pointing-Pointer Hypoxia decreases Acanthamoeba-induced inflammatory response via TLR4 signaling. Black-Right-Pointing-Pointer LPS-induced the secretion of IL-6 and IL-8 is abated by hypoxia via TLR4 signaling. -- Abstract: Acanthamoeba keratitis (AK) is a vision-threatening corneal infection that is intimately associated with contact lens use which leads to hypoxic conditions on the corneal surface. However, the effect of hypoxia on the Acanthamoeba-induced host inflammatory response of corneal epithelial cells has not been studied. In the present study, we investigated the effect of hypoxia on the Acanthamoeba-induced production of inflammatory mediators interleukin-8 (IL-8) and interferon-{beta} (IFN-{beta}) in human corneal epithelial cells and then evaluated its effects on the Toll-like receptor 4 (TLR4) signaling, including TLR4 and myeloid differentiation primary response gene (88) (MyD88) expression as well as the activation of nuclear factor kappa-light-chain-enhancer of activated B cells (NF-{kappa}B) and extracellular signal-regulated kinases 1/2 (ERK1/2). We then studied the effect of hypoxia on a TLR4-specific inflammatory response triggered by the TLR4 ligand lipopolysaccharide (LPS). Our data showed that hypoxia significantly decreased the production of IL-8 and IFN-{beta}. Furthermore, hypoxia attenuated Acanthamoeba-triggered TLR4 expression as well as the activation of NF-{kappa}B and ERK1/2, indicating that hypoxia abated Acanthamoeba-induced inflammatory responses by affecting TLR4 signaling. Hypoxia also inhibited LPS-induced IL-6 and IL-8 secretion, myeloid differentiation primary response gene (88

  13. Reporting of T4 Genotype of Acanthamoeba Isolates in Recreational Water Sources of Gilan Province, Northern Iran

    Maryam Niyyati

    2015-02-01

    Full Text Available Background: Acanthamoeba spp. is the causative agent of blindness keratitis and fatal encephalaitis. Presence of Acanthamoeba spp. in a wide variety of niches such as different water types can lead to exposure of high risk people such as contact lens wearers. The main aim of the present study was to explore the occurrence of Acanthamoeba genotypes in the recreational water sources using both morphological and molecular approaches in Gilan province, Iran.Materials and Methods: Overall, 50 samples were collected from recreational water sources including man- made and natural waters in Gilan province. Filtration and cultivation of samples was performed using non-nutrient agar. Cloning of Acanthamoeba spp. was done to eliminate bacterial and fungi contamination. PCR amplification and sequencing were performed using genus-specific primer pair. Genotype identification was based on homology analysis of 18S rRNA gene (DF3 of the obtained sequences with the available genes in the gene bank data base.Results: Out of 50 water samples, 15 (30% were positive for Acanthamoeba trophozoites and cysts according to morphological criteria. Cloning of 13 isolates (26% was done successfully. Molecular analysis of 13 Acanthamoeba strain revealed that all isolates were belonged to potentially pathogenic T4 genotype.Conclusion: T4 genotype is the main cause of Acanthamoeba-related infections. Presence of Acanthamoeba belonged to T4 genotype in recreational water sources is of concern for high risk people. Alarming sign and education to high risk people is of utmost importance to prevent such infections.

  14. Acanthamoeba produces disseminated infection in locusts and traverses the locust blood-brain barrier to invade the central nervous system

    Kirk Ruth; Goldsworthy Graham; Mortazavi Parisa N; Khan Naveed A.

    2010-01-01

    Abstract Background Many aspects of Acanthamoeba granulomatous encephalitis remain poorly understood, including host susceptibility and chronic colonization which represent important features of the spectrum of host-pathogen interactions. Previous studies have suggested locusts as a tractable model in which to study Acanthamoeba pathogenesis. Here we determined the mode of parasite invasion of the central nervous system (CNS). Results Using Acanthamoeba isolates belonging to the T1 and T4 gen...

  15. Acanthamoeba produces disseminated infection in locusts and traverses the locust blood-brain barrier to invade the central nervous system

    Kirk Ruth

    2010-07-01

    Full Text Available Abstract Background Many aspects of Acanthamoeba granulomatous encephalitis remain poorly understood, including host susceptibility and chronic colonization which represent important features of the spectrum of host-pathogen interactions. Previous studies have suggested locusts as a tractable model in which to study Acanthamoeba pathogenesis. Here we determined the mode of parasite invasion of the central nervous system (CNS. Results Using Acanthamoeba isolates belonging to the T1 and T4 genotypes, the findings revealed that amoebae induced sickness behaviour in locusts, as evidenced by reduced faecal output and weight loss and, eventually, leading to 100% mortality. Significant degenerative changes of various tissues were observed by histological sectioning. Both isolates produced disseminated infection, with viable amoebae being recovered from various tissues. Histological examination of the CNS showed that Acanthamoeba invaded the locust CNS, and this is associated with disruption of the perineurium cell/glial cell complex, which constitutes the locust blood-brain barrier. Conclusions This is the first study to demonstrate that Acanthamoeba invades locust brain by modulating the integrity of the insect's blood-brain barrier, a finding that is consistent with the human infection. These observations support the idea that locusts provide a tractable model to study Acanthamoeba encephalitis in vivo. In this way the locust model may generate potentially useful leads that can be tested subsequently in mammalian systems, thus replacing the use of vertebrates at an early stage, and reducing the numbers of mammals required overall.

  16. Macronuclear chromatin structure dynamics in Colpoda inflata (Protista, Ciliophora) resting encystment.

    Tiano, L; Chessa, M G; Carrara, S; Tagliafierro, G; Delmonte Corrado, M U

    1999-01-01

    The chromatin structure dynamics of the Colpoda inflata macronucleus have been investigated in relation to its functional condition, concerning chromatin body extrusion regulating activity. Samples of 2- and 25-day-old resting cysts derived from a standard culture, and of 1-year-old resting cysts derived from a senescent culture, were examined by means of histogram analysis performed on acquired optical microscopy images. Three groups of histograms were detected in each sample. Histogram classification, clustering and matching were assessed in order to obtain the mean histogram of each group. Comparative analysis of the mean histogram showed a similarity in the grey level range of 25-day- and 1-year-old cysts, unlike the wider grey level range found in 2-day-old cysts. Moreover, the respective mean histograms of the three cyst samples appeared rather similar in shape. All this implies that macronuclear chromatin structural features of 1-year-old cysts are common to both cyst standard cultures. The evaluation of the acquired images and their respective histograms evidenced a dynamic state of the macronuclear chromatin, appearing differently condensed in relation to the chromatin body extrusion regulating activity of the macronucleus. The coexistence of a chromatin-decondensed macronucleus with a pycnotic extrusion body suggests that chromatin unable to decondense, thus inactive, is extruded. This finding, along with the presence of chromatin structural features common to standard and senescent cyst populations, supports the occurrence of 'rejuvenated' cell lines from 1-year-old encysted senescent cells, a phenomenon which could be a result of accomplished macronuclear renewal. PMID:10439214

  17. Citologia de impressão no diagnóstico de infecção corneana por Acanthamoeba: relato de caso Diagnosis of Acanthamoeba corneal infection by impression cytology: case report

    Jeison de Nadai Barros

    2007-03-01

    Full Text Available Relatamos três casos de infecção corneana por Acanthamoeba sp em que foi possível detectar cistos do microorganismo com a técnica de citologia de impressão. Três pacientes encaminhados ao Laboratório de Doenças Externas Oculares em 2004 com alterações superficiais da córnea foram submetidos ao exame de citologia de impressão para investigação da presença de cistos de Acanthamoeba sp. Duas amostras foram obtidas da córnea de cada paciente e coradas com PAS, hematoxilina e Papanicolaou. Investigação microbiológica de rotina e cultura também foram realizadas após raspado da córnea. O cultivo das amostras e a citologia de impressão foram positivas para Acanthamoeba sp em todos os pacientes, ao passo que os raspados corados com Giemsa foram positivos em dois casos. A citologia de impressão revelou cistos de Acanthamoeba sp entre feixe de células epiteliais corneanas e como células isoladas. Foram observados cistos no epitélio de um dos pacientes com a citologia de impressão após três meses de tratamento, enquanto o raspado foi negativo. No exame anatomopatológico observaram-se cistos no epitélio e estroma de uma córnea receptora de um dos pacientes após transplante. Neste estudo, a citologia de impressão detectou com sucesso cistos de Acanthamoeba sp em pacientes com acometimento epitelial. Por tratar-se de método não invasivo, a técnica pode ser usada para facilitar o diagnóstico mais precoce da infecção por Acanthamoeba, sendo útil também no acompanhamento do tratamento da doença.To describe three cases of corneal infection due to Acanthamoeba sp in which was possible to detect Acanthamoeba sp cysts by the corneal impression cytology technique. Three patients referred to the External Eye Disease Laboratory in 2004 with superficial corneal alterations were submitted to corneal specimen collection by impression cytology filter paper to investigate the presence of Acanthamoeba sp cysts. Two impression

  18. Stress Management in Cyst-Forming Free-Living Protists: Programmed Cell Death and/or Encystment

    Naveed Ahmed Khan

    2015-01-01

    Full Text Available In the face of harsh conditions and given a choice, a cell may (i undergo programmed cell death, (ii transform into a cancer cell, or (iii enclose itself into a cyst form. In metazoans, the available evidence suggests that cellular machinery exists only to execute or avoid programmed cell death, while the ability to form a cyst was either lost or never developed. For cyst-forming free-living protists, here we pose the question whether the ability to encyst was gained at the expense of the programmed cell death or both functions coexist to counter unfavorable environmental conditions with mutually exclusive phenotypes.

  19. Stress Management in Cyst-Forming Free-Living Protists: Programmed Cell Death and/or Encystment

    Naveed Ahmed Khan; Junaid Iqbal; Ruqaiyyah Siddiqui

    2015-01-01

    In the face of harsh conditions and given a choice, a cell may (i) undergo programmed cell death, (ii) transform into a cancer cell, or (iii) enclose itself into a cyst form. In metazoans, the available evidence suggests that cellular machinery exists only to execute or avoid programmed cell death, while the ability to form a cyst was either lost or never developed. For cyst-forming free-living protists, here we pose the question whether the ability to encyst was gained at the expense of the ...

  20. Stress Management in Cyst-Forming Free-Living Protists: Programmed Cell Death and/or Encystment

    Khan, Naveed Ahmed; Iqbal, Junaid

    2015-01-01

    In the face of harsh conditions and given a choice, a cell may (i) undergo programmed cell death, (ii) transform into a cancer cell, or (iii) enclose itself into a cyst form. In metazoans, the available evidence suggests that cellular machinery exists only to execute or avoid programmed cell death, while the ability to form a cyst was either lost or never developed. For cyst-forming free-living protists, here we pose the question whether the ability to encyst was gained at the expense of the programmed cell death or both functions coexist to counter unfavorable environmental conditions with mutually exclusive phenotypes. PMID:25648302

  1. Distribution of Acanthamoeba Genotypes Isolated from Recreational and Therapeutic Geothermal Water Sources in Southwestern Iran.

    Niyyati, Maryam; Saberi, Reza; Latifi, Alireza; Lasjerdi, Zohreh

    2016-01-01

    A comprehensive survey was conducted along 10 km of geothermal rivers in southwestern Iran. A total of 40 water samples were tested for the presence of Acanthamoeba spp., and genotypes were determined by targeting the diagnostic fragment 3 region of the 18S rRNA gene. The pathogenic potential of all positive isolates was also identified using tolerance ability test. High occurrences of Acanthamoeba (50%) were detected in the sampling areas. Based on sequencing analysis, isolates belonging to T4 (93.7%) and T2 (6.25%) genotypes were reported. Thermo- and osmotolerance tests revealed that five strains are highly pathogenic. Since every collection site of this study was associated with high human activity, posting of warning signs, monitoring of recreational water sources, and awareness of high-risk people are of utmost importance. To the best of our knowledge, the present research is the first to report T2 genotype from geothermal water sources in Iran. PMID:27127409

  2. Identification and properties of proteases from an Acanthamoeba isolate capable of producing granulomatous encephalitis

    Sissons, James; Alsam, Selwa; Goldsworthy, Graham; Lightfoot, Mary; Jarroll, Edward L; Khan, Naveed Ahmed

    2006-01-01

    Background Granulomatous amoebic encephalitis due to Acanthamoeba is often a fatal human disease. However, the pathogenesis and pathophysiology of Acanthamoeba encephalitis remain unclear. In this study, the role of extracellular Acanthamoeba proteases in central nervous system pathogenesis and pathophysiology was examined. Results Using an encephalitis isolate belonging to T1 genotype, we observed two major proteases with approximate molecular weights of 150 KD and 130 KD on SDS-PAGE gels using gelatin as substrate. The 130 KD protease was inhibited with phenylmethylsulfonyl fluoride (PMSF) suggesting that it is a serine protease, while the 150 KD protease was inhibited with 1, 10-phenanthroline suggesting that it is a metalloprotease. Both proteases exhibited maximal activity at neutral pH and over a range of temperatures, indicating their physiological relevance. These proteases degrade extracellular matrix (ECM), which provide structural and functional support to the brain tissue, as shown by the degradation of collagen I and III (major components of collagenous ECM), elastin (elastic fibrils of ECM), plasminogen (involved in proteolytic degradation of ECM), as well as casein and haemoglobin. The proteases were purified partially using ion-exchange chromatography and their effects were tested in an in vitro model of the blood-brain barrier using human brain microvascular endothelial cells (HBMEC). Neither the serine nor the metalloprotease exhibited HBMEC cytotoxicity. However, the serine protease exhibited HBMEC monolayer disruptions (trypsin-like) suggesting a role in blood-brain barrier perturbations. Conclusion Overall, these data suggest that Acanthamoeba proteases digest ECM, which may play crucial role(s) in invasion of the brain tissue by amoebae. PMID:16672059

  3. Identification and properties of proteases from an Acanthamoeba isolate capable of producing granulomatous encephalitis

    Jarroll Edward L

    2006-05-01

    Full Text Available Abstract Background Granulomatous amoebic encephalitis due to Acanthamoeba is often a fatal human disease. However, the pathogenesis and pathophysiology of Acanthamoeba encephalitis remain unclear. In this study, the role of extracellular Acanthamoeba proteases in central nervous system pathogenesis and pathophysiology was examined. Results Using an encephalitis isolate belonging to T1 genotype, we observed two major proteases with approximate molecular weights of 150 KD and 130 KD on SDS-PAGE gels using gelatin as substrate. The 130 KD protease was inhibited with phenylmethylsulfonyl fluoride (PMSF suggesting that it is a serine protease, while the 150 KD protease was inhibited with 1, 10-phenanthroline suggesting that it is a metalloprotease. Both proteases exhibited maximal activity at neutral pH and over a range of temperatures, indicating their physiological relevance. These proteases degrade extracellular matrix (ECM, which provide structural and functional support to the brain tissue, as shown by the degradation of collagen I and III (major components of collagenous ECM, elastin (elastic fibrils of ECM, plasminogen (involved in proteolytic degradation of ECM, as well as casein and haemoglobin. The proteases were purified partially using ion-exchange chromatography and their effects were tested in an in vitro model of the blood-brain barrier using human brain microvascular endothelial cells (HBMEC. Neither the serine nor the metalloprotease exhibited HBMEC cytotoxicity. However, the serine protease exhibited HBMEC monolayer disruptions (trypsin-like suggesting a role in blood-brain barrier perturbations. Conclusion Overall, these data suggest that Acanthamoeba proteases digest ECM, which may play crucial role(s in invasion of the brain tissue by amoebae.

  4. Effect of Parachlamydia acanthamoebae on pulmonary function parameters in a bovine respiratory model.

    Lohr, M; Prohl, A; Ostermann, C; Diller, R; Greub, G; Reinhold, P

    2016-07-01

    The aim of this study was to evaluate pulmonary dysfunction induced by experimental infection with Parachlamydia acanthamoebae in calves. Intrabronchial inoculation with P. acanthamoebae was performed in 31 calves aged 2-3 months old at two different challenge doses of 10(8) and 10(10) inclusion-forming units (IFU) per animal. Control animals received heat inactivated bacteria. The effects on pulmonary gas exchange were determined by arterial blood gas analysis and haemoximetry during the 7 days post inoculation (DPI). For pulmonary function testing (PFT), impulse oscillometry, capnography, and measurement of O2 uptake were undertaken in spontaneously breathing animals 7 and 3 days before inoculation and were repeated until 10 DPI. In the early phase after challenge (1-3 DPI), mild hypoxaemia occurred, which was accompanied by a significant reduction in both tidal and alveolar volumes (each related to bodyweight, BW). In parallel, expiratory flow rate and specific ventilation (i.e. minute ventilation related to O2 uptake) were significantly increased. Minute and alveolar ventilations (each related to metabolic BW) increased significantly due to higher respiratory rates, lasting until 4 and 5 DPI, respectively. Oxygen uptake was slightly reduced during the first 2 days after challenge, but increased significantly during the recovery phase, from 4 to 8 DPI. No deterioration in respiratory mechanics or acid-base balance was observed. Respiratory infection with 10(10) IFU P. acanthamoebae per calf induced mild respiratory dysfunction, mainly characterised by hypoxaemia. The study's findings do not indicate severe pathophysiological consequences of P. acanthamoebae infection on pulmonary function in the bovine host. PMID:27240907

  5. Isolation of Acanthamoeba Spp. from Drinking Waters in Several Hospitals of Iran

    HR Bagheri

    2010-06-01

    Full Text Available Background: Acanthamoeba is an opportunistic amphizoic protozoan found in different wa­ter sources including swimming pool as well as in sewage. The aim of this study was to in­vestigate the prevalence of Acanthamoeba in tap-water samples in Iran.Method: In this descriptive cross-sectional study, 94 samples of cold and warm tap-water were collected from different wards of hospitals in 13 cities of Iran in 2007-2008. Free resi­dual chlorine, pH, and temperature of samples were measured. After filtration through multi­pore nylon membrane, samples were cultured on non-nutrient agar. Then we investigated ex­istence of Acanthamoeba by reverse contrast phase microscope.Results: Acanthamoeba was found in 45 samples (48%. Thirty-four and 11 positive samples were collected from cold and warm tap water, respectively. The samples belonged to the cate­gory of 20-30 °C temperature with 0-2 ppm free residual chlorine and pH 6-7.4 showed the most coincidence to the positive cases. The greatest proportion of positive samples was ob­tained from Mashhad hospitals, while all samples collected from Arak and Semnan hospitals were negative.Conclusion: considering the results of this study and the pathogenic role of this protozoan on pa­tients with immunodeficiency, as well as capability of this microorganism in carrying other pathogens such as Legionella, further studies are needed. What is more important, potable water in hospitals should follow the procedure of treatment and sanitation, in order to prevent the relevant nosocomial infections.

  6. Prevalence of Acanthamoeba and other naked amoebae in South Florida domestic water.

    Shoff, M E; Rogerson, A; Kessler, K; Schatz, S; Seal, D V

    2008-03-01

    The purpose was to identify the prevalence of naked amoebae in tap water in south Florida to ascertain the risk of amoebal infections of the cornea in contact lens wearers. Over the course of a 2-year period, water samples were collected from sites throughout Broward, Palm Beach, and Dade counties, Florida. The presence of amoebae in samples was based on an enrichment cultivation method appropriate for Acanthamoeba. Amoebae were identified using diagnostic features discernable by light microscopy. A total of 283 water samples were processed and amoebae were noted in 80 of these. Acanthamoeba were found on 8 occasions (2.8%). The genera Hartmannella and Vahlkampfia, rarely involved in keratitis cases, were found in 3.5% and 2.8% of samples, respectively. A total of 19 different naked amoebae were recorded and amoebae (regardless of genus) were present in 19.4% of all samples. Previous surveys in England and Korea have shown that acanthamoebae are found in 15 to 30% of tap water samples in the home and have been associated with corneal infection in contact lens wearers. The incidence of acanthamoebae infection in the USA (2.8%) has been found to be lower than that in the UK and it has been postulated that this is related to the lack of a storage water tank in the roof loft space. However, the level of treatment of municipal water is clearly not effective at killing amoebal cysts (or trophozoites) as evidenced by the high occurrence of amoebae (19.4%) in this study. PMID:17998610

  7. Experimental infection of T4 Acanthamoeba genotype determines the pathogenic potential.

    Alves, Daniella de Sousa Mendes Moreira; Moraes, Aline Silva; Alves, Luciano Moreira; Gurgel-Gonçalves, Rodrigo; Lino Junior, Ruy de Souza; Cuba-Cuba, César Augusto; Vinaud, Marina Clare

    2016-09-01

    T4 is the Acanthamoeba genotype most related to cases of granulomatous amoebic encephalitis (GAE) in immunocompromised patients and of keratitis in contact lens wearers. The determination of the pathogenic potential of Acanthamoeba clinical and environmental isolates using experimental models is extremely important to elucidate the capacity of free-living organisms to establish and cause disease in hosts. The aim of this study was to compare and evaluate the histopathology and culture between two different routes of experimental infection of T4 Acanthamoeba isolated from environmental and clinical source in mice (intracranial and intraperitoneal). Swiss isogenic healthy mice were inoculated with 10(4) trophozoites by intracranial (IC) and intraperitoneal (IP) routes and observed during 21 days. The brains from animals inoculated by the IC route were collected and from the animals of the IP inoculation group, the brains, livers, kidneys, spleens, and lungs were removed. The organs were prepared and appropriately divided to be evaluated with histopathology and culture. There was no significant difference between the inoculation routes in terms of isolates recovery (χ(2) = 0.09; p = 0.76). In the IC group, isolate recovery rate was significantly higher in histopathology than the one achieved by culture (χ(2) = 6.45; p protozoa, improving the knowledge on free-living amoebae isolates. PMID:27164833

  8. High occurrence of Acanthamoeba genotype T4 in soil sources from Bolívar State, Venezuela.

    Wagner, Carolina; Reyes-Batlle, María; Hernán, Aurora; Rojas, Elsy; Pérez, Gladymar; López-Arencibia, Atteneri; Sifaoui, Ines; Martínez-Carretero, Enrique; Piñero, José E; Valladares, Basilio; Lorenzo-Morales, Jacob

    2016-09-01

    Pathogenic strains of Acanthamoeba are causative agents of keratitis and encephalitis that often may end fatal in humans and other animals. In the present study, twenty-seven soil samples were collected in the Bolivar State in Venezuela and checked for the presence of Acanthamoeba. Samples were cultivated onto 2% non-nutrient agar plates seeded with a layer of heat killed E. coli. Amplification by PCR and sequencing of the DF3 region of the 18S rDNA of Acanthamoeba was carried out in order to confirm morphological identification of the amoebae. Furthermore, Acanthamoeba spp. was isolated from 51.8% of soil samples. Sequencing of the DF3 region of the 18S rDNA resulted in the identification of genotype T4 in all samples. To the best of our knowledge, this is the first report of genotype T4 in soil sources from Venezuela. Further studies should be carried out in this State and in the country in order to determine the current occurrence of Acanthamoeba in Venezuelan environments. PMID:27447209

  9. Incidence and histopathology of encysted progenetic metacercaria of Clinostomum complanatum (Digenea:Clinostomidae) in Channa punctatus and its development in experimental host

    PA Ahammed Shareef; SMA Abidi

    2012-01-01

    Objective: To study the incidence of encysted progenetic metacercariae of Clinostomum complanatum (C. complanatum) in Channa punctatus (C. punctatus), associated histopathology and the experimental infection to laboratory chicken to obtain ovigerous adult worms. Methods:Live C. punctatus were brought from local fish market of Aligarh, India, dissected and examined on a monthly basis for the presence of C. complanatum cysts. For histochemistry, infected tissue sections with attached cysts were processed for haematoxylene and eosin staining. Cysts were aseptically fed to 4 day old leghorn chicken to obtain adult worms. Mechanically excysted metacercaria and the ovigerous adult worms were stained in carmine to prepare permanent slides. Results: One year survey for the infection of encysted progenetic metacercaria of C. complanatum in C. punctatus revealed the prevalence, intensity and abundance of 24.7%, 2.27 and 0.608, respectively. Histopathology showed heavy infiltration of immune cells at the site of cyst attachment and some tissue damage was also evident. Following feeding to experimental chicken, about 41.07%of the encysted metacercariae were able to excyst and migrate back to bucco-pharyngeal region where they tenaciously attached and fed on blood, and transformed into ovigerous adult worms from 62 hours onwards of post infection. Conclusions:The parasite is potentially pathogenic to the host, and the availability of a suitable intermediate host can be a contributing factor for the occurrence of C. complanatum metacercaria either in the excysted or encysted form, indicating loose host specificity and zoonotic potential.

  10. Direct photoaffinity labeling by nucleotides of the apparent catalytic site on the heavy chains of smooth muscle and Acanthamoeba myosins

    The heavy chains of Acanthamoeba myosins, IA, IB and II, turkey gizzard myosin, and rabbit skeletal muscle myosin subfragment-1 were specifically labeled by radioactive ATP, ADP, and UTP, each of which is a substrate or product of myosin ATPase activity, when irradiated with uv light at 00C. With UTP, as much as 0.45 mol/mol of Acanthamoeba myosin IA heavy chain and 1 mol/mol of turkey gizzard myosin heavy chain was incorporated. Evidence that the ligands were associated with the catalytic site included the observations that reaction occurred only with nucleotides that are substrates or products of the ATPase activity; that the reaction was blocked by pyrophosphate which is an inhibitor of the ATPase activity; that ATP was bound as ADP; and that label was probably restricted to a single peptide following limited subtilisin proteolysis of labeled Acanthamoeba myosin IA heavy chain and extensive cleavage with CNBr and trypsin of labeled turkey gizzard myosin heavy chain

  11. Isolation, morphologic, serologic and molecular identification of Acanthamoeba T4 genotype from the liver of a Temminck's tragopan (Tragopan temminckii).

    Visvesvara, Govinda S; Shoff, Megan E; Sriram, Rama; Booton, Gregory C; Crary, Monica; Fuerst, Paul A; Hanley, Christopher S; Garner, Michael M

    2010-06-24

    Members of the genus Acanthamoeba are usually free-living amoebae that are found in a variety of ecological niches including soil, fresh and brackish water, dust in the air, heating, ventilating, and air conditioning filters, swimming pools and hot tubs. Occasionally they are also known to cause central nervous system infections in humans and animals. We isolated into culture an amoeba from the liver of a Temminck's tragopan (horned pheasant) (Tragopan temminckii) that died of amoebic infection. We identified the infecting amoeba as Acanthamoeba sp. based on culture characteristics, cyst morphology and immunofluorescence assays. Additionally, we identified the amoeba as Acanthamoeba, genotype T4, by sequencing a diagnostic region of the nuclear small subunit ribosomal RNA gene. PMID:20347228

  12. Random amplified polymorphic DNA profiles as a tool for the characterization of Brazilian keratitis isolates of the genus Acanthamoeba

    Alves J.M.P.

    2000-01-01

    Full Text Available The genus Acanthamoeba comprises free-living amebae identified as opportunistic pathogens of humans and other animal species. Morphological, biochemical and molecular approaches have shown wide genetic diversity within the genus. In an attempt to determine the genetic relatedness among isolates of Acanthamoeba we analyzed randomly amplified polymorphic DNA (RAPD profiles of 11 Brazilian isolates from cases of human keratitis and 8 American type culture collection (ATCC reference strains. We found that ATCC strains belonging to the same species present polymorphic RAPD profiles whereas strains of different species show very similar profiles. Although most Brazilian isolates could not be assigned with certainty to any of the reference species, they could be clustered according to pattern similarities. The results show that RAPD analysis is a useful tool for the rapid characterization of new isolates and the assessment of genetic relatedness of Acanthamoeba spp. A comparison between RAPD analyses and morphological characteristics of cyst stages is also discussed.

  13. In vitro anti-Acanthamoeba synergistic effect of chlorhexidine and cationic carbosilane dendrimers against both trophozoite and cyst forms.

    Heredero-Bermejo, I; Sánchez-Nieves, J; Soliveri, J; Gómez, R; de la Mata, F J; Copa-Patiño, J L; Pérez-Serrano, J

    2016-07-25

    Acanthamoeba sp. are the causative agents of severe illnesses in humans such as Acanthamoeba keratitis (AK) and granulomatous amoebic encephalitis (GAE). Medical therapy is not yet well established. Treatments of AK last for several months and generate toxicity, resistances appear due to the cysts stage and recurrences can occur. In this study has been demonstrated that the combination of chlorhexidine digluconate (CLX) and carbosilane dendrimers containing ammonium or guanidine moieties has in vitro synergistic effect against Acanthamoeba polyphaga. This synergy provokes an important reduction in the minimal trophozoite amoebicidal concentration (MTAC) of CLX, which means a reduction of their toxic effects on human cells. Moreover, some CLX/dendrimer combinations show important activity against the cyst resistance stage. PMID:27173821

  14. Isolation and molecular characterization of Acanthamoeba strains isolated from the oral cavity of immunosuppressed individuals in Tehran, Iran.

    Memari, Fatemeh; Niyyati, Maryam; Lorenzo-Morales, Jacob; Jonaydi, Zaynab

    2016-09-01

    Acanthamoeba spp. is an opportunistic protozoan parasite which is the causative agent of granulomatous amoebic encephalitis (GAE) and Acanthamoeba keratitis (AK). GAE usually occurs in immunocompromised patients which in most cases is fatal. The present study was conducted to determine the genotypes of Acanthamoeba isolated from patients with compromised immunological status. For this purpose, 90 samples from the oral cavity of these individuals were collected in different hospitals of Tehran, Iran using sterile cotton swabs. Samples were cultured in 2% Non-Nutrient Agar (NNA) plates in order to check for the presence of amoebae. Identification of isolates was carried out using both morphological and molecular tools. The pathogenic potential of the obtained strains was assessed by performing osmo- and thermotolerance assays as previously described. Genotyping of the isolates was carried out by PCR/sequencing of the DF3 region of the 18S rDNA gene of Acanthamoeba. From the 90 collected samples, 11 (13.4%) were positive for Acanthamoeba genus. Molecular analysis revealed the presence of genotypes T3, T4 and T11, although most of the isolates belonged to genotype T4. Only 3 of the isolates genotyped as T4 were positive for the pathogenic potential assays. To this end if the immunological status is considered as one of the key factors for the development of GAE due to Acanthamoeba in the previous reported cases, individuals suffering from the conditions mentioned in this study should be considered as a high risk group of population in Iran and worldwide. PMID:27447206

  15. Compositions and methods for pathogen transport

    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.

  16. Results of case-control studies support the association between contact lens use and Acanthamoeba keratitis

    Pacella E

    2013-05-01

    Full Text Available Elena Pacella,1 Giuseppe La Torre,2 Maria De Giusti,2 Chiara Brillante,1 Anna Maria Lombardi,2 Gianpaolo Smaldone,1 Tommaso Lenzi,1 Fernanda Pacella11Department of Sense Organs, Faculty of Medicine and Dentistry, Sapienza University of Rome, Italy; 2Department of Public Health and Infectious Diseases, Faculty of Pharmacy and Medicine, Sapienza University of Rome, ItalyBackground: Acanthamoeba keratitis (AK is ever more frequently reported in industrialized countries. The loss of the corneal surface integrity consequent to secondary microtrauma produced by the use of contact lens (CL favors the penetration of the parasite into the corneal tissue.Objectives: A scientific review was performed to investigate the association of CL wear as an Acanthamoeba keratitis (AK risk factor.Methods: A computerized screening of 7834 Medline articles (4623 from PubMed; 3211 from Scopus used a strict selection criteria of case-control studies involving CL wear and/or trauma.Results: The search yielded five case-control studies published from 1995 to 2012. All studies included showed a statistically significant positive association between AK and CL use, with a combined odds ratio (OR of 10.21 (95%, confidence intervals [CI]; 3.57–27.64.Statistical analysis: All studies included showed a statistically significant positive association between AK and CL use, though with differing OR values.Conclusion: Though rare, AK should be held in higher consideration when ophthalmologists are faced with CL users exhibiting simplex-like lesions associated with circular stromal infiltrates and disproportionate ocular pain in respect to the objective clinical picture.Keywords: keratitis, contact lens, Acanthamoeba

  17. A novel antiamoebic agent against Acanthamoeba sp. - A causative agent for eye keratitis infection

    Kusrini, Eny; Hashim, Fatimah; Azmi, Wan Nor Nadhirah Wan Noor; Amin, Nakisah Mat; Estuningtyas, Ari

    2016-01-01

    The terbium trinitrate.trihydrate.18-crown ether-6, Tb(NO3)3(OH2)3.(18C6) complex has been characterized by elemental analysis, photoluminescence and single X-ray diffraction. The IC50 values were determined based on MTT assay while light and fluorescence microscopy imaging were employed to evaluate the cellular morphological changes. Alkaline comet assay was performed to analyze the DNA damage. The photoluminescence spectrum of the Tb complex excited at 325 nm displayed seven luminescence peaks corresponding to the 5D4 → 7F0, 1, 2, 3, 4, 5, 6 transitions. The cytotoxicity and genotoxicity studies indicated that the Tb(NO3)3(OH2)3.(18C6) complex and its salt form as well as the 18C6 molecule have excellent anti-amoebic activity with very low IC50 values are 7, 2.6 and 1.2 μg/mL, respectively, with significant decrease (p < 0.05) in Acanthamoeba viability when the concentration was increased from 0 to 30 μg/mL. The mode of cell death in Acanthamoeba cells following treatment with the Tb complex was apoptosis. This is in contrast to the Tb(NO3)3.6H2O salt- and 18C6 molecule-treated Acanthamoeba, which exhibited necrotic type cells. The percentage of DNA damage following treatment with all the compounds at the IC25 values showed high percentage of type 1 with the % nuclei damage are 14.15 ± 2.4; 46.00 ± 4.2; 36.36 ± 2.4; 45.16 ± 0.6%, respectively for untreated, treated with Tb complex, Tb salt and 18C6 molecule. The work features promising potential of Tb(NO3)3(OH2)3.(18C6) complex as anti-amoebic agent, representing a therapeutic option for Acanthamoeba keratitis infection.

  18. Acanthamoeba Species Keratitis in a Soft Contact Lens Wearer Molecularly Linked to Well Water

    Samira Mubareka

    2006-01-01

    Full Text Available Acanthamoeba species keratitis has been associated with soft contact lens wear. In the present report, an epidemiological link was established between the patient's isolate and well water from the home using molecular methods. To the authors' knowledge, this is the first case in Canada where such a link has been established. Primary care practitioners and specialists, including ophthalmologists and infectious diseases specialists, must maintain a high degree of clinical suspicion in soft contact lens wearers with keratitis unresponsive to conventional topical and systemic treatment.

  19. In vitro cytotoxicity of Acanthamoeba spp. isolated from contact lens containers in Korea by crystal violet staining and LDH release assay

    Shin, Ho-Joon; Cho, Myung-Soo; Jung, Suk-Yul; Kim, Hyung-Il; Im, Kyung-il

    2000-01-01

    In order to observe the cytotoxicity of Acanthamoeba spp., which were isolated from contact lens containers as ethiological agents for the probable amoebic keratitis in Korea, the crystal violet staining method and LDH release assay were carried out. In the crystal violet staining method, among eight contact lens container isolates, isolate 3 (Acanthamoeba KA/LS5) showed 83.6% and 81.8% of cytotoxicity, and isolate 7 (Acanthamoeba KA/LS37) showed 28.2% and 25.1% of cytotoxicity, in 1 mg/ml an...

  20. Eukar yotic cell encystation and cancer cell dormancy:is a greater devil veiled in the details of a lesser evil?

    Abdul Mannan Baig; Naveed Ahmed Khan; Farhat Abbas

    2015-01-01

    Cancer cell dormancy is the main cause of cancer recurrence and failure of therapy as dormant cells evade not only the anticancer drugs but also the host immune system. hTese dormant cells veil themselves from detection by imaging and/or using biomarkers, which imposes an additional problem in targeting such cells. A similar form of hibernation process known as encystation is studied in detail for pathogenic unicellular eukaryotic microorganisms. By examination using microarray gene expression proifles, immunocytochemistry tools, and siRNAs during the process of encystation, understanding the covert features of cancer cell dormancy as proposed could be possible. hTis knowledge can be extended to dormant cancer cells to uncover the mechanisms that underlie this ghost, yet dangerous state of human cancers. We propose a strategy to induce dormancy and exit this state by application of knowledge gained from the encystation induction and retrieval processes in pathogenic eukaryotic microorganisms. Given that early detection and characterization of dormant malignant tumor cells is important as a general strategy to monitor and prevent the development of overt metastatic disease, this homology may enable the design of therapies that could either awake the dormant cell from dormancy to make it available for therapies or prolong such a phase to make cancer appear as a chronic disease.

  1. A Case of Medication-Resistant Acanthamoeba Keratitis Treated by Corneal Crosslinking in Turkey

    Goktug Demirci

    2013-01-01

    Full Text Available Purpose. To report a case of medication-resistant acanthamoeba keratitis (AK treated successfully by corneal crosslinking (CXL. Methods. A 26-year-old male with medication-resistant AK underwent a standard CXL procedure with local anesthesia, followed by central corneal epithelial debridement, application of riboflavin 0.1%, and UV-A irradiation. Results. The patient experienced a dramatic symptomatic improvement within 24 hours. At two months, keratitis was healed with a semitransparent paracentral scar that did not affect visual acuity. Conclusions. Our experience, considered in the context of recent studies, suggests that CXL may be an option for selected patients with medication-resistant AK and corneal melting. CXL allows patients to avoid emergency keratoplasty and experience rapid symptomatic relief.

  2. Progress on excretory-secretory antigens of Trichinella spiralis pre-encysted larva%旋毛虫成囊前期幼虫排泄分泌抗原的研究进展

    董明治; 申丽洁

    2009-01-01

    在旋毛虫感染过程中,成囊前期幼虫(PEL)是旋毛虫侵入宿主肌肉的侵入期.旋毛虫PEL比成囊期幼虫(EL)在宿主体内约早2周出现.成囊前期幼虫抗原(PELA)对旋毛虫病的早期免疫学诊断具有较高的敏感性.而旋毛虫排泄分泌抗原具双重免疫功能,即有良好的抗原性和免疫原性.因此,本文就旋毛虫成囊前期幼虫ES抗原研究进展做一综述.%The pre-encysted larvae is the period of invasive hosts muscle in the development of Trichinella spiralis. T. spiralis pre-encysted larvae than encysted larva appeared about two weeks early in hosts. The pre-encysted larva antigens of T. spiralis have high sensitivity in the eary diagnosis of immunization. And excretory-secretory antigen of T. spiralis has double immune function, that is good antigenicity and immunogenicity. Therefore, this article introduced the progress on excretory-secretory antigens of T. spiralis pre-encysted larva.

  3. Lens Epithelial Cell Death Secondary to Acanthamoeba Keratitis: Absence of Capsular Bag Opacification Six Years after Cataract Surgery

    Moreno-Montañes, J. (Javier); Barrio-Barrio, J. (Jesús); E. Nova; Werner, L.

    2011-01-01

    Purpose: To show the evolution of anterior chamber structures 6 years after cataract surgery in a case with Acanthamoeba keratitis (AK). Methods: A 37-year-old woman with AK receiving long-term treatment with chlorhexidine, propamidine isethionate and steroids developed a white cataract and iris atrophy. Penetrating keratoplasty and cataract surgery were performed with subsequent intraocular pressure elevation requiring Molteno shunt implantation. Two years after the last surgery, endothel...

  4. A study of the spectrum of Acanthamoeba keratitis: A three-year study at a tertiary eye care referral center in South India

    Bharathi Jayahar

    2007-01-01

    Full Text Available Purpose: To determine the epidemiological and clinical characteristics of Acanthamoeba keratitis and also to determine the sensitivity and specificity of smears in the detection of Acanthamoeba . Materials and Methods: A retrospective review of all culture-positive cases of Acanthamoeba keratitis seen between October 1999 and August 2002 was performed. Corneal scrapes were subjected to culture and microscopy using standard protocols. Results: Out of 3183 consecutive patients with clinically diagnosed corneal ulcers evaluated, 33 (1.04% were found to be due to Acanthamoeba . Twenty-four out of 33 (72.72% were less than 51 years of age ( P < 0.001. All patients were from rural areas ( P < 0.001 and 26 (78.79% of them were agricultural workers ( P = 0.031. All 33 had history of corneal injury ( P < 0.001 and 28 (84.85% patients had injury with mud ( P < 0.001. All 33 (100% patients had previous medical treatment ( P =0.009 and 10 (30.3% had used traditional eye medicines ( P =0.183. A clinical pattern of ring infiltrate was characteristic in 15 (45.45% patients. The diameter of the corneal ulcer was more than 6mm in 27 (81.82% eyes ( P < 0.001. Twenty-six (78.79% patients had visual acuity of perception of light on initial presentation ( P < 0.001 and 24 (72.73% had the same as their final visual outcome. The sensitivity of 10% potassium hydroxide (KOH preparation was found to be higher ( P < 0.001 in the detection of Acanthamoeba cysts. Conclusion: The incidence of Acanthamoeba keratitis amongst the corneal ulcer patients was 1% in this setting and it was mainly due to corneal injury by mud. The KOH preparation is a sensitive diagnostic tool for the detection of Acanthamoeba . Delayed diagnosis or misdiagnosis and inappropriate antimicrobial therapy results in poor visual outcome.

  5. Comparison of moxidectin oral gel and ivermectin oral paste against a spectrum of internal parasites of ponies with special attention to encysted cyathostome larvae.

    Monahan, C M; Chapman, M R; Taylor, H W; French, D D; Klei, T R

    1996-06-01

    Two dosages of moxidectin oral gel were evaluated and compared to a therapeutic dose of ivermectin oral paste in the control of a spectrum of gastrointestinal parasites of ponies naturally infected in southern Louisiana or Mississippi. Thirty-two mixed-breed ponies ranging in age from one to 21 years were used in this controlled test. Eight weeks prior to the experiment, ponies grazing on contaminated pasture were moved to a paddock and fed a pelleted ration, thus reducing or eliminating the potential for additional infection and ensuring the existence of a population of encysted larvae. Ponies were then allocated to replicates of four animals based on values of fecal strongyle egg counts and percent strongyle larvae composition determined from Baermann sedimentations of fecal cultures. Members of replicates were allocated to one of four treatment groups: moxidectin oral gel administered at 300 micrograms kg-1 body weight, moxidectin oral gel at 400 micrograms kg-1, the oral gel vehicle as negative control, and ivermectin oral paste at 200 micrograms kg-1. Prior to treatment, ponies were confined in pairs to covered concrete runs by treatment group. Two weeks following treatment, necropsy examinations of all animals were performed. Parasites were recovered from the lumen of the stomach, the intestinal tract, the cranial mesenteric artery and its major branches, the peritoneal body wall and from pepsin digests of mucosal scrapings taken from the cecum and large colon. Encysted cyathostome larval burdens were also compared using mural transillumination of segments of the large colon for visualization of the encysted forms. Control ponies were not uniformly infected with the spectrum of parasites; however, moxidectin, at either dosage, compared favorably with ivermectin in the control of the adults of Strongylus vulgaris, Strongylus edentatus, Triodontophorus spp., Oesophagodontus robustus, Trichostrongylus axei, Oxyuris equi, Parascaris equorum, Habronema muscae, as

  6. Collection of pre-encysted larvae of Trichinella spiralis%旋毛虫成囊前期幼虫的收集

    董明治; 申丽洁

    2010-01-01

    目的 探索旋毛虫成囊前期幼虫的收集时间与方法.方法 20只成年大鼠,每只大鼠经口感染3000条旋毛虫脱囊幼虫,分别于第14、15、16、17、18、19、20天将大鼠处死,用人工胃蛋白酶消化肌肉收集旋毛虫.结果 感染后第14、15、16大膈肌均未见旋毛虫,第17、18、19、20天膈肌有旋毛虫的侵入并逐渐增多;第14、15、16天均未收集到旋毛虫,第17、18、19、20天平均每只大鼠分别收集5000、8000、10000、30000条旋毛虫.结论 旋毛虫在感染后第20天是旋毛虫成囊前期幼虫收集的最佳时期.人工胃蛋白酶消化肌肉可以收集成囊前期幼虫.%Objective To explore the collection time and method of the pre-encysted larvae of Trichinella spiralis. Methods Twenty adult rats were inoculated with 3 000 encysted larva of Trichinella spir-alis per rat by the oral route. At the 14th, 15th, 16th, 17th, 18th, 19th, 20th day after infection, the rats were killed and the muscles were digested with pepsin to collect Trichinella spiralis. Results At the 14th,15th, 16th day after infection, no Trichinella spiralis was found in diaphragm. At the 17th, 18th, 19th, 20th day after infection, Trichinella spiralis could be seen in the diaphragm and the number of Trichinella spiralis in-creased with time passing. At the 14th, 15th, 16th day, no Trichinella spiralis was obtained. At the 17th,18th, 19th, 20th day, the mean number of Trichinella spirulis collected were 5000, 8000, 10000, 30000per rat respectively. Conclusion The 20th day after Trichinella spiralis infection is the best time to collect the pre-encysted larvae of Trichinella spiralis. Pepsin digestion of muscles can be the choice of methods for col-lecting the pre-encysted larvae of Trichinella spirulis.

  7. Isolation and Genotyping of Acanthamoeba Strains from Envi-ron¬mental Sources in Ahvaz City, Khuzestan Province, South-ern Iran

    M Rahdar

    2012-12-01

    Full Text Available Background: Acanthamoeba spp. are free-living amoebae commonly found in the environmental sources such as water, soil, and air. This ubiquitous amoeba is the causative agent of amoebic kerati­tis (AK. The objective of the present study was to investigate the presence of Acanthamoeba spp. in water and soil sources in Ahvaz City, Khuzestan Province, southern Iran.Methods: In general, 110 samples of water and soil were taken from different localities of Ahvaz including agricultural canals, rivers, and swimming pools. Filtration and cultivation were carried out on non-nutrient agar medium (NNA. Axenic cultivation was performed for all of positive isolates. PCR analysis was conducted on positive samples. Sequencing was done for 15 PCR products. Geno­types were identified by Blast search and homology analysis.Result: Acanthamoeba spp. was found in 43 (71.6% of samples of water and 13 (26% soil samples. Genotyping of 15 samples proved that Acanthamoeba belonged to T4 (86.6%, T2 (6.6%, and T5 (6.6% genotypes.Conclusion: TYI-S-33 medium could be better than PYG medium for Acanthamoeba axenic culture.

  8. In vitro culture, serologic and molecular analysis of Acanthamoeba isolated from the liver of a keel-billed toucan (Ramphastos sulfuratus).

    Visvesvara, Govinda S; Booton, Gregory C; Kelley, Darryl J; Fuerst, Paul; Sriram, Rama; Finkelstein, Ariana; Garner, Michael M

    2007-01-19

    Members of the genus Acanthamoeba are usually free-living amebae and are found in a variety of ecological niches including soil, fresh and brackish water, dust in air, filters of heating, ventilating, and air conditioning units, swimming pools and hot tubs, etc. Occasionally, they are also known to cause central nervous system infections in humans and other animals. We isolated into culture an amoeba from the liver tissue of a keel-billed toucan and identified it as Acanthamoeba sp. based on culture characteristics and immunofluorescent analysis. Further, we characterized the cultured amoeba and also the amoeba in the liver tissue as Acanthamoeba, genotype T4, by sequencing a diagnostic region of the nuclear small subunit ribosomal RNA gene. PMID:16962705

  9. Acanthamoeba and other free-living amoebae in bat guano, an extreme habitat.

    Mulec, Janez; Dietersdorfer, Elisabeth; Üstüntürk-Onan, Miray; Walochnik, Julia

    2016-04-01

    Several representatives of the so-called free-living amoebae (FLA) are of medical relevance, not only as facultative pathogens but also as vehicles for pathogenic bacteria. Some FLA can survive and even grow under extreme environmental conditions. Bat guano is an exceptional habitat, the conditions becoming gradually more extreme with aging. In the current study, samples of bat guano of different ages from five caves in Slovenia were screened for the presence of FLA. FLA were isolated from almost all guano samples, including guano with a pH of 3.5. Only the two samples that had been drawn from >20-year-old guano were negative for FLA. Generally, FLA diversity correlated to high concentrations of cultivable bacteria (∼10(8) CFU/g) and fungi (∼10(5) CFU/g). Interestingly, the absence of FLA in seasoned guanos was mirrored by the presence of dictyostelid slime moulds. The isolated amoebae were identified as belonging to the genera Acanthamoeba, Copromyxa, Naegleria, Sappinia, Tetramitus, Thecamoeba, Vahlkampfia, Vannella and Vermamoeba. To the best of our knowledge, this is the first study on the diversity of FLA in guano. PMID:26678653

  10. Comparison of a PCR-Based Method with Culture and Direct Examination for Diagnosis of Acanthamoeba keratitis

    S Farnia

    2009-05-01

    Full Text Available "nBackground: The aim was to compare three different methods (direct examination, culture and PCR meth­ods for the diagnosis of Acanthamoeba keratitis (AK in corneal scrapes."nMethods: Twenty eight corneal scrapes and contact lenses were collected from keratitis patients and re­ferred to the De­partment of Medical Parasitology and Mycology, School of Public Health, Tehran Univer­sity of Medical Sci­ences. Corneal scrapes were divided in three parts for direct examination, culture on non-nutrient agar and PCR analysis. PCR analysis was also performed using a 18S rRNA gene primer pair (DF3 region. DF3 (Diagnostic frag­ment 3 is a region of the nuclear small subunit ribosomal RNA gene which is specific for detecting Acan­thamoeba strains."nResults:  Acanthamoeba was the causative agent of keratitis in 50% of the patients. Direct smear of all pre­pared corneal scrapes in AK patients was negative and culture was positive in only 14.3% of the isolates. PCR analysis was positive in 71.4% of AK patients. These three methods were negative in corneal scrapes of non-AK patients. The sensitivity and specificity of PCR technique for the detection of Acanthamoeba sp. were calculated as 71.4% and 100%, respectively."nConclusion: According to high sensitivity and specificity of PCR-based method, this study confirmed that PCR using 18S rRNA gene primers (DF3 region is more useful for detecting AK cases compare to culture and direct microscopy methods.

  11. Ether-type moieties in the lipid part of glycoinositolphospholipids of Acanthamoeba rhysodes.

    Karaś, Magdalena A; Russa, Ryszard

    2014-04-01

    Ether lipids were identified among components liberated with HF and nitrous acid deamination from Acanthamoeba rhysodes whole cells and its membrane glycoinositolphospholipids (GIPL). Liberated ether glycerols were converted to various derivatives that served characterization thereof. These included TMS and isopropylidene derivatives, oxidation with sodium periodate to aldehyde followed by reduction with NaBH4 to alcohol, and reaction of the alcohol with acetic anhydrite to form acetate derivatives. Periodate sensitivity demonstrated that the alkyl side chains were linked to the sn-1 position of glycerol. Combined information from TLC, GC-MS analysis, MALDI-TOF spectrometry, and chemical degradation experiments indicated the presence of ether-linked saturated normal and branched hydrocarbons with a length of C20-23 in the phospholipid fraction, C20-24 in free GPI, and C21-23 in the LPG polymer. The distribution of particular classes of alkylglycerols was similar for phospholipid and GPI fractions, and amounted to 2.62% (±0.04-0.28) 1-O-eicosanyl-sn-glycerol, 16.66% (±0.32-1.1) 1-O-uncosanyl-sn-glycerol, 9.18% (±0.33-1.37) anteiso-1-O-docosanyl-sn-glycerol, 47.56% (±0.32-2.14) 1-O-docosanyl-sn-glycerol, 20.56% (±0.58-1.67) anteiso-1-O-tricosanyl-sn-glycerol, and 2.34% (±0.12-0.63) 1-O-tricosanyl-sn-glycerol. For LPG preparation, the most abundant were anteiso-1-O-tricosanyl-sn-glycerol (57.26%) and 1-O-docosanyl-sn-glycerol (30.12%). The data from TLC and GC-MS analysis showed that ether lipids from phospholipids probably represent the lyso-alkylglycerol type, while those derived from GIPL are alkylacylglycerol moieties. PMID:24535098

  12. The fate of Helicobacter pylori phagocytized by Acanthamoeba polyphaga demonstrated by fluorescent in situ hybridization and quantitative polymerization chain reaction tests

    Helicobacter pylori able to express green fluorescent protein, as well as an ATCC strain, and a clinical isolate of this pathogen were evaluated for their ability to survive predation by Acanthamoeba polyphaga. Ingestion was evaluated by microscopic observation of the GFP-H. pyl...

  13. The GacS/A-RsmA Signal Transduction Pathway Controls the Synthesis of Alkylresorcinol Lipids that Replace Membrane Phospholipids during Encystment of Azotobacter vinelandii SW136

    Romero, Yanet; Guzmán, Josefina; Moreno, Soledad; Cocotl-Yañez, Miguel; Vences-Guzmán, Miguel Ángel; Castañeda, Miguel; Espín, Guadalupe; Segura, Daniel

    2016-01-01

    Azotobacter vinelandii is a soil bacterium that undergoes a differentiation process that forms cysts resistant to desiccation. During encystment, a family of alkylresorcinols lipids (ARs) are synthesized and become part of the membrane and are also components of the outer layer covering the cyst, where they play a structural role. The synthesis of ARs in A. vinelandii has been shown to occur by the activity of enzymes encoded in the arsABCD operon. The expression of this operon is activated by ArpR, a LysR-type transcriptional regulator whose transcription occurs during encystment and is dependent on the alternative sigma factor RpoS. In this study, we show that the two component response regulator GacA, the small RNA RsmZ1 and the translational repressor protein RsmA, implicated in the control of the synthesis of other cysts components (i.e., alginate and poly-ß-hydroxybutyrate), are also controlling alkylresorcinol synthesis. This control affects the expression of arsABCD and is exerted through the regulation of arpR expression. We show that RsmA negatively regulates arpR expression by binding its mRNA, repressing its translation. GacA in turn, positively regulates arpR expression through the activation of transcription of RsmZ1, that binds RsmA, counteracting its repressor activity. This regulatory cascade is independent of RpoS. We also show evidence suggesting that GacA exerts an additional regulation on arsABCD expression through an ArpR independent route. PMID:27055016

  14. Efficacy of hand held, inexpensive UV light sources on Acanthamoeba, causative organism in amoebic keratitis

    Ivan Cometa

    2010-01-01

    Full Text Available Ivan Cometa1, Andrew Rogerson1, Scott Schatz21Department of Biology, California State University Fresno, Fresno, CA, USA; 2Arizona College of Optometry, Midwestern University, Glendale, AZ, USAAbstract: Multipurpose lens cleaning solutions (MPS fail to consistently kill or inactivate Acanthamoeba cysts and UV irradiation, while effective at high doses, can damage contact lenses. The present study considered synergy of action between MPS and hand-held inexpensive (ie, relatively weak UV irradiation units. Regardless of disinfection method recently formed cysts (<10 days were far more susceptible to treatment than mature cysts (>14 days. This has important implications for future protocols on testing methods for killing amoebae. The study also showed that cysts of different strains (two tested, FLA2 and P120 are variable in their response to MPS, presumably reflecting differences in cyst wall structure and thus permeability to the disinfectant. On the other hand, the effect of UV irradiation was not wall structure dependent. A 6-hour treatment with MPS alone killed trophic amoebae but failed to kill any mature cysts. Cysts of strain FLA2 were killed after 24 hours with MPS but cysts of strain P120 survived. UV irradiation with the larger 4 W unit killed all cysts after 7 minutes and was more effective than the smaller battery-powered unit (after 10 minutes about 50% of cysts were killed. When the larger unit was used with the MPS disinfection, all trophozoites were killed using UV for 3 minutes and MPS for 1 hour. The resistant P120 cysts remained a challenge but a 2- to 4-minute UV treatment followed by MPS for 3 or 6 hours reduced mature cyst survival by about 50%. The small unit in combination with MPS was less effective but did reduce the time required to kill trophic amoebae in MPS (6 hours MPS alone versus 3 hours MPS with a 1-minute UV treatment. In short, inexpensive UV units do enhance MPS disinfection and future lens cleaning systems

  15. Acanthamoeba feature a unique backpacking strategy to trap and feed on Listeria monocytogenes and other motile bacteria

    Doyscher, Dominik; Fieseler, Lars; Dons, Lone Elisabet;

    2013-01-01

    , forming large aggregates of densely packed bacteria that we termed backpacks. While the assembly of backpacks is dependent on bacterial motility, flagellation alone is not sufficient. Electron micrographs showed that the aggregates are held together by filaments of likely amoebal origin. Time......-lapse microscopy revealed that shortly after the bacteria are collected, the amoeba can change direction of movement, phagocytose the backpack and continue to repeat the process. The phenomenon was also observed with avirulent L.?monocytogenes mutants, non-pathogenic Listeria, and other motile bacteria, indicating...... that formation of backpacks is not specific for L.?monocytogenes, and independent of bacterial pathogenicity or virulence. Hence, backpacking appears to represent a unique and highly effective strategy of Acanthamoeba to trap and feed on motile bacteria....

  16. Comparison of molecular diagnostic methods for the detection of Acanthamoeba spp. from clinical specimens submitted for keratitis.

    Khairnar, Krishna; Tamber, Gurdip S; Ralevski, Filip; Pillai, Dylan R

    2011-08-01

    Acanthamoeba spp. are responsible for a significant annual number of keratitis (AK) cases leading to vision-threatening disease worldwide. Current methods rely on direct examination of specimens by microscopy and/or culture. The former lacks sensitivity and the latter suffers from a poor turnaround time. We undertook a comparison of all published molecular methods, evaluating performance characteristics such as analytical sensitivity, specificity, limit of detection (LOD), reproducibility, accuracy, and cost of test. The study population comprised 128 patients. Eligible specimens were tested prospectively between April 2007 and May 2010 by microscopy and/or culture. Eleven different specimen types were used including corneal scrapings (51.5%), corneal swab (17.9%), and contact lens material (10.9%). Results of 2 published gel-based polymerase chain reaction (PCR) and 2 published real-time quantitative (Q) PCR methods were compared in a blinded manner to direct microscopic examination and/or culture for the detection of Acanthamoeba in clinical specimens. QPCR (Riviere method) had the highest sensitivity at 89.3%, excellent accuracy using ROC analysis (AUC ∼0.90), lowest LOD down to 0.1 organism per microliter, and superior linear correlation with parasite density (R(2) = 0.9965) when compared with microscopy, culture, and other molecular methods. Phylogenetic analysis using a sequence-based typing method revealed that clinical isolates in this population with AK were genetically distinct from granulomatous amebic encephalitis or environmental isolates. The QPCR method was more expensive ($14.80) than traditional methods such as culture ($2.50) or microscopy ($2.50). However, 13 culture- and microscopy-negative specimens were positive by QPCR during the study period, suggesting that detection using QPCR may result in reduced complications and health care costs associated with misdiagnosed AK. PMID:21658877

  17. Selective Requirement of the Shikimate Pathway of Legionella pneumophila for Intravacuolar Growth within Human Macrophages but Not within Acanthamoeba

    Jones, Snake C.; Price, Christopher T. D.; Santic, Marina

    2015-01-01

    Legionella pneumophila utilizes the Dot/Icm type IV translocation system to proliferate within a vacuole in a wide variety of natural amoebal hosts and in alveolar macrophages of the human accidental host. Although L. pneumophila utilizes host amino acids as the main sources of carbon and energy, it is not known whether de novo synthesis of amino acids by intravacuolar L. pneumophila contributes to its nutrition. The aroB and aroE genes encode enzymes for the shikimate pathway that generates the aromatic amino acids Phe, Trp, and Tyr. Here we show the aroB and aroE mutants of L. pneumophila to be defective in growth in human monocyte-derived macrophages (hMDMs) but not in Acanthamoeba spp. The aroB and aroE mutants are severely attenuated in intrapulmonary proliferation in the A/J mouse model of Legionnaires' disease, and the defect is fully complemented by the respective wild-type alleles. The two mutants grow normally in rich media but do not grow in defined media lacking aromatic amino acids, and the growth defect is rescued by inclusion of the aromatic amino acids, which are essential for production of the pyomelanin pigment. Interestingly, supplementation of infected hMDMs with the three aromatic amino acids or with Trp alone rescues the intramacrophage defect of the aroE but not the aroB mutant. Therefore, the shikimate pathway of L. pneumophila is differentially required for optimal growth within human macrophages, which are auxotrophic for Trp and Phe, but is dispensable for growth within the Acanthamoeba spp. that synthesize the aromatic amino acids. PMID:25847958

  18. Outbreeding lethality between toxic Group I and nontoxic Group III Alexandrium tamarense spp. isolates: Predominance of heterotypic encystment and implications for mating interactions and biogeography

    Brosnahan, Michael L.; Kulis, David M.; Solow, Andrew R.; Erdner, Deana L.; Percy, Linda; Lewis, Jane; Anderson, Donald M.

    2010-02-01

    We report the zygotic encystment of geographically dispersed isolates in the dinoflagellate species complex Alexandrium tamarense, in particular, successful mating of toxic Group I and nontoxic Group III isolates. However, hypnozygotes produced in Group I/III co-cultures complete no more than three divisions after germinating. Previous reports have suggested a mate recognition mechanism whereby hypnozygotes produced in co-cultures could arise from either homotypic (inbred) or heterotypic (outbred) gamete pairs. To determine the extent to which each occurs, a nested PCR assay was developed to determine parentage of individual hypnozygotes. The vast majority of hypnozygotes from pairwise Group I/III co-cultures were outbred, so that inviability was a result of hybridization, not inbreeding. These findings support the assertion that complete speciation underlies the phylogenetic structure of the Alexandrium tamarense species complex. Additionally, the ribosomal DNA (rDNA) copy numbers of both hybrid and single ribotype hypnozygotes were reduced substantially from those of haploid motile cells. The destruction of rDNA loci may be crucial for the successful mating of genetically distant conjugants and appears integral to the process of encystment. The inviability of Group I/III hybrids is important for public health because the presence of hybrid cysts may indicate ongoing displacement of a nontoxic population by a toxic one (or vice versa). Hybrid inviability also suggests a bloom control strategy whereby persistent, toxic Group I blooms could be mitigated by introduction of nontoxic Group III cells. The potential for hybridization in nature was investigated by applying the nested PCR assay to hypnozygotes from Belfast Lough, Northern Ireland, a region where Group I and III populations co-occur. Two hybrid cysts were identified in 14 successful assays, demonstrating that Group I and III populations do interbreed in that region. However, an analysis of mating data

  19. Reevaluating the Role of Acanthamoeba Proteases in Tissue Invasion: Observation of Cytopathogenic Mechanisms on MDCK Cell Monolayers and Hamster Corneal Cells

    Maritza Omaña-Molina; Arturo González-Robles; Lizbeth Iliana Salazar-Villatoro; Jacob Lorenzo-Morales; Ana Ruth Cristóbal-Ramos; Verónica Ivonne Hernández-Ramírez; Patricia Talamás-Rohana; Adolfo René Méndez Cruz; Adolfo Martínez-Palomo

    2013-01-01

    The morphological analysis of the cytopathic effect on MDCK cell monolayers and hamster cornea and qualitative and quantitative analyses of conditioned medium and proteases were evaluated and compared between two strains of Acanthamoeba genotype T4. Further than highlighting the biological differences found between both strains, the most important observation in this study was the fact that proteases both in total extracts and in conditioned medium are apparently not determinant in tissue d...

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

    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

  1. Emerging Threats for Human Health in Poland: Pathogenic Isolates from Drug Resistant Acanthamoeba Keratitis Monitored in terms of Their In Vitro Dynamics and Temperature Adaptability

    Lidia Chomicz

    2015-01-01

    Full Text Available Amphizoic amoebae generate a serious human health threat due to their pathogenic potential as facultative parasites, causative agents of vision-threatening Acanthamoeba keratitis (AK. Recently, AK incidences have been reported with increasing frequency worldwide, particularly in contact lens wearers. In our study, severe cases of AK in Poland and respective pathogenic isolates were assessed at clinical, morphological, and molecular levels. Misdiagnoses and the unsuccessful treatment in other ophthalmic units delayed suitable therapy, and resistance to applied chemicals resulted in severe courses and treatment difficulties. Molecular assessment indicated that all sequenced pathogenic corneal isolates deriving from Polish patients with AK examined by us showed 98–100% homology with Acanthamoeba genotype T4, the most prevalent genotype in this human ocular infection worldwide. In vitro assays revealed that the pathogenic strains are able to grow at elevated temperature and have a wide adaptive capability. This study is our subsequent in vitro investigation on pathogenic Acanthamoeba strains of AK originating from Polish patients. Further investigations designed to foster a better understanding of the factors leading to an increase of AK observed in the past years in Poland may help to prevent or at least better cope with future cases.

  2. The role of contact lenses and ocular TRAUMA in determining Acanthamoeba Keratitis: a case-control study in Italy.

    Elena Pacella

    2012-03-01

    Full Text Available

    Abstract:
    Background: acanthamoeba keratitis (a.k. is a rare secondary infection due to acanthamoeba (a invading the cornea. cofactors of risk may include TrauMa and contact lenses (cL. The objectives of this observational study are to evaluate the occurrence of a.k. in patients accessing the department of Special Senses of the Teaching Hospital “Policlinico umberto I" of rome with ocular infections dur- ing the period from 2005 to 2011, and the role played by risk factors which included contact lenses, TrauMa, sex and age.
    Methods: a case-control study involving 714 subjects was conducted (350 male and 364 female. all of the patients underwent a thorough ocular examination with confocal microscopy and a review of their medical history to outline relevant variables (contact lens and TrauMa. Statistical analysis calculated the frequency distribution and the odds ratio (or (95%cI for the risk factors "contact lenses" (cL and "TrauMa". The odds ratio (95%cI was adjusted for age and sex trough Logistic regression. Results: out of 714 patients, 11 patients (7 male and 4 female with a mean age of 34.09 years (19 to 51 years old were found to be affected by a.k. The age group most afflicted ranged from 30 to 51 years old. Statistical analysis produced a crude or equal to 17.68 (95%cI 2.25-138.89 for cL-a.k. and or equal to 7.71 (95%cI 0.89-66.75 for ocular TrauMa.-a.k. Logistic regression performed to adjust or for age and sex showed the following values, respectively: or*=82.54 (95%cI 3.04-2239.58 and or*=11.52 (95%cI 1.19-111.76.
    Conclusions: the data highlights an increase in a.k. cases during the 6 year study period. The association between this pathology and the use of contact lenses and/or TrauMa has been statisti- cally proven. The strength of this association increases when the odds ratio (or* is adjusted for the confounding risk factors of

  3. Host-dependent variations in the seasonal prevalence and intensity of heterophyid encysted metacercariae (Digenea: Heterophyidea) in brackish water fish in Egypt.

    Elsheikha, Hany M; Elshazly, Attef M

    2008-05-01

    The prevalence of heterophyid (Digenea: Heterophyidae) encysted metacercariae (EMC) in its second intermediate host, the fish Mugil spp. and Tilapia spp. was studied in a subtropical permanent Lake in northeastern Egypt. Seasonal changes in the occurrence of the EMC in different fish hosts were monitored in a longitudinal field survey lasting 12 months from June 2006 to May 2007. This study tested two hypotheses; (i) prevalence and intensity of heterophyid EMC fluctuate seasonally throughout the year and (ii) variation in the prevalence and intensity of heterophyid EMC is host-dependent. A total of 832 fish specimens comprising 5 species collected from Manzala Lake, northeastern Egypt were examined by artificial gastric juice digestion for EMC. All five species of brackish water fish examined were found to harbor the EMC of the family Heterophyidae in their muscles. The overall infection prevalence of EMC over 12 months was 23.2%. The adult flukes recovered from puppies experimentally infected with morphologically different metacercariae from different fish species were compatible with six species belong to five genera of Heterophyidae, namely, Heterophyes heterophyes, Heterophyes aequalis, Pygidiopsis genata, Phagicola sp., Haplorchis sp. and Stictodora sp. EMC of H. heterophyes were most abundant, detected in 56% of the total fish examined. P. genata was ranked second, followed by Phagicola sp., H. aequalis, Haplorchis sp., and Stictodora sp., respectively. Seasonal differences in infection were observed for all heterophyid species studied in all fish species examined. Heterophyid infections reached peak prevalences during the summer season 38.2% followed by spring 26.6% and autumn 19.3% seasons, whereas the lowest prevalence was recorded in the winter 8.7%. Intensity of heterophyid EMC followed the same seasonal pattern, being high during summer months and low in winter months. All fish species were infected with all the heterophyid digeneans, but with different

  4. Morphological and Molecular Identification of Acanthamoeba Spp from Surface Waters in Birjand, Iran, During 2014-2015

    Mahmoodreza Behravan

    2016-04-01

    Full Text Available Background & Aims of the Study: Free-living amoebae (FLA are opportunistic and ubiquitous protozoa that are widely found in various environmental sources. They are known to cause serious human infections including a fatal encephalitis, a blinding keratitis, and pneumonia. So, due to their medical importance, the identification of free living amoeba in water resources, as a source of human infection, is necessary. The objective of this study was to isolate the Acanthamoebaspp from the surface waters of Birjand, Iran, during 2014-2015 by Morphological and molecular method. Materials and Methods:  In a cross-sectional study, 50 samples were collected from different localities of Birjand city including the surface waters, pools and fountains in parks,squares and water stations from the October 2014 to the January 2015.Each sample was filtered through a nitrocellulose membrane filters and cultured on non-nutrient agar (NNA with Escherichia coli suspension and incubated for 1 week to 2 months at room temperature.The plates were examined by the microscopy to morphologically identify Acanthamoeba species. Following DNA extraction, PCR specific primers was used to confirm the identification morphologically. Results:  Out of 50 water samples, 19 (38% were positive for Acanthamoebatrophozoites and cysts according to the morphological criteria. In addition, Acanthamoebaspp was identified by PCR method, using genus specific primers pairs in 15 (78.9% cases of positive cultures, showing anearly 500bp band. Conclusion: According to the prevalent of Acanthamoebaspp in the surface stagnant waters of Birjand, more attention to the potential role of such waters in transmission of infection by the regional clinicians and health practitioners is necessary.

  5. Queratitis por Acanthamoeba sp.: primer caso confirmado por aislamiento y tipificación molecular en Bahía Blanca, provincia de Buenos Aires, Argentina Acanthamoeba sp. keratitis: first case confirmed by isolation and molecular typification in Bahía Blanca, Buenos Aires Province, Argentina

    M. L. Gertiser; E. Giagante; E. Sgattoni; N. Basabe; Rivero, F.; H. Luján; M. Occhionero; L. Paniccia; E. Visciarelli; S.R. Costamagna

    2010-01-01

    Algunas especies del género Acanthamoeba provocan queratitis amebiana (QA), una infección de la córnea muy dolorosa, generalmente unilateral, con compromiso importante de la visión. Se presenta el caso de una mujer de 31 años usuaria de lentes de contacto con hábitos inadecuados de higiene y conservación de las lentes. La paciente consultó por enrojecimiento y dolor intenso en el ojo derecho, de dos meses de evolución. Luego del examen oftalmológico y ante la sospecha de una etiología parasit...

  6. Protozoa: a novel Campylobacter reservoir?

    In previous in vitro studies we found that Campylobacter jejuni remained viable for longer periods of time when they were cultivated in the presence of Tetrahymena pyriformis (ciliate) and Acanthamoeba castellanii (amoeba) than when they were in an independent planktonic state. Increased survival t...

  7. AcEST: BP911838 [AcEST

    Full Text Available (Hsp70/Hs... 65 2e-09 tr|P90647|P90647_ACACA Transformation-sensitive protein homolog ... 64 4e-09 tr|B0JN3... 107 >tr|P90647|P90647_ACACA Transformation-sensitive protein homolog OS=Acanthamoeba castellanii PE=2 SV=1

  8. The killing efficiency of arilin and contact lens solutions on Acanthamoeba cultured in vitro%甲硝唑与护理液对棘阿米巴原虫体外杀伤效果的评估

    车成业; 赵桂秋; 张丽丽

    2008-01-01

    目的:分析6种角膜接触镜多功能护理液和加入甲硝唑滴跟液的护理液对自生生活性棘阿米巴原虫的杀伤效果.方法:将6种多功能护理液分别加入96孔板中,每种护理液占用48孔,其中24孔滴入阿米巴悬液,另外24孔先滴入甲硝唑滴眼液后再滴入阿米巴悬液,室温静置8h后在倒置显微镜下观察残存阿米巴的形态变化和数量.将残存的棘阿米巴原虫分别在PYG培养液中培养5d,观察其形态、活性与增殖能力的变化.结果:单纯护理液组1-6号阿米巴检出率分别为0%、80.3%、29.1%、41.7%、62.5%、79.2%,加入甲硝唑滴眼液后护理液1-6号阿米巴检出率分别为0%、0%、4.2%、8.3%、16.7%、16.7%,3-6号护理液加与不加甲硝唑滴眼液杀伤阿米巴的效力差异有统计学意义(3-6组的X2值分别为3.75、7.11、10.54和18.78,P<0.05).残存的棘阿米巴原虫经培养后活力与增殖力减弱.结论:部分多功能护理液对棘阿米巴原虫的杀伤效果不佳,添加甲硝唑滴眼液后杀伤效果明显提高.%AIM:To analyze the killing efficiency of six kinds of contact lens solutions and solutions with arilin on free living Acanthamoeba culturedin vitroMETHODS:Six kinds of contact lens solutions were added into 96-well microtiter plates,respectively,with each care solutions used 48 holes of them.Suspension of Acanthamoeba were added into 24 of these holes.and arilin gutta and suspension of Acanthamoeba were added into the other 24 holes.After standing in room ternperature for 8 hours,the morphologic change and quantity of the remnant Acanthamoeba were observed under the jnverted microscope.The remnant Acanthamoeba were cultivanted in peptone-yeast extract-glucose (PYG)-culture medium for 5 days.Their variation of appearance,activity and reproductive activity were observed.RESULTS:In the six experimental groups using contact lens solutions only.the detection rate of Acanthamoeba of were 0%,80.3%,29.1%,41.7%,62.5% and

  9. Bilateral Acanthamoeba ulcer in a user of disposable soft contact lenses: a tragic incident or a consequence of the aggressive policy of soft contact lens trading? Úlcera bilateral por Acanthamoeba em usuária de lentes de contato gelatinosas descartáveis: um incidente trágico ou uma conseqüência da política agressiva de venda de lentes de contato gelatinosas?

    Sidney Júlio de Faria e Sousa; Vanderson Glerian Dias; Luís Antonio Gorla Marcomini

    2008-01-01

    This is the report of a case of bilateral Acanthamoeba keratitis in a 19-year-old woman who bought a pair of disposable soft contact lenses in a boutique. She wore this same pair of lenses for 3 months daily without the appropriate care. This led to bilateral corneal transplantation with cataract extraction and also trabeculectomy in the right eye. When last seen, both grafts were crystal clear but the visual acuities were far from satisfactory. She also had bilateral secondary glaucoma, bare...

  10. Temperature stress: reacting and adapting: lessons from poikilotherms.

    Harwood, John L

    2007-10-01

    Acanthamoeba castellanii (A. castellanii) is a common soil- or water-borne protozoon that feeds on bacteria by phagocytosis. A. castellanii can grow between 4 and 32 degrees C and has to adapt quickly to chilling in order to survive. We have identified a Delta12-fatty acid desaturase as key to low temperature adaptation. The activity of this enzyme is mainly increased through gene expression and new protein synthesis. Interestingly, the activity can also be altered independently by dissolved oxygen levels. In addition, we have identified a gene for the Delta12-desaturase, which, when expressed in yeast, catalyses Delta15-desaturation also. Moreover, it is also capable of producing very unusual n-1 polyunsaturated products. PMID:17584990

  11. Plasma membrane biogenesis in eukaryotic cells: translocation of newly synthesized lipid.

    Mills, J T; Furlong, S T; Dawidowicz, E A

    1984-01-01

    We examined the transfer of sterols and phospholipids from their site of synthesis to the plasma membrane of Acanthamoeba castellanii. Cells were labeled with [3H]acetate, and plasma membrane fractions were isolated under conditions that minimize the nonspecific exchange of lipids between subcellular membrane fractions. Sterols and phospholipids were purified from both whole-cell homogenates and isolated plasma membrane. In whole cells, 3H-labeled lipids were formed, with no apparent time lag...

  12. The free-living amoeba Willaertia magna, is particularly resistant to infection by the pathogenic bacteria Legionella pneumophila

    Dey, Rafik; Cavalié, Laurent; Vernet, Christine; Bodennec, Jacques; Pernin, Pierre

    2008-01-01

    Legionella pneumophila, the causative agent of Legionnaire's disease, is well characterized as a bacteria surviving and developing, almost exclusively, as intracellular parasite within freshwater protozoa. Several species of protozoa and ciliae have been shown to support the growth of the pathogenic bacteria. In the present study, we report for the first time the behaviour of the protozoan Willaertia magna towards L. pneumophila and compared it with Acanthamoeba castellanii and Hartmannella v...

  13. Legionella Pneumophila Transcriptome during Intracellular Multiplication in Human Macrophages

    Faucher, Sébastien P.; Mueller, Catherine A.; Shuman, Howard A.

    2011-01-01

    Legionella pneumophila is the causative agent of Legionnaires’ disease, an acute pulmonary infection. L. pneumophila is able to infect and multiply in both phagocytic protozoa, such as Acanthamoeba castellanii, and mammalian professional phagocytes. The best-known L. pneumophila virulence determinant is the Icm/Dot type IVB secretion system, which is used to translocate more than 150 effector proteins into host cells. While the transcriptional response of Legionella to the intracellular envir...

  14. Characterization of giardin protein expression during encystation of Giardia duodenalis

    Giardia duodenalis trophozoites attach to the gut surface by means of a ventral disk that contains various giardin proteins that appear to be important to VD structural integrity. One approach to preventing giardiasis is to stimulate giardin-specific antibodies and thereby block trophozoite attachme...

  15. Dicty_cDB: AFF514 [Dicty_cDB

    Full Text Available arhtkasneaaeyaklvaqrqqakakrfrqsfkqkscxpsk Frame B: *verwyivyyylkks**tllkqfnkrkfieveh*mvqpkvii*tsntinklttrqtmsak...*kscssh*sqq*srrir*trcpktts*sqeipskfqakkllxk* Homology vs CSM-cDNA Score E Sequences producing significant...own update 2004.12.25 Homology vs DNA Score E Sequences producing significant alignment...80 4e-26 9 U03732 |U03732.1 Acanthamoeba castellanii Neff mitochondrion rRNA larg...9, full insert sequence. 58 5e-22 5 AF416765 |AF416765.1 Ehrlichia chaffeensis strain Arkansas 23S ribosomal RNA an

  16. Interaction between Mycobacterium avium subsp. paratuberculosis and environmental protozoa

    Rowe Michael T

    2006-07-01

    Full Text Available Abstract Background Interactions between Mycobacterium avium subsp. paratuberculosis (Map and free-living protozoa in water are likely to occur in nature. The potential impact of ingestion of Map by two naturally occurring Acanthamoeba spp. on this pathogen's survival and chlorine resistance was investigated. Results Between 4.6 and 9.1% of spiked populations of three Map strains (NCTC 8578, B2 and ATCC 19698, which had been added at a multiplicity of infection of 10:1, were ingested by Acanthamoeba castellanii CCAP 1501/1B and A. polyphaga CCAP 1501/3B during co-culture for 3 h at 25°C. Map cells were observed to be present within the vacuoles of the amoebae by acid-fast staining. During extended co-culture of Map NCTC 8578 at 25°C for 24 d with both A. castellanii and A. polyphaga Map numbers did not change significantly during the first 7 days of incubation, however a 1–1.5 log10 increase in Map numbers was observed between days 7 and 24 within both Acanthamoeba spp. Ingested Map cells were shown to be more resistant to chlorine inactivation than free Map. Exposure to 2 μg/ml chlorine for 30 min resulted in a log10 reduction of 0.94 in ingested Map but a log10 reduction of 1.73 in free Map (p Conclusion This study demonstrated that ingestion of Map by and survival and multiplication of Map within Acanthamoeba spp. is possible, and that Map cells ingested by amoebae are more resistant to inactivation by chlorine than free Map cells. These findings have implications with respect to the efficacy of chlorination applied to Map infected surface waters.

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

    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.

  18. Free-living freshwater amoebae differ in their susceptibility to the pathogenic bacterium Legionella pneumophila.

    Dey, Rafik; Bodennec, Jacques; Mameri, Mouh Oulhadj; Pernin, Pierre

    2009-01-01

    Legionella pneumophila is known as a facultative intracellular parasite of free-living soil and freshwater amoebae, of which several species have been shown to support the growth of the pathogenic bacteria. We report for the first time the behaviour of two strains (c2c and Z503) of the amoeba Willaertia magna towards different strains of L. pneumophila serogroup 1 and compared it with Acanthamoeba castellanii and Hartmannella vermiformis, known to be L. pneumophila permissive. In contrast to the results seen with other amoebae, W. magna c2c inhibited the growth of one strain of Legionella (L. pneumophila, Paris), but not of others belonging to the same serogroup (L. pneumophila, Philadelphia and L. pneumophila, Lens). Also, the different L. pneumophila inhibited cell growth and induced cell death in A. castellanii, H. vermiformis and W. magna Z503 within 3-4 days while W. magna c2c strain remained unaffected even up to 7 days. Electron microscopy demonstrated that the formation of numerous replicative phagosomes observed within Acanthamoeba and Hartmannella is rarely seen in W. magna c2c cocultured with L. pneumophila. Moreover, the morphological differences were observed between L. pneumophila cultured either with Willaertia or other amoebae. These observations show that amoebae are not all equally permissive to L. pneumophila and highlight W. magna c2c as particularly resistant towards some strains of this bacterium. PMID:19016880

  19. Sterility and the disinfection potential of Indian contact lens solutions

    Gopinathan Usha

    1994-01-01

    Full Text Available Ocular infection associated with microbial contamination of contact lens care products is a major problem in contact lens wearers. The sterility and the antimicrobial activity of contact lens care systems reflect their suitability for disinfection of contact lenses. These factors remain to be evaluated for the various newer contact lens care products manufactured in India. In this study, 35 bottles of contact lens solutions marketed by different manufacturing units in India were tested for sterility. Seven solutions were tested for antimicrobial effectiveness employing the D value method of analysis. The D value is defined as the time required to reduce a population of organisms by 90% (one log unit. A standard inoculum of the ocular isolates of Staphylococcus aureus, Pseudomonas aeruginosa, Serratia marcescens, Aspergillus fumigatus, Fusarium solani, and Acanthamoeba castellanii were used as challenge organisms. Bacterial contamination was detected in 20 (57.1% solution bottles and none yielded fungus or Acanthamoeba. Pseudomonas species were the most commonly encountered contaminant (11/20; 55%. Only sterile solutions were analyzed for antimicrobial activity. D values ranging between 12 and 20 minutes were demonstrated by six of the seven solutions against bacterial challenge. Good antifungal activity was noticed in five solutions against Fusarium solani though results varied with Aspergillus flavus and Candida albicans. All solutions were adequately effective against Acanthamoeba.

  20. Acanthamoeba and bacteria produce antimicrobials to target their counterpart

    Iqbal, Junaid; Siddiqui, Ruqaiyyah; Khan, Naveed Ahmed

    2014-01-01

    Background In the microbial ecosystem, microbes compete for space and nutrients. Consequently, some have developed the ability to kill or inhibit the growth of other competing microbes by producing antimicrobial substances. As the ‘producer’ species are generally immune to these substances, their compounds act on the competing microbial species and give the producer more space and access to nutrients for growth. Many currently used antibiotics were developed by exploiting this potential of ce...

  1. Species specificity of a monoclonal antibody produced to Naegleria fowleri and partial characterization of its antigenic determinant.

    Réveiller, F L; Marciano-Cabral, F; Pernin, P; Cabanes, P A; Legastelois, S

    2000-08-01

    Monoclonal antibody (Mab) 5D12 against Naegleria fowleri was analyzed for species specificity. Mab 5D12 reacted with a ubiquitous epitope present on the membrane of N. fowleri but not with soluble antigens. The Mab did not react with N. lovaniensis, N. gruberi, N. australiensis, or Acanthamoeba castellanii. The decreased reactivity of Mab 5D12 with N. fowleri observed after periodate oxidation, after digestion of carbohydrate moieties by three glycosidases, or after treatment of amebas with tunicamycin strongly suggests that the antigenic determinant has a polysaccharide component. Inhibition of the reactivity of Mab 5D12 by soluble saccharides supports the idea that N-acetyl or amino groups may play an important role in the recognition of the carbohydrate component of the epitope by the Mab. The specificity of Mab 5D12 makes this an ideal reagent for the identification of N. fowleri in environmental samples or in clinical specimens. PMID:10952262

  2. Isolation and complete genome sequencing of Mimivirus bombay, a Giant Virus in sewage of Mumbai, India.

    Chatterjee, Anirvan; Ali, Farhan; Bange, Disha; Kondabagil, Kiran

    2016-09-01

    We report the isolation and complete genome sequencing of a new Mimiviridae family member, infecting Acanthamoeba castellanii, from sewage in Mumbai, India. The isolated virus has a particle size of about 435 nm and a 1,182,200-bp genome. A phylogeny based on the DNA polymerase sequence placed the isolate as a new member of the Mimiviridae family lineage A and was named as Mimivirus bombay. Extensive presence of Mimiviridae family members in different environmental niches, with remarkably similar genome size and genetic makeup, point towards an evolutionary advantage that needs to be further investigated. The complete genome sequence of Mimivirus bombay was deposited at GenBank/EMBL/DDBJ under the accession number KU761889. PMID:27330993

  3. Survival and Virulence of Campylobacter spp. in the Environment

    Bui, Thanh Xuan

    : Escherichia coli and Enterococcus spp., in raw slurry, in the liquid fraction of separated slurry, and in the liquid fraction after ozonation to ground water using intact soil columns models. I observed that solid-liquid separation of slurry increased the redistribution of contaminants in liquid fraction...... in the soil compared to raw slurry, and the recovery of E. coli and Enterococcus spp. was higher for liquid fraction after the four leaching events. The liquid fraction also resulted in a higher leaching of all contaminants except Enterococcus spp. than raw slurry while the Ozonation reduced E. coli leaching....... Interestingly, I identified the depletion of dissolved oxygen by A. castellanii as the major contributor for the observed amoeba-mediated growth enhancement. To test whether another protozoan rather than Acanthamoeba has similar impacts on survival of C. jejuni as well as other food-borne pathogens S...

  4. srRNA evolution and phylogenetic relationships of the genus Naegleria (Protista: Rhizopoda).

    Baverstock, P R; Illana, S; Christy, P E; Robinson, B S; Johnson, A M

    1989-05-01

    A rapid RNA sequencing technique was used to partially sequence the small-subunit ribosomal RNA (srRNA) of four species of the amoeboid genus Naegleria. The extent of nucleotide sequence divergence between the two most divergent species was roughly similar to that found between mammals and frogs. However, the pattern of variation among the Naegleria species was quite different from that found for those species of tetrapods characterized to date. A phylogenetic analysis of the consensus Naegleria sequence showed that Naegleria was not monophyletic with either Acanthamoeba castellanii or Dictyostelium discoideum, two other amoebas for which sequences were available. It was shown that the semiconserved regions of the srRNA molecule evolve in a clocklike fashion and that the clock is time dependent rather than generation dependent. PMID:2622334

  5. UV inactivation of pathogenic and indicator microorganisms

    Survival was measured as a function of the dose of germicidal UV light for the bacteria Escherichia coli, Salmonella typhi, Shigella sonnei, Streptococcus faecalis, Staphylococcus aureus, and Bacillus subtilis spores, the enteric viruses poliovirus type 1 and simian rotavirus SA11, the cysts of the protozoan Acanthamoeba castellanii, as well as for total coliforms and standard plate count microorganisms from secondary effluent. The doses of UV light necessary for a 99.9% inactivation of the cultured vegetative bacteria, total coliforms, and standard plate count microorganisms were comparable. However, the viruses, the bacterial spores, and the amoebic cysts required about 3 to 4 times, 9 times, and 15 times, respectively, the dose required for E. coli. These ratios covered a narrower relative dose range than that previously reported for chlorine disinfection of E. coli, viruses, spores, and cysts

  6. UV inactivation of pathogenic and indicator microorganisms

    Chang, J.C.; Ossoff, S.F.; Lobe, D.C.; Dorfman, M.H.; Dumais, C.M.; Qualls, R.G.; Johnson, J.D.

    1985-06-01

    Survival was measured as a function of the dose of germicidal UV light for the bacteria Escherichia coli, Salmonella typhi, Shigella sonnei, Streptococcus faecalis, Staphylococcus aureus, and Bacillus subtilis spores, the enteric viruses poliovirus type 1 and simian rotavirus SA11, the cysts of the protozoan Acanthamoeba castellanii, as well as for total coliforms and standard plate count microorganisms from secondary effluent. The doses of UV light necessary for a 99.9% inactivation of the cultured vegetative bacteria, total coliforms, and standard plate count microorganisms were comparable. However, the viruses, the bacterial spores, and the amoebic cysts required about 3 to 4 times, 9 times, and 15 times, respectively, the dose required for E. coli. These ratios covered a narrower relative dose range than that previously reported for chlorine disinfection of E. coli, viruses, spores, and cysts.

  7. Fate of pathogenic Bacillus cereus spores after ingestion by protist grazers

    Winding, Anne; Santos, Susana; Hendriksen, Niels Bohse; Jakobsen, Hans

    evolution of Bacillus cereus group bacteria (e.g. B. cereus, B. anthracis, B. thuringiensis) as a pathogen. It has been hypothesized that the spore stage protects against digestion by predating protists. Indeed, B. thuringiensis spores have been shown to be readily ingested by ciliated protists but failed...... to be digested (Manasherob et al 1998 AEM 64:1750-). Here we report how diverse protist grazers grow on both vegetative cells and spores of B. cereus and how the bacteria survive ingestion and digestion, and even proliferate inside the digestive vacuoles of ciliated protists. The survival ability of...... B. cereus was initially investigated in microcosms inoculated with pure cultures of the protists Acanthamoeba castellanii, Tetrahymena pyriformis and Cercomonas sp. as grazers. Individual protist cultures were fed with fluorescently labelled (CellTracker™RedCMTPX) B. cereus spores or vegetative...

  8. Black cobra (Naja naja karachiensis) lysates exhibit broad-spectrum antimicrobial activities

    Sagheer, Mehwish; Siddiqui, Ruqaiyyah; Iqbal, Junaid; Khan, Naveed Ahmed

    2014-01-01

    It is hypothesized that animals living in polluted environments possess antimicrobials to counter pathogenic microbes. The fact that snakes feed on germ-infested rodents suggests that they encounter pathogenic microbes and likely possess antimicrobials. The venom is used only to paralyze the rodent, but the ability of snakes to counter potential infections in the gut due to disease-ridden rodents requires robust action of the immune system against a broad range of pathogens. To test this hypothesis, crude lysates of different organs of Naja naja karachiensis (black cobra) were tested for antimicrobial properties. The antimicrobial activities of extracts were tested against selected bacterial pathogens (neuropathogenic Escherichia coli K1, methicillin-resistant Staphylococcus aureus (MRSA), Pseudomonas aeruginosa, and Streptococcus pneumonia), protist (Acanthamoeba castellanii), and filamentous fungus (Fusarium solani). The findings revealed that plasma and various organ extracts of N. n. karachiensis exhibited antimicrobial activity against E. coli K1, MRSA, P. aeruginosa, S. pneumoniae, A. castellanii, and F. solani in a concentration-dependent manner. The results of this study are promising for the development of new antimicrobials. PMID:24625321

  9. Optimized DNA extraction methods for encysted embryos of the endangered fairy shrimp, Branchinecta sandiegonensis

    Steele, A.N.; Simovich, M.A.; Pepino, D.; Schroeder, K.M.; Vandergast, A.G.; Bohonak, A.J.

    2009-01-01

    The San Diego fairy shrimp Branchinecta sandiegonensis is a federally endangered species endemic to vernal pools in southern California, USA. Filling events in these habitats are highly variable, with some pools failing to hold water long enough for reproduction over many successive years. Studies of this species are thus hindered by the relatively rare appearance of aquatically active life history phases. Because diapausing cysts are abundant and present at all times, they provide an underutilized opportunity for both species identification and genetic studies. However, methods for extracting DNA from cysts are technically challenging because of their structure and size. Here we present a protocol for extracting DNA from B. sandiegonensis cysts in sufficient quantities for "DNA Barcoding", microsatellite analysis and other genotyping and sequencing applications. The technique will aid in population genetic studies and species identification (since taxonomic keys only distinguish among adults), and will be applicable to other crustaceans with similar diapausing cysts. ?? Springer Science+Business Media B.V. 2008.

  10. Encysted Tenia solium larva of oral cavity: Case report with review of literature

    Bhuvana Krishnamoorthy

    2012-01-01

    Full Text Available Cysticercosis is caused by the larvae of the pig tapeworm, Tenia solium. Oral cysticercosis is a rare event and is often a diagnostic challenge to the clinician. We report a 12-year-old girl who presented with a single, painless, nodule on the lower lip that was diagnosed as cysticercosis. Current literature on the clinical presentations, investigations, and treatment of the condition has been reviewed in this article. We have also proposed a set of criteria for the diagnosis of oral cysticercosis.

  11. Encysted Tenia solium larva of oral cavity: Case report with review of literature

    Bhuvana Krishnamoorthy; N Suma Gundareddy; Manu Dhillon; Siddharth Srivastava; Manisha Lakhanpal Sharma; Sangeeta Singh Malik

    2012-01-01

    Cysticercosis is caused by the larvae of the pig tapeworm, Tenia solium. Oral cysticercosis is a rare event and is often a diagnostic challenge to the clinician. We report a 12-year-old girl who presented with a single, painless, nodule on the lower lip that was diagnosed as cysticercosis. Current literature on the clinical presentations, investigations, and treatment of the condition has been reviewed in this article. We have also proposed a set of criteria for the diagnosis of oral cysticer...

  12. Sequencing illustrates the transcriptional response of Legionella pneumophila during infection and identifies seventy novel small non-coding RNAs.

    Weissenmayer, Barbara A

    2011-01-01

    Second generation sequencing has prompted a number of groups to re-interrogate the transcriptomes of several bacterial and archaeal species. One of the central findings has been the identification of complex networks of small non-coding RNAs that play central roles in transcriptional regulation in all growth conditions and for the pathogen\\'s interaction with and survival within host cells. Legionella pneumophila is a gram-negative facultative intracellular human pathogen with a distinct biphasic lifestyle. One of its primary environmental hosts in the free-living amoeba Acanthamoeba castellanii and its infection by L. pneumophila mimics that seen in human macrophages. Here we present analysis of strand specific sequencing of the transcriptional response of L. pneumophila during exponential and post-exponential broth growth and during the replicative and transmissive phase of infection inside A. castellanii. We extend previous microarray based studies as well as uncovering evidence of a complex regulatory architecture underpinned by numerous non-coding RNAs. Over seventy new non-coding RNAs could be identified; many of them appear to be strain specific and in configurations not previously reported. We discover a family of non-coding RNAs preferentially expressed during infection conditions and identify a second copy of 6S RNA in L. pneumophila. We show that the newly discovered putative 6S RNA as well as a number of other non-coding RNAs show evidence for antisense transcription. The nature and extent of the non-coding RNAs and their expression patterns suggests that these may well play central roles in the regulation of Legionella spp. specific traits and offer clues as to how L. pneumophila adapts to its intracellular niche. The expression profiles outlined in the study have been deposited into Genbank\\'s Gene Expression Omnibus (GEO) database under the series accession GSE27232.

  13. The type II secretion system of Legionella pneumophila elaborates two aminopeptidases, as well as a metalloprotease that contributes to differential infection among protozoan hosts.

    Rossier, Ombeline; Dao, Jenny; Cianciotto, Nicholas P

    2008-02-01

    Legionella pneumophila, the agent of Legionnaires' disease, is an intracellular parasite of aquatic amoebae and human macrophages. A key factor for L. pneumophila in intracellular infection is its type II protein secretion system (Lsp). In order to more completely define Lsp output, we recently performed a proteomic analysis of culture supernatants. Based upon the predictions of that analysis, we found that L. pneumophila secretes two distinct aminopeptidase activities encoded by the genes lapA and lapB. Whereas lapA conferred activity against leucine, phenylalanine, and tyrosine aminopeptides, lapB was linked to the cleavage of lysine- and arginine-containing substrates. To assess the role of secreted aminopeptidases in intracellular infection, we examined the relative abilities of lapA and lapB mutants to infect human U937 cell macrophages as well as Hartmannella vermiformis and Acanthamoeba castellanii amoebae. Although these experiments identified a dispensable role for LapA and LapB, they uncovered a previously unrecognized role for the type II-dependent ProA (MspA) metalloprotease. Whereas proA mutants were not defective for macrophage or A. castellanii infection, they (but not their complemented derivatives) were impaired for growth upon coculture with H. vermiformis. Thus, ProA represents the first type II effector implicated in an intracellular infection event. Furthermore, proA represents an L. pneumophila gene that shows differential importance among protozoan infection models, suggesting that the legionellae might have evolved some of its factors to especially target certain of their protozoan hosts. PMID:18083880

  14. RELATIONSHIP OF TADPOLE STAGE TO LOCATION OF ECHINOSTOME CERCARIAE ENCYSTMENT AND THE CONSEQUENCES FOR TADPOLE SURVIVAL. (R825867)

    The perspectives, information and conclusions conveyed in research project abstracts, progress reports, final reports, journal abstracts and journal publications convey the viewpoints of the principal investigator and may not represent the views and policies of ORD and EPA. Concl...

  15. Experimental study on pathogenic potential of six Acanthamoeba strains isolated from fish

    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

  16. Acanthamoeba polyphaga mimivirus stability in environmental and clinical substrates: implications for virus detection and isolation.

    Fábio P Dornas

    Full Text Available Viruses are extremely diverse and abundant and are present in countless environments. Giant viruses of the Megavirales order have emerged as a fascinating research topic for virologists around the world. As evidence of their ubiquity and ecological impact, mimiviruses have been found in multiple environmental samples. However, isolation of these viruses from environmental samples is inefficient, mainly due to methodological limitations and lack of information regarding the interactions between viruses and substrates. In this work, we demonstrate the long-lasting stability of mimivirus in environmental (freshwater and saline water and hospital (ventilator plastic device tube substrates, showing the detection of infectious particles after more than 9 months. In addition, an enrichment protocol was implemented that remarkably increased mimivirus detection from all tested substrates, including field tests. Moreover, biological, morphological and genetic tests revealed that the enrichment protocol maintained mimivirus particle integrity. In conclusion, our work demonstrated the stability of APMV in samples of environmental and health interest and proposed a reliable and easy protocol to improve giant virus isolation. The data presented here can guide future giant virus detection and isolation studies.

  17. Acanthamoeba polyphaga mimivirus Stability in Environmental and Clinical Substrates: Implications for Virus Detection and Isolation

    Dornas, Fábio P.; Silva, Lorena C. F.; de Almeida, Gabriel M.; Rafael K Campos; Boratto, Paulo V.M.; Ana P M Franco-Luiz; La Scola, Bernard; Ferreira, Paulo C. P.; Kroon, Erna G.; Abrahão, Jônatas S.

    2014-01-01

    Viruses are extremely diverse and abundant and are present in countless environments. Giant viruses of the Megavirales order have emerged as a fascinating research topic for virologists around the world. As evidence of their ubiquity and ecological impact, mimiviruses have been found in multiple environmental samples. However, isolation of these viruses from environmental samples is inefficient, mainly due to methodological limitations and lack of information regarding the interactions betwee...

  18. Structure and function of a unique pore-forming protein from a pathogenic acanthamoeba

    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 acantha

  19. A conserved OmpA-like protein in Legionella pneumophila required for efficient intracellular replication.

    Goodwin, Ian P; Kumova, Ogan K; Ninio, Shira

    2016-08-01

    The OmpA-like protein domain has been associated with peptidoglycan-binding proteins, and is often found in virulence factors of bacterial pathogens. The intracellular pathogen Legionella pneumophila encodes for six proteins that contain the OmpA-like domain, among them the highly conserved uncharacterized protein we named CmpA. Here we set out to characterize the CmpA protein and determine its contribution to intracellular survival of L. pneumophila Secondary structure analysis suggests that CmpA is an inner membrane protein with a peptidoglycan-binding domain at the C-teminus. A cmpA mutant was able to replicate normally in broth, but failed to compete with an isogenic wild-type strain in an intracellular growth competition assay. The cmpA mutant also displayed significant intracellular growth defects in both the protozoan host Acanthamoeba castellanii and in primary bone marrow-derived macrophages, where uptake into the cells was also impaired. The cmpA phenotypes were completely restored upon expression of CmpA in trans The data presented here establish CmpA as a novel virulence factor of L. pneumophila that is required for efficient intracellular replication in both mammalian and protozoan hosts. PMID:27421957

  20. Amoebozoa possess lineage-specific globin gene repertoires gained by individual horizontal gene transfers.

    Dröge, Jasmin; Buczek, Dorota; Suzuki, Yutaka; Makałowski, Wojciech

    2014-01-01

    The Amoebozoa represent a clade of unicellular amoeboid organisms that display a wide variety of lifestyles, including free-living and parasitic species. For example, the social amoeba Dictyostelium discoideum has the ability to aggregate into a multicellular fruiting body upon starvation, while the pathogenic amoeba Entamoeba histolytica is a parasite of humans. Globins are small heme proteins that are present in almost all extant organisms. Although several genomes of amoebozoan species have been sequenced, little is known about the phyletic distribution of globin genes within this phylum. Only two flavohemoglobins (FHbs) of D. discoideum have been reported and characterized previously while the genomes of Entamoeba species are apparently devoid of globin genes. We investigated eleven amoebozoan species for the presence of globin genes by genomic and phylogenetic in silico analyses. Additional FHb genes were identified in the genomes of four social amoebas and the true slime mold Physarum polycephalum. Moreover, a single-domain globin (SDFgb) of Hartmannella vermiformis, as well as two truncated hemoglobins (trHbs) of Acanthamoeba castellanii were identified. Phylogenetic evidence suggests that these globin genes were independently acquired via horizontal gene transfer from some ancestral bacteria. Furthermore, the phylogenetic tree of amoebozoan FHbs indicates that they do not share a common ancestry and that a transfer of FHbs from bacteria to amoeba occurred multiple times. PMID:25013378

  1. The CpxRA two-component system contributes to Legionella pneumophila virulence.

    Tanner, Jennifer R; Li, Laam; Faucher, Sébastien P; Brassinga, Ann Karen C

    2016-06-01

    The bacterium Legionella pneumophila is capable of intracellular replication within freshwater protozoa as well as human macrophages, the latter of which results in the serious pneumonia Legionnaires' disease. A primary factor involved in these host cell interactions is the Dot/Icm Type IV secretion system responsible for translocating effector proteins needed to establish and maintain the bacterial replicative niche. Several regulatory factors have been identified to control the expression of the Dot/Icm system and effectors, one of which is the CpxRA two-component system, suggesting essentiality for virulence. In this study, we generated cpxR, cpxA and cpxRA in-frame null mutant strains to further delineate the role of the CpxRA system in bacterial survival and virulence. We found that cpxR is essential for intracellular replication within Acanthamoeba castellanii, but not in U937-derived macrophages. Transcriptome analysis revealed that CpxRA regulates a large number of virulence-associated proteins including Dot/Icm effectors as well as Type II secreted substrates. Furthermore, the cpxR and cpxRA mutant strains were more sodium resistant than the parental strain Lp02, and cpxRA expression reaches maximal levels during postexponential phase. Taken together, our findings suggest the CpxRA system is a key contributor to L. pneumophila virulence in protozoa via virulence factor regulation. PMID:26934669

  2. Short-Term and Long-Term Survival and Virulence of Legionella pneumophila in the Defined Freshwater Medium Fraquil.

    Nilmini Mendis

    Full Text Available Legionella pneumophila (Lp is the etiological agent responsible for Legionnaires' disease, a potentially fatal pulmonary infection. Lp lives and multiplies inside protozoa in a variety of natural and man-made water systems prior to human infection. Fraquil, a defined freshwater medium, was used as a highly reproducible medium to study the behaviour of Lp in water. Adopting a reductionist approach, Fraquil was used to study the impact of temperature, pH and trace metal levels on the survival and subsequent intracellular multiplication of Lp in Acanthamoeba castellanii, a freshwater protozoan and a natural host of Legionella. We show that temperature has a significant impact on the short- and long-term survival of Lp, but that the bacterium retains intracellular multiplication potential for over six months in Fraquil. Moreover, incubation in Fraquil at pH 4.0 resulted in a rapid decline in colony forming units, but was not detrimental to intracellular multiplication. In contrast, variations in trace metal concentrations had no impact on either survival or intracellular multiplication in amoeba. Our data show that Lp is a resilient bacterium in the water environment, remaining infectious to host cells after six months under the nutrient-deprived conditions of Fraquil.

  3. The ClpP protease homologue is required for the transmission traits and cell division of the pathogen Legionella pneumophila

    Zhang Qin-fen

    2010-02-01

    Full Text Available Abstract Background Legionella pneumophila, the intracellular bacterial pathogen that causes Legionnaires' disease, exhibit characteristic transmission traits such as elevated stress tolerance, shortened length and virulence during the transition from the replication phase to the transmission phase. ClpP, the catalytic core of the Clp proteolytic complex, is widely involved in many cellular processes via the regulation of intracellular protein quality. Results In this study, we showed that ClpP was required for optimal growth of L. pneumophila at high temperatures and under several other stress conditions. We also observed that cells devoid of clpP exhibited cell elongation, incomplete cell division and compromised colony formation. Furthermore, we found that the clpP-deleted mutant was more resistant to sodium stress and failed to proliferate in the amoebae host Acanthamoeba castellanii. Conclusions The data present in this study illustrate that the ClpP protease homologue plays an important role in the expression of transmission traits and cell division of L. pneumophila, and further suggest a putative role of ClpP in virulence regulation.

  4. Short-Term and Long-Term Survival and Virulence of Legionella pneumophila in the Defined Freshwater Medium Fraquil

    Mendis, Nilmini; McBride, Peter; Faucher, Sébastien P.

    2015-01-01

    Legionella pneumophila (Lp) is the etiological agent responsible for Legionnaires’ disease, a potentially fatal pulmonary infection. Lp lives and multiplies inside protozoa in a variety of natural and man-made water systems prior to human infection. Fraquil, a defined freshwater medium, was used as a highly reproducible medium to study the behaviour of Lp in water. Adopting a reductionist approach, Fraquil was used to study the impact of temperature, pH and trace metal levels on the survival and subsequent intracellular multiplication of Lp in Acanthamoeba castellanii, a freshwater protozoan and a natural host of Legionella. We show that temperature has a significant impact on the short- and long-term survival of Lp, but that the bacterium retains intracellular multiplication potential for over six months in Fraquil. Moreover, incubation in Fraquil at pH 4.0 resulted in a rapid decline in colony forming units, but was not detrimental to intracellular multiplication. In contrast, variations in trace metal concentrations had no impact on either survival or intracellular multiplication in amoeba. Our data show that Lp is a resilient bacterium in the water environment, remaining infectious to host cells after six months under the nutrient-deprived conditions of Fraquil. PMID:26406895

  5. Host - parasite interactions: : the relationship between encystment load of the freshwater pearl mussel (Margaritifera margaritifera) and genetic diversity of its host (Salmo trutta)

    Winser, Helena

    2015-01-01

    The freshwater pearl mussel (FPM, Margaritifera margaritifera) is an endangered bivalve mollusc with an obligatory larval parasitic phase on brown trout (Salmo trutta). The FPM has declined throughout its entire range due to causes such as habitat degradation, eutrophication, acidification, changed hydrology and lack of host fish. This study aimed to investigate if heterozygosity, allelic richness, number of alleles, inbreeding and differentiation of brown trout (Salmo trutta) are related to ...

  6. Surviving within the amoebal exocyst: the Mycobacterium avium complex paradigm

    Drancourt Michel

    2010-04-01

    Full Text Available Abstract Background Most of environmental mycobacteria have been previously demonstrated to resist free-living amoeba with subsequent increased virulence and resistance to antibiotics and biocides. The Mycobacterium avium complex (MAC comprises of environmental organisms that inhabit a wide variety of ecological niches and exhibit a significant degree of genetic variability. We herein studied the intra-ameobal location of all members of the MAC as model organisms for environmental mycobacteria. Results Type strains for M. avium, Mycobacterium intracellulare, Mycobacterium chimaera, Mycobacterium colombiense, Mycobacterium arosiense, Mycobacterium marseillense, Mycobacterium timonense and Mycobacterium bouchedurhonense were co-cultivated with the free-living amoeba Acanthamoeba polyphaga strain Linc-AP1. Microscopic analyses demonstrated the engulfment and replication of mycobacteria into vacuoles of A. polyphaga trophozoites. Mycobacteria were further entrapped within amoebal cysts, and survived encystment as demonstrated by subculturing. Electron microscopy observations show that, three days after entrapment into A. polyphaga cysts, all MAC members typically resided within the exocyst. Conclusions Combined with published data, these observations indicate that mycobacteria are unique among amoeba-resistant bacteria, in residing within the exocyst.

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

    Olofsson, Jenny; Griekspoor Berglund, Petra; Olsen, Björn; Ellström, Patrik; Axelsson-Olsson, Diana

    2015-01-01

    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. co...

  8. Genetic Characterization of Clinical Acanthamoeba Isolates Using Gene Loci of Nuclear and Mitochondrial Small Subunit Ribosomal RNA

    ラフマン, モハメド, モシウル

    2013-01-01

    以下に掲載:The Korean Journal of Parasitology 51(4) pp.401-411 2013. Korean Society for Parasitology and Tropical Medicine. 共著者:Md Moshiur Rahman, Kenji Yagita, Akira Kobayashi, Yosaburo Oikawa, Amjad I.A. Hussein, Takahiro Matsumura and Masaharu Tokoro

  9. Genetic Characterization of Clinical Acanthamoeba Isolates Using Gene Loci of Nuclear and Mitochondrial Small Subunit Ribosomal RNA

    ラフマン, モハメド, モシウル

    2013-01-01

    要旨Abstract 以下に掲載:The Korean Journal of Parasitology. 51(4) pp.401-411 2013. Korean Society for Parasitology and Tropical Medicine. 共著者:Md Moshiur Rahman, Kenji Yagita, Akira Kobayashi, Yosaburo Oikawa, Amjad I.A. Hussein, Takahiro Matsumura and Masaharu Tokoro

  10. The Rising Dominance of Shigella sonnei: An Intercontinental Shift in the Etiology of Bacillary Dysentery.

    Corinne N Thompson

    Full Text Available Shigellosis is the major global cause of dysentery. Shigella sonnei, which has historically been more commonly isolated in developed countries, is undergoing an unprecedented expansion across industrializing regions in Asia, Latin America, and the Middle East. The precise reasons underpinning the epidemiological distribution of the various Shigella species and this global surge in S. sonnei are unclear but may be due to three major environmental pressures. First, natural passive immunization with the bacterium Plesiomonas shigelloides is hypothesized to protect populations with poor water supplies against S. sonnei. Improving the quality of drinking water supplies would, therefore, result in a reduction in P. shigelloides exposure and a subsequent reduction in environmental immunization against S. sonnei. Secondly, the ubiquitous amoeba species Acanthamoeba castellanii has been shown to phagocytize S. sonnei efficiently and symbiotically, thus allowing the bacteria access to a protected niche in which to withstand chlorination and other harsh environmental conditions in temperate countries. Finally, S. sonnei has emerged from Europe and begun to spread globally only relatively recently. A strong selective pressure from localized antimicrobial use additionally appears to have had a dramatic impact on the evolution of the S. sonnei population. We hypothesize that S. sonnei, which exhibits an exceptional ability to acquire antimicrobial resistance genes from commensal and pathogenic bacteria, has a competitive advantage over S. flexneri, particularly in areas with poorly regulated antimicrobial use. Continuing improvement in the quality of global drinking water supplies alongside the rapid development of antimicrobial resistance predicts the burden and international distribution of S. sonnei will only continue to grow. An effective vaccine against S. sonnei is overdue and may become one of our only weapons against this increasingly dominant and

  11. cDNA sequence analysis of a 29-kDa cysteine-rich surface antigen of pathogenic Entamoeba histolytica

    A λgt11 cDNA library was constructed from poly(U)-Spharose-selected Entamoeba histolytica trophozoite RNA in order to clone and identify surface antigens. The library was screened with rabbit polyclonal anti-E. histolytica serum. A 700-base-pair cDNA insert was isolated and the nucleotide sequence was determined. The deduced amino acid sequence of the cDNA revealed a cysteine-rich protein. DNA hybridizations showed that the gene was specific to E. histolytica since the cDNA probe reacted with DNA from four axenic strains of E. histolytica but did not react with DNA from Entamoeba invadens, Acanthamoeba castellanii, or Trichomonas vaginalis. The insert was subcloned into the expression vector pGEX-1 and the protein was expressed as a fusion with the C terminus of glutathione S-transferase. Purified fusion protein was used to generate 22 monoclonal antibodies (mAbs) and a mouse polyclonal antiserum specific for the E. histolytica portion of the fusion protein. A 29-kDa protein was identified as a surface antigen when mAbs were used to immunoprecipitate the antigen from metabolically 35S-labeled live trophozoites. The surface location of the antigen was corroborated by mAb immunoprecipitation of a 29-kDa protein from surface-125I-labeled whole trophozoites as well as by the reaction of mAbs with live trophozoites in an indirect immunofluorescence assay performed at 4 degree C. Immunoblotting with mAbs demonstrated that the antigen was present on four axenic isolates tested. mAbs recognized epitopes on the 29-kDa native antigen on some but not all clinical isolates tested

  12. Analysis of the Legionella longbeachae genome and transcriptome uncovers unique strategies to cause Legionnaires' disease.

    Christel Cazalet

    2010-02-01

    Full Text Available Legionella pneumophila and L. longbeachae are two species of a large genus of bacteria that are ubiquitous in nature. L. pneumophila is mainly found in natural and artificial water circuits while L. longbeachae is mainly present in soil. Under the appropriate conditions both species are human pathogens, capable of causing a severe form of pneumonia termed Legionnaires' disease. Here we report the sequencing and analysis of four L. longbeachae genomes, one complete genome sequence of L. longbeachae strain NSW150 serogroup (Sg 1, and three draft genome sequences another belonging to Sg1 and two to Sg2. The genome organization and gene content of the four L. longbeachae genomes are highly conserved, indicating strong pressure for niche adaptation. Analysis and comparison of L. longbeachae strain NSW150 with L. pneumophila revealed common but also unexpected features specific to this pathogen. The interaction with host cells shows distinct features from L. pneumophila, as L. longbeachae possesses a unique repertoire of putative Dot/Icm type IV secretion system substrates, eukaryotic-like and eukaryotic domain proteins, and encodes additional secretion systems. However, analysis of the ability of a dotA mutant of L. longbeachae NSW150 to replicate in the Acanthamoeba castellanii and in a mouse lung infection model showed that the Dot/Icm type IV secretion system is also essential for the virulence of L. longbeachae. In contrast to L. pneumophila, L. longbeachae does not encode flagella, thereby providing a possible explanation for differences in mouse susceptibility to infection between the two pathogens. Furthermore, transcriptome analysis revealed that L. longbeachae has a less pronounced biphasic life cycle as compared to L. pneumophila, and genome analysis and electron microscopy suggested that L. longbeachae is encapsulated. These species-specific differences may account for the different environmental niches and disease epidemiology of these

  13. A survey of PPR proteins identifies DYW domains like those of land plant RNA editing factors in diverse eukaryotes.

    Schallenberg-Rüdinger, Mareike; Lenz, Henning; Polsakiewicz, Monika; Gott, Jonatha M; Knoop, Volker

    2013-01-01

    The pentatricopeptide repeat modules of PPR proteins are key to their sequence-specific binding to RNAs. Gene families encoding PPR proteins are greatly expanded in land plants where hundreds of them participate in RNA maturation, mainly in mitochondria and chloroplasts. Many plant PPR proteins contain additional carboxyterminal domains and have been identified as essential factors for specific events of C-to-U RNA editing, which is abundant in the two endosymbiotic plant organelles. Among those carboxyterminal domain additions to plant PPR proteins, the so-called DYW domain is particularly interesting given its similarity to cytidine deaminases. The frequency of organelle C-to-U RNA editing and the diversity of DYW-type PPR proteins correlate well in plants and both were recently identified outside of land plants, in the protist Naegleria gruberi. Here we present a systematic survey of PPR protein genes and report on the identification of additional DYW-type PPR proteins in the protists Acanthamoeba castellanii, Malawimonas jakobiformis, and Physarum polycephalum. Moreover, DYW domains were also found in basal branches of multi-cellular lineages outside of land plants, including the alga Nitella flexilis and the rotifers Adineta ricciae and Philodina roseola. Intriguingly, the well-characterized and curious patterns of mitochondrial RNA editing in the slime mold Physarum also include examples of C-to-U changes. Finally, we identify candidate sites for mitochondrial RNA editing in Malawimonas, further supporting a link between DYW-type PPR proteins and C-to-U editing, which may have remained hitherto unnoticed in additional eukaryote lineages. PMID:23899506

  14. Virulence phenotypes of Legionella pneumophila associated with noncoding RNA lpr0035.

    Jayakumar, Deepak; Early, Julie V; Steinman, Howard M

    2012-12-01

    The Philadelphia-1 strain of Legionella pneumophila, the causative organism of Legionnaires' disease, contains a recently discovered noncoding RNA, lpr0035. lpr0035 straddles the 5' chromosomal junction of a 45-kbp mobile genetic element, pLP45, which can exist as an episome or integrated in the bacterial chromosome. A 121-bp deletion was introduced in strain JR32, a Philadelphia-1 derivative. The deletion inactivated lpr0035, removed the 49-bp direct repeat at the 5' junction of pLP45, and locked pLP45 in the chromosome. Intracellular multiplication of the deletion mutant was decreased by nearly 3 orders of magnitude in Acanthamoeba castellanii amoebae and nearly 2 orders of magnitude in J774 mouse macrophages. Entry of the deletion mutant into amoebae and macrophages was decreased by >70%. The level of entry in both hosts was restored to that in strain JR32 by plasmid copies of two open reading frames immediately downstream of the 5' junction and plasmid lpr0035 driven by its endogenous promoter. When induced from a tac promoter, plasmid lpr0035 completely reversed the intracellular multiplication defect in macrophages but was without effect in amoebae. These data are the first evidence of a role for noncoding RNA lpr0035, which has homologs in six other Legionella genomes, in entry of L. pneumophila into amoebae and macrophages and in host-specific intracellular multiplication. The data also demonstrate that deletion of a direct-repeat sequence restricts the mobility of pLP45 and is a means of studying the role of pLP45 mobility in Legionella virulence phenotypes. PMID:22966048

  15. The unusual 23S rRNA gene of Coxiella burnetii: two self-splicing group I introns flank a 34-base-pair exon, and one element lacks the canonical omegaG.

    Raghavan, Rahul; Miller, Scott R; Hicks, Linda D; Minnick, Michael F

    2007-09-01

    We describe the presence and characteristics of two self-splicing group I introns in the sole 23S rRNA gene of Coxiella burnetii. The two group I introns, Cbu.L1917 and Cbu.L1951, are inserted at sites 1917 and 1951 (Escherichia coli numbering), respectively, in the 23S rRNA gene of C. burnetii. Both introns were found to be self-splicing in vivo and in vitro even though the terminal nucleotide of Cbu.L1917 is adenine and not the canonical conserved guanine, termed OmegaG, found in Cbu.L1951 and all other group I introns described to date. Predicted secondary structures for both introns were constructed and revealed that Cbu.L1917 and Cbu.L1951 were group IB2 and group IA3 introns, respectively. We analyzed strains belonging to eight genomic groups of C. burnetii to determine sequence variation and the presence or absence of the elements and found both introns to be highly conserved (>/=99%) among them. Although phylogenetic analysis did not identify the specific identities of donors, it indicates that the introns were likely acquired independently; Cbu.L1917 was acquired from other bacteria like Thermotoga subterranea and Cbu.L1951 from lower eukaryotes like Acanthamoeba castellanii. We also confirmed the fragmented nature of mature 23S rRNA in C. burnetii due to the presence of an intervening sequence. The presence of three selfish elements in C. burnetii's 23S rRNA gene is very unusual for an obligate intracellular bacterium and suggests a recent shift to its current lifestyle from a previous niche with greater opportunities for lateral gene transfer. PMID:17644584

  16. Role of pathogens in multiple sclerosis.

    Libbey, Jane E; Cusick, Matthew F; Fujinami, Robert S

    2014-01-01

    Multiple sclerosis (MS) is an inflammatory demyelinating autoimmune disease of the central nervous system (CNS). Although the etiology of MS is unknown, genetic and environmental factors play a role. Infectious pathogens are the likely environmental factors involved in the development of MS. Pathogens associated with the development or exacerbation of MS include bacteria, such as Mycoplasma pneumoniae and Chlamydia pneumoniae, the Staphylococcus aureus-produced enterotoxins that function as superantigens, viruses of the herpes virus (Epstein-Barr virus and human herpesvirus 6) and human endogenous retrovirus (HERV) families and the protozoa Acanthamoeba castellanii. Evidence, from studies with humans and animal models, supporting the association of these various pathogens with the development and/or exacerbation of MS will be discussed along with the potential mechanisms including molecular mimicry, epitope spreading and bystander activation. In contrast, infection with certain parasites such as helminthes (Schistosoma mansoni, Fasciola hepatica, Hymenolepis nana, Trichuris trichiura, Ascaris lumbricoides, Strongyloides stercolaris, Enterobius vermicularis) appears to protect against the development or exacerbation of MS. Evidence supporting the ability of parasitic infections to protect against disease will be discussed along with a brief summary of a recent Phase I clinical trial testing the ability of Trichuris suis ova treatment to improve the clinical course of MS. A complex interaction between the CNS (including the blood-brain barrier), multiple infections with various infectious agents (occurring in the periphery or within the CNS), and the immune response to those various infections may have to be deciphered before the etiology of MS can be fully understood. PMID:24266364

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

    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.

  18. cDNA sequence analysis of a 29-kDa cysteine-rich surface antigen of pathogenic Entamoeba histolytica

    Torian, B.E.; Stroeher, V.L.; Stamm, W.E. (Univ. of Washington, Seattle (USA)); Flores, B.M. (Louisiana State Univ. Medical Center, New Orleans (USA)); Hagen, F.S. (Zymogenetics Incorporated, Seattle, WA (USA))

    1990-08-01

    A {lambda}gt11 cDNA library was constructed from poly(U)-Spharose-selected Entamoeba histolytica trophozoite RNA in order to clone and identify surface antigens. The library was screened with rabbit polyclonal anti-E. histolytica serum. A 700-base-pair cDNA insert was isolated and the nucleotide sequence was determined. The deduced amino acid sequence of the cDNA revealed a cysteine-rich protein. DNA hybridizations showed that the gene was specific to E. histolytica since the cDNA probe reacted with DNA from four axenic strains of E. histolytica but did not react with DNA from Entamoeba invadens, Acanthamoeba castellanii, or Trichomonas vaginalis. The insert was subcloned into the expression vector pGEX-1 and the protein was expressed as a fusion with the C terminus of glutathione S-transferase. Purified fusion protein was used to generate 22 monoclonal antibodies (mAbs) and a mouse polyclonal antiserum specific for the E. histolytica portion of the fusion protein. A 29-kDa protein was identified as a surface antigen when mAbs were used to immunoprecipitate the antigen from metabolically {sup 35}S-labeled live trophozoites. The surface location of the antigen was corroborated by mAb immunoprecipitation of a 29-kDa protein from surface-{sup 125}I-labeled whole trophozoites as well as by the reaction of mAbs with live trophozoites in an indirect immunofluorescence assay performed at 4{degree}C. Immunoblotting with mAbs demonstrated that the antigen was present on four axenic isolates tested. mAbs recognized epitopes on the 29-kDa native antigen on some but not all clinical isolates tested.

  19. Transcriptional down-regulation and rRNA cleavage in Dictyostelium discoideum mitochondria during Legionella pneumophila infection.

    Chenyu Zhang

    Full Text Available Bacterial pathogens employ a variety of survival strategies when they invade eukaryotic cells. The amoeba Dictyostelium discoideum is used as a model host to study the pathogenic mechanisms that Legionella pneumophila, the causative agent of Legionnaire's disease, uses to kill eukaryotic cells. Here we show that the infection of D. discoideum by L. pneumophila results in a decrease in mitochondrial messenger RNAs, beginning more than 8 hours prior to detectable host cell death. These changes can be mimicked by hydrogen peroxide treatment, but not by other cytotoxic agents. The mitochondrial large subunit ribosomal RNA (LSU rRNA is also cleaved at three specific sites during the course of infection. Two LSU rRNA fragments appear first, followed by smaller fragments produced by additional cleavage events. The initial LSU rRNA cleavage site is predicted to be on the surface of the large subunit of the mitochondrial ribosome, while two secondary sites map to the predicted interface with the small subunit. No LSU rRNA cleavage was observed after exposure of D. discoideum to hydrogen peroxide, or other cytotoxic chemicals that kill cells in a variety of ways. Functional L. pneumophila type II and type IV secretion systems are required for the cleavage, establishing a correlation between the pathogenesis of L. pneumophila and D. discoideum LSU rRNA destruction. LSU rRNA cleavage was not observed in L. pneumophila infections of Acanthamoeba castellanii or human U937 cells, suggesting that L. pneumophila uses distinct mechanisms to interrupt metabolism in different hosts. Thus, L. pneumophila infection of D. discoideum results in dramatic decrease of mitochondrial RNAs, and in the specific cleavage of mitochondrial rRNA. The predicted location of the cleavage sites on the mitochondrial ribosome suggests that rRNA destruction is initiated by a specific sequence of events. These findings suggest that L. pneumophila specifically disrupts mitochondrial

  20. Nuclease Activity of Legionella pneumophila Cas2 Promotes Intracellular Infection of Amoebal Host Cells

    Gunderson, Felizza F.; Mallama, Celeste A.; Fairbairn, Stephanie G.

    2014-01-01

    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. PMID:25547789

  1. Iron Limitation Triggers Early Egress by the Intracellular Bacterial Pathogen Legionella pneumophila.

    O'Connor, Tamara J; Zheng, Huaixin; VanRheenen, Susan M; Ghosh, Soma; Cianciotto, Nicholas P; Isberg, Ralph R

    2016-08-01

    Legionella pneumophila is an intracellular bacterial pathogen that replicates in alveolar macrophages, causing a severe form of pneumonia. Intracellular growth of the bacterium depends on its ability to sequester iron from the host cell. In the L. pneumophila strain 130b, one mechanism used to acquire this essential nutrient is the siderophore legiobactin. Iron-bound legiobactin is imported by the transport protein LbtU. Here, we describe the role of LbtP, a paralog of LbtU, in iron acquisition in the L. pneumophila strain Philadelphia-1. Similar to LbtU, LbtP is a siderophore transport protein and is required for robust growth under iron-limiting conditions. Despite their similar functions, however, LbtU and LbtP do not contribute equally to iron acquisition. The Philadelphia-1 strain lacking LbtP is more sensitive to iron deprivation in vitro Moreover, LbtP is important for L. pneumophila growth within macrophages while LbtU is dispensable. These results demonstrate that LbtP plays a dominant role over LbtU in iron acquisition. In contrast, loss of both LbtP and LbtU does not impair L. pneumophila growth in the amoebal host Acanthamoeba castellanii, demonstrating a host-specific requirement for the activities of these two transporters in iron acquisition. The growth defect of the ΔlbtP mutant in macrophages is not due to alterations in growth kinetics. Instead, the absence of LbtP limits L. pneumophila replication and causes bacteria to prematurely exit the host cell. These results demonstrate the existence of a preprogrammed exit strategy in response to iron limitation that allows L. pneumophila to abandon the host cell when nutrients are exhausted. PMID:27185787

  2. Seasonal variation and incidence of Stictodora tridactyla in fish from lake Manzala.

    Abou-Zakham, A; el-Shazley, A; el-Ganayni, F; Romia, S A; Abou-Shady, A F; el-Kholy, E I

    1990-06-01

    Stictodora tridactyla encysted metacercariae were detected in Mugil and Tilapia collected only from brackish water fish from lake Manzala. The prevalence and intensity of encysted metacercariae were higher in summer than in winter. PMID:2332641

  3. Effect of mutating the regulatory phosphoserine and conserved threonine on the activity of the expressed catalytic domain of Acanthamoeba myosin I heavy chain kinase

    Szczepanowska, Joanna; Ramachandran, Umamaheswari; Herring, Christopher J.; Gruschus, James M.; Qin, Jun; Korn, Edward D.; Brzeska, Hanna

    1998-01-01

    Phosphorylation of Ser-627 is both necessary and sufficient for full activity of the expressed 35-kDa catalytic domain of myosin I heavy chain kinase (MIHCK). Ser-627 lies in the variable loop between highly conserved residues DFG and APE at a position at which a phosphorylated Ser/Thr also occurs in many other Ser/Thr protein kinases. The variable loop of MIHCK contains two other hydroxyamino acids: Thr-631, which is conserved in almost all Ser/Thr kinases, and Thr-632, which is not conserve...

  4. Effects of drinking water in Acanthamoeba contamination of contact lens and contact lens storage cases%饮用水对角膜接触镜保存盒棘阿米巴污染的影响

    王月华; 崔春权; 郑善子

    2006-01-01

    [目的]观察饮用水的棘阿米巴污染情况,研究其与角膜接触镜保存盒棘阿米巴污染的关系[方法]对延吉市居民饮用水棘阿米巴污染情况及大学生角膜接触镜保存盒和保存液的棘阿米巴污染情况进行调查.将103份饮用水和93份角膜接触镜保存液及保存盒擦拭液接种于有灭活大肠杆菌的无营养琼脂培养基中在28℃条件下培养1周,观察棘阿米巴生长情况.[结果]93份角膜接触镜保存盒擦拭液及保存液中有1份(1%)检出棘阿米巴污染,103份饮用水中有41份(40%)检出棘阿米巴污染[结论]角膜接触镜保存液和保存盒棘阿米巴污染可能与不正确地使用饮用水清洗和保存角膜接触镜有关.

  5. Use of Phage Antibodies to Distinguish Closely Related Species of Protozoan Parasites

    Paget, Timothy; Khan, Naveed; Temple, Graham; Hough, Victoria; Greenman, John

    2000-01-01

    Acanthamoeba are typically identified in the laboratory using culture and microscopic observation. In this paper we describe the isolation and specificity of antibody fragments that can be used for the identification of Acanthamoeba. A phage library expressing a large repertoire (approx. 5×109) of antibody fragments was used to generate two libraries one enriched for bacteriophage that exhibit genus specific binding and the other containing bacteriophage that bind specifically to pathogenic Acanthamoeba. Individual clones were isolated on the basis of binding by ELISA, and then flowcytometry and immunofluorescence were used for further characterisation. Four monoclonal antibodies were isolated, specific for Acanthamoeba at the generic level with clone HPPG6 exhibiting the highest level of binding. Furthermore clone HPPG55 was specific for pathogenic species of Acanthamoeba. PMID:11360833

  6. Use of Phage Antibodies to Distinguish Closely Related Species of Protozoan Parasites

    Timothy Paget

    2000-01-01

    Full Text Available Acanthamoeba are typically identified in the laboratory using culture and microscopic observation. In this paper we describe the isolation and specificity of antibody fragments that can be used for the identification of Acanthamoeba. A phage library expressing a large repertoire (approx. 5×109 of antibody fragments was used to generate two libraries one enriched for bacteriophage that exhibit genus specific binding and the other containing bacteriophage that bind specifically to pathogenic Acanthamoeba. Individual clones were isolated on the basis of binding by ELISA, and then flowcytometry and immunofluorescence were used for further characterisation. Four monoclonal antibodies were isolated, specific for Acanthamoeba at the generic level with clone HPPG6 exhibiting the highest level of binding. Furthermore clone HPPG55 was specific for pathogenic species of Acanthamoeba.

  7. Population dynamics of active and total ciliate populations in arable soil amended with wheat

    Ekelund, F.; Frederiksen, Helle B.; Ronn, R.

    2002-01-01

    population may be encysted. The factors governing the dynamics of active and encysted cells in the soil are not well understood. Our objective was to determine the dynamics of active and encysted populations of ciliates during the decomposition of freshly added organic material. We monitored, in soil...... mathematical modeling. Following the addition of fresh organic material, bacterial numbers increased more than 1,400-fold. There was a temporary increase in the number of active ciliates, followed by a rapid decline, although the size of the bacterial prey populations remained high. During this initial burst...

  8. Dicty_cDB: Contig-U06541-1 [Dicty_cDB

    Full Text Available cyclase... 44 7.5 1 ( EF152287 ) Strongylocentrotus purpuratus BRCA1 (brca1) mRNA,... 44 7.5 1 ( AC131169 ) ...1 ( CW927554 ) EDCBM45TF A. castellanii, 6-8 kb library from tot... 44 7.5 1 ( CL534328 ) trib011xa13.g1 T. reesei HindIII BAC libr...us lewisii genomic, ge... 44 7.5 1 ( CW927722 ) EDCBN39TR A. castellanii, 6-8 kb library from tot... 44 7.5 ...ce (All Frames) Frame A: t*idtnkkmkllfilliltitiknclsinindnnnqiinlkindtsysmvpyqdeaca...E Sequences producing significant alignments: (bits) Value Contig-U06541-1 (Conti

  9. Аcanthamoeba keratitis and outcomes of the treatment (clinical cases

    M. M. Bikbov

    2016-01-01

    Full Text Available Acanthamoeba keratitis (AK — corneal inflammation caused by protozoa — Acanthamoeba. The disease is related mainly with wearing of contact lens, pathogen — Acanthamoeba penetrates under the contact lenses. The cases of AK after Lasik are described. Corneal epithelium microtrauma and contact with sources of environmental pollution are the risk factors of the invasion of Acanthamoeba into the cornea.Acanthamoebas affect soil, stagnant reservoirs, swimming pools, drain pipes, tap water, etc. Diagnosis can be confirmed only when cysts are detected in cornea material, plate count, smear, and the containers where such material is stored. Long life cycle including cyst stage helps keratitis relapsing course last for months. It is not always possible to detect cysts. Assessment of etiology presents certain difficulties.Such aspects as history taking, monitoring the clinical course of disease, confocal microscopy that enables to reveal acanthamoebas in vivo help to determine a diagnosis.Disease is characterized by a long chronic course with the development descemetocele and cornea perforation. The sequence of remissions and exacerbations of inflammation, apparently, occurs due to peculiarities of Acanthamoeba life cycle. Acanthamoeba keratitis is difficult to treat, it’s resistant to antibiotics. Untimely diagnosed disease and delayed initiation of treatment in patients with AK promote corneal perforation and inflammatory process in deep eye structures what often leads to eye enucleation.Severe pain is typical for AK in onset of disease. It is caused by emergence of subepithelial infiltrates and defects along the nerve fibers located in the most sensitive surface layer of the cornea. Conservative treatment is conducted with the use of antiseptics. The most effective one against cysts is 0.02% chlorhexidine which is prepared ex. temporae.The article presents the peculiarities of clinical course of Acanthamoeba keratitis, clinical cases of the

  10. Pumilio Homologue from Saprolegnia parasitica Specifically Expressed in Undifferentiated Spore Cysts

    Andersson, M. Gunnar; Cerenius, Lage

    2002-01-01

    The expression of spore-specific marker transcripts at different stages of the asexual life cycle of Saprolegnia parasitica was analyzed. One of the markers, designated puf1, was found to be expressed transiently upon each of several cycles of zoospore encystment and reemergence. The transcript is induced immediately upon zoospore encystment and is rapidly lost when a cyst is triggered to germinate. In nongerminating cysts, puf1 is maintained until a time point when the cysts can no longer be...

  11. Chapter 42. Waterborne and Foodborne Parasites

    This chapter identifies the most prominent parasites in North America that are acquired through contaminated food and water including protozoa (Acanthamoeba, Naegleria, Entamoeba, Giardia, Cryptosporidium, Cystoisospora, Cyclospora, Toxoplasma, and Balantidium), nematodes (Trichinella, Angiostrongyl...

  12. Effect of disinfectants on pathogenic free-living amoebae: in axenic conditions.

    Cursons, R T; Brown, T J; Keys, E A

    1980-01-01

    The amoebicidal properties of chlorine, chlorine dioxide, ozone, and deciquam 222 were examined in axenic conditions. Naegleria spp. were found to be more sensitive to chlorine and chlorine dioxide than Acanthamoeba spp. No marked difference in sensitivity to ozone or deciquam 222 could be detected between the pathogenic (A-1) and nonpathogenic (1501) strains of Acanthamoeba and the pathogenic (MsT) and nonpathogenic (P1200f) strains of Naegleria. Methods of disinfection are discussed with re...

  13. Marine mimivirus relatives are probably large algal viruses

    Claverie Jean-Michel; Sandaa Ruth-Anne; Bratbak Gunnar; Larsen Jens; Monier Adam; Ogata Hiroyuki

    2008-01-01

    Abstract Background Acanthamoeba polyphaga mimivirus is the largest known ds-DNA virus and its 1.2 Mb-genome sequence has revealed many unique features. Mimivirus occupies an independent lineage among eukaryotic viruses and its known hosts include only species from the Acanthamoeba genus. The existence of mimivirus relatives was first suggested by the analysis of the Sargasso Sea metagenomic data. Results We now further demonstrate the presence of numerous "mimivirus-like" sequences using a l...

  14. Marine mimivirus relatives are probably large algal viruses

    Monier, Adam; Borggaard Larsen, Jens; Sandaa, Ruth-Anne; Bratbak, Gunnar; Claverie, Jean-Michel; Ogata, Hiroyuki

    2008-01-01

    Background Acanthamoeba polyphaga mimivirus is the largest known ds-DNA virus and its 1.2 Mb-genome sequence has revealed many unique features. Mimivirus occupies an independent lineage among eukaryotic viruses and its known hosts include only species from the Acanthamoeba genus. The existence of mimivirus relatives was first suggested by the analysis of the Sargasso Sea metagenomic data. Results We now further demonstrate the presence of numerous "mimivirus-like" sequences using a ...

  15. Use of Phage Antibodies to Distinguish Closely Related Species of Protozoan Parasites

    Timothy Paget; Naveed Khan; Graham Temple; Victoria Hough; John Greenman

    2002-01-01

    Acanthamoeba are typically identified in the laboratory using culture and microscopic observation. In this paper we describe the isolation and specificity of antibody fragments that can be used for the identification of Acanthamoeba. A phage library expressing a large repertoire (approx. 5×109) of antibody fragments was used to generate two libraries one enriched for bacteriophage that exhibit genus specific binding and the other containing bacteriophage that bind specifically to pathogenic A...

  16. Treatment of Granulomatous Amoebic Encephalitis with Voriconazole and Miltefosine in an Immunocompetent Soldier

    Webster, Duncan; Umar, Imram; Kolyvas, George; Bilbao, Juan; Guiot, Marie-Christine; Duplisea, Kevin; Qvarnstrom, Yvonne; Visvesvara, Govinda S.

    2012-01-01

    A 38-year-old male immunocompetent soldier developed generalized seizures. He underwent surgical debulking and a progressive demyelinating pseudotumor was identified. Serology and molecular testing confirmed a diagnosis of granulomatous amoebic encephalitis caused by Acanthamoeba sp. in this immunocompetent male. The patient was treated with oral voriconazole and miltefosine with Acanthamoeba titers returning to control levels and serial imaging demonstrating resolution of the residual lesion.

  17. Nye diagnostiske tiltag ved akantamøbekeratitis

    Nielsen, Esben Tranholm; Heegaard, Steffen; Hjortdal, Jesper Østergaard

    2011-01-01

    Acanthamoeba keratitis is a rare but sight threatening condition. A major problem is that the disease is difficult to diagnose and often mistaken for herpes infection in its early stages. We present an update in diagnostics with the presentation of five recent cases of Acanthamoeba keratitis wher...... the use of confocal microscopy and PCR based DNA analysis of epithelial scrapings played a pivotal role. An early diagnose is crucial for achieving a successful outcome....

  18. Role of Contact Lens Wear, Bacterial Flora, and Mannose-Induced Pathogenic Protease in the Pathogenesis of Amoebic Keratitis

    Alizadeh, Hassan; Neelam, Sudha; Hurt, Michael; Niederkorn, Jerry Y.

    2005-01-01

    The ocular surface is continuously exposed to potential pathogens, including free-living amoebae. Acanthamoeba species are among the most ubiquitous amoebae, yet Acanthamoeba keratitis is remarkably rare. The pathogenesis of Acanthamoeba keratitis is a complex, sequential process. Here we show that Acanthamoeba keratitis is profoundly affected by mannosylated proteins on the ocular surface, which stimulate the amoebae to elaborate a 133-kDa pathogenic protease. The mannose-induced protease (MIP133) mediates apoptosis of the corneal epithelium, facilitates corneal invasion, and degrades the corneal stroma. We show that contact lens wear upregulates mannosylated proteins on the corneal epithelium, stimulates MIP133 secretion, and exacerbates corneal disease. Corynebacterium xerosis, a constituent of the ocular flora, contains large amounts of mannose and is associated with Acanthamoeba keratitis. The present results show that amoebae exposed to C. xerosis produce increased amounts of MIP133 and more severe corneal disease. Oral immunization with MIP133 mitigates Acanthamoeba keratitis and demonstrates the feasibility of antidisease vaccines for pathogens that resist immune elimination. PMID:15664950

  19. The role of an organic matrix during the formation of siliceous scales in the heliozoon Actinophrys sol (actinophryida, protista).

    Newman, P J; Patterson, D J

    1993-07-25

    Actinophrys sol is a freshwater heliozoon which has trophic and encysted body forms. During encystment, siliceous scales are laid down in silica deposition vesicles. The scales form one layer of a multi-layered cyst wall. Scale production is described using light microscopy, transmission electron microscopy, scanning electron microscopy, and X-ray microanalysis. Silica is laid down on an organic matrix which is visible prior to silicification and after removal of silica with hydrofluoric acid. Actinophrys sol can be cultured under silica impoverished conditions, with the result that the siliceous plates are absent. The cysts continue to form but are fragile. Silica is not a prerequisite for the processes of encystment and cyst formation. PMID:23195646

  20. Incorporation of 3H-thymidine during early developmental stages of ovoviviparous and oviparous embryos in the brine

    DNA synthesis in the embryos during various developmental stages in the brood pouch of the Artemia female as well as the encysted eggs produced by oviparity was studied by autoradiography using 3H-thymidine. Nuclei of the embryos through the blastula stage in the brood pouch were labelled, regardless of the existence of the thick shell surrounding the embryos. However, incorporation of 3H-thymidine into nuclei was absent in the stages between gastrula and early nauplii which had developed in the brood pouch or were delivered from encysted dry eggs. (author)

  1. ROLE OF CA2+ AND CAMP IN A CELL SIGNALING PATHWAY FOR RESTING CYST FORMATION OF CILIATED PROTOZOAN COLPODA CUCULLUS

    Matsuoka, Tatsuomi; Kondoh, Asuka; Sabashi, Kunihisa; Nagano, Nobuaki; Akematsu, Takahiko; Kida, Akemi; Iino, Ryota

    2009-01-01

    Resting cyst formation (encystment) of Colpoda cucullus is caused by an increase in an external Ca2+ concentration or overpopulation of Colpoda vegetative cells. The Ca2+-mediated or overpopulation-mediated encystment was suppressed by Ca2+ channel blockers (Cd2+, La3+, Ni2+), Ca2+-chelating reagents (EGTA, BAPTA), calmodulin antagonists (W-7, trifluoperazine), Rp-cAMPS (an cAMP analog antagonist) and 2-deoxyadenosine (a P-site inhibitor of adenylate cyclase). On the other hand, by the additi...

  2. Dicty_cDB: Contig-U15722-1 [Dicty_cDB

    Full Text Available ium discoideum chromosome 2 map 2234041... 38 0.38 18 ( CW932140 ) EDCCB74TF A. castellanii, 6-8 kb libra...ent US 6258559. 46 7.1 1 ( AY392431 ) Dictyostelium discoideum DNA ligase gene, complet... 46 7.1 1 ( AC171134 ) Helobdella...:none) Dictyostelium discoideum chromoso... 59 1e-06 X08030_1( X08030 |pid:none) Chicken mRNA frag...cdndsgn ckcdtqtkgltceesrlliesldsinskggtiniigyfgnttslltikigesqcknikvf nettlkcdigegkgihnitiidddlsftainkfqyldektkiskisrgaiagi...cdndsgn ckcdtqtkgltceesrlliesldsinskggtiniigyfgnttslltikigesqcknikvf nettlkcdigegkgihnitiidddlsftainkfqyldektkiskisrgaiagi

  3. Vannellid Species Isolated from Freshwater Source in a Park in Jamaica, West Indies.

    Todd, Cheridah D; Reyes-Batlle, María; Valladares, Basilio; Lindo, John F; Lorenzo-Morales, Jacob

    2015-01-01

    Free-living amoebae (FLA) occupy a wide range of freshwater, marine, and soil habitats, and are opportunistic pathogens in human beings. While Acanthamoeba spp., Naegleria fowleri, and Balamuthia mandrillaris are well-known opportunistic organisms, Vannella epipetala is nonpathogenic. Sediments were collected from a freshwater source from a park in Jamaica to investigate the presence of FLA. Acanthamoeba and Naegleria spp. were not recovered; however, a Vannellid species identified by microscopy and PCR analysis as V. epipetala was isolated. These nonpathogens pose a threat to human beings as they may act as Trojan horses for microsporidian parasites and other pathogens, thereby facilitating their transmission to human beings. PMID:26512204

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

    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

  5. Characterization of IgG monoclonal antibody targeted to both tissue cyst and sporocyst walls of Toxoplasma gondii

    Toxoplasma gondii infects approximately one third of the human population and animals habiting terrestrial and aquatic environments. Its environmentally resistant oocysts are excreted by felids, and the stage encysted in tissues (tissue cysts), are important in the horizontal transmission of T. gon...

  6. Molecular Characterization of a New Tetratrichomonas Species in a Patient with Empyema▿

    Mantini, Cléa; Souppart, Laetitia; Noël, Christophe; Duong, Thanh Hai; Mornet, Michel; Carroger, Guy; Dupont, Pierre; Masseret, Elodie; Goustille, Julien; Capron, Monique; Duboucher, Christophe; Dei-Cas, Eduardo; Viscogliosi, Eric

    2009-01-01

    A new Tetratrichomonas species was identified by molecular and phylogenetic approaches in the pleural fluid from a patient with encysted empyema leading to dyspnea. This observation raised the questions of the real prevalence of pulmonary trichomonosis in humans, the zoonotic potential of trichomonads, and the existence of human-host-adapted strains. PMID:19420167

  7. Molecular Characterization of a New Tetratrichomonas Species in a Patient with Empyema▿

    Mantini, Cléa; Souppart, Laetitia; Noël, Christophe; Duong, Thanh Hai; Mornet, Michel; Carroger, Guy; Dupont, Pierre; Masseret, Elodie; Goustille, Julien; Capron, Monique; Duboucher, Christophe; Dei-Cas, Eduardo; Viscogliosi, Eric

    2009-01-01

    A new Tetratrichomonas species was identified by molecular and phylogenetic approaches in the pleural fluid from a patient with encysted empyema leading to dyspnea. This observation raised the questions of the real prevalence of pulmonary trichomonosis in humans, the zoonotic potential of trichomonads, and the existence of human-host-adapted strains.

  8. A new plain film finding in cysticercosis

    Ruiz, M.E.; Haughton, V.M. (Medical Coll. of Wisconsin, Milwaukee, WI (USA). Dept. of Radiology)

    1991-02-01

    Lytic lesions of the skull due to cysticercosis are described. Encysted larvae in the subarachnoid space eroded the inner table of the skull in a 10-year-old Guatemalan male. In an area with endemic cysticercosis infections, this diagnosis should be considered when round, lytic defects are demonstrated in a skull radiograph. (orig.).

  9. A new plain film finding in cysticercosis

    Lytic lesions of the skull due to cysticercosis are described. Encysted larvae in the subarachnoid space eroded the inner table of the skull in a 10-year-old Guatemalan male. In an area with endemic cysticercosis infections, this diagnosis should be considered when round, lytic defects are demonstrated in a skull radiograph. (orig.)

  10. What's in a frog stomach? Solving a 150 year old mystery (Diptera: Calliphoridae)

    The taxon Acanthosoma chrysalis Mayer, 1844, described from Germany on a number of alleged parasites encysted in the peritoneal wall of the stomach of edible frogs, is revised and shown to be first instar larvae of blow flies (Calliphoridae). Based on the shape of mouthhooks and abdominal cuticular ...

  11. The neurotropic parasite Toxoplasma gondii increases dopamine metabolism

    The common parasite Toxoplasma gondii induces behavioral alterations in its hosts including phenotypes increasing the likelihood of its transmission in rodents and reports of psychobehavioral alterations in humans. We have found that elevated levels of dopamine are associated with the encysted stage...

  12. The Ciliate Colpoda: "Instant" Protozoan

    Smith, Anne Muller; Giese, Arthur C.

    1973-01-01

    Describes the characteristics of Colpoda, a ciliated protozoan which is able to survive in a dry, encysted state for long periods of time. Outlines the procedures for culturing the organism and producing cyst preparations, and recommends its use in the high school biology laboratory. (JR)

  13. Substratum preferences in two notocotylid (Digenea, Notocotylidae) cercariae from Hydrobia ventrosa at the White Sea

    Gonchar, Anna; Galaktionov, Kirill V.

    2016-07-01

    Notocotylids are digeneans with a two-host life cycle. Their cercariae encyst on underwater substrata, and final hosts (mostly water birds) get infected by consuming encysted metacercariae. The aim of this study was to assess whether notocotylid cercariae have encystment substratum preferences. We used Cercaria Notocotylidae sp. No 11 Deblock, 1980 (presumably Paramonostomum alveatum Mehlis 1846) and C. Notocotylidae sp. No 12 Deblock, 1980 associated with mudsnails Hydrobia ventrosa at the White Sea. Three series of experiments were performed in which distribution of cysts across different combinations of substrata was measured. The results suggest that C. Notocotylidae sp. No 11 cercariae encyst almost exclusively on the leaves of green plants without any plant species preferences. C. Notocotylidae sp. No 12 cercariae use shells of living molluscs and plant substrata equally often but avoid empty shells. These preferences are probably adapted to the feeding habits of the final hosts and this may enhance the transmission of the studied notocotylid species. Mechanisms of the observed preferences might be associated with the ability of cercariae to recognize substrata features: either chemical cues or surface structure. Substratum selectivity in cercariae is considered as a first step towards downward incorporation of the second intermediate host into the life cycle.

  14. Delayed tension pneumothorax complicating staphylococcal pneumonia.

    Yates, S. P.; Morcos, S. K.

    1988-01-01

    A case is described in which a tension pneumothorax complicated staphylococcal pneumonia 11 months after its onset. The delayed and subacute/chronic nature of the tension pneumothorax is unusual. The case also highlights the difficult differential diagnosis between subpleural lung cysts and encysted pneumothorax.

  15. Redescription and life cycle of the monorchiid Postmonorcheides maclovini Szidat, 1950 (Digenea) from the Southwestern Atlantic Ocean: Morphological and molecular data.

    Bagnato, Estefanía; Gilardoni, Carmen; Pina, Susana; Rodrigues, Pedro; Cremonte, Florencia

    2016-02-01

    The adult monorchiid, Postmonorcheides maclovini Szidat, 1950, digenean parasite of the Patagonian blennie Eleginops maclovinus (Cuvier) (Eleginopidae) from Puerto Deseado (47° 45' S, 65° 55' W), Argentina, was characterized and its life cycle elucidated. P. maclovinus is the only species of the genus Postmonorcheides, proposed by Szidat (1950) from Tierra del Fuego province (~54° S), Argentina. This digenean uses the Patagonian blennie as definitive host, and the intertidal bivalve Lasaea adansoni (Gmelin) (Lasaeidae) as both first and second intermediate hosts (metacercariae encyst inside sporocysts), being the first record of this clam as intermediate host of trematode parasites. The cercaria may, in addition to encysting in the sporocyst, emerge and presumably infect other intermediate hosts. This is the second report of a monorchiid species with metacercariae encysting inside the sporocyst. Adults were found parasitizing the fish stomach, pyloric caeca and intestine with a prevalence of 100%; sporocysts with cercariae and/or metacercariae were found parasitizing the gonad of the bivalve with a prevalence of 2.78%. The cercariae possess a well-developed tail and eye-spots are absent. The ITS1 sequence from the adult digeneans found in the Patagonian blennie, identified as P. maclovini, was found to be identical to the ITS1 sequences obtained both from sporocysts containing cercariae and encysted metacercariae found in L. adansoni. PMID:26423199

  16. In vitro induction of Entamoeba histolytica cyst-like structures from trophozoites.

    Aguilar-Díaz, Hugo; Díaz-Gallardo, Martha; Laclette, Juan P; Carrero, Julio C

    2010-01-01

    Inhibition of encystment can be conceived as a potentially useful mechanism to block the transmission of Entamoeba histolytica under natural conditions. Unfortunately, amoeba encystment has not been achieved in vitro and drugs inhibiting the formation of cysts are not available. Luminal conditions inducing encystment in vivo are also unknown, but cellular stress such as exposure to reactive oxygen species from immune cells or intestinal microbiota could be involved. A role for certain divalent cations as cofactors of enzymes involved in excystment has also been described. In this study, we show that trophozoite cultures, treated with hydrogen peroxide in the presence of trace amounts of several cations, transform into small-sized spherical and refringent structures that exhibit resistance to different detergents. Ultrastructural analysis under scanning and transmission electron microscopy revealed multinucleated structures (some with four nuclei) with smooth, thick membranes and multiple vacuoles. Staining with calcofluor white, as well as an ELISA binding assay using wheat germ agglutinin, demonstrated the presence of polymers of N-acetylglucosamine (chitin), which is the primary component of the natural cyst walls. Over-expression of glucosamine 6-phosphate isomerase, likely to be the rate-limiting enzyme in the chitin synthesis pathway, was also confirmed by RT-PCR. These results suggest that E. histolytica trophozoites activated encystment pathways when exposed to our treatment. PMID:20169067

  17. In vitro induction of Entamoeba histolytica cyst-like structures from trophozoites.

    Hugo Aguilar-Díaz

    Full Text Available Inhibition of encystment can be conceived as a potentially useful mechanism to block the transmission of Entamoeba histolytica under natural conditions. Unfortunately, amoeba encystment has not been achieved in vitro and drugs inhibiting the formation of cysts are not available. Luminal conditions inducing encystment in vivo are also unknown, but cellular stress such as exposure to reactive oxygen species from immune cells or intestinal microbiota could be involved. A role for certain divalent cations as cofactors of enzymes involved in excystment has also been described. In this study, we show that trophozoite cultures, treated with hydrogen peroxide in the presence of trace amounts of several cations, transform into small-sized spherical and refringent structures that exhibit resistance to different detergents. Ultrastructural analysis under scanning and transmission electron microscopy revealed multinucleated structures (some with four nuclei with smooth, thick membranes and multiple vacuoles. Staining with calcofluor white, as well as an ELISA binding assay using wheat germ agglutinin, demonstrated the presence of polymers of N-acetylglucosamine (chitin, which is the primary component of the natural cyst walls. Over-expression of glucosamine 6-phosphate isomerase, likely to be the rate-limiting enzyme in the chitin synthesis pathway, was also confirmed by RT-PCR. These results suggest that E. histolytica trophozoites activated encystment pathways when exposed to our treatment.

  18. Phosphonoxins: Rational design and discovery of a potent nucleotide anti-Giardia agent

    Suk, D. H.; Rejman, Dominik; Dykstra, Ch.; Pohl, Radek; Pankiewicz, K. W.; Patterson, S. E.

    2007-01-01

    Roč. 17, č. 10 (2007), s. 2811-2816. ISSN 0960-894X Institutional research plan: CEZ:AV0Z40550506 Keywords : Giardia * encystation * cyst wall synthase * nucleotide Subject RIV: CC - Organic Chemistry Impact factor: 2.604, year: 2007

  19. Granulomatous amebic encephalitis following hematopoietic stem cell transplantation

    Ninh Doan

    2015-01-01

    Conclusion: The authors report the third case of GAE after autologous stem cell transplant, and the ninth case overall after HSCT. This case is unusual due to its rapid clinical presentation after HSCT compared to prior literature. The case highlights the need for high suspicion of Acanthamoeba infection in this patient population.

  20. 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

    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...

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

    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.

  2. Counting Legionella cells within single amoeba host cells

    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...

  3. [THERAPEUTIC ACTIVITY OF MICRONIZED MEBENDAZOLE IN THE MUSCULAR PHASE OF EXPERIMENTAL TRICHINELLA SPIRALIS INVASION IN ALBINO MICE].

    Kukhaleva, I V; Kovalenko, F P; Shkolyar, N A; Legonkov, Yu A; Musaev, A Kh; Bulanova, T E; Samochatova, E I

    2015-01-01

    The incidence of trichinosis in Russia was 0.07 per 100,000 population in 2014, which was 2.9-fold higher than that in 2013. Two WHO recommended medications mebendazole and albendazole are now used to treat humari trichinosis. The drugs are active against only mature helminths and non-encysted muscle larvae. The original oil suspension of micronized mebendazole was.found to have 100% efficacy against trichinosis in albino mice in the late muscular phase (encysted larvae) of hyperinvasion after intensive therapy under lifetime diagnostic guidance during and after a treatment cycle. The lifetime diagnostic method used to evaluate the larvicidal activity of anti-trichinosis agents in animals with experimental trichinosis revealed the signs of viaility, established a trend for deatih of Trichinella larvae, and determined their destructive changes. PMID:26827586

  4. Scanning electron microscope observations of brine shrimp larvae from space shuttle experiments

    DeBell, L.; Paulsen, A.; Spooner, B.

    1992-01-01

    Brine shrimp are encysted as gastrula stage embryos, and may remain dehydrated and encysted for years without compromising their viability. This aspect of brine shrimp biology is desirable for studying development of animals during space shuttle flight, as cysts placed aboard a spacecraft may be rehydrated at the convenience of an astronaut, guaranteeing that subsequent brine shrimp development occurs only on orbit and not on the pad during launch delays. Brine shrimp cysts placed in 5 ml syringes were rehydrated with salt water and hatched during a 9 day space shuttle mission. Subsequent larvae developed to the 8th larval stage in the sealed syringes. We studied the morphogenesis of the brine shrimp larvae and found the larvae from the space shuttle experiments similar in rate of growth and extent of development, to larvae grown in sealed syringes on the ground. Extensive differentiation and development of embryos and larvae can occur in a microgravity environment.

  5. Homologous recombination occurs in Entamoeba and is enhanced during growth stress and stage conversion.

    Nishant Singh

    Full Text Available Homologous recombination (HR has not been demonstrated in the parasitic protists Entamoeba histolytica or Entamoeba invadens, as no convenient method is available to measure it. However, HR must exist to ensure genome integrity, and possible genetic exchange, especially during stage conversion from trophozoite to cyst. Here we show the up regulation of mitotic and meiotic HR genes in Entamoeba during serum starvation, and encystation. To directly demonstrate HR we use a simple PCR-based method involving inverted repeats, which gives a reliable read out, as the recombination junctions can be determined by sequencing the amplicons. Using this read out, we demonstrate enhanced HR under growth stress in E. histolytica, and during encystation in E. invadens. We also demonstrate recombination between chromosomal inverted repeats. This is the first experimental demonstration of HR in Entamoeba and will help future investigations into this process, and to explore the possibility of meiosis in Entamoeba.

  6. Successful treatment of multi-focal XDR tuberculous osteomyelitis.

    Alshukairi, Abeer N; Alrajhi, Abdulrahman A; Alamri, Abdulfattah W; Alothman, Adel F

    2016-01-01

    We herein describe the nosocomial transmission of a pre-XDR or MDR case of pulmonary tuberculosis in a HIV-negative health care worker in an area endemic for MDR and XDR tuberculosis. Following inadequate therapy and non-compliance, he presented with extra-pulmonary XDR tuberculosis in the form of multi-focal osteomyelitis and encysted pleural effusion. He was cured after two years of treatment with various anti-tuberculous drugs in addition to interferon gamma. PMID:26631433

  7. A rare gigantic solitary cysticercosis pseudotumour of the neck

    Uledi, Seif J

    2010-01-01

    Cysticercosis is one of the ancient parasitic infections and endemic in many parts of the developing world. Humans acquire cysticercosis when they ingest pork tapeworm eggs either through faecal-oral route by eating faecally contaminated food or by auto infection. The clinical picture largely depends on the location of larval encystment. Neurocysticercosis is the most common form of presentation. Solitary extra neural lesions are quite rare and fairly small in size. We present a very unique c...

  8. Experimental Toxoplasmosis in Rats Induced Orally with Eleven Strains of Toxoplasma gondii of Seven Genotypes: Tissue Tropism, Tissue Cyst Size, Neural Lesions, Tissue Cyst Rupture without Reactivation, and Ocular Lesions.

    Dubey, JP; Ferreira, LR; Alsaad, M; Verma, SK; Alves, DA; Holland, GN; McConkey, GA

    2016-01-01

    The protozoan parasite Toxoplasma gondii is one of the most widely distributed and successful parasites. Toxoplasma gondii alters rodent behavior such that infected rodents reverse their fear of cat odor, and indeed are attracted rather than repelled by feline urine. The location of the parasite encysted in the brain may influence this behavior. However, most studies are based on the highly susceptible rodent, the mouse.Latent toxoplasmosis was induced in rats (10 rats per T. gondii strains) ...

  9. Vesical nephrogenic adenoma: an unusual presentation of a bladder tumour

    Martínez-Sanchíz, Carlos; Martínez-Ruiz, Jesús; Anguita-Fernandez, Pedro J.; Giménez-Bachs, José M.; Atiénzar-Tobarra, Manuel; Rodríguez, Julio Antonio Virseda; Salinas-Sánchez, Antonio S

    2011-01-01

    Vesical nephrogenic adenoma is a rare, benign entity that appears most commonly in middle-aged males. Its etiology is unknown, but it has been linked to chronic irritating factors, such as infection, trauma, urological surgery, kidney stones, foreign bodies and chemical agents, such as Bacille Calmette-Guerin. We report 2 new cases with a history of transurethral resection of the bladder and the prostate and a history of prolonged voiding symptoms. In both cases, the findings of encysted tubu...

  10. Persistence of the emerging pathogen Batrachochytrium dendrobatidis outside the amphibian host greatly increases the probability of host extinction

    Mitchell, Kate M.; Thomas S Churcher; Trenton W J Garner; Fisher, Matthew C

    2007-01-01

    Pathogens do not normally drive their hosts to extinction; however, Batrachochytrium dendrobatidis, which causes amphibian chytridiomycosis, has been able to do so. Theory predicts that extinction can be caused by long-lived or saprobic free-living stages. The hypothesis that such a stage occurs in B. dendrobatidis is supported by the recent discovery of an apparently encysted form of the pathogen. To investigate the effect of a free-living stage of B. dendrobatidis on host population dynamic...

  11. Studies on Cercariae from Kuwait Bay. XI. Description and surface topography of Cercaria kuwaitae XI sp.n. (Digenea: Echinostomatidae)

    1999-01-01

    A new echinostome cercaria, Cercaria kuwaitae XI sp.n., from the prosobranch gastropod Cerithidea cingulata (Gmelin) from Kuwait Bay is described. The new cercaria is characterized by 23 collar spines and primary excretory tubules with distinct diverticula. The cercaria encysts in the snail host and is similar to those of Acanthoparyphium sp. The surface topography of the redia, cercaria and metacercarial cyst wall is studied by scanning electron microscopy. This is the first echinostome cerc...

  12. Oral Immunization of BALB/c Mice with Giardia duodenalis Recombinant Cyst Wall Protein Inhibits Shedding of Cysts

    Larocque, R; Nakagaki, K.; Lee, P.; Abdul-Wahid, A.; Faubert, G M

    2003-01-01

    The process of encystation is a key step in the Giardia duodenalis life cycle that allows this intestinal protozoan to survive between hosts during person-to-person, animal-to-person, waterborne, or food-borne transmission. The release of cysts from infected persons and animals is the main contributing factor to contamination of the environment. Genes coding for cyst wall proteins (CWPs), which could be used for developing a transmission-blocking vaccine, have been cloned. Since the immunogen...

  13. The Effects of Salinity, Phosphate Deprivation and Eosin on Growth and Astaxanthin Production in Unicellular Green Alga, Haematococcus Pluvialis

    M. Mahdiyeh; S. Farrahi-Ashtiani; I. Nahvi

    2002-01-01

    In order to increase ketocarotenoid astaxanthin production by Haematococcus pluvialis, the effects of salinity, phosphate deprivation and eosin on growth and astaxanhin production were examined. The experiments were carried out in the growth chamber at 20oC under sterile conditions. The results showed that under salinity, phosphate deprivation and eosin supply, encystment was induced, and at the same time, astaxanthin formation and dry weight increased. Furthermore, by adding histidine to cul...

  14. Self-Mating in the Definitive Host Potentiates Clonal Outbreaks of the Apicomplexan Parasites Sarcocystis neurona and Toxoplasma gondii

    Wendte, Jered M.; Miller, Melissa A.; Lambourn, Dyanna M.; Magargal, Spencer L.; Jessup, David A.; Michael E Grigg

    2010-01-01

    Tissue-encysting coccidia, including Toxoplasma gondii and Sarcocystis neurona, are heterogamous parasites with sexual and asexual life stages in definitive and intermediate hosts, respectively. During its sexual life stage, T. gondii reproduces either by genetic out-crossing or via clonal amplification of a single strain through self-mating. Out-crossing has been experimentally verified as a potent mechanism capable of producing offspring possessing a range of adaptive and virulence potentia...

  15. Assessing zoospore Phytophthora activity to enhance disease management and promote ecological surveillance of chestnut ink disease

    Gouveia, Eugénia; Nunes, Luís

    2013-01-01

    Phytophthora cinnamomi and P. cambivora are soil borne Oomycetes that cause Chestnut Ink Disease, a lethal and widespread disease of the European chestnut (Castanea saliva Mill. Zoospores are the main infective propagules that actively swimming in liquid environments, reach the roots, encyst and infect the host. Gathering these biological characteristics we studied, on potting mix previously infested with P. cinnamomi, zoospore re lease and environmental conditions that prom...

  16. Assessing phytophtora zoospore a to enhance disease management and promote ecological surveillance of chestnut ink disease

    Gouveia, Eugénia; Nunes, Luís

    2014-01-01

    Phytophthora cinnamomi and P. cambivora are soil borne oomycetes that cause Chestnut Ink Disease, a lethal and widespread disease of the European chestnut (Castanea sativa Mill.). Soil moisture is a key factor for the onset of Phytophthora root rot epidemics. Zoospores are the main infective propagules that reach the roots by swimming in liquid environments, become encysted and after that infect the host. Considering this biological uniqueness, we studied zoospore release an...

  17. Description and proposed life cycle of Maritrema novaezealandensis n. sp. (Microphallidae) parasitic in red-billed gulls, Larus novaehollandiae scopulinus, from Otago Harbor, South Island, New Zealand

    Martorelli, Sergio R; Fredensborg, Brian Lund; Mouritsen, Kim Nørgaard;

    2004-01-01

    Maritrema novaezealandensis n. sp. is described from Otago Harbor, South Island, New Zealand, on the basis of adult specimens collected from the Red-billed gull, Larus novaehollandiae scopulinus, and excysted metacercariae obtained from crabs. It belongs to the "eroliae group" and differs from...... metacercariae encysted in the body cavity of the crustacean host. Finally, the definitive host, the gull, L. n. scopulinus, harbors the adult worms in its intestine....

  18. Contributions to the Understanding of the Bloom Dynamics of Pyrodinium bahamense var. compressum: A Toxic Red Tide Causative Organism

    Rhodora Azanza

    1997-01-01

    Pyrodinium bahamense var. compressum has been the primary organism responsible for the toxic red tide episodes which have been recurring in Manila Bay, Philippines since 1988. The life history of the species has been elucidated through encystment-excystment studies in vitro, from which its obligatory dormancy has been demonstrated. Cyst-mapping studies have shown that this life stage of the organism occurs relatively higher in the Bataan and Cavite areas where the greater number of red tide o...

  19. The first human case of Trichinella spiralis infection in Korea

    Sohn, Woon-Mok; Kim, Han-Mo; Chung, Dong-Il; YEE, SUNG TAE

    2000-01-01

    Three cases of human infection by Trichinella spiralis were first confirmed by detecting encysted larvae in the biopsied muscle in December 1997, in Korea. The patients were one 35- and two 39-year-old males residing in Kochang-gun, Kyongsangnam-do. They had a common past history of eating raw liver, spleen, blood and muscle of a badger, Meles meles melanogenys, and complained of high fever, facial and periorbital edema, and myalgia. Hematologic and biochemical examinations revealed leukocyto...

  20. Ectopic fascioliasis mimicking a colon tumor

    Makay, Ozer; Gurcu, Baris; Caliskan, Cemil; Nart, Deniz; Tuncyurek, Muge; KORKUT, Mustafa

    2007-01-01

    Fasciola hepatica, a leaf shaped trematode that is common in cattle, sheep and goats, is acquired by eating raw water plants like watercress or drinking water infected with the encysted form of the parasite. The varied clinical presentations of fascioliasis still make a high index of suspicion mandatory. Besides having a wide spectrum of hepatobiliary symptoms like obstructive jaundice, cholangitis and liver cirrhosis, the parasitic infection also has extrabiliary manifestations. Until recent...

  1. First report of fossilized cysts produced by the benthic Bysmatrum subsalsum (Dinophyceae) from a shallow Mexican lagoon in the Gulf of Mexico

    Limoges, A.; K. N. Mertens; Ruiz-Fernandez, C; de Vernal, A.

    2015-01-01

    Cysts belonging to the benthic dinoflagellate Bysmatrum subsalsum were recovered from palynologically treated sediments collected in the Alvarado Lagoon (southwestern Gulf of Mexico). The cysts are proximate, reflecting the features of the parent thecal stage, and their autofluorescence implies a dinosporin composition similar to the cyst walls of phototrophic species. This finding is important for our understanding of B. subsalsum life cycle transitions and ecology. Encystment may play an im...

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

    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

  3. A comparison of the Giardia lamblia trophozoite and cyst transcriptome using microarrays

    Widmer Giovanni

    2011-05-01

    Full Text Available Abstract Background Compared with many protists, Giardia lamblia has a simple life cycle alternating between cyst and trophozoite. Most research on the molecular biology of Giardia parasites has focused on trophozoites and the processes of excystation and encystation, whereas cysts have attracted less interest. The striking morphological differences between the dormant cyst and the rapidly dividing and motile trophozoite implies profound changes in the metabolism as the parasite encysts in the host's intestine and excysts upon ingestion by a new host. Results To investigate the magnitude of the transcriptional changes occurring during the G. lamblia life cycle we compared the transcriptome of G. lamblia trophozoites and cysts using single-color oligonucleotide microarrays. Cysts were found to possess a much smaller transcriptome, both in terms of mRNA diversity and abundance. Genes encoding proteins related to ribosomal functions are highly over-represented. The comparison of the transcriptome of cysts generated in culture or extracted from feces revealed little overlap, raising the possibility of significant biological differences between the two types of cysts. Conclusions The comparison of the G. lamblia cyst and trophozoite transcriptome showed that transcripts of most genes are present at a lower level in cysts. This global view of the cyst and trophozoite transcriptome complements studies focused on the expression of selected genes during trophozoite multiplication, encystation and excystation.

  4. Survival of sylvatic Trichinella spiralis isolates in frozen tissue and processed meat products.

    Worley, D E; Seesee, F M; Espinosa, R H; Sterner, M C

    1986-11-01

    The ability of Trichinella spiralis larvae to survive at subfreezing temperatures encysted in the musculature of wild carnivorous mammals was assessed by evaluating motility and infectivity (to rodents) of trichinae at various intervals after storage in frozen skeletal muscle. Fifty to 60% of the larvae in grizzly bear meat were alive after storage for 27 months at -6.5 to -20 C, and 30% to 50% were still alive at 34 months. However, none survived for 38 months, on the basis of infectivity in mice and larval motility. Trichinella larvae survived up to 4 months in frozen (-6.5 to -20 C) wolverine tissue. Viable larvae were not recovered from mountain lion or fisher muscle frozen for 1 month. The effect of postslaughter processing on Trichinella larvae encysted in bear meat was evaluated by use of a similar bioassay procedure. Viability of larvae recovered from black bear meat that had been processed into ham or jerky was not affected by dry curing with a commercial salt mixture. Trichinae from both preparations induced infections in mice (58 to 90 larvae/g of tissue). However, a combination of curing and smoking was consistently lethal to encysted larvae. Viable trichinae were not recovered from ground bear meat preparations (pepperoni, salami, or sausage) processed according to commercial standards. PMID:3505925

  5. The life cycle of Hexangium sigani Goto & Ozaki, 1929 (Digenea: Microscaphidiidae) from the Red Sea.

    Hassanine, R M El-Said; Al-Zahrani, D A; Touliabah, H El-S; Youssef, E M

    2016-09-01

    The microscaphidiid Hexangium sigani Goto & Ozaki, 1929 was found in the intestine of Siganus rivulatus, a siganid fish permanently resident in a lagoon within the mangrove swamps on the Egyptian coast of the Gulf of Aqaba. Intra-molluscan stages of this trematode (mother sporocysts, rediae and cercariae) were found in the gonads and digestive gland of Nassarius pullus (Gastropoda: Nassariidae), a common snail in the same lagoon. Consequently, the life cycle of H. sigani was elucidated under natural conditions: eggs are directly ingested by the snail; mother sporocysts and rediae reach maturity 5-7 and 16-17 weeks post-infection; rediae contain 18-26 developing cercariae; fully developed cercariae are monostome, without penetration glands, emerge from the snail during the night 18-19 weeks post-infection and rapidly encyst on aquatic vegetation (there is no second intermediate host); encysted metacercariae are not progenetic; 2-day-old metacercariae encysted on filamentous algae fed to S. rivulatus developed into fully mature worms 5-6 weeks post-infection. The cycle was completed in about 24 weeks. The intra-molluscan stages are very similar to those of Dictyangium chelydrae Stunkard, 1943, the only described intra-molluscan stages of any microscaphidiid. However, they are also similar to those of the family Mesometridae. The present study of H. sigani describes the first complete microscaphidiid life cycle, and implicitly supports the phylogenetic relationship of this family with the Mesometridae inferred from molecular phylogenetic studies. PMID:26279005

  6. Bioavailability and efficacy characteristics of two different oral liquid formulations of albendazole.

    García, Juan José; Bolás, Francisco; Torrado, Juan José

    2003-01-16

    The oral bioavailability and anthelmintic efficacy in mice of a new formulation of albendazole (ABZ) dissolved in a solution of hydroxypropyl-beta-cyclodextrin (HPCD) are compared with a conventional ABZ suspension of carboxymethylcellulose. Plasma concentrations of ABZ and albendazole sulphoxide (ABZ-SO), its active and main metabolite, were assayed by HPLC. The AUC(0- infinity ) and C(max) values obtained for both ABZ and ABZ-SO, after administration of the ABZ-HPCD solution were significantly higher (Psuspension. Although, the differences between the ABZ and ABZ-SO-T(max) values were found not to be significant, regardless of the formulation. The anthelmintic activities against enteral (pre-adult) and parenteral (migrating and encysted larvae) stages of Trichinella spiralis were studied in mice. The ABZ solution was more efficient against pre-adult and encysted larvae than the ABZ suspension. The efficacy differences between both formulations against the migrating larvae, were found to be not significant (P<0.05). For the migrating parasite stage, there was a linear correlation between the anthelmintic activity and pharmacokinetical parameters with respect to the ABZ-AUC(0- infinity ) value. Meanwhile, for the muscular encysted parasite stage, better relationships were obtained for AUC(0- infinity ) and C(max) values from ABZ-SO, which had correlation coefficients of 0.996 and 0.987, respectively. PMID:12527162

  7. Development of a recombinant protein-based ELISA for diagnosis of larval cyathostomin infection.

    Mitchell, Mairi C; Tzelos, Thomas; Handel, Ian; McWilliam, Hamish E G; Hodgkinson, Jane E; Nisbet, Alasdair J; Kharchenko, Vitaliy O; Burgess, Stewart T G; Matthews, Jacqueline B

    2016-07-01

    Cyathostomins are ubiquitous nematodes of horses. Once ingested, they can spend a substantial time as encysted larvae in the intestinal wall. The larvae can comprise up to 90% of the total burden, with up to several million worms reported in individuals. These stages can emerge in large numbers to cause life-threatening colitis. Direct methods for detection of encysted larval burdens in live horses do not exist. Previously, two antigen complexes were identified as promising markers for infection. A component of these, cyathostomin gut associated larval antigen-1 (Cy-GALA-1), was identified following immunoscreening of a complementary DNA library. Serum immunoglobulin G(T) (IgG(T)) responses to Cy-GALA-1 were shown to inform on larval infection. Sequence analysis of polymerase chain reaction products amplified from individual worms indicated that Cy-GALA-1 was derived from Cyathostomum pateratum. As cyathostomin infections always comprise multiple species, a diagnostic test must account for this. Here, segments of the Cy-gala gene were isolated from four common species, Cyathostomum catinatum, Cylicocyclus ashworthi, Cylicostephanus goldi and Cylicostephanus longibursatus, and the associated proteins expressed in recombinant form. The specificity and immunogenicity of each protein was confirmed. Each protein was assessed by enzyme linked immuno sorbent assay (ELISA) for its ability for informing on the presence of encysted larval infection and the level of burden. PMID:27174468

  8. Parasite infections (Trematoda, Digenea) of Sagitta friderici (Chaetognatha) from the southwestern Atlantic Ocean: prevalence and distribution.

    Daponte, María C; Gil de Pertierra, Alicia A; Palmieri, Mónica A; Ostrowskide Núñez, Margarita

    2006-08-30

    The following chaetognaths were found in the Atlantic Ocean between 34 to 40 degrees S and 52 degrees 20' to 62 degrees 00' W: Sagitta friderici, S. tasmanica, S. minima, S. gazellae, and S. enflata (in order of abundance). Of these, only S. friderici was parasitised by unencysted metacercariae of the families Derogenidae (Derogenes sp.), Hemiuridae (Ectenurus sp.) and Fellodistomidae (Monascus filiformis), and encysted metacercariae of Lepocreadiidae. The percentage of infection for each sampling station varied from 0.033 to 0.001 in August and from 0.02 to 0.001 in October 1996, with the highest values occurring at stations closer to the coast. The intensity of infection (worms per host) varied from 1 to 3 for Ectenurus sp. and was 1 for Derogenes sp., Monascus filiformis and Lepocreadiidae. Unencysted metacercariae were found in mature developmental stages of chaetognaths, whereas encysted ones occurred mainly in juveniles. The size and length of the ovaries of parasitised and unparasitised chaetognaths did not differ significantly. This is the first report of encysted Lepocreadiidae metacercariae and a progenetic metacercaria of Ectenurus sp. in Chaetognatha from the SW Atlantic Ocean. PMID:17058603

  9. Amebas de vida libre en muestras de agua de piscinas del Departamento de Lima

    María Beltrán de Estrada

    1997-01-01

    Full Text Available Las amebas de vida libre son numerosas y se encuentran en la naturaleza, en medios como: agua, suelos y vegetación. Los géneros Naegleria, Acanthamoeba, Hartmannella y Vahlkampfia, tienen importancia en salud pública, pues son oportunistas y afectan al hombre (enfermedad de los nadadores. Del estudio de 68 muestras de agua en 34 piscinas, 3 zonas de un río y 7 estanques o lagunas, resultaron el 40,9% con amebas de vida libre tales como: Naegleria, Acanthamoeba, Hartmannella y Vahlkampfia. La virulencia de las amebas de vida libre fue estudiada en los animales de laboratorio por inoculación craneal, nasal y ocular. Los resultados obtenidos nos permiten recomendar y considerar el mantenimiento y tratamiento periódico y/o permanente de las piscinas con productos químicos, eficaces y eficientes sin que perjudiquen la salud humana.

  10. Dicty_cDB: Contig-U10367-1 [Dicty_cDB

    Full Text Available ino Acid sequence trcvnkkkemlkskisfisrctykiieaqkfeaqyltkikpsnqninprkskflkffkdq pkitnntnktktkakanakatpsetteat...rcvnkkkemlkskisfisrctykiieaqkfeaqyltkikpsnqninprkskflkffkdq pkitnntnktktkakanakatpsetteattkeknkvmtflgkpfefnr...e E Sequences producing significant alignments: (bits) Value Contig-U10367-1 (Contig-U10367-1Q) /CSM_Contig/Conti... 1489 0.0 Cont.................................................done Score E Sequences producing significant alignments: (bits....b1 GMW2 Glycine max genomic, genomic ... 48 0.77 1 ( AY653733 ) Acanthamoeba pol

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

    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. PMID:26412057

  12. Mimivirus relatives in the Sargasso sea

    Claverie Jean-Michel; Ghedin Elodie

    2005-01-01

    Abstract The discovery and genome analysis of Acanthamoeba polyphaga Mimivirus, the largest known DNA virus, challenged much of the accepted dogma regarding viruses. Its particle size (>400 nm), genome length (1.2 million bp) and huge gene repertoire (911 protein coding genes) all contribute to blur the established boundaries between viruses and the smallest parasitic cellular organisms. Phylogenetic analyses also suggested that the Mimivirus lineage could have emerged prior to the individual...

  13. Diagnostic accuracy of microbial keratitis with in vivo scanning laser confocal microscopy

    Hau, Scott; Dart, John; Vesaluoma, Minna; Parmar, Dipak; Claerhout, Ilse; Bibi, Kanom; Larkin, Frank

    2010-01-01

    Abstract Aims To determine the accuracy of diagnosing microbial keratitis by masked medical and non-medical observers using the Heidelberg Retina Tomograph II / Rostock Cornea Module in vivo confocal microscope. Methods Confocal images were selected for 62 eyes with culture or biopsy proven infections. The cases comprised of 26 Acanthamoeba, 12 fungus, 3 Microsporidia, 2 Nocardia, and 19 bacterial infections (controls). The reference standard for comparison was a pos...

  14. Molecular comparison of cultivable protozoa from a pristine and a polycyclic aromatic hydrocarbon polluted site

    Lara, Enrique; Berney, Cédric; Ekelund, Flemming; Harms, Hauke; Chatzinotas, Antonis

    2010-01-01

    We compared the abundance and diversity of cultivable protozoa (flagellates and amoebae) in a polycyclic aromatic hydrocarbon (PAH) polluted soil and an unpolluted control, by isolating and cultivating clonal strains. The number of cultivable protozoa was higher in the polluted soil; however, the polluted soil displayed an impoverished community, dominated by certain taxa, such as Acanthamoeba sp. We isolated a total of 31 protozoan strains to characterize them morphologically and by 18S rRNA...

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

    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

    BACKGROUND: 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. METHODOLOGY/PRINCIPAL FINDINGS: 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 ...

  16. Footprinting of ribosomal RNA genes by transcription initiation factor and RNA polymerase I.

    Bateman, E.; Iida, C T; Kownin, P; Paule, M R

    1985-01-01

    The binding of a species-specific transcription initiation factor (TIF) and purified RNA polymerase I to the promoter region of the 39S ribosomal RNA gene from Acanthamoeba were studied by using DNase I "footprinting." Conditions were chosen such that the footprints obtained could be correlated with the transcriptional activity of the TIF-containing fractions used and that the labeled DNA present would itself serve as a template for transcription. The transcription factor binds upstream from ...

  17. Survey of Infectious Etiologies of Bovine Abortion during Mid- to Late Gestation in Dairy Herds

    Barkallah, Mohamed; Gharbi, Yaakoub; Ben Hassena, Amal; Ben Slima, Ahlem; Mallekh, Zouhir; Gautier, Michel; Greub, Gilbert; Gdoura, Radhouane

    2014-01-01

    Bovine abortion of unknown infectious etiology still remains a major economic problem. Thus, we investigated whether Brucella spp., Listeria monocytogenes, Salmonella spp., Campylobacter spp. and Coxiella burnetii are associated with abortion and/or stillbirth in Tunisian dairy cattle. Using a pan-Chlamydiales PCR, we also investigated the role of Chlamydiaceae, Waddlia chondrophila, Parachlamydia acanthamoebae and other members of the Chlamydiales order in this setting. Veterinary samples ta...

  18. Survey of Infectious Etiologies of Bovine Abortion during Mid- to Late Gestation in Dairy Herds.

    Barkallah M.; Gharbi Y.; Hassena A.B.; Slima A.B.; Mallek Z.; Gautier M.; Greub G; Gdoura R.; Fendri I.

    2014-01-01

    Bovine abortion of unknown infectious etiology still remains a major economic problem. Thus, we investigated whether Brucella spp., Listeria monocytogenes, Salmonella spp., Campylobacter spp. and Coxiella burnetii are associated with abortion and/or stillbirth in Tunisian dairy cattle. Using a pan-Chlamydiales PCR, we also investigated the role of Chlamydiaceae, Waddlia chondrophila, Parachlamydia acanthamoebae and other members of the Chlamydiales order in this setting. Veterinary samples ta...

  19. Microscopic isolation and characterization of free living amoebae (FLA from surface water sources.in Birjand, the capital city of the South Khorasan

    Mahmoodreza Behravan

    2015-07-01

    Conclusion: The results indicated that a significant percentage of surface water sources in Birjand city was contaminated with Acanthamoeba spp. It is necessary for physicians, therefore, to take into account the diseases caused by these infectious agents. Besides, local regional health professionals should take into consideration the potential role of surface stagnant water sources in transferring these infectious agents. Placing warning signs in areas contaminated with these infectious agents seems a useful measure.

  20. First Isolation of a Giant Virus from Wild Hirudo medicinalis Leech: Mimiviridae isolation in Hirudo medicinalis

    Mondher Boughalmi; Isabelle Pagnier; Sarah Aherfi; Philippe Colson; Didier Raoult; Bernard La Scola

    2013-01-01

    Giant viruses and amoebae are common in freshwater, where they can coexist with other living multicellular organisms. We screened leeches from the species Hirudo medicinalis for giant viruses. We analyzed five H. medicinalis obtained from Tunisia (3) and France (2). The leeches were decontaminated and then dissected to remove internal parts for co-culture with Acanthamoeba polyphaga. The genomes of isolated viruses were sequenced on a 454 Roche instrument, and a comparative genomics analysis ...

  1. Occurrence and Control of Legionella in Recycled Water Systems

    Jjemba, Patrick K.; William Johnson; Zia Bukhari; 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...

  2. Balamuthia mandrillaris, Free-Living Ameba and Opportunistic Agent of Encephalitis, Is a Potential Host for Legionella pneumophila Bacteria

    Shadrach, Winlet Sheba; Rydzewski, Kerstin; Laube, Ulrike; Holland, Gudrun; Özel, Muhsin; Kiderlen, Albrecht F; Flieger, Antje

    2005-01-01

    Balamuthia mandrillaris is a free-living ameba and an opportunistic agent of granulomatous encephalitis in humans and other mammalian species. Other free-living amebas, such as Acanthamoeba and Hartmannella, can provide a niche for intracellular survival of bacteria, including the causative agent of Legionnaires' disease, Legionella pneumophila. Infection of amebas by L. pneumophila enhances the bacterial infectivity for mammalian cells and lung tissues. Likewise, the pathogenicity of amebas ...

  3. Detection of Balamuthia mandrillaris DNA in the storage case of contact lenses in Germany.

    Balczun, Carsten; Scheid, Patrick L

    2016-05-01

    Acanthamoeba spp. are frequently the etiological agents of a severe form of sight-threatening keratitis, called Acanthamoeba keratitis. The contact lens storage solution of a patient with keratitis of unknown genesis was screened using our diagnostic tools to detect potentially pathogenic free-living amoebae (FLA). Culture methods and a triplex quantitative real-time polymerase chain reaction (qPCR) targeting Acanthamoeba spp., Naegleria fowleri, and Balamuthia mandrillaris were used in context of this routine screening. While no amoebae were detected by culture, qPCR specifically detected DNA of B. mandrillaris. This FLA is known as the etiological agent of a fatal form of encephalitis in humans and other mammals, Balamuthia amoebic encephalitis (BAE). A fragment of the 18S rDNA gene was amplified from the sample and showed 99 % sequence identity to B. mandrillaris sequences from GenBank. To the best of our knowledge, this is the first report of B. mandrillaris found in association with contact lenses. Although no viable amoeba was obtained by culturing efforts, the verification of B. mandrillaris DNA in the contact lens storage solution demonstrates how easily this pathogen might come into close contact with humans. PMID:26965426

  4. Survey of infectious etiologies of bovine abortion during mid- to late gestation in dairy herds.

    Barkallah, Mohamed; Gharbi, Yaakoub; Hassena, Amal Ben; Slima, Ahlem Ben; Mallek, Zouhir; Gautier, Michel; Greub, Gilbert; Gdoura, Radhouane; Fendri, Imen

    2014-01-01

    Bovine abortion of unknown infectious etiology still remains a major economic problem. Thus, we investigated whether Brucella spp., Listeria monocytogenes, Salmonella spp., Campylobacter spp. and Coxiella burnetii are associated with abortion and/or stillbirth in Tunisian dairy cattle. Using a pan-Chlamydiales PCR, we also investigated the role of Chlamydiaceae, Waddlia chondrophila, Parachlamydia acanthamoebae and other members of the Chlamydiales order in this setting. Veterinary samples taken from mid to late-term abortions from twenty dairy herds were tested. From a total of 150 abortion cases collected, infectious agents were detected by PCR in 73 (48.66%) cases, 13 (8.66%) of which represented co-infections with two infectious agents. Detected pathogens include Brucella spp (31.3%), Chlamydiaceae (4.66%), Waddlia chondrophila (8%), Parachlamydia acanthamoebae (5.33%), Listeria monocytogenes (4.66%) and Salmonella spp. (3.33%). In contrast, Campylobacter spp. and Coxiella burnetii DNA were not detected among the investigated veterinary samples. This demonstrates that different bacterial agents may cause bovine abortion in Tunisia. This is the first report suggesting the role of Parachlamydia acanthamoebae in bovine abortion in Africa. Further studies with a larger number of samples are necessary to confirm whether this emerging pathogen is directly linked to abortion in cattle. PMID:24662769

  5. Survey of infectious etiologies of bovine abortion during mid- to late gestation in dairy herds.

    Mohamed Barkallah

    Full Text Available Bovine abortion of unknown infectious etiology still remains a major economic problem. Thus, we investigated whether Brucella spp., Listeria monocytogenes, Salmonella spp., Campylobacter spp. and Coxiella burnetii are associated with abortion and/or stillbirth in Tunisian dairy cattle. Using a pan-Chlamydiales PCR, we also investigated the role of Chlamydiaceae, Waddlia chondrophila, Parachlamydia acanthamoebae and other members of the Chlamydiales order in this setting. Veterinary samples taken from mid to late-term abortions from twenty dairy herds were tested. From a total of 150 abortion cases collected, infectious agents were detected by PCR in 73 (48.66% cases, 13 (8.66% of which represented co-infections with two infectious agents. Detected pathogens include Brucella spp (31.3%, Chlamydiaceae (4.66%, Waddlia chondrophila (8%, Parachlamydia acanthamoebae (5.33%, Listeria monocytogenes (4.66% and Salmonella spp. (3.33%. In contrast, Campylobacter spp. and Coxiella burnetii DNA were not detected among the investigated veterinary samples. This demonstrates that different bacterial agents may cause bovine abortion in Tunisia. This is the first report suggesting the role of Parachlamydia acanthamoebae in bovine abortion in Africa. Further studies with a larger number of samples are necessary to confirm whether this emerging pathogen is directly linked to abortion in cattle.

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

    Sílvia Cervero-Aragó

    Full Text Available 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.

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

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

    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

  9. Export of cyst wall material and Golgi organelle neogenesis in Giardia lamblia depend on endoplasmic reticulum exit sites.

    Faso, Carmen; Konrad, Christian; Schraner, Elisabeth M; Hehl, Adrian B

    2013-04-01

    Giardia lamblia parasitism accounts for the majority of cases of parasitic diarrheal disease, making this flagellated eukaryote the most successful intestinal parasite worldwide. This organism has undergone secondary reduction/elimination of entire organelle systems such as mitochondria and Golgi. However, trophozoite to cyst differentiation (encystation) requires neogenesis of Golgi-like secretory organelles named encystation-specific vesicles (ESVs), which traffic, modify and partition cyst wall proteins produced exclusively during encystation. In this work we ask whether neogenesis of Golgi-related ESVs during G. lamblia differentiation, similarly to Golgi biogenesis in more complex eukaryotes, requires the maintenance of distinct COPII-associated endoplasmic reticulum (ER) subdomains in the form of ER exit sites (ERES) and whether ERES are also present in non-differentiating trophozoites. To address this question, we identified conserved COPII components in G. lamblia cells and determined their localization, quantity and dynamics at distinct ERES domains in vegetative and differentiating trophozoites. Analogous to ERES and Golgi biogenesis, these domains were closely associated to early stages of newly generated ESV. Ectopic expression of non-functional Sar1 GTPase variants caused ERES collapse and, consequently, ESV ablation, leading to impaired parasite differentiation. Thus, our data show how ERES domains remain conserved in G. lamblia despite elimination of steady-state Golgi. Furthermore, the fundamental eukaryotic principle of ERES to Golgi/Golgi-like compartment correspondence holds true in differentiating Giardia presenting streamlined machinery for secretory organelle biogenesis and protein trafficking. However, in the Golgi-less trophozoites ERES exist as stable ER subdomains, likely as the sole sorting centres for secretory traffic. PMID:23094658

  10. Influence of site, season, silvering stage, and length on the parasites of the European eel Anguilla anguilla in two Mediterranean coastal lagoons of the island of Corsica, France using indicator species method.

    Filippi, Jean-José; Quilichini, Yann; Foata, Joséphine; Marchand, Bernard

    2013-08-01

    The parasites of 425 European eels, Anguilla anguilla, were studied between 2009 and 2012 in two Mediterranean coastal lagoons of the island of Corsica, France. An indicator value (IndVal) method was used for analysis, which combines measures of fidelity and specificity. Because of its resilience to detect changes in abundance, IndVal is an effective ecological bioindicator. The IndVal method demonstrated that site, season, silvering stage, and length could influence the occurrence of parasite species in European eel. A randomization test identified ten parasite species as having a significant indicator value for site (lagoons differed principally in salinity: oligohaline to polyhaline for the Biguglia lagoon and polyhaline to euhaline for the Urbino lagoon; the digeneans Bucephalus anguillae and Lecithochirium musculus, the cestodes Bothriocephalus claviceps, Proteocephalus macrocephalus, and larvae of Myzophyllobothrium sp., the nematodes Anguillicoloides crassus, and encysted larvae of Contracaecum sp., the acanthocephalan Acanthocephaloides incrassatus, the monogenean Pseudodactyogyrus anguillae, and the copepod Ergasilus gibbus); one parasite species for the spring season (the acanthocephalan A. incrassatus); six parasite species for silvering stage (yellow, pre-silver, silver; the trematodes B. anguillae and Deropristis inflata, encysted larvae of the nematode Contracaecum sp., the acanthocephalan A. incrassatus, the monogenean P. anguillae, and the copepod E. gibbus); and three parasite species for some of the five length classes (the cestode P. macrocephalus, encysted larvae of the nematode Contracaecum sp., and the monogenean P. anguillae). Data for species composition and infection levels should help to improve the management of parasitism in the populations of European eels. PMID:23739809

  11. Self-mating in the definitive host potentiates clonal outbreaks of the apicomplexan parasites Sarcocystis neurona and Toxoplasma gondii.

    Jered M Wendte

    Full Text Available Tissue-encysting coccidia, including Toxoplasma gondii and Sarcocystis neurona, are heterogamous parasites with sexual and asexual life stages in definitive and intermediate hosts, respectively. During its sexual life stage, T. gondii reproduces either by genetic out-crossing or via clonal amplification of a single strain through self-mating. Out-crossing has been experimentally verified as a potent mechanism capable of producing offspring possessing a range of adaptive and virulence potentials. In contrast, selfing and other life history traits, such as asexual expansion of tissue-cysts by oral transmission among intermediate hosts, have been proposed to explain the genetic basis for the clonal population structure of T. gondii. In this study, we investigated the contributing roles self-mating and sexual recombination play in nature to maintain clonal population structures and produce or expand parasite clones capable of causing disease epidemics for two tissue encysting parasites. We applied high-resolution genotyping against strains isolated from a T. gondii waterborne outbreak that caused symptomatic disease in 155 immune-competent people in Brazil and a S. neurona outbreak that resulted in a mass mortality event in Southern sea otters. In both cases, a single, genetically distinct clone was found infecting outbreak-exposed individuals. Furthermore, the T. gondii outbreak clone was one of several apparently recombinant progeny recovered from the local environment. Since oocysts or sporocysts were the infectious form implicated in each outbreak, the expansion of the epidemic clone can be explained by self-mating. The results also show that out-crossing preceded selfing to produce the virulent T. gondii clone. For the tissue encysting coccidia, self-mating exists as a key adaptation potentiating the epidemic expansion and transmission of newly emerged parasite clones that can profoundly shape parasite population genetic structures or cause

  12. Self-mating in the definitive host potentiates clonal outbreaks of the apicomplexan parasites Sarcocystis neurona and Toxoplasma gondii.

    Wendte, Jered M; Miller, Melissa A; Lambourn, Dyanna M; Magargal, Spencer L; Jessup, David A; Grigg, Michael E

    2010-01-01

    Tissue-encysting coccidia, including Toxoplasma gondii and Sarcocystis neurona, are heterogamous parasites with sexual and asexual life stages in definitive and intermediate hosts, respectively. During its sexual life stage, T. gondii reproduces either by genetic out-crossing or via clonal amplification of a single strain through self-mating. Out-crossing has been experimentally verified as a potent mechanism capable of producing offspring possessing a range of adaptive and virulence potentials. In contrast, selfing and other life history traits, such as asexual expansion of tissue-cysts by oral transmission among intermediate hosts, have been proposed to explain the genetic basis for the clonal population structure of T. gondii. In this study, we investigated the contributing roles self-mating and sexual recombination play in nature to maintain clonal population structures and produce or expand parasite clones capable of causing disease epidemics for two tissue encysting parasites. We applied high-resolution genotyping against strains isolated from a T. gondii waterborne outbreak that caused symptomatic disease in 155 immune-competent people in Brazil and a S. neurona outbreak that resulted in a mass mortality event in Southern sea otters. In both cases, a single, genetically distinct clone was found infecting outbreak-exposed individuals. Furthermore, the T. gondii outbreak clone was one of several apparently recombinant progeny recovered from the local environment. Since oocysts or sporocysts were the infectious form implicated in each outbreak, the expansion of the epidemic clone can be explained by self-mating. The results also show that out-crossing preceded selfing to produce the virulent T. gondii clone. For the tissue encysting coccidia, self-mating exists as a key adaptation potentiating the epidemic expansion and transmission of newly emerged parasite clones that can profoundly shape parasite population genetic structures or cause devastating disease

  13. 北海道江別市内のペットショップで市販されていたカメ類の寄生線虫類

    木本, 有子; 浅川, 満彦

    1998-01-01

    For the sake of an epidemiological analysis on the helminth infection with pet tortoises sold in Ebetsu, Japan, 23 individuals belonging to 11 tortoise species were examined helminthologically. From Chelus fimbriatus(abbreviated to Cf), Chelodina siebenrocki(Cs), Testudo horsfieldi(Th) and Kinixys sp.(K), 6 nematodes, namely Thaparia sp.(host:Th), Mehdiella sp.(Th), Tachygonetria sp.(Cs, Th), Labiduris sp.(K), Capillariidae gen.sp.(Cf)and encysted larvae(Cf), were obtained. ペット用カメ類における寄生蠕虫の感染...

  14. The Histopathology of the Infection of Tilapia rendalli and Hypostomus regani (Osteichthyes by Lasidium Larvae of Anodontites trapesialis (Mollusca, Bivalvia

    Silva-Souza Ângela Teresa

    2002-01-01

    Full Text Available It is described the histopathology of the infection of Tilapia rendalli (Osteichthyes, Perciformes, Cichlidae and Hypostomus regani (Osteichthyes, Siluriformes, Loricariidae by lasidium larvae of Anodontites trapesialis (Mollusca, Bivalvia, Mycetopodidae. The larvae were encysted within the epidermis of the host, being surrounded by a thin hyaline membrane, 3-6 µm thick, of parasite origin. A proliferative host cell reaction did not occur. The histopathology of the infection shows that the lesions induced by the parasites are minimal. However, the numerous small lesions produced by the release of the larvae may provide optimal conditions for the infection by opportunistic pathogens, namely fungus, which may eventually cause the death of the host.

  15. The histopathology of the infection of Tilapia rendalli and Hypostomus regani (Osteichthyes) by lasidium larvae of Anodontites trapesialis (Mollusca, Bivalvia).

    Silva-Souza, Angela Teresa; Eiras, Jorge C

    2002-04-01

    It is described the histopathology of the infection of Tilapia rendalli (Osteichthyes, Perciformes, Cichlidae) and Hypostomus regani (Osteichthyes, Siluriformes, Loricariidae) by lasidium larvae of Anodontites trapesialis (Mollusca, Bivalvia, Mycetopodidae). The larvae were encysted within the epidermis of the host, being surrounded by a thin hyaline membrane, 3-6 microm thick, of parasite origin. A proliferative host cell reaction did not occur. The histopathology of the infection shows that the lesions induced by the parasites are minimal. However, the numerous small lesions produced by the release of the larvae may provide optimal conditions for the infection by opportunistic pathogens, namely fungus, which may eventually cause the death of the host. PMID:12048579

  16. Import of exotic and zoonotic trematodes (Heterophyidae: Centrocestus sp.) in Xiphophorus maculatus

    Mehrdana, Foojan; Jensen, Hannah M.; Kania, Per Walter;

    2014-01-01

    Ornamental fish, Xiphophorus maculatus, were imported from Singapore to Denmark for distribution to local aquarists. Importers observed lethargic and erratic swimming patterns among fish and forwarded a total of 30 fish for pathological examination to a university diagnostic service. All fish were...... diagnosed infected with encysted Centrocestus sp. metacercariae in gills (prevalence of 100% and mean intensity of 454.5 ± 161.9 parasites per fish). Metacercariae were identified by morphological and molecular methods. Cysts (mean length 163.3 ± 13.7 μm and mean width 113.3 ± 10.6 μm) contained a bent...

  17. Identification of cAMP-dependent phosphorylated proteins involved in the formation of environment-resistant resting cysts by the terrestrial ciliate Colpoda cucullus

    Y Sogame

    2014-07-01

    Full Text Available In the terrestrial ciliate Colpoda cucullus, an elevation of the intracellular cAMP concentration was reported to be involved in environment-resistant resting cyst formation. In the present study, cAMP-dependently phosphorylated proteins of encystment-induced C. cucullus were isolated with Phos-tag agarose phosphate-affinity beads and subsequent SDS-PAGE. In a liquid chromatography/tandem mass spectrometry analysis of these phosphoproteins, 27-, 37- and 43-kDa proteins (p27, p37 and p43 were identified as Rieske iron-sulfur protein, histone H4 (hyperacetylated form, and actin, respectively.

  18. Fine needle aspiration cytology of subcutaneous cysticercosis of the breast. Case report and pathogenic discussion.

    Vuong, P N

    1989-01-01

    A case of subcutaneous cysticercosis of the breast diagnosed by fine needle aspiration cytology is reported. The cytologic preparations showed the presence of many inflammatory cells mixed with spiked spherules resembling the rambutan tropical fruit. These spherules arose from the degenerated integument of the parasite found in the histologically excised cyst. They are suggestive of the presence of an encysted flat worm, but are not specific for cysticercosis, whose identification is based on the study of the cephalic extremity of the parasite. PMID:2781970

  19. Studies on Cercariae from Kuwait Bay. XI. Description and surface topography of Cercaria kuwaitae XI sp.n. (Digenea: Echinostomatidae

    Abdul-Salam J

    1999-01-01

    Full Text Available A new echinostome cercaria, Cercaria kuwaitae XI sp.n., from the prosobranch gastropod Cerithidea cingulata (Gmelin from Kuwait Bay is described. The new cercaria is characterized by 23 collar spines and primary excretory tubules with distinct diverticula. The cercaria encysts in the snail host and is similar to those of Acanthoparyphium sp. The surface topography of the redia, cercaria and metacercarial cyst wall is studied by scanning electron microscopy. This is the first echinostome cercaria to be recorded in a gastropod from the Arabian Gulf region.

  20. The life history of Pygidiopsis macrostomum Travassos, 1928 (Digenea: Heterophyidae

    Susana Balmant Emerique Simões

    2009-02-01

    Full Text Available The life history of the trematode Pygidiopsis macrostomum Travassos, 1928 is described for the first time. Rediae and cercariae were obtained from naturally infected snails Heleobia australis (d´Orbigny, a new first intermediate host. Metacercariae were found encysted in the mesenteries of three naturally infected guppies, Phalloptychus januarius (Hensel, Jenynsia multidentata (Jenyns (new host records and Poecilia vivipara Bloch and Schneider. Experimental infections were successfully completed in the intermediate hosts H. australis and Poe. vivipara reared in the laboratory and hamsters Mesocricetus auratus Waterhouse were utilised as a definitive host.

  1. Life history studies of heterophyid Trematodes in the neotropical region: Ascocotyle (Leighia hadra sp.n

    Margarita Ostrowski de Nunez

    1992-12-01

    Full Text Available The life cycle of Ascocotyle (Leighia hadra n.sp. was experimentally reproduced, starting from cercariae from naturally infected Littoridina parchappei, collected from Los Ranchos stream, near Mercedes city, Buenos Aires Province, Argentina. Metacercariae were found encysted in the liver and mesentery of experimentally and naturally infected fishes Cnesterodon decemmaculatus and Jenynsia lineata. Adults were obtained experimentally in chicks and mice. The natural host is unknown. The new species is compared with Ascocotyle (Leighia mcintoshi Price 1936 as described by Leigh, 1974, differing in behavior and morphology of cercarial, metacercarial and adult stages

  2. Improving bioavailability and anthelmintic activity of albendazole by preparing albendazole-cyclodextrin complexes.

    García-Rodriguez, J J; Torrado, J; Bolás, F

    2001-06-01

    The bioavailability and anthelmintic activity of albendazole-cyclodextrin complexes (ABZ-CDC) compared to albendazole suspensions in carboxymethylcellulose (ABZ-CMC) was assessed in a mouse model for Trichinella infections. Swiss CD-1 mice experimentally infected with T. spiralis were treated with both formulations against enteral (adult worms) and parenteral (migrating and encysted larvae). Oral bioavailability was assessed in age matched mice treated with 50 mg/kg of both formulations. The anthelmintic effects and plasma concentration of the active metabolite albendazole-sulphoxide (ABZSO) enantiomer (-) were significantly increased following administration of ABZ-CDC in relation to ABZ-CMC. PMID:11484352

  3. Galactose/N-acetylgalactosamine lectin: the coordinator of host cell killing

    Douglas R Boettner; Christopher Huston; William A Petri Jr

    2002-11-01

    Entamoeba histolytica is an enteric parasite that can kill host cells via a contact-dependent mechanism. This killing involves the amoebic surface protein referred to as the Gal/GalNAc lectin. 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 encystation. There is also possible involvement of the lectin in phagocytosis and caspase activation in host cells.

  4. Intracoelomic parasitic Sporozoa in the burrowing spatangoid echinoid Echinocardium cordatum: coelomocyte reaction and formation of brown bodies

    de Ridder, C.; Jangoux, M.

    1984-03-01

    Echinocardium cordatum frequently harbours in its coelomic cavity the protozoan parasite Lithocystis schneideri. Motile stages of this gregarine (trophozoites and gamonts) may be surrounded by echinoid coelomocytes which show a peculiar transformation of their shape (i.e. each cell develops a single spine-like extension giving a pincushion aspect to the parasite). Encysted stages of the gregarine (gametocysts and sporocysts) are found mostly within brown bodies. Brown bodies are particular mesothelium-covered formations occurring usually in the coelomic cavity of E. cordatum. It is suggested that brown bodies naturally originate from detached fragments of mesenteries.

  5. Improving bioavailability and anthelmintic activity of albendazole by preparing albendazole-cyclodextrin complexes

    García-Rodriguez J.J.

    2001-06-01

    Full Text Available The bioavailability and anthelmintic activity of albendazole-cyclodextrin complexes (ABZ-CDC compared to albendazole suspensions in carboxymethylcellulose (ABZ-CMC was assessed in a mouse model for Trichinella infections. Swiss CD-1 mice experimentally infected with T. spiralis were treated with both formulations against enteral (adult worms and parenteral (migrating and encysted larvae. Oral bioavailability was assessed in age matched mice treated with 50 mg/kg of both formulations. The anthelmintic effects and plasma concentration of the active metabolite albendazole-sulphoxide (ABZSO enantiomer (– were significantly increased following administration of ABZ-CDC in relation to ABZ-CMC.

  6. Comparison between Two Methods for Diagnosis of Trichinellosis: Trichinoscopy and Artificial Digestion

    María Laura Vignau

    1997-09-01

    Full Text Available Two direct methods for the diagnosis of trichinellosis were compared: trichinoscopy and artificial digestion. Muscles from 17 wistar rats, orally infected with 500 Trichinella spiralis encysted larvae were examined. From each of the following muscles: diaphragm, tongue, masseters, intercostals, triceps brachialis and cuadriceps femoralis, 648,440 larvae from 1 g samples were recovered. The linear correlation between trichinoscopy and artificial digestion was very high and significant (r=0.94, p< 0.0001, showing that both methods for the detection of muscular larvae did not differ significantly. In both methods, significant differences were found in the distribution of larvae per gramme of muscle

  7. Trichinella spiralis: Mere Co-Existence or Carcinogenic Parasite For Oral Squamous Cell Carcinoma?

    Shirazi, Nadia; Bist, Sampan Singh; Ahmad, Sohaib; Harsh, Meena

    2015-10-01

    Trichinella spiralis is a parasite which is usually seen in pork-eaters. Most of the trichinosis infections cause little or no symptoms. We report a rare case of a middle aged North Indian male who presented with a painless ulcer in right buccal mucosa which was biopsied and reported as squamous cell carcinoma. Wide local excision was done subsequently which showed encysted larvae of Trichinella spiralis in the deeper skeletal muscle bundles. This article supports the carcinogenic potential of trichinosis and suggests timely work-up and treatment of the parasite. PMID:26557527

  8. Thraustochytrium gaertnerium sp. nov.: A new Thraustochytrid Stramenopilan protist from mangroves of Goa, India

    Bongiorni, L.; Jain, R.; Raghukumar, S.; Aggarwal, R.K.

    in the first type directly becoming zoosporangia; zoosporangia eucarpic, dividing directly into 10—75 zoospores. Zoospores re- leased initially by a rupture in the cell wall, the latter disintegrating totally afterwards. A single proliferation body, 5—10mm.... Zoospores measuring 5.5—7.3mm in length and 3—4.5mm wide. Life cycle and Taxonomy Young vegetative cells resulting from freshly encysted zoospores measured around 4—5mm in diameter (Fig. 1). In pine pollen cultures, such starting from the time of zoospore...

  9. Monitoring of Waterborne Parasites in Two Drinking Water Treatment Plants: A Study in Sarawak, Malaysia

    Reena Leeba Richard

    2016-06-01

    Full Text Available The occurrence of waterborne parasites coupled with water parameters at various processing sites of two drinking water treatment plants (A and B and seven distribution system (DS sites in Sarawak, Malaysia were studied. Ten liters of water underwent immunomagnetic separation (IMS technique to detect the presence of Giardia and Cryptosporidium (oocysts. The remaining supernatant was used to detect other parasites whilst 50 mL of water sample was each used in the detection of free-living amoebae and fecal coliforms. Sampled water was positive for Giardia (32.9%; 28/85, Cryptosporidium (18.8%; 16/85 followed by Spirometra ova-like (25.9%; 22/85, Blastocystis-like (25.9%; 22/85, nematode larvae-like (8.2%; 7/85 and Taenia ova-like (1.2%; 1/85. Meanwhile, 90.2% (55/61 samples were positive for Acanthamoeba and Naegleria via cultivation and of these, 11 isolates were confirmed as Acanthamoeba genotype T3 (5/7 and T4 (2/7 followed by Naegleria sp. (4/11, Naegleria italica (2/11, Naegleria australiensis (1/11, Naegleria angularis (1/11 and Vahlkampfia sp. (3/11. Cryptosporidium, Acanthamoeba and Naegleria were also detected in one of the seven tested DS sites. Only Giardia and Cryptosporidium showed significant correlations with fluoride and fecal coliforms. These results describe the occurrence of waterborne parasites that will assist key stakeholders in mitigating contamination at the specific sites.

  10. Pathogenesis of amoebic encephalitis: Are the amoebae being credited to an 'inside job' done by the host immune response?

    Baig, Abdul Mannan

    2015-08-01

    Pathogenic free living amoeba like Naegleria fowleri, Acanthamoeba spp., and Balamuthia mandrillaris are known to cause fatal "amoebic meningoencephalitis" by acquiring different route of entries to the brain. The host immune response to these protist pathogens differs from each another, as evidenced by the postmortem gross and microscopic findings from the brains of the affected patients. Cited with the expression of 'brain eating amoeba' when the infection is caused by N. fowleri, this expression is making its way into parasitology journals and books. The impression that it imparts is, as if the brain damage is substantially due to the enzymes and toxins produced by this amoeba. A detailed review of the literature, analysis of archived specimens and with our experimental assays, here we establish that with N. fowleri, Acanthamoeba and Balamuthia spp., the infections result in an extensive brain damage that in fact is substantially caused by the host immune response rather than the amoeba. Due to the comparatively larger sizes of these pathogens and the prior exposure of the amoebal antigen to the human body, the host immune system launches an amplified response that not only breaches the blood brain barrier (BBB), but also becomes the major cause of brain damage in Amoebic meningoencephalitis. It is our understanding that for N. fowleri the host immune response is dominated by acute inflammatory cytokines and that, in cases of Acanthamoeba and Balamuthia spp., it is the type IV hypersensitivity reaction that fundamentally not only contributes to disruption and leakiness of the blood brain barrier (BBB) but also causes the neuronal damage. The further intensification of brain damage is done by toxins and enzymes secreted by the amoeba, which causes the irreversible brain damage. PMID:25930186

  11. Monitoring of Waterborne Parasites in Two Drinking Water Treatment Plants: A Study in Sarawak, Malaysia

    Richard, Reena Leeba; Ithoi, Init; Abd Majid, Mohamad Azlan; Wan Sulaiman, Wan Yusoff; Tan, Tian Chye; Nissapatorn, Veeranoot; Lim, Yvonne Ai Lian

    2016-01-01

    The occurrence of waterborne parasites coupled with water parameters at various processing sites of two drinking water treatment plants (A and B) and seven distribution system (DS) sites in Sarawak, Malaysia were studied. Ten liters of water underwent immunomagnetic separation (IMS) technique to detect the presence of Giardia and Cryptosporidium (oo)cysts. The remaining supernatant was used to detect other parasites whilst 50 mL of water sample was each used in the detection of free-living amoebae and fecal coliforms. Sampled water was positive for Giardia (32.9%; 28/85), Cryptosporidium (18.8%; 16/85) followed by Spirometra ova-like (25.9%; 22/85), Blastocystis-like (25.9%; 22/85), nematode larvae-like (8.2%; 7/85) and Taenia ova-like (1.2%; 1/85). Meanwhile, 90.2% (55/61) samples were positive for Acanthamoeba and Naegleria via cultivation and of these, 11 isolates were confirmed as Acanthamoeba genotype T3 (5/7) and T4 (2/7) followed by Naegleria sp. (4/11), Naegleria italica (2/11), Naegleria australiensis (1/11), Naegleria angularis (1/11) and Vahlkampfia sp. (3/11). Cryptosporidium, Acanthamoeba and Naegleria were also detected in one of the seven tested DS sites. Only Giardia and Cryptosporidium showed significant correlations with fluoride and fecal coliforms. These results describe the occurrence of waterborne parasites that will assist key stakeholders in mitigating contamination at the specific sites. PMID:27367710

  12. ASPECTOS BIOLÓGICOS DAS PRINCIPAIS AMEBAS DE VIDA-LIVRE DE IMPORTÂNCIA MÉDICA

    Paulo Henrique Matayoshi Calixto

    2014-08-01

    Full Text Available O objetivo desta revisão é de compilar informações sobre os principais aspectos das amebas de vida-livre (AVL. Para tal, foram consultados artigos depositados no PubMed, entre os anos de 1947 a 2014. As AVL são protozoários amplamente distribuídos no ambiente. Apesar de não serem considerados parasitos clássicos, tais como o Trypanosoma cruzi, as AVL são responsáveis por infecções oportunistas e não oportunistas em seres humanos e animais. Entre todas as AVL, apenas três gêneros são conhecidamente capazes de infectar o homem e outros animais, são eles Balamuthia, Naegleria e Acanthamoeba. Felizmente, as infecções por AVL são extremamente raras. Espécies desses gêneros são responsáveis por infecções fatais, tais como: i Acanthamoeba causa a encefalite amebiana granulomatosa (EAG, além de causar ceratites e infecções de pele, rins e pulmões; ii Balamuthia mandrillaris também provoca o mesmo tipo de encefalite; iii Naegleria fowleri é o agente etiológico da meningoencefalite amebiana primária (MAP. Palavras-chave: Amebas de vida-livre; Balamuthia mandrillaris; Naegleria fowleri; Acanthamoeba spp.; encefalite amebiana. DOI: http://dx.doi.org/10.18561/2179-5746/biotaamazonia.v4n2p124-129

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

    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. PMID:25306486

  14. Monitoring of Waterborne Parasites in Two Drinking Water Treatment Plants: A Study in Sarawak, Malaysia.

    Richard, Reena Leeba; Ithoi, Init; Abd Majid, Mohamad Azlan; Wan Sulaiman, Wan Yusoff; Tan, Tian Chye; Nissapatorn, Veeranoot; Lim, Yvonne Ai Lian

    2016-01-01

    The occurrence of waterborne parasites coupled with water parameters at various processing sites of two drinking water treatment plants (A and B) and seven distribution system (DS) sites in Sarawak, Malaysia were studied. Ten liters of water underwent immunomagnetic separation (IMS) technique to detect the presence of Giardia and Cryptosporidium (oo)cysts. The remaining supernatant was used to detect other parasites whilst 50 mL of water sample was each used in the detection of free-living amoebae and fecal coliforms. Sampled water was positive for Giardia (32.9%; 28/85), Cryptosporidium (18.8%; 16/85) followed by Spirometra ova-like (25.9%; 22/85), Blastocystis-like (25.9%; 22/85), nematode larvae-like (8.2%; 7/85) and Taenia ova-like (1.2%; 1/85). Meanwhile, 90.2% (55/61) samples were positive for Acanthamoeba and Naegleria via cultivation and of these, 11 isolates were confirmed as Acanthamoeba genotype T3 (5/7) and T4 (2/7) followed by Naegleria sp. (4/11), Naegleria italica (2/11), Naegleria australiensis (1/11), Naegleria angularis (1/11) and Vahlkampfia sp. (3/11). Cryptosporidium, Acanthamoeba and Naegleria were also detected in one of the seven tested DS sites. Only Giardia and Cryptosporidium showed significant correlations with fluoride and fecal coliforms. These results describe the occurrence of waterborne parasites that will assist key stakeholders in mitigating contamination at the specific sites. PMID:27367710

  15. AcEST: DK957280 [AcEST

    Full Text Available ia with plantlets Developmental stage gametophytes with sporophytes...12 Definition Adiantum capillus-veneris mRNA. clone: TST39A01NGRL0027_P07. 5' end sequence. Accession DK957280 Tissue type prothall...in R34 OS=Acanthamoeba polyphaga mimivirus Align length 33 Score (bit) 29.6 E-value ...rized protein OS=Ajellomyces capsulata (strain NAm1 / WU24) GN=HCAG_01495 PE=4 SV=1 Length = ... sp|Q5UPB0|YR034_MIMIV Uncharacterized protein R34 OS=Acanthamoeb... 30 7.4 >sp|Q5UPB0|YR034_MIMIV Uncharacterized prote

  16. Molecular comparison of cultivable protozoa from a pristine and a polycyclic aromatic hydrocarbon polluted site

    Lara, E; Berney, C; Ekelund, Flemming;

    2007-01-01

    We compared the abundance and diversity of cultivable protozoa (flagellates and amoebae) in a polycyclic aromatic hydrocarbon (PAH) polluted soil and an unpolluted control, by isolating and cultivating clonal strains. The number of cultivable protozoa was higher in the polluted soil; however, the...... polluted soil displayed an impoverished community, dominated by certain taxa, such as Acanthamoeba sp. We isolated a total of 31 protozoan strains to characterize them morphologically and by 18S rRNA gene sequence analysis. This approach, i.e. combining morphological and molecular information had the...

  17. Dicty_cDB: Contig-U11160-1 [Dicty_cDB

    Full Text Available -115 AB032945_1( AB032945 |pid:none) Homo sapiens mRNA for KIAA1119 pro... 419 e-115 ( P47808 ) RecName: Full=High molecular weight...23662( A23662 )myosin I, high molecular weight - Acanthamoeba sp. 416 e-114 AB209957_1( AB209957 |pid:none) ...*nrsk*iyyaiyckyyw**yrgrks*knhfr v*sfigsiw*cknit*q*fksfw*iygdsiqfgr*srrw*nyklplgkvtcnqsnsr*t* fsyflstfkrfirggkeni...*iqrrfnlyinrtsisfnesi*tirnlwk*sn*fi*r*t*i*nstty l*yc**si*sikirg*esmyyyirrirsr*nrsk*iyyaiyckyyw**yrgrks*knhfr v*sfigsiw*ckni...kcliq* Frame B: *lfnrke*iirffflflnlyf*inpxk*kikkyknkn**kveksknilkkk--- ---cwptgvlvv*ifyipyliiylfiflffiyifiy*skmatfkrdltkn

  18. Characterization of protein expression levels with label-free detected reverse phase protein arrays.

    Guo, Xuexue; Deng, Yihong; Zhu, Chenggang; Cai, Junlong; Zhu, Xiangdong; Landry, James P; Zheng, Fengyun; Cheng, Xunjia; Fei, Yiyan

    2016-09-15

    In reverse-phase protein arrays (RPPA), one immobilizes complex samples (e.g., cellular lysate, tissue lysate or serum etc.) on solid supports and performs parallel reactions of antibodies with immobilized protein targets from the complex samples. In this work, we describe a label-free detection of RPPA that enables quantification of RPPA data and thus facilitates comparison of studies performed on different samples and on different solid supports. We applied this detection platform to characterization of phosphoserine aminotransferase (PSAT) expression levels in Acanthamoeba lysates treated with artemether and the results were confirmed by Western blot studies. PMID:27372609

  19. Dicty_cDB: Contig-U16028-1 [Dicty_cDB

    Full Text Available ochromis chilotes isolate... 140 3e-44 AF410881_1( AF410881 |pid:none) Frankliniella occidentalis cys...150 2e-44 AC140034_6( AC140034 |pid:none) Medicago truncatula clone mth2-17g... 150 2e-44 AF410883_1( AF410883 |pid:none) Fra...=Cysteine proteinase 2; EC=3.4.22... 231 4e-78 EF053509_1( EF053509 |pid:none) Acanthamoeba castella...49 FJ349098_1( FJ349098 |pid:none) Acanthodendrilla sp. Vietnam cathe... 141 2e-49 DQ280314_1( DQ280314 |pid:none) Hymeniacid...tylgtpfdasslemt psekvfggvqansvdwrakgavtpiknqgecggcwsfsatgategaqyiangdsdltsvs eqqlidcsgsygnngcegglmtlafeyiinnkgid

  20. Dicty_cDB: CHP689 [Dicty_cDB

    Full Text Available smsalliivkqnsqsvglfth*qgt*agfrp s*drlvlpyccqlfa***hdlvreelscritgl*vyltk**paatirwimaerl*vrihs rnanq...ml*svnvvgnirllvgdhfql*ti*liavtlqfpsgycggwlnsnlyprsaqllr l*ttik*trccklpssvqlrg Frame C: cwptggkqnwrcglnqi... 13 unordered pieces. 343 0.0 13 AL158197 |AL158197.26 Human DNA sequence *** SEQ...fiktagrwpwksvsakecvtthlpng lalkmddassgwsmpnr*kk**yf*rvgrregnvea*m*irvelslvqilmvvaniqknl l*rpkwgrvp*qwnslmges...54 8e-06 U12823_1( U12823 |pid:none) Acanthamoeba polyphaga CDC:0187:1 hemo... 45 3e-05 EU955547_1( EU955547 |pid:none) Zea ma

  1. Dicty_cDB: Contig-U11874-1 [Dicty_cDB

    Full Text Available ne) Aspergillus niger contig An11c027... 57 8e-07 AP007171_26( AP007171 |pid:none) Aspergillus oryzae RIB40 geno...modium falciparum 3D7 chromosome 14 section 3... 38 0.17 13 ( AY653733 ) Acanthamoeba polyphaga mimivirus, complete geno...CAAGCCTATAATAAAATCAACATTCCATTACTCAGGTC ATCAACTATGTCACCTTTTATTAGNAAACATCNAGGTTCAT Gap gap included Contig length 881 Chromos...MRC20-573A12... 32 1.4 9 ( CP000102 ) Methanosphaera stadtmanae DSM 3091, complete geno...me. 38 1.4 19 ( AC115599 ) Dictyostelium discoideum chromosome 2 map 4229098... 32 1.4 8 ( AM438418 ) Vitis vinifera, whole gen

  2. Role for RpoS but Not RelA of Legionella pneumophila in Modulation of Phagosome Biogenesis and Adaptation to the Phagosomal Microenvironment

    Abu-Zant, Alaeddin; Asare, Rexford; Graham, James E.; Abu Kwaik, Yousef

    2006-01-01

    The induction of virulence traits by Legionella pneumophila at the post-exponential phase has been proposed to be triggered by the stringent response mediated by RelA, which triggers RpoS. We show that L. pneumophila rpoS but not relA is required for early intracellular survival and replication within human monocyte-derived macrophages and Acanthamoeba polyphaga. In addition, L. pneumophila rpoS but not relA is required for expression of the pore-forming activity. We provide evidence that Rpo...

  3. Occurrence and Control of Legionella in Recycled Water Systems

    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 its protozoa symbionts in recycled water with the aim of developing a risk management strategy. The review considered the intricate ecological relationships between Legionella and protozoa, methods for detecting both symbionts, and the efficacy of various disinfectants. PMID:26140674

  4. Glucosylceramide synthesis inhibition affects cell cycle progression, membrane trafficking, and stage differentiation in Giardia lamblia[S

    Štefanić, Saša; Spycher, Cornelia; Morf, Laura; Fabriàs, Gemma; Casas, Josefina; Schraner, Elisabeth; Wild, Peter; Hehl, Adrian B.; Sonda, Sabrina

    2010-01-01

    Synthesis of glucosylceramide via glucosylceramide synthase (GCS) is a crucial event in higher eukaryotes, both for the production of complex glycosphingolipids and for regulating cellular levels of ceramide, a potent antiproliferative second messenger. In this study, we explored the dependence of the early branching eukaryote Giardia lamblia on GCS activity. Biochemical analyses revealed that the parasite has a GCS located in endoplasmic reticulum (ER) membranes that is active in proliferating and encysting trophozoites. Pharmacological inhibition of GCS induced aberrant cell division, characterized by arrest of cytokinesis, incomplete cleavage furrow formation, and consequent block of replication. Importantly, we showed that increased ceramide levels were responsible for the cytokinesis arrest. In addition, GCS inhibition resulted in prominent ultrastructural abnormalities, including accumulation of cytosolic vesicles, enlarged lysosomes, and clathrin disorganization. Moreover, anterograde trafficking of the encystations-specific protein CWP1 was severely compromised and resulted in inhibition of stage differentiation. Our results reveal novel aspects of lipid metabolism in G. lamblia and specifically highlight the vital role of GCS in regulating cell cycle progression, membrane trafficking events, and stage differentiation in this parasite. In addition, we identified ceramide as a potent bioactive molecule, underscoring the universal conservation of ceramide signaling in eukaryotes. PMID:20335568

  5. Glucosylceramide synthesis inhibition affects cell cycle progression, membrane trafficking, and stage differentiation in Giardia lamblia.

    Stefanić, Sasa; Spycher, Cornelia; Morf, Laura; Fabriàs, Gemma; Casas, Josefina; Schraner, Elisabeth; Wild, Peter; Hehl, Adrian B; Sonda, Sabrina

    2010-09-01

    Synthesis of glucosylceramide via glucosylceramide synthase (GCS) is a crucial event in higher eukaryotes, both for the production of complex glycosphingolipids and for regulating cellular levels of ceramide, a potent antiproliferative second messenger. In this study, we explored the dependence of the early branching eukaryote Giardia lamblia on GCS activity. Biochemical analyses revealed that the parasite has a GCS located in endoplasmic reticulum (ER) membranes that is active in proliferating and encysting trophozoites. Pharmacological inhibition of GCS induced aberrant cell division, characterized by arrest of cytokinesis, incomplete cleavage furrow formation, and consequent block of replication. Importantly, we showed that increased ceramide levels were responsible for the cytokinesis arrest. In addition, GCS inhibition resulted in prominent ultrastructural abnormalities, including accumulation of cytosolic vesicles, enlarged lysosomes, and clathrin disorganization. Moreover, anterograde trafficking of the encystations-specific protein CWP1 was severely compromised and resulted in inhibition of stage differentiation. Our results reveal novel aspects of lipid metabolism in G. lamblia and specifically highlight the vital role of GCS in regulating cell cycle progression, membrane trafficking events, and stage differentiation in this parasite. In addition, we identified ceramide as a potent bioactive molecule, underscoring the universal conservation of ceramide signaling in eukaryotes. PMID:20335568

  6. Epigenetic mechanisms regulate stage differentiation in the minimized protozoan Giardia lamblia.

    Sonda, Sabrina; Morf, Laura; Bottova, Iveta; Baetschmann, Hansruedi; Rehrauer, Hubert; Caflisch, Amedeo; Hakimi, Mohamed-Ali; Hehl, Adrian B

    2010-04-01

    Histone modification is an important mechanism regulating both gene expression and the establishment and maintenance of cellular phenotypes during development. Regulation of histone acetylation via histone acetylases and deacetylases (HDACs) appears to be particularly crucial in determining gene expression patterns. In this study we explored the effect of HDAC inhibition on the life cycle of the human pathogen Giardia lamblia, a highly reduced parasitic protozoan characterized by minimized cellular processes. We found that the HDAC inhibitor FR235222 increased the level of histone acetylation and induced transcriptional regulation of approximately 2% of genes in proliferating and encysting parasites. In addition, our analyses showed that the levels of histone acetylation decreased during differentiation into cysts, the infective stage of the parasite. Importantly, FR235222 treatment during encystation reversed this histone hypo-acetylation and potently blocked the formation of cysts. These results provide the first direct evidence for epigenetic regulation of gene expression in this simple eukaryote. This suggests that regulation of histone acetylation is involved in the control of Giardia stage differentiation, and identifies epigenetic mechanisms as a promising target to prevent Giardia transmission. PMID:20132448

  7. The single dynamin family protein in the primitive protozoan Giardia lamblia is essential for stage conversion and endocytic transport.

    Gaechter, Verena; Schraner, Elisabeth; Wild, Peter; Hehl, Adrian B

    2008-01-01

    Dynamins are universally conserved large guanosine triphosphatases, which function as mechanoenzymes in membrane scission. The primitive protozoan Giardia lamblia has a single dynamin-related protein (GlDRP) with an unusual domain structure. Giardia lacks a Golgi apparatus but generates transient Golgi-like delay compartments dubbed encystation-specific vesicles (ESVs), which serve to accumulate and mature cyst wall proteins during differentiation to infectious cyst forms. Here, we analyze the function of GlDRP during growth and encystation and demonstrate that it relocalizes from peripheral endosomal-lysosomal compartments to nascent ESVs. We show that GlDRP is necessary for secretion of the cyst wall material and ESV homeostasis. Expression of a dominant-negative GlDRP variant does not interfere with ESV formation but blocks cyst formation completely prior to regulated exocytosis. GlDRP colocalizes with clathrin at the cell periphery and is necessary for endocytosis of surface proteins to endosomal-lysosomal organelles in trophozoites. Electron microscopy and live cell imaging reveal gross morphological changes as well as functional impairment of the endocytic system in cells expressing the dominant-negative GlDRP. Thus, giardial DRP plays a key role in two distinct trafficking pathways and in organelle homeostasis, both essential functions for the proliferation of the parasite in the gut and its transmission to a new host. PMID:17892527

  8. High Prevalence of Porocephalus crotali Infection on a Barrier Island (Cumberland Island) off the Coast of Georgia, with Identification of Novel Intermediate Hosts.

    Yabsley, Michael J; Ellis, Angela E; Cleveland, Christopher A; Ruckdeschel, Carol

    2015-10-01

    Porocephalus crotali is a pentastomid parasite that uses crotaline snakes as definitive hosts and a variety of rodents as intermediate hosts. A study of definitive and intermediate pentastome hosts on Cumberland Island, Georgia, revealed high prevalence of P. crotali infection in crotalid snakes as well as several mammalian species. Despite the presence of numerous nymphs in some animals, clinical signs of disease were not observed. In intermediate hosts, the liver, mesentery, and reproductive organs were most commonly infected. No gross evidence of tissue damage was noted in association with the numerous encysted nymphal pentastomes, and histopathology demonstrated minimal reaction to the encysted nymphs. Partial 18S rRNA gene sequences confirmed the parasites were P. crotali. In contrast to many previous reports in rodents, the prevalence on this barrier island was high, and this is the first report of Virginia opossums (Didelphis virginiana) and any insectivore species as intermediate hosts. Although generally not considered pathogenic, the long-term consequences of high nymph intensities on individuals deserve attention. PMID:26042344

  9. Pumilio Homologue from Saprolegnia parasitica Specifically Expressed in Undifferentiated Spore Cysts

    Andersson, M. Gunnar; Cerenius, Lage

    2002-01-01

    The expression of spore-specific marker transcripts at different stages of the asexual life cycle of Saprolegnia parasitica was analyzed. One of the markers, designated puf1, was found to be expressed transiently upon each of several cycles of zoospore encystment and reemergence. The transcript is induced immediately upon zoospore encystment and is rapidly lost when a cyst is triggered to germinate. In nongerminating cysts, puf1 is maintained until a time point when the cysts can no longer be triggered to germinate and thus have become determined for zoospore reemergence. The results show that the cyst stage has two phases, of about equal duration, which are physiologically and transcriptionally distinct and that the transcriptional machinery of oomycetes is also active in nongerminating spores. puf1 encodes a putative mRNA binding protein belonging to a conserved class of proteins including the Drosophila melanogaster Pumilio protein, Caenorhabditis elegans FBF, and Saccharomyces cerevisiae Puf5, all of which are involved in regulation of gene expression by posttranscriptional mechanisms. PMID:12455976

  10. Proteomic and ultrastructural analysis of the effect of a new nitazoxanide-N-methyl-1H-benzimidazole hybrid against Giardia intestinalis.

    Matadamas-Martínez, Félix; Castillo, Rafael; Hernández-Campos, Alicia; Méndez-Cuesta, Carlos; de Souza, Wanderley; Gadelha, Ana Paula; Nogueda-Torres, Benjamín; Hernández, José Manuel; Yépez-Mulia, Lilián

    2016-04-01

    In an effort to develop alternative drugs for the treatment of giardiasis our research group has synthesized and evaluated a novel nitazoxanide and N-methyl-1H-benzimidazole hybrid molecule, named CMC-20. It showed an IC50 of 0.010μM on Giardia intestinalis, lower than the IC50 values of 0.015, 0.037 and 1.224μM for nitazoxanide, albendazole and metronidazole, respectively. In addition, we report studies carried out on its mechanism of action and effect at the ultrastructural level on G. intestinalis. The proteomic analysis of trophozoites treated with CMC-20 revealed significant changes in the expression level of proteins of the cytoskeleton, alpha and beta tubulin, alpha-1, beta giardin and axoneme-associated protein, among other molecules. Ultrastructural studies demonstrated that CMC-20 induces morphological changes on the parasite that loses its characteristic pear shape. Uncommon large bulbous structure at the flagella end, and parasites showing flange membrane bending and a concave depression in the ventral region, resembling an encystation process, were also observed. In addition, some apoptotic and autophagic-like features, such as membrane blebbing, intense vacuolation, chromatin condensation and multilamellar bodies were detected. Phosphatidylserine externalization was determined as an apoptotic marker by flow cytometry and immunofluorescence microscopy; however, a typical ladder-like DNA fragmentation profile was not detected. Although it was found that CMC-20 triggers the encystation process, damage to the cyst wall indicates loss of viability. PMID:27033928

  11. Small-sized euryhaline fish as intermediate hosts of the digenetic trematode Cryptocotyle concavum

    Zander, C. D.; Kollra, H.-G.; Antholz, B.; Meyer, W.; Westphal, D.

    1984-03-01

    Cercariae of the trematode Cryptocotyle concavum, which encyst in skin and/or kidney of sticklebacks and gobies, were studied in the Schlei Fjord (western Baltic Sea). Mean incidence of dermal cysts was 48 % in Gasterosteus aculeatus and 37 % in Pungitius pungitius. No cysts were found in the kidneys of sticklebacks. While 97 % of Pomatoschistus microps had encysted metacercariae in the kidneys, only 2 % had cysts in the skin. Pomatoschistus minutus, however, showed hardly any cyst infestation of either skin or kidney. In P. microps the intensity of infestation by metacercariae was frequently more than 50 cysts; in contrast, sticklebacks rarely exhibited more than 5 dermal cysts. Infested fish were larger than 10 mm in total length, the incidence rate increasing with growth. Parasitic infestation depends on ambient salinity: C. concavum was not found at salinities below 4 ‰. In contrast to the high incidence in fish, the first hosts — the snails Hydrobia stagnalis and H. neglecta — showed remarkably low infection rates (3 to 5 %). The findings reported are related to the distribution of C. concavum, the mode of life of infested fish, the feeding habits of the final hosts and the infestation of P. microps by other parasites. Evidently, P. microps represents an optimal second host for C. concavum.

  12. Wild Rodents as Experimental Intermediate Hosts of Lagochilascaris minor Leiper, 1909

    Julieta Machado Paçô

    1999-07-01

    Full Text Available A total of 25 specimens of Cavia porcellus (guinea pig, 5 Dasyprocta agouti (agouti, and 22 Calomys callosus (vesper mice were inoculated with infective eggs of Lagochilascaris minor. The inoculum was prepared with embryonated eggs and orally administered to each individual animal through an esophagus probe. In parallel, 100 specimens of Felis catus domesticus were individually fed with 55-70 nodules containing 3rd-stage larvae encysted in tissues of infected rodents. Animals were examined and necropsied at different time intervals. The migration and encystment of L3 larva was observed in viscera, skeletal muscle, adipose and subcutaneous tissues from all rodents. Adult worms localized at abscesses in the cervical region, rhino, and oropharynx were recovered from domestic cats inoculated with infected rodent tissues. Through this study we can conclude that: (1 wild rodents act as intermediate hosts, characterizing this ascarid heteroxenic cycle; (2 in natural conditions rodents could possibly act as either intermediate hosts or paratenic hosts of Lagochilascaris minor; (3 despite the occurrence of an auto-infecting cycle, in prime-infection of felines (definite hosts the cycle is only completed when intermediate hosts are provided; and (4 in the wild, rodents could serve as a source of infection for humans as they are frequently used as food in regions with the highest incidence of human lagochilascariasis.

  13. The Role of the ncRNA RgsA in the Oxidative Stress Response and Biofilm Formation in Azotobacter vinelandii.

    Huerta, Jesús Manuel; Aguilar, Israel; López-Pliego, Liliana; Fuentes-Ramírez, Luis Ernesto; Castañeda, Miguel

    2016-06-01

    Azotobacter vinelandii is a soil bacterium that forms desiccation-resistant cysts, and the exopolysaccharide alginate is essential for this process. A. vinelandii also produces alginate under vegetative growth conditions, and this production has biotechnological significance. Poly-β-hydroxybutyrate (PHB) is another polymer synthetized by A. vinelandii that is of biotechnological interest. The GacS/A two-component signal transduction system plays an important role in regulating alginate production, PHB synthesis, and encystment. GacS/A in turn controls other important regulators such as RpoS and the ncRNAs that belong to the Rsm family. In A. vinelandii, RpoS is necessary for resisting oxidative stress as a result of its control over the expression of the catalase Kat1. In this work, we characterized a new ncRNA in A. vinelandii that is homologous to the P16/RsgA reported in Pseudomonas. We found that the expression of rgsA is regulated by GacA and RpoS and that it was essential for oxidative stress resistance. However, the activity of the catalase Kat1 is unaffected in rgsA mutants. Unlike those reported in Pseudomonas, RgsA in A. vinelandii regulates biofilm formation but not polymer synthesis or the encystment process. PMID:26858204

  14. Activity, stability and folding analysis of the chitinase from Entamoeba histolytica.

    Muñoz, Patricia L A; Minchaca, Alexis Z; Mares, Rosa E; Ramos, Marco A

    2016-02-01

    Human amebiasis, caused by the parasitic protozoan Entamoeba histolytica, remains as a significant public health issue in developing countries. The life cycle of the parasite compromises two main stages, trophozoite and cyst, linked by two major events: encystation and excystation. Interestingly, the cyst stage has a chitin wall that helps the parasite to withstand harsh environmental conditions. Since the amebic chitinase, EhCHT1, has been recognized as a key player in both encystation and excystation, it is plausible to consider that specific inhibition could arrest the life cycle of the parasite and, thus, stop the infection. However, to selectively target EhCHT1 it is important to recognize its unique biochemical features to have the ability to control its cellular function. Hence, to gain further insights into the structure-function relationship, we conducted an experimental approach to examine the effects of pH, temperature, and denaturant concentration on the enzymatic activity and protein stability. Additionally, dependence on in vivo oxidative folding was further studied using a bacterial model. Our results attest the potential of EhCHT1 as a target for the design and development of new or improved anti-amebic therapeutics. Likewise, the potential of the oxidoreductase EhPDI, involved in oxidative folding of amebic proteins, was also confirmed. PMID:26526675

  15. UV irradiation responses in Giardia intestinalis.

    Einarsson, Elin; Svärd, Staffan G; Troell, Karin

    2015-07-01

    The response to ultraviolet light (UV) radiation, a natural stressor to the intestinal protozoan parasite Giardia intestinalis, was studied to deepen the understanding of how the surrounding environment affects the parasite during transmission. UV radiation at 10 mJ/cm(2) kills Giardia cysts effectively whereas trophozoites and encysting parasites can recover from UV treatment at 100 mJ/cm(2) and 50 mJ/cm(2) respectively. Staining for phosphorylated histone H2A showed that UV treatment induces double-stranded DNA breaks and flow cytometry analyses revealed that UV treatment of trophozoites induces DNA replication arrest. Active DNA replication coupled to DNA repair could be an explanation to why UV light does not kill trophozoites and encysting cells as efficiently as the non-replicating cysts. We also examined UV-induced gene expression responses in both trophozoites and cysts using RNA sequencing (RNA seq). UV radiation induces small overall changes in gene expression in Giardia but cysts show a stronger response than trophozoites. Heat shock proteins, kinesins and Nek kinases are up-regulated, whereas alpha-giardins and histones are down-regulated in UV treated trophozoites. Expression of variable surface proteins (VSPs) is changed in both trophozoites and cysts. Our data show that Giardia cysts have limited ability to repair UV-induced damage and this may have implications for drinking- and waste-water treatment when setting criteria for the use of UV disinfection to ensure safe water. PMID:25825252

  16. Consequence of temperature changes on cercarial shedding from Galba truncatula infected with Fasciola hepatica or Paramphistomum daubneyi

    Rondelaud Daniel

    2013-01-01

    Full Text Available Experimental infections of Galba truncatula (two populations with Fasciola hepatica or Paramphistomum daubneyi were carried out to study the effect of water temperature changes (3 h at a mean of 12 °C every week on cercarial shedding during the patent period. The results were compared with those of control snails infected according to the same protocol and always maintained at 20 °C. Compared to controls, a significant increase in the number of cercariae-shedding snails, a significantly longer patent period and significantly greater cercarial production were noted in temperature-challenged snails, regardless of the type of digenean infection. In contrast, the number of incompletely formed metacercariae was significantly higher in temperature-challenged snails than in controls. Incompletely formed metacercariae of F. hepatica consisted of cysts whose colour remained whitish after shedding (25.4% for temperature-challenged snails or whose dome was flattened after encystment (74.6%. Those of P. daubneyi were totally dark brown or blackish after formation. These incomplete metacercariae might originate from young differentiating cercariae within the snail body (F. hepatica or from cercariae which died just after encystment (P. daubneyi. The use of regular temperature changes for snails infected with F. hepatica or P. daubneyi must be monitored carefully during collection of metacercariae to select completely formed cysts for infecting definitive hosts.

  17. Free-living protozoa in the gastrointestinal tract and feces of pigs: Exploration of an unknown world and towards a protocol for the recovery of free-living protozoa.

    Chavatte, N; Lambrecht, E; Van Damme, I; Sabbe, K; Houf, K

    2016-07-30

    Associations with free-living protozoa (FLP) have been implicated in the persistence of foodborne pathogenic bacteria in food-related environments. To date however no information is available on the presence of FLP in the gastrointestinal tract (GIT) of pigs, which represents an important reservoir for zoonotic foodborne bacteria and hence a potential location for associations with FLP. This is at least partly due to the lack of adequate protocols to recover FLP from intestinal content and feces. In the present study different protocols to recover FLP from the porcine GIT and feces were tested. The most effective protocols were then applied to explore the presence of live FLP in the pig GIT and feces. A filtration based protocol was identified as the most suitable method to recover viable FLP from the porcine GIT and feces. Cultivable FLP were recovered from different parts of the GIT, suggesting at least a transient presence of FLP in this habitat. Free-living amoebae species (Acanthamoeba spp., Hyperamoeba sp., Vannella sp., Vermamoeba vermiformis, hartmannellids and vahlkampfiids) but also ciliates (Colpoda sp. and Tetrahymena/Glaucoma lookalike) and flagellates (cercomonads, bodonids and glissomonads) were recovered and cultured from pig intestinal content. Acanthamoeba hatchetti and Filamoeba sinensis were isolated for the first time from pig intestinal content. Despite high gastric acidity, non-cyst forming amoeba species were also detected which suggests survival of their trophozoites in the animal GIT. PMID:27369581

  18. Marine mimivirus relatives are probably large algal viruses

    Claverie Jean-Michel

    2008-01-01

    Full Text Available Abstract Background Acanthamoeba polyphaga mimivirus is the largest known ds-DNA virus and its 1.2 Mb-genome sequence has revealed many unique features. Mimivirus occupies an independent lineage among eukaryotic viruses and its known hosts include only species from the Acanthamoeba genus. The existence of mimivirus relatives was first suggested by the analysis of the Sargasso Sea metagenomic data. Results We now further demonstrate the presence of numerous "mimivirus-like" sequences using a larger marine metagenomic data set. We also show that the DNA polymerase sequences from three algal viruses (CeV01, PpV01, PoV01 infecting different marine algal species (Chrysochromulina ericina, Phaeocystis pouchetii, Pyramimonas orientalis are very closely related to their homolog in mimivirus. Conclusion Our results suggest that the numerous mimivirus-related sequences identified in marine environments are likely to originate from diverse large DNA viruses infecting phytoplankton. Micro-algae thus constitute a new category of potential hosts in which to look for new species of Mimiviridae.

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

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

    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.

  1. Distribuição das ceratites infecciosas em hospital terciário no Brasil

    Fábio Iglesias Marujo

    2013-12-01

    Full Text Available OBJETIVO: Investigar sazonalidade de ceratities infecciosas em serviço terciário no Brasil. MÉTODOS: Estudo retrospectivo de dados coletados do Laboratório de Microbiologia Ocular do Departamento de Oftalmologia da Universidade Federal de São Paulo de 2005 a 2009. Foram definidas como ceratites infecciosas aquelas com cultura positiva para o agente. Dados foram distribuídos de acordo com os meses do ano e modelos de regressão linear com variáveis "dummy" foram utilizados para testar sazonalidade. RESULTADOS: Foram diagnosticados 1.468 casos de ceratite infecciosa durante o período do estudo. Bactérias foram responsáveis por 80,3% dos casos seguidas por fungos (7%, Acanthamoeba (6% e casos mistos (6,7%. Testes estatísticos não mostraram sazonalidade para bactérias (p=0,064, fungos (p=0,379, Acanthamoeba (p=0,062 ou casos mistos (p=0,441. CONCLUSÃO: Não foi observada sazonalidade nas ceratites infecciosas em nossa amostra.

  2. Local and systemic inflammatory and immunologic reactions to cyathostomin larvicidal therapy in horses.

    Nielsen, M K; Loynachan, A T; Jacobsen, S; Stewart, J C; Reinemeyer, C R; Horohov, D W

    2015-12-15

    Encysted cyathostomin larvae are ubiquitous in grazing horses. Arrested development occurs in this population and can lead to an accumulation of encysted larvae. Large numbers of tissue larvae place the horse at risk for developing larval cyathostominosis. This disease complex is caused by mass emergence of these larvae and is characterized by a generalized acute typhlocolitis and manifests itself as a profuse protein-losing watery diarrhea with a reported case-fatality rate of about 50%. Two anthelmintic formulations have a label claim for larvicidal therapy of these encysted stages; moxidectin and a five-day regimen of fenbendazole. There is limited knowledge about inflammatory and immunologic reactions to larvicidal therapy. This study was designed to evaluate blood acute phase reactants as well as gene expression of pro-inflammatory cytokines, both locally in the large intestinal walls and systemically. Further, mucosal tissue samples were evaluated histopathologically as well as analyzed for gene expression of pro- and anti-inflammatory cytokines, cluster of differentiation (CD) cell surface proteins, and select transcription factors. Eighteen juvenile horses with naturally acquired cyathostomin infections were randomly assigned to three treatment groups; one group served as untreated controls (Group 1), one received a five-day regimen of fenbendazole (10mg/kg) (Group 2), and one group received moxidectin (0.4mg/kg) (Group 3). Horses were treated on day 0 and euthanatized on days 18-20. Serum and whole blood samples were collected on days 0, 5, and 18. All horses underwent necropsy with collection of tissue samples from the ventral colon and cecum. Acute phase reactants measured included serum amyloid A, iron and fibrinogen, and the cytokines evaluated included interferon γ, tumor necrosis factor α, transforming growth factor (TGF)-β, and interleukins 1β, 4, 5, 6, and 10. Transcription factors evaluated were FoxP3, GATA3 and tBet, and CD markers included

  3. An unusual case of intracystic papillary carcinoma of breast with invasive component

    Suryawanshi Kishor H, Nikumbh Dhiraj B, Damle Rajshri P, Dravid NV, Tayde Yogesh

    2014-07-01

    Full Text Available Papillary carcinoma of the breast is a rare malignant tumor, constituting 1-2 % of breast neoplasms mostly affecting elderly postmenopausal women. Intracystic (Encysted papillary carcinoma (IPC is a rare distinct entity with slow growth rate and overall favourable prognosis regardless of whether it is in situ alone or associated with invasive component. Treatment modalities vary from conservative surgery to radical surgery with or without adjuvant therapy depending upon the associated component (DCIS or invasive of the tumor. Herein, we report a case of 55-year-old female presented with a painless lump in the right breast. FNAC yielded haemorrhagic fluid with scanty cellularity of atypical ductal epithelial cells. Patient underwent wide local excision. The final histopathological diagnosis revealed intracystic papillary carcinoma associated with invasive ductal carcinoma, NOS type.

  4. First report of fossilized cysts produced by the benthic Bysmatrum subsalsum (Dinophyceae) from a shallow Mexican lagoon in the Gulf of Mexico.

    Limoges, Audrey; Mertens, Kenneth Neil; Ruíz-Fernández, Ana-Carolina; de Vernal, Anne

    2015-02-01

    Cysts belonging to the benthic dinoflagellate Bysmatrum subsalsum were recovered from palynologically treated sediments collected in the Alvarado Lagoon (southwestern Gulf of Mexico). The cysts are proximate, reflecting the features of the parent thecal stage, and their autofluorescence implies a dinosporin composition similar to the cyst walls of phototrophic species. This finding is important for our understanding of B. subsalsum life cycle transitions and ecology. Encystment may play an important role in the bloom dynamics of this species as it can enable the formation of a sediment cyst bank that allows reinoculation of the water column when conditions become favorable. This is the first report of a fossilized cyst produced by a benthic dinoflagellate recovered from sub-recent sediments. PMID:26986270

  5. Disseminated cysticercosis in a child: whole-body MR diagnosis with the use of parallel imaging

    Cysticercosis is a parasitic disease caused by infestation with the encysted larval stage of the pork tapeworm, Taenia solium. Disseminated cysticercosis is an exceptional expression of this disease characterized by high morbidity due to massive symptomatic parasite burden in the central nervous system, striated muscles, subcutaneous tissues and other organs. Less than 50 such cases have been reported worldwide, and fewer than 10 children. We report on the whole-body MR diagnosis of extensively disseminated cysticercosis in a child. The critical role of whole-body MR as a stand-alone modality in the diagnosis and management of this pleomorphic disease is highlighted. Whole-body MR diagnosis of an infectious disease is unique. (orig.)

  6. Effect of ultraviolet-A radiation on the production of Leptolegnia chapmanii (Saprolegniales: Saprolegniaceae) zoospores on dead Aedes aegypti (Diptera: Culicidae) larvae and their larvicidal activity.

    Rueda Páramo, Manuel E; López Lastra, Claudia C; García, Juan J; Fernandes, Éverton K K; Marreto, Ricardo N; Luz, Christian

    2015-09-01

    Impact of UV-radiation in entomopathogens in aquatic environments remains little investigated. The present study reports on the effect of UV-A on the larvicidal activity of Leptolegnia chapmanii zoospores in Aedes aegypti; on the production of zoospores in larvae killed by the pathogen and then exposed to UV-A; and on the activity of these zoospores against healthy larvae. Whereas the virulence of free zoospores in A. aegypti larvae was affected by a UV-A exposure time longer than 10min, production of zoospores in larvae and their virulence were not hampered at a maximal 8h exposure of dead larvae to UV-A. Findings suggest that dead larvae and zoosporangia provide a certain protection to zoospores against UV-A and emphasize the susceptibility of free encysted zoospores to such radiation. PMID:26259676

  7. INVESTIGATION ON MODEL OF INFLAMMATORY PLEURAL EFFUSION IN GUINEA PIGS

    冯源; 殷凯生; 王祥

    2002-01-01

    Objective To establish an animal model of inflammatory pleural effusion.Methods Forty guinea pigs were divided into two groups: experimental group with 7 subgroups and control group. In the experimental group the right chest cavity of each guinea pig was injected with 0.8~1.0 ml of 1% carrageenan, and guinea pigs of each subgroup were killed and observed respectively on day 1, 2, 3, 5, 7, 10 and day 14 after injection.Results Occurring on day 1(within 24 hours), pleural effusion reached the maximum on day 2~3 after injection, so did the neutrophil count in pleural effusion and inflammation of both pleura and lungs and then gradually decreased. The fibrosis and adhesion of pleura appeared on day 7 and were obvious on day 10. The encysted pleurisy was formed on day 14.Conclusion The carrageenan is an ideal pleural inflammatory inducer. This animal model is useful for studying pleural effusion.

  8. New paradigms for understanding and step changes in treating active and chronic, persistent apicomplexan infections.

    McPhillie, Martin; Zhou, Ying; El Bissati, Kamal; Dubey, Jitender; Lorenzi, Hernan; Capper, Michael; Lukens, Amanda K; Hickman, Mark; Muench, Stephen; Verma, Shiv Kumar; Weber, Christopher R; Wheeler, Kelsey; Gordon, James; Sanders, Justin; Moulton, Hong; Wang, Kai; Kim, Taek-Kyun; He, Yuqing; Santos, Tatiana; Woods, Stuart; Lee, Patty; Donkin, David; Kim, Eric; Fraczek, Laura; Lykins, Joseph; Esaa, Farida; Alibana-Clouser, Fatima; Dovgin, Sarah; Weiss, Louis; Brasseur, Gael; Wirth, Dyann; Kent, Michael; Hood, Leroy; Meunieur, Brigitte; Roberts, Craig W; Hasnain, S Samar; Antonyuk, Svetlana V; Fishwick, Colin; McLeod, Rima

    2016-01-01

    Toxoplasma gondii, the most common parasitic infection of human brain and eye, persists across lifetimes, can progressively damage sight, and is currently incurable. New, curative medicines are needed urgently. Herein, we develop novel models to facilitate drug development: EGS strain T. gondii forms cysts in vitro that induce oocysts in cats, the gold standard criterion for cysts. These cysts highly express cytochrome b. Using these models, we envisioned, and then created, novel 4-(1H)-quinolone scaffolds that target the cytochrome bc1 complex Qi site, of which, a substituted 5,6,7,8-tetrahydroquinolin-4-one inhibits active infection (IC50, 30 nM) and cysts (IC50, 4 μM) in vitro, and in vivo (25 mg/kg), and drug resistant Plasmodium falciparum (IC50, toxoplasmosis, malaria, and ~2 billion persons chronically infected with encysted bradyzoites. PMID:27412848

  9. Disseminated cysticercosis in a child: whole-body MR diagnosis with the use of parallel imaging

    Kumar, Atin; Goenka, Ajit Harishkumar [All India Institute of Medical Sciences, Department of Radiology, New Delhi, Delhi (India); Choudhary, Anita; Sahu, Jitendra Kumar; Gulati, Sheffali [All India Institute of Medical Sciences, Department of Paediatrics, New Delhi, Delhi (India)

    2010-02-15

    Cysticercosis is a parasitic disease caused by infestation with the encysted larval stage of the pork tapeworm, Taenia solium. Disseminated cysticercosis is an exceptional expression of this disease characterized by high morbidity due to massive symptomatic parasite burden in the central nervous system, striated muscles, subcutaneous tissues and other organs. Less than 50 such cases have been reported worldwide, and fewer than 10 children. We report on the whole-body MR diagnosis of extensively disseminated cysticercosis in a child. The critical role of whole-body MR as a stand-alone modality in the diagnosis and management of this pleomorphic disease is highlighted. Whole-body MR diagnosis of an infectious disease is unique. (orig.)

  10. An unusual case of cycticercosis of the tendon sheath of the tendoachilles.

    Sharat Agarwal

    2010-04-01

    Full Text Available Cysticercosis is an infection by the larval stage (cystcercus cellulosae of the cestode, Taenia Solium (pork tape worm, especially in those individuals who live in the endemic areas. After gaining entry into the body, the larvae become encysted and may lie in subcutaneous tissue, striated muscle, the vitreous humor, or other tissues. We report an unusual case of cysticercosis of the the tendon sheath of the tendoachilles that presented as a swelling of the tendoachilles. Upon Fine Needle Aspiration and Cytology (FNAC that were conducted preoperatively, the possibility of villonodular synovitis was identified. However, the cysticercosis diagnosis was confirmed later after an excisional biopsy was performed. We could find no reports in the literature concerning an occurrence of cysticercosis in the tendon sheath of tendoachilles.

  11. Modeling effective transmission pathways and control of the world's most successful parasite.

    Turner, Matthew; Lenhart, Suzanne; Rosenthal, Benjamin; Zhao, Xiaopeng

    2013-06-01

    Toxoplasma gondii(T. gondii) is a single-celled, intracellular protozoan responsible for the disease toxoplasmosis. The parasite is prevalent worldwide, and it infects all warm-blooded vertebrates. Consumption of meats in which this parasite has encysted confers risk of infection to people and other animals, as does ingestion of water or foods contaminated with environmentally resistant oocysts excreted by cats. Vertical transmission (from mother to offspring) is also possible, leading to disease risk and contributing additional means of ensuring perpetuation of transmission. In this work, we adopt a differential equation model to investigate the effective transmission pathways of T. gondii, as well as potential control mechanisms. Detailed analyses are carried out to examine the significance of transmission routes, virulence, vertical transmission, parasite-induced changes in host behavior, and controls based on vaccination and harvesting. Modeling and analysis efforts may shed insights into understanding the complex life cycle of T. gondii. PMID:23624067

  12. Histological identification of muscular sarcocystis: A report of two cases

    Mani Makhija

    2012-01-01

    Full Text Available Sarcocystis is an apicomplexan protozoan belonging to same phylum as toxoplasma. The parasite encysts inside striated muscles of its intermediate host. Humans are accidental host infected by eating food or water contaminated with oocysts or sporocysts of an infected definitive host. The infection is increasing in Southeast Asia and may be overlooked in histological sections if one is not aware of the histomorphological features. The size and shape of the bradyzoites and the appearance of the cyst wall are the reliable features to distinguish this parasite from other parasites of the same phylum. The incidence of human infection is rising in Southeast Asia and histopathology is an important method for the diagnosis of muscular infection. It is important to recognize the histomorphology of this parasite and its differentiation from similar parasites.

  13. Helminths Parasite Larvae collected from Arabian Gulf Fish. 4. Description of four Larvae including two Metacercarae, one Didymozoid and one Acanthocephalan from Emirati Coasts

    Four helminth larvae from different fish hosts caught from Emirate coasts are described. Two are metacercariae related to genus Southampton's, type (I) and (II). Type (I) was found encysted in body cavities of the Indian halibut Psettodes erumei and the areolate grouper Epinephelus areolatusu and type (II) in the golden stripped goatfish Mulloides flavolineatus. The third larva is related to Acanthocephala and identified as Serrasentis sagittifer. It was collected from the body cavity of the spotted lizard fish Saurida undosquamius. The fourth larva belongs to the didymozoid trematodes, and was found infecting the kidneys of different hosts, such as Saurida undosquamis, the Jack pomfret Parastromateus niger and mackerel tuna Euthynnus affins. The larva related to Stephanostomum type (I) and type (II) and the dydimozoid type are described for the first time in the Arabian Gulf. (author)

  14. Contributions to the Understanding of the Bloom Dynamics of Pyrodinium bahamense var. compressum: A Toxic Red Tide Causative Organism

    Rhodora Azanza

    1997-12-01

    Full Text Available Pyrodinium bahamense var. compressum has been the primary organism responsible for the toxic red tide episodes which have been recurring in Manila Bay, Philippines since 1988. The life history of the species has been elucidated through encystment-excystment studies in vitro, from which its obligatory dormancy has been demonstrated. Cyst-mapping studies have shown that this life stage of the organism occurs relatively higher in the Bataan and Cavite areas where the greater number of red tide occurrences have also been reported. A cyst-based model has been developed as an initial step in understanding the role of physical processes in the development, occurrence/recurrence, and ultimately, advancement of Pyrodinium red tides in Manila Bay.To help mitigate or prevent the negative impacts of toxic red tides, particularly in Manila Bay, regular closure of shellfish harvest in areas affected by Pyrodinium bloom should be considered, based on long term monitoring and research data sets.

  15. Effect of infection by irradiated Trichinella Spirals larvae on mice and assessment the role of Al bendazole in treating them

    The present study was carried out to investigate the effect of infection with irradiated Trichinella Spiralis larvae on mice and to asses the role of albendazole in treating them. This study included parasitological and histopathological studies on mice infected with irradiated Trichinella Spiralis larvae in comparison with mice infected with non-irradiated Trichinella Spiralis only or with mice treated after infection by albendazole. The obtained data revealed that, in mice infected with irradiated Trichinella Spiralis larvae (50 Krad or 80 Krad), the number and length of worms in the small intestine, as well as, the number of encysted larvae in muscles of mice, especially diaphragm and tongue, were significantly decreased. Also, using al bendazole 24 hours after infection with irradiated larvae lead to high significant decrease in all the previously mentioned parameters

  16. Ectopic fascioliasis mimicking a colon tumor

    Ozer Makay; Baris Gurcu; Cemil Caliskan; Deniz Nart; Muge Tuncyurek; Mustafa Korkut

    2007-01-01

    Fasciola hepatica, a leaf shaped trematode that is common in cattle, sheep and goats, is acquired by eating raw water plants like watercress or drinking water infected with the encysted form of the parasite.The varied clinical presentations of fascioliasis still make a high index of suspicion mandatory. Besides having a wide spectrum of hepatobiliary symptoms like obstructive jaundice, cholangitis and liver cirrhosis, the parasitic infection also has extrabiliary manifestations.Until recently, extrahepatic fascioliasis has been reported in the subcutaneous tissue, brain, lungs, epididymis,inguinal lymph nodes, stomach and the cecum. In this report, a strange manifestation of the fasciola infection in a site other than the liver, a colonic fascioliasis, is presented.

  17. Infectious ileocecitis--appendicitis mimicking syndrome.

    Zganjer, M; Roic, G; Cizmic, A; Pajic, A

    2005-01-01

    The purpose of our study is to emphasize the central role of ultrasound (US) in finding the cause of abdominal pain in children. Ultrasound of the lower abdomen quadrant should be considered in all cases in which the clinical signs and symptoms are not diagnostic of appendicitis. There is a wide range of clinical syndromes and diseases which can easily be diagnosed using a high resolution ultrasound with adjunct of color and power Doppler. The spectrum of abnormalities includes appendicitis, mesenteric lymphadenitis, infectious ileocecitis, Crohn's disease, intussusception, ovarian cysts, and encysted cerebrospinal fluid. One of the most common causes of acute abdominal pain in children is acute terminal ileitis (infectious ileocecitis) with mesenteric lymphadenitis. Ultrasound is the best tool to rapidly differentiate this disease from acute appendicitis, and prevent unnecessary laparotomy (Ref. 12). PMID:16201735

  18. Isolation and identification of N-mercapto-4-formylcarbostyril, an antibiotic produced by Pseudomonas fluorescens.

    Fakhouri, W; Walker, F; Vogler, B; Armbruster, W; Buchenauer, H

    2001-12-01

    Pseudomonas fluorescens strain G308 isolated from barley leaves produces a novel antibiotic substance that was purified by preparative TLC and HPLC and identified as N-mercapto-4-formylcarbostyril (Cbs) by LC/DAD, IR, LC-ES(+)/MS, LC-ES(-)/MS, GC-EI/MS, LC-HRES(+)/MS, mass isotope ratios analysis, 1H NMR and 13C NMR analysis. The purified new antibiotic compound is effective against many phytopathogenic fungi in vitro. The compound inhibited at 25 ppm spore germination and germ tube growth of the following fungi; Fusarium oxysporum f. sp. lycopersici, Fusarium culmorum, Cladosporium cucumerinum and Colletotrichum lagenarium. At concentrations up to 125 ppm, the compound did not interfere with release of zoospores from sporangia and germination of encysted zoospores of Phytophthora infestans. PMID:11738425

  19. Proteomics of secretory and endocytic organelles in Giardia lamblia.

    Petra B Wampfler

    Full Text Available Giardia lamblia is a flagellated protozoan enteroparasite transmitted as an environmentally resistant cyst. Trophozoites attach to the small intestine of vertebrate hosts and proliferate by binary fission. They access nutrients directly via uptake of bulk fluid phase material into specialized endocytic organelles termed peripheral vesicles (PVs, mainly on the exposed dorsal side. When trophozoites reach the G2/M restriction point in the cell cycle they can begin another round of cell division or encyst if they encounter specific environmental cues. They induce neogenesis of Golgi-like organelles, encystation-specific vesicles (ESVs, for regulated secretion of cyst wall material. PVs and ESVs are highly simplified and thus evolutionary diverged endocytic and exocytic organelle systems with key roles in proliferation and transmission to a new host, respectively. Both organelle systems physically and functionally intersect at the endoplasmic reticulum (ER which has catabolic as well as anabolic functions. However, the unusually high degree of sequence divergence in Giardia rapidly exhausts phylogenomic strategies to identify and characterize the molecular underpinnings of these streamlined organelles. To define the first proteome of ESVs and PVs we used a novel strategy combining flow cytometry-based organelle sorting with in silico filtration of mass spectrometry data. From the limited size datasets we retrieved many hypothetical but also known organelle-specific factors. In contrast to PVs, ESVs appear to maintain a strong physical and functional link to the ER including recruitment of ribosomes to organelle membranes. Overall the data provide further evidence for the formation of a cyst extracellular matrix with minimal complexity. The mass spectrometry proteomics data have been deposited to the ProteomeXchange Consortium with the dataset identifier PXD000694.

  20. Selective condensation drives partitioning and sequential secretion of cyst wall proteins in differentiating Giardia lamblia.

    Christian Konrad

    2010-04-01

    Full Text Available Controlled secretion of a protective extracellular matrix is required for transmission of the infective stage of a large number of protozoan and metazoan parasites. Differentiating trophozoites of the highly minimized protozoan parasite Giardia lamblia secrete the proteinaceous portion of the cyst wall material (CWM consisting of three paralogous cyst wall proteins (CWP1-3 via organelles termed encystation-specific vesicles (ESVs. Phylogenetic and molecular data indicate that Diplomonads have lost a classical Golgi during reductive evolution. However, neogenesis of ESVs in encysting Giardia trophozoites transiently provides basic Golgi functions by accumulating presorted CWM exported from the ER for maturation. Based on this "minimal Golgi" hypothesis we predicted maturation of ESVs to a trans Golgi-like stage, which would manifest as a sorting event before regulated secretion of the CWM. Here we show that proteolytic processing of pro-CWP2 in maturing ESVs coincides with partitioning of CWM into two fractions, which are sorted and secreted sequentially with different kinetics. This novel sorting function leads to rapid assembly of a structurally defined outer cyst wall, followed by slow secretion of the remaining components. Using live cell microscopy we find direct evidence for condensed core formation in maturing ESVs. Core formation suggests that a mechanism controlled by phase transitions of the CWM from fluid to condensed and back likely drives CWM partitioning and makes sorting and sequential secretion possible. Blocking of CWP2 processing by a protease inhibitor leads to mis-sorting of a CWP2 reporter. Nevertheless, partitioning and sequential secretion of two portions of the CWM are unaffected in these cells. Although these cysts have a normal appearance they are not water resistant and therefore not infective. Our findings suggest that sequential assembly is a basic architectural principle of protective wall formation and requires

  1. Proteomics of secretory and endocytic organelles in Giardia lamblia.

    Wampfler, Petra B; Tosevski, Vinko; Nanni, Paolo; Spycher, Cornelia; Hehl, Adrian B

    2014-01-01

    Giardia lamblia is a flagellated protozoan enteroparasite transmitted as an environmentally resistant cyst. Trophozoites attach to the small intestine of vertebrate hosts and proliferate by binary fission. They access nutrients directly via uptake of bulk fluid phase material into specialized endocytic organelles termed peripheral vesicles (PVs), mainly on the exposed dorsal side. When trophozoites reach the G2/M restriction point in the cell cycle they can begin another round of cell division or encyst if they encounter specific environmental cues. They induce neogenesis of Golgi-like organelles, encystation-specific vesicles (ESVs), for regulated secretion of cyst wall material. PVs and ESVs are highly simplified and thus evolutionary diverged endocytic and exocytic organelle systems with key roles in proliferation and transmission to a new host, respectively. Both organelle systems physically and functionally intersect at the endoplasmic reticulum (ER) which has catabolic as well as anabolic functions. However, the unusually high degree of sequence divergence in Giardia rapidly exhausts phylogenomic strategies to identify and characterize the molecular underpinnings of these streamlined organelles. To define the first proteome of ESVs and PVs we used a novel strategy combining flow cytometry-based organelle sorting with in silico filtration of mass spectrometry data. From the limited size datasets we retrieved many hypothetical but also known organelle-specific factors. In contrast to PVs, ESVs appear to maintain a strong physical and functional link to the ER including recruitment of ribosomes to organelle membranes. Overall the data provide further evidence for the formation of a cyst extracellular matrix with minimal complexity. The mass spectrometry proteomics data have been deposited to the ProteomeXchange Consortium with the dataset identifier PXD000694. PMID:24732305

  2. Selective condensation drives partitioning and sequential secretion of cyst wall proteins in differentiating Giardia lamblia.

    Konrad, Christian; Spycher, Cornelia; Hehl, Adrian B

    2010-04-01

    Controlled secretion of a protective extracellular matrix is required for transmission of the infective stage of a large number of protozoan and metazoan parasites. Differentiating trophozoites of the highly minimized protozoan parasite Giardia lamblia secrete the proteinaceous portion of the cyst wall material (CWM) consisting of three paralogous cyst wall proteins (CWP1-3) via organelles termed encystation-specific vesicles (ESVs). Phylogenetic and molecular data indicate that Diplomonads have lost a classical Golgi during reductive evolution. However, neogenesis of ESVs in encysting Giardia trophozoites transiently provides basic Golgi functions by accumulating presorted CWM exported from the ER for maturation. Based on this "minimal Golgi" hypothesis we predicted maturation of ESVs to a trans Golgi-like stage, which would manifest as a sorting event before regulated secretion of the CWM. Here we show that proteolytic processing of pro-CWP2 in maturing ESVs coincides with partitioning of CWM into two fractions, which are sorted and secreted sequentially with different kinetics. This novel sorting function leads to rapid assembly of a structurally defined outer cyst wall, followed by slow secretion of the remaining components. Using live cell microscopy we find direct evidence for condensed core formation in maturing ESVs. Core formation suggests that a mechanism controlled by phase transitions of the CWM from fluid to condensed and back likely drives CWM partitioning and makes sorting and sequential secretion possible. Blocking of CWP2 processing by a protease inhibitor leads to mis-sorting of a CWP2 reporter. Nevertheless, partitioning and sequential secretion of two portions of the CWM are unaffected in these cells. Although these cysts have a normal appearance they are not water resistant and therefore not infective. Our findings suggest that sequential assembly is a basic architectural principle of protective wall formation and requires minimal Golgi sorting

  3. EVALUATION OF THE THERAPEUTIC EFFICACY OF LEVAMISOLE HYDROCHLORIDE ON THIRD-STAGE LARVAE OF Lagochilascaris minor IN EXPERIMENTALLY INFECTED MICE

    CAMPOS, Dulcinéa Maria Barbosa; BARBOSA, Alverne Passos; OLIVEIRA, Jayrson Araújo; BARBOSA, Carlos Augusto Lopes; LOBO, Tamara Flavia Correa; SILVA, Luana Gabriella; THOMAZ, Douglas Vieira; PEIXOTO, Josana de Castro

    2016-01-01

    Lagochilascariosis, a disease caused by Lagochilascaris minor, affects the neck, sinuses, tonsils, lungs, the sacral region, dental alveoli, eyeballs and the central nervous system of humans. A cycle of autoinfection may occur in human host tissues characterized by the presence of eggs, larvae and adult worms. This peculiarity of the cycle hinders therapy, since there are no drugs that exhibit ovicidal, larvicidal and vermicidal activity. Given these facts, we studied the action of levamisole hydrochloride on third-stage larvae in the migration phase (G1) and on encysted larvae (G3) of L. minor. To this end, 87 inbred mice of the C57BL/6 strain were divided into test groups comprising 67 animals (G1-37; G3-30) and a control group (G2-10; G4-10) with 20 animals. Each animal was inoculated orally with 2,000 infective eggs of the parasite. The animals of the test groups were treated individually with a single oral dose of levamisole hydrochloride at a concentration of 0.075 mg. The drug was administered either 30 minutes prior to the parasite inoculation (G1 animals) or 120 days after the inoculation (G3 animals). The mice in the control groups were not treated with the drug. After the time required for the migration and the encysting of L. minor larvae, all the animals were euthanized and their tissues examined. The data were analyzed using the Student's unpaired t-test and the Levene test. The groups showed no statistically significant difference. Levamisole hydrochloride was ineffective on third-stage larvae of L. minor. These findings explain the massive expulsion of live adult worms, as well as the use of long treatment schemes, owing to the persistence of larvae and eggs in human parasitic lesions. PMID:27253745

  4. EVALUATION OF THE THERAPEUTIC EFFICACY OF LEVAMISOLE HYDROCHLORIDE ON THIRD-STAGE LARVAE OF Lagochilascaris minor IN EXPERIMENTALLY INFECTED MICE

    Dulcinéa Maria Barbosa CAMPOS

    2016-01-01

    Full Text Available Lagochilascariosis, a disease caused by Lagochilascaris minor, affects the neck, sinuses, tonsils, lungs, the sacral region, dental alveoli, eyeballs and the central nervous system of humans. A cycle of autoinfection may occur in human host tissues characterized by the presence of eggs, larvae and adult worms. This peculiarity of the cycle hinders therapy, since there are no drugs that exhibit ovicidal, larvicidal and vermicidal activity. Given these facts, we studied the action of levamisole hydrochloride on third-stage larvae in the migration phase (G1 and on encysted larvae (G3 of L. minor. To this end, 87 inbred mice of the C57BL/6 strain were divided into test groups comprising 67 animals (G1-37; G3-30 and a control group (G2-10; G4-10 with 20 animals. Each animal was inoculated orally with 2,000 infective eggs of the parasite. The animals of the test groups were treated individually with a single oral dose of levamisole hydrochloride at a concentration of 0.075 mg. The drug was administered either 30 minutes prior to the parasite inoculation (G1 animals or 120 days after the inoculation (G3 animals. The mice in the control groups were not treated with the drug. After the time required for the migration and the encysting of L. minor larvae, all the animals were euthanized and their tissues examined. The data were analyzed using the Student's unpaired t-test and the Levene test. The groups showed no statistically significant difference. Levamisole hydrochloride was ineffective on third-stage larvae of L. minor. These findings explain the massive expulsion of live adult worms, as well as the use of long treatment schemes, owing to the persistence of larvae and eggs in human parasitic lesions.

  5. A fast and reliable procedure for spore collection from anaerobic fungi: Application for RNA uptake and long-term storage of isolates.

    Calkins, Shelby; Elledge, Nicole C; Hanafy, Radwa A; Elshahed, Mostafa S; Youssef, Noha

    2016-08-01

    Anaerobic gut fungi (AGF) represent a basal fungal lineage (phylum Neocallimastigomycota) that resides in the rumen and alimentary tracts of herbivores. The AGF reproduce asexually, with a life cycle that involves flagellated zoospores released from zoosporangia followed by encystment, germination and the subsequent development of rhizomycelia. A fast and reliable approach for AGF spore collection is critical not only for developmental biology studies, but also for molecular biological (e.g. AMT-transformation and RNAi) approaches. Here, we developed and optimized a simple and reliable procedure for the collection of viable, competent, and developmentally synchronized AGF spores under strict anaerobic conditions. The approach involves growing AGF on agar medium in serum bottles under anaerobic conditions, and flooding the observed aerial growth to promote spore release from sporangia into the flooding suspension. The released spores are gently collected using a wide bore sterile needle. Process optimization resulted in the recovery of up to 7×10(9) spores per serum bottle. Further, the released spores exhibited synchronized development from flagellated spores to encysted spores and finally to germinating spores within 90min from the onset of flooding. At the germinating spore stage, the obtained spores were competent, and readily uptook small interfering RNA (siRNA) oligonucleotides. Finally, using multiple monocentric and polycentric AGF isolates, we demonstrate that AGF grown on agar surface could retain viability for up to 16weeks at 39°C, and hence this solid surface growth procedure represents a simple, cryopreservative- and freezing temperature-free approach for AGF storage. PMID:27288952

  6. MSCs: The Sentinel and Safe-Guards of Injury.

    Caplan, Arnold I

    2016-07-01

    Mesenchymal stem cells (MSCs) were originally named because they could differentiate in a variety of mesenchymal phenotypes in culture. Evidence indicates that MSCs arise from perivascular cells, pericytes, when the blood vessels are broken or inflamed. These pericyte/MSCs are situated on every blood vessel in the body. The MSCs sense the micro-environment of the injury site and secrete site-specific factors that serve several important reparative functions: first, a curtain of molecules from the front of the MSCs provide a barrier from the interrogation of the over-aggressive immune system. Second, from the back of the MSCs, a different set of bioactive agents inhibit scar formation and establish a regenerative micro-environment. Third, if bacteria are sensed by the MSCs, they produce powerful protein antibiotics that kill the bacteria on contact. Last, the MSCs surround and encyst intruding solid objects like a piece of wood (a "splinter") or other foreign objects. The MSCs act as a combination paramedic and emergency room (ER) staff to survey the damage, isolate foreign components, stabilize the injured tissues, provide antibiotics and encysting protection before a slower, medicinal sequence can be initiated to regenerate the damaged tissue. The MSCs, thus, act as sentinels to safeguard the individual from intrusion and chronic injury. A societal treatment system has evolved, paramedics and ER procedures, which mirror in a macro-sense what MSCs orchestrate in a micro-sense. Key to this new understanding is that MSCs are not "stem cells," but rather as Medicinal Signaling Cells as the therapeutic agents. J. Cell. Physiol. 231: 1413-1416, 2016. © 2015 Wiley Periodicals, Inc. PMID:26565391

  7. Development of RNA Interference Trigger-Mediated Gene Silencing in Entamoeba invadens.

    Suresh, Susmitha; Ehrenkaufer, Gretchen; Zhang, Hanbang; Singh, Upinder

    2016-04-01

    Entamoeba histolytica, a protozoan parasite, is an important human pathogen and a leading parasitic cause of death. The organism has two life cycle stages, trophozoites, which are responsible for tissue invasion, and cysts, which are involved in pathogen transmission. Entamoeba invadens is the model system to study Entamoeba developmental biology, as high-grade regulated encystation and excystation are readily achievable. However, the lack of gene-silencing tools in E. invadens has limited the molecular studies that can be performed. Using the endogenous RNA interference (RNAi) pathway in Entamoeba, we developed an RNAi-based trigger gene-silencing approach inE. invadens We demonstrate that a gene's coding region that has abundant antisense small RNAs (sRNAs) can trigger silencing of a gene that is fused to it. The trigger fusion leads to the generation of abundant antisense sRNAs that map to the target gene, with silencing occurring independently of trigger location at the 5' or 3' end of a gene. Gene silencing is stably maintained during development, including encystation and excystation. We have used this approach to successfully silence two E. invadens genes: a putative rhomboid protease gene and a SHAQKY family Myb gene. The Myb gene is upregulated during oxidative stress and development, and its downregulation led, as predicted, to decreased viability under oxidative stress and decreased cyst formation. Thus, the RNAi trigger silencing method can be used to successfully investigate the molecular functions of genes inE. invadens Dissection of the molecular basis of Entamoeba stage conversion is now possible, representing an important technical advance for the system. PMID:26787723

  8. Monitoring of Legionella pneumophila viability after chlorine dioxide treatment using flow cytometry.

    Mustapha, Pascale; Epalle, Thibaut; Allegra, Séverine; Girardot, Françoise; Garraud, Olivier; Riffard, Serge

    2015-04-01

    The viability of three Legionella pneumophila strains was monitored after chlorine dioxide (ClO2) treatment using a flow cytometric assay. Suspensions of L. pneumophila cells were submitted to increasing concentrations of ClO2. Culturable cells were still detected when using 4 mg/L, but could no longer be detected after exposure to 6 mg/L of ClO2, although viable but not culturable (VBNC) cells were found after exposure to 4-5 mg/L of ClO2. When testing whether these VBNC were infective, two of the strains were resuscitated after co-culture with Acanthamoeba polyphaga, but neither of them could infect macrophage-like cells. PMID:25725384

  9. Dicty_cDB: Contig-U01941-1 [Dicty_cDB

    Full Text Available Frame C: rikiqqqqqqqqqqhqqqq*lqqlrlqmlwliilkikiiiiliiiiiiiinnliiimnn* mkflkmlk**lermkvilwklkv*ivimv*piqilvil...ne CH240-394N1, WORKING DRAFT SEQUE... 42 0.68 5 ( AY653733 ) Acanthamoeba polyphaga mimivirus, complete gen...to callus, TAMU Solanum lycopersicu... 34 5.5 2 ( AW625601 ) EST319508 tomato radicle, 5 d post-im...34 6.3 2 ( BI423219 ) EST533885 tomato callus, TAMU Solanum lycopersicu... 34 6.3 2 ( FH551127 ) CHO_OF4501x... ( EJ973847 ) 1093022101464 Global-Ocean-Sampling_GS-30-02-01-1... 34 6.4 2 ( FH551041 ) CHO_OF4501xf19f1.ab1 CHO_OF4 Nicotiana

  10. Occurrence and Control of Legionella in Recycled Water Systems

    Patrick K. Jjemba

    2015-07-01

    Full Text Available 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 its protozoa symbionts in recycled water with the aim of developing a risk management strategy. The review considered the intricate ecological relationships between Legionella and protozoa, methods for detecting both symbionts, and the efficacy of various disinfectants.

  11. Dicty_cDB: Contig-U16527-1 [Dicty_cDB

    Full Text Available S... 1285 0.0 3 ( AM168032 ) Dictyostelium capitatum 18S rRNA gene, isolate...ne, co... 254 0.0 17 ( AF316545 ) Acanthamoeba sp. KA/E12 18S ribosomal RNA gene, c... 254 0.0 17 ( EF025003 ) Uncultured eukaryote...ar... 262 0.0 18 ( EF025035 ) Uncultured eukaryote clone Elev_18S_7101 18S ribo... 254 0.0 17 ( AF333608 ) Aca.... 89 1e-15 AB116235_1( AB116235 |pid:none) Rosellinia sp. PF1022 tef1 gene fo... 89 1e-15 D45837_1( D45837 |pid:none) Neur...tavvyrsewpi*rgsftgsvvagwrapnrhshllyceq whespdvasa*ktcatn*swsvrrpflgmhalshprrrqqcrlsrrvkatrpmq*vteeq *gdllakc

  12. Dicty_cDB: Contig-U09543-1 [Dicty_cDB

    Full Text Available discoideum ORF DG1040 gene, partial cds. 44 0.026 3 ( AP006852 ) Candida albicans genomic DNA, chromosome 7, com..._1( AX972104 |pid:none) Sequence 2907 from Patent EP1104808. 43 0.029 CP001185_1028( CP001185 |pid:none) Thermosipho african... 7 cl... 91 1e-16 AF514806_1( AF514806 |pid:none) Leishmania mexicana mexicana mu ad... 91...01_1( EU021001 |pid:none) Narceus americanus clone Nam69f2 p... 37 1.6 AM494966_153( AM494966 |pid:none) Leishmania braziliens...tyostelium discoideum strain AX4 chromosome 2 ... 36 0.006 11 ( AY653733 ) Acanthamoeba polyphaga mimivirus, com

  13. Presence of pathogenic microorganisms in power-plant cooling waters. Report for October 1, 1979-September 30, 1981

    Tyndall, R.L.

    1982-10-01

    Cooling waters from eleven geographically disparate power plants were tested for the presence of Naegleria fowleri and Legionella pneumophila (LDB). Control source waters for each plant were also tested for these pathogens. Water from two of the eleven plants contained pathogenic Naegleria, and infectious Legionella were found in seven of the test sites. Pathogenic Naegleria were not found in control waters, but infectious Legionella were found in five of the eleven control source water sites. Concentrations of nitrite, sulfate, and total organic carbon correlated with the concentrations of LDB. A new species of Legionella was isolated from one of the test sites. In laboratory tests, both Acanthamoeba and Naegleria were capable of supporting the growth of Legionella pneumophila.

  14. Dicty_cDB: SLB439 [Dicty_cDB

    Full Text Available SL (Link to library) SLB439 (Link to dictyBase) - - - Contig-U16455-1 SLB439Z (Link... to Original site) - - SLB439Z 709 - - - - Show SLB439 Library SL (Link to library) Clone ID SLB439 (Link to...ycdb.biol.tsukuba.ac.jp/CSM/SL/SLB4-B/SLB439Q.Seq.d/ Representative seq. ID SLB43...9Z (Link to Original site) Representative DNA sequence >SLB439 (SLB439Q) /CSM/SL/SLB4-B/SLB439Q.Seq.d/ XXXXX...r 1-alpha; Short... 307 2e-82 AY582829_1( AY582829 |pid:none) Acanthamoeba culber

  15. Dicty_cDB: SLB144 [Dicty_cDB

    Full Text Available SL (Link to library) SLB144 (Link to dictyBase) - - - Contig-U16455-1 SLB144Z (Link... to Original site) - - SLB144Z 710 - - - - Show SLB144 Library SL (Link to library) Clone ID SLB144 (Link to...ycdb.biol.tsukuba.ac.jp/CSM/SL/SLB1-B/SLB144Q.Seq.d/ Representative seq. ID SLB14...4Z (Link to Original site) Representative DNA sequence >SLB144 (SLB144Q) /CSM/SL/SLB1-B/SLB144Q.Seq.d/ XXXXX...29 |pid:none) Acanthamoeba culbertsoni ef1a gene... 321 2e-86 AY643820_1( AY643820 |pid:none) Arcyria denuda

  16. Dicty_cDB: SLB344 [Dicty_cDB

    Full Text Available SL (Link to library) SLB344 (Link to dictyBase) - - - Contig-U16455-1 SLB344Z (Link... to Original site) - - SLB344Z 620 - - - - Show SLB344 Library SL (Link to library) Clone ID SLB344 (Link to...ycdb.biol.tsukuba.ac.jp/CSM/SL/SLB3-B/SLB344Q.Seq.d/ Representative seq. ID SLB34...4Z (Link to Original site) Representative DNA sequence >SLB344 (SLB344Q) /CSM/SL/SLB3-B/SLB344Q.Seq.d/ XXXXX...RNA for elongation factor ... 263 3e-69 AY582829_1( AY582829 |pid:none) Acanthamoeba culbertsoni ef1a gene..

  17. Dicty_cDB: SLB518 [Dicty_cDB

    Full Text Available SL (Link to library) SLB518 (Link to dictyBase) - - - Contig-U16455-1 SLB518Z (Link... to Original site) - - SLB518Z 694 - - - - Show SLB518 Library SL (Link to library) Clone ID SLB518 (Link to...ycdb.biol.tsukuba.ac.jp/CSM/SL/SLB5-A/SLB518Q.Seq.d/ Representative seq. ID SLB51...8Z (Link to Original site) Representative DNA sequence >SLB518 (SLB518Q) /CSM/SL/SLB5-A/SLB518Q.Seq.d/ XXXXX...29_1( AY582829 |pid:none) Acanthamoeba culbertsoni ef1a gene... 313 2e-84 protein

  18. Dicty_cDB: SLB811 [Dicty_cDB

    Full Text Available SL (Link to library) SLB811 (Link to dictyBase) - - - Contig-U16455-1 SLB811Z (Link... to Original site) - - SLB811Z 553 - - - - Show SLB811 Library SL (Link to library) Clone ID SLB811 (Link to...ycdb.biol.tsukuba.ac.jp/CSM/SL/SLB8-A/SLB811Q.Seq.d/ Representative seq. ID SLB81...1Z (Link to Original site) Representative DNA sequence >SLB811 (SLB811Q) /CSM/SL/SLB8-A/SLB811Q.Seq.d/ XXXXX...:none) Axinella verrucosa mRNA for elonga... 225 7e-58 AY582829_1( AY582829 |pid:none) Acanthamoeba culberts

  19. Dicty_cDB: SLB739 [Dicty_cDB

    Full Text Available SL (Link to library) SLB739 (Link to dictyBase) - - - Contig-U16455-1 SLB739Z (Link... to Original site) - - SLB739Z 685 - - - - Show SLB739 Library SL (Link to library) Clone ID SLB739 (Link to...ycdb.biol.tsukuba.ac.jp/CSM/SL/SLB7-B/SLB739Q.Seq.d/ Representative seq. ID SLB73...9Z (Link to Original site) Representative DNA sequence >SLB739 (SLB739Q) /CSM/SL/SLB7-B/SLB739Q.Seq.d/ XXXXX...303 2e-81 ( Q00251 ) RecName: Full=Elongation factor 1-alpha; Short... 303 2e-81 AY582829_1( AY582829 |pid:none) Acanthamoeba culb

  20. Dicty_cDB: SLB883 [Dicty_cDB

    Full Text Available SL (Link to library) SLB883 (Link to dictyBase) - - - Contig-U16455-1 SLB883Z (Link... to Original site) - - SLB883Z 554 - - - - Show SLB883 Library SL (Link to library) Clone ID SLB883 (Link to...ycdb.biol.tsukuba.ac.jp/CSM/SL/SLB8-D/SLB883Q.Seq.d/ Representative seq. ID SLB88...3Z (Link to Original site) Representative DNA sequence >SLB883 (SLB883Q) /CSM/SL/SLB8-D/SLB883Q.Seq.d/ XXXXX...alpha; Short... 232 4e-60 AY582829_1( AY582829 |pid:none) Acanthamoeba culbertson

  1. Dicty_cDB: SLB265 [Dicty_cDB

    Full Text Available SL (Link to library) SLB265 (Link to dictyBase) - - - Contig-U16455-1 SLB265Z (Link... to Original site) - - SLB265Z 632 - - - - Show SLB265 Library SL (Link to library) Clone ID SLB265 (Link to...ycdb.biol.tsukuba.ac.jp/CSM/SL/SLB2-C/SLB265Q.Seq.d/ Representative seq. ID SLB26...5Z (Link to Original site) Representative DNA sequence >SLB265 (SLB265Q) /CSM/SL/SLB2-C/SLB265Q.Seq.d/ XXXXX...AY643816 |pid:none) Lycogala epidendrum elongation fac... 261 2e-68 AY582829_1( AY582829 |pid:none) Acanthamoeba culb

  2. Dicty_cDB: SLB508 [Dicty_cDB

    Full Text Available SL (Link to library) SLB508 (Link to dictyBase) - - - Contig-U16455-1 SLB508Z (Link... to Original site) - - SLB508Z 629 - - - - Show SLB508 Library SL (Link to library) Clone ID SLB508 (Link to...ycdb.biol.tsukuba.ac.jp/CSM/SL/SLB5-A/SLB508Q.Seq.d/ Representative seq. ID SLB50...8Z (Link to Original site) Representative DNA sequence >SLB508 (SLB508Q) /CSM/SL/SLB5-A/SLB508Q.Seq.d/ XXXXX... |pid:none) Acanthamoeba culbertsoni ef1a gene... 267 2e-70 AJ581004_1( AJ581004 |pid:none) Axinella verruco

  3. Dicty_cDB: SLB693 [Dicty_cDB

    Full Text Available SL (Link to library) SLB693 (Link to dictyBase) - - - Contig-U16455-1 SLB693Z (Link... to Original site) - - SLB693Z 615 - - - - Show SLB693 Library SL (Link to library) Clone ID SLB693 (Link to...ycdb.biol.tsukuba.ac.jp/CSM/SL/SLB6-D/SLB693Q.Seq.d/ Representative seq. ID SLB69...3Z (Link to Original site) Representative DNA sequence >SLB693 (SLB693Q) /CSM/SL/SLB6-D/SLB693Q.Seq.d/ XXXXX...gation factor ... 263 3e-69 AY582829_1( AY582829 |pid:none) Acanthamoeba culbertsoni ef1a gene... 263 4e-69

  4. Dicty_cDB: SLB314 [Dicty_cDB

    Full Text Available SL (Link to library) SLB314 (Link to dictyBase) - - - Contig-U16455-1 SLB314Z (Link... to Original site) - - SLB314Z 660 - - - - Show SLB314 Library SL (Link to library) Clone ID SLB314 (Link to...ycdb.biol.tsukuba.ac.jp/CSM/SL/SLB3-A/SLB314Q.Seq.d/ Representative seq. ID SLB31...4Z (Link to Original site) Representative DNA sequence >SLB314 (SLB314Q) /CSM/SL/SLB3-A/SLB314Q.Seq.d/ XXXXX...ha; Short... 289 6e-77 AY582829_1( AY582829 |pid:none) Acanthamoeba culbertsoni ef1a gene... 289 6e-77 prote

  5. Dicty_cDB: SLB890 [Dicty_cDB

    Full Text Available SL (Link to library) SLB890 (Link to dictyBase) - - - Contig-U16455-1 SLB890Z (Link... to Original site) - - SLB890Z 556 - - - - Show SLB890 Library SL (Link to library) Clone ID SLB890 (Link to...ycdb.biol.tsukuba.ac.jp/CSM/SL/SLB8-D/SLB890Q.Seq.d/ Representative seq. ID SLB89...0Z (Link to Original site) Representative DNA sequence >SLB890 (SLB890Q) /CSM/SL/SLB8-D/SLB890Q.Seq.d/ XXXXX...lpha; Short... 224 1e-57 AY582829_1( AY582829 |pid:none) Acanthamoeba culbertsoni

  6. Dicty_cDB: SLB431 [Dicty_cDB

    Full Text Available SL (Link to library) SLB431 (Link to dictyBase) - - - Contig-U16455-1 SLB431Z (Link... to Original site) - - SLB431Z 694 - - - - Show SLB431 Library SL (Link to library) Clone ID SLB431 (Link to...ycdb.biol.tsukuba.ac.jp/CSM/SL/SLB4-B/SLB431Q.Seq.d/ Representative seq. ID SLB43...1Z (Link to Original site) Representative DNA sequence >SLB431 (SLB431Q) /CSM/SL/SLB4-B/SLB431Q.Seq.d/ XXXXX...29_1( AY582829 |pid:none) Acanthamoeba culbertsoni ef1a gene... 309 5e-83 protein

  7. Dicty_cDB: SLB409 [Dicty_cDB

    Full Text Available SL (Link to library) SLB409 (Link to dictyBase) - - - Contig-U16455-1 SLB409Z (Link... to Original site) - - SLB409Z 687 - - - - Show SLB409 Library SL (Link to library) Clone ID SLB409 (Link to...ycdb.biol.tsukuba.ac.jp/CSM/SL/SLB4-A/SLB409Q.Seq.d/ Representative seq. ID SLB40...9Z (Link to Original site) Representative DNA sequence >SLB409 (SLB409Q) /CSM/SL/SLB4-A/SLB409Q.Seq.d/ XXXXX...pid:none) Acanthamoeba culbertsoni ef1a gene... 303 2e-81 protein update 2009. 6.

  8. Dicty_cDB: SLB109 [Dicty_cDB

    Full Text Available SL (Link to library) SLB109 (Link to dictyBase) - - - Contig-U16455-1 SLB109Z (Link... to Original site) - - SLB109Z 564 - - - - Show SLB109 Library SL (Link to library) Clone ID SLB109 (Link to...ycdb.biol.tsukuba.ac.jp/CSM/SL/SLB1-A/SLB109Q.Seq.d/ Representative seq. ID SLB10...9Z (Link to Original site) Representative DNA sequence >SLB109 (SLB109Q) /CSM/SL/SLB1-A/SLB109Q.Seq.d/ XXXXX...me: Full=Elongation factor 1-alpha; Short... 246 2e-64 AY582829_1( AY582829 |pid:none) Acanthamoeba culb

  9. Dicty_cDB: SLB118 [Dicty_cDB

    Full Text Available SL (Link to library) SLB118 (Link to dictyBase) - - - Contig-U16455-1 SLB118Z (Link... to Original site) - - SLB118Z 573 - - - - Show SLB118 Library SL (Link to library) Clone ID SLB118 (Link to...ycdb.biol.tsukuba.ac.jp/CSM/SL/SLB1-A/SLB118Q.Seq.d/ Representative seq. ID SLB11...8Z (Link to Original site) Representative DNA sequence >SLB118 (SLB118Q) /CSM/SL/SLB1-A/SLB118Q.Seq.d/ XXXXX...-62 AY582829_1( AY582829 |pid:none) Acanthamoeba culbertsoni ef1a gene... 241 1e-62 AB326305_1( AB326305 |pi

  10. Dicty_cDB: SLB838 [Dicty_cDB

    Full Text Available SL (Link to library) SLB838 (Link to dictyBase) - - - Contig-U16455-1 SLB838Z (Link... to Original site) - - SLB838Z 531 - - - - Show SLB838 Library SL (Link to library) Clone ID SLB838 (Link to...ycdb.biol.tsukuba.ac.jp/CSM/SL/SLB8-B/SLB838Q.Seq.d/ Representative seq. ID SLB83...8Z (Link to Original site) Representative DNA sequence >SLB838 (SLB838Q) /CSM/SL/SLB8-B/SLB838Q.Seq.d/ XXXXX...AY582829_1( AY582829 |pid:none) Acanthamoeba culbertsoni ef1a gene... 223 2e-57 AY643820_1( AY643820 |pid:no

  11. Dicty_cDB: SLB572 [Dicty_cDB

    Full Text Available SL (Link to library) SLB572 (Link to dictyBase) - - - Contig-U16455-1 SLB572Z (Link... to Original site) - - SLB572Z 574 - - - - Show SLB572 Library SL (Link to library) Clone ID SLB572 (Link to...ycdb.biol.tsukuba.ac.jp/CSM/SL/SLB5-C/SLB572Q.Seq.d/ Representative seq. ID SLB57...2Z (Link to Original site) Representative DNA sequence >SLB572 (SLB572Q) /CSM/SL/SLB5-C/SLB572Q.Seq.d/ XXXXX... 1e-62 AY582829_1( AY582829 |pid:none) Acanthamoeba culbertsoni ef1a gene... 241 1e-62 AB326305_1( AB326305

  12. Dicty_cDB: SLB583 [Dicty_cDB

    Full Text Available SL (Link to library) SLB583 (Link to dictyBase) - - - Contig-U16455-1 SLB583Z (Link... to Original site) - - SLB583Z 457 - - - - Show SLB583 Library SL (Link to library) Clone ID SLB583 (Link to...ycdb.biol.tsukuba.ac.jp/CSM/SL/SLB5-D/SLB583Q.Seq.d/ Representative seq. ID SLB58...3Z (Link to Original site) Representative DNA sequence >SLB583 (SLB583Q) /CSM/SL/SLB5-D/SLB583Q.Seq.d/ XXXXX... P18624 ) RecName: Full=Elongation factor 1-alpha; Short... 231 4e-60 AY582829_1( AY582829 |pid:none) Acanthamoeba culb

  13. Dicty_cDB: SLB363 [Dicty_cDB

    Full Text Available SL (Link to library) SLB363 (Link to dictyBase) - - - Contig-U16455-1 SLB363Z (Link... to Original site) - - SLB363Z 582 - - - - Show SLB363 Library SL (Link to library) Clone ID SLB363 (Link to...ycdb.biol.tsukuba.ac.jp/CSM/SL/SLB3-C/SLB363Q.Seq.d/ Representative seq. ID SLB36...3Z (Link to Original site) Representative DNA sequence >SLB363 (SLB363Q) /CSM/SL/SLB3-C/SLB363Q.Seq.d/ XXXXX...lpha; Short... 244 9e-64 AY582829_1( AY582829 |pid:none) Acanthamoeba culbertsoni ef1a gene... 244 1e-63 AB3

  14. Dicty_cDB: SLB102 [Dicty_cDB

    Full Text Available SL (Link to library) SLB102 (Link to dictyBase) - - - Contig-U16455-1 SLB102Z (Link... to Original site) - - SLB102Z 575 - - - - Show SLB102 Library SL (Link to library) Clone ID SLB102 (Link to...ycdb.biol.tsukuba.ac.jp/CSM/SL/SLB1-A/SLB102Q.Seq.d/ Representative seq. ID SLB10...2Z (Link to Original site) Representative DNA sequence >SLB102 (SLB102Q) /CSM/SL/SLB1-A/SLB102Q.Seq.d/ XXXXX...9 4e-62 AY582829_1( AY582829 |pid:none) Acanthamoeba culbertsoni ef1a gene... 239 5e-62 AB326305_1( AB326305

  15. Dicty_cDB: SLB334 [Dicty_cDB

    Full Text Available SL (Link to library) SLB334 (Link to dictyBase) - - - Contig-U16455-1 SLB334Z (Link... to Original site) - - SLB334Z 546 - - - - Show SLB334 Library SL (Link to library) Clone ID SLB334 (Link to...ycdb.biol.tsukuba.ac.jp/CSM/SL/SLB3-B/SLB334Q.Seq.d/ Representative seq. ID SLB33...4Z (Link to Original site) Representative DNA sequence >SLB334 (SLB334Q) /CSM/SL/SLB3-B/SLB334Q.Seq.d/ XXXXX.... 224 1e-57 AY582829_1( AY582829 |pid:none) Acanthamoeba culbertsoni ef1a gene...

  16. Evaluation of military field-water quality: Volume 6, Infectious organisms of military concern associated with nonconsumptive exposure: Assessment of health risks and recommendations for establishing related standards

    Cooper, R.C.; Olivieri, A.W.; Danielson, R.E.; Badger, P.G.

    1986-02-01

    This study is an assessment of the risk of illness due to exposure to water-related (i.e., water-based, water-washed) infectious organisms. The organisms under consideration are Aeromonas spp., Leptospira spp., Pseudomonas spp., Staphylococcus spp., non-cholerae Vibrio spp., Acanthamoeba spp., Balantidium coli, Naegleria spp., Ascaris lumbricoides, Dracunculus medinesis, Schistosoma spp., and the agents responsible for cercarial dermatitis (i.e., Trichobilharzia, Gigantobilharzia, and Austrobilharzia). Evaluation of the risk to disease associated with the above pathogens requires information in specific areas such as dose response, concentration of agents in the environment, and environmental persistence. The existing body of knowledge concerning these agents ranges from speculation to established fact. Unfortunately, areas of information critical to risk assessment are frequently unavailable. Because of this lack of data, the risk assessment presented is semiquantitative and limited to the presentation of an environmental classification scheme. 14 refs., 2 figs., 57 tabs.

  17. Giant viruses come of age.

    Fischer, Matthias G

    2016-06-01

    Viruses with genomes up to a few megabases in length are a common occurrence in nature, even though they have escaped our notice until recently. These giant viruses infect mainly single-celled eukaryotes and isolation efforts concentrating on amoebal hosts alone have spawned hundreds of viral isolates, featuring viruses with previously unseen virion morphologies and the largest known viral genomes and particles. One of the challenges that lie ahead is to analyze and categorize the available data and to establish an approved classification system that reflects the evolutionary relationships and biological properties of these viruses. Extensive sampling of Acanthamoeba-infecting mimiviruses and initial characterization of their virophage parasites have provided a first blueprint of the genetic diversity and composition of a giant virus clade that will facilitate the taxonomic grouping of these fascinating microorganisms. PMID:26999382

  18. [Investigation of potentially pathogenic free-living amoebae and their in vivo pathogenicity in water supplies of Turkey].

    Yazar, Süleyman; Gürbüz, Esra; Sönmez, Mehmet Fatih; Çetinkaya, Ülfet; Kuk, Salih

    2016-07-01

    Free-living amoebae (FLA) are found widely in soil and water in the nature. Among them in which potentially pathogenic for humans and animals are known as "potential pathogenic free-living amoebae (PPFLA)". PPFLA are characterized as the causes of clinical manifestations leading to death especially in immunosuppressed people. Four genus of PPFLA (Acanthamoeba, Naegleria, Balamuthia and Sappinia) are known to be pathogenic to humans. The aims of this study were to investigate the presence of PPFLA in the water supplies in Turkey and to determine their in vivo pathogenicity. A total of 664 water samples were collected from the ponds, rivers, streams and wells found in provinces located at different regions (central, western, eastern and southeastern regions) of Turkey. These samples were initially inoculated in the monoxenic culture media and evaluated by both microscopy and polymerase chain reaction (PCR) in terms of the presence of FLA. The samples identified as positive were then cultured in axenic media, the growth of amoebae that were confirmed microscopically, were than studied with PCR for molecular characterization. The isolates that were found positive by PCR from axenic cultures were inoculated intranasally to immunocompetent and immunodeficient (athymic) [BALB/c Rag2(-/-) gamma(c)(-/-)] BALB/c mice followed by the evaluation on the 21st day by histopathological and molecular methods to investigate their in vivo pathogenicity. In our study, 143 water samples were detected as positive in monoxenic cultures and 41 of them were detected as positive in axenic cultures. Twenty of 41 samples detected as positive in axenic culture could be continued in culture for three months. As a result of PCR using primers common to SYA, only nine have been identified from 20 samples as positive. According to the result of the PCR with specific primers, all (n= 9) were positive for Acanthamoeba sp., eight for Sappini sp. and five for Balamuthia mandrillaris, while none was

  19. Dicty_cDB: Contig-U15469-1 [Dicty_cDB

    Full Text Available :none) Oncorhynchus mykiss procathepsin L... 120 1e-25 AF410883_1( AF410883 |pid:none) Frankliniella occide...ulis japonicus aCatL mRNA for... 118 5e-25 AF410881_1( AF410881 |pid:none) Frankliniella occide...ium discoideum chromoso... 171 2e-43 EF053509_1( EF053509 |pid:none) Acanthamoeba castella...s clone KtCL53 cathe... 81 6e-14 X03930_1( X03930 |pid:none) Dictyostelium discoideum 3'gene frag...ney cDN... 111 4e-23 FJ349098_1( FJ349098 |pid:none) Acanthodendrilla sp. Vietnam cathe... 71

  20. Mimiviruses: Replication, Purification, and Quantification.

    Abrahão, Jônatas Santos; Oliveira, Graziele Pereira; Ferreira da Silva, Lorena Christine; Dos Santos Silva, Ludmila Karen; Kroon, Erna Geessien; La Scola, Bernard

    2016-01-01

    The aim of this protocol is to describe the replication, purification, and titration of mimiviruses. These viruses belong to the Mimiviridae family, the first member of which was isolated in 1992 from a cooling tower water sample collected during an outbreak of pneumonia in a hospital in Bradford, England. In recent years, several new mimiviruses have been isolated from different environmental conditions. These giant viruses are easily replicated in amoeba of the Acanthamoeba genus, its natural host. Mimiviruses present peculiar features that make them unique viruses, such as the particle and genome size and the genome's complexity. The discovery of these viruses rekindled discussions about their origin and evolution, and the genetic and structural complexity opened up a new field of study. Here, we describe some methods utilized for mimiviruses replication, purification, and titration. © 2016 by John Wiley & Sons, Inc. PMID:27153385

  1. Contact lens disinfection by ultraviolet light

    A 253.7-nm ultraviolet light with an intensity of 1,100 microW/cm2 was tested for its germicidal activity against contact lenses and storage solutions contaminated with various corneal pathogens. The exposure time necessary to reduce a concentration of organisms from 10(6)/ml to less than 10/ml was 30 seconds for Staphylococcus aureus, 60 seconds for Pseudomonas aeruginosa, and 84 seconds for Candida albicans. The time necessary to sterilize a suspension of 10(4)/ml Acanthamoeba polyphaga was less than three minutes with this technique. Four brands of soft contact lenses were exposed to ultraviolet light for over eight hours without changing their appearance, comfort, or refraction

  2. Contact lens disinfection by ultraviolet light

    Dolman, P.J.; Dobrogowski, M.J. (Univ. of British Columbia, Vancouver (Canada))

    1989-12-15

    A 253.7-nm ultraviolet light with an intensity of 1,100 microW/cm2 was tested for its germicidal activity against contact lenses and storage solutions contaminated with various corneal pathogens. The exposure time necessary to reduce a concentration of organisms from 10(6)/ml to less than 10/ml was 30 seconds for Staphylococcus aureus, 60 seconds for Pseudomonas aeruginosa, and 84 seconds for Candida albicans. The time necessary to sterilize a suspension of 10(4)/ml Acanthamoeba polyphaga was less than three minutes with this technique. Four brands of soft contact lenses were exposed to ultraviolet light for over eight hours without changing their appearance, comfort, or refraction.

  3. Systemic acanthamoebiasis associated with canine distemper in dogs in the semiarid region of Paraíba, Brazil

    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

  4. Revised Mimivirus major capsid protein sequence reveals intron-containing gene structure and extra domain

    Suzan-Monti Marie

    2009-05-01

    Full Text Available Abstract Background Acanthamoebae polyphaga Mimivirus (APM is the largest known dsDNA virus. The viral particle has a nearly icosahedral structure with an internal capsid shell surrounded with a dense layer of fibrils. A Capsid protein sequence, D13L, was deduced from the APM L425 coding gene and was shown to be the most abundant protein found within the viral particle. However this protein remained poorly characterised until now. A revised protein sequence deposited in a database suggested an additional N-terminal stretch of 142 amino acids missing from the original deduced sequence. This result led us to investigate the L425 gene structure and the biochemical properties of the complete APM major Capsid protein. Results This study describes the full length 3430 bp Capsid coding gene and characterises the 593 amino acids long corresponding Capsid protein 1. The recombinant full length protein allowed the production of a specific monoclonal antibody able to detect the Capsid protein 1 within the viral particle. This protein appeared to be post-translationnally modified by glycosylation and phosphorylation. We proposed a secondary structure prediction of APM Capsid protein 1 compared to the Capsid protein structure of Paramecium Bursaria Chlorella Virus 1, another member of the Nucleo-Cytoplasmic Large DNA virus family. Conclusion The characterisation of the full length L425 Capsid coding gene of Acanthamoebae polyphaga Mimivirus provides new insights into the structure of the main Capsid protein. The production of a full length recombinant protein will be useful for further structural studies.

  5. Pathogen filtration to control plant disease outbreak in greenhouse production

    Jeon, Sangho; Krasnow, Charles; Bhalsod, Gemini; Granke, Leah; Harlan, Blair; Hausbeck, Mary; Zhang, Wei

    2016-04-01

    Previous research has been extensively focused on understanding the fate and transport of human microbial pathogens in soil and water environments. However, little is known about the transport of plant pathogens, although these pathogens are often found in irrigation waters and could cause severe crop damage and economical loss. Water mold pathogens including Phytophthora spp. and Pythium spp. are infective to a wide range of vegetable and floriculture crops, and they are primarily harbored in soils and disseminated through water flow. It is challenging to control these pathogens because they often quickly develop resistance to many fungicides. Therefore, this multi-scale study aimed to investigate physical removal of plant pathogens from water by filtration, thus reducing the pathogen exposure risks to crops. In column-scale experiments, we studied controlling factors on the transport and retention of Phytophthora capsici zoospores in saturated columns packed with iron oxide coated-sand and uncoated-sand under varying solution chemistry. Biflagellate zoospores were less retained than encysted zoospores, and lower solution pH and greater iron oxide content increased the retention of encysted zoospores. These results provided insights on environmental dispersal of Phytophthora zoospores in natural soils as well as on developing cost-effective engineered filtration systems for pathogen removal. Using small-scale greenhouse filtration systems, we further investigated the performance of varying filter media (i.e., granular sand, iron oxide coated ceramic porous media, and activated carbon) in mitigating disease outbreaks of Phytophthora and Pythium for greenhouse-grown squash and poinsettia, respectively, in comparison with fungicide treatment. For squash, filtration by iron oxide coated media was more effective in reducing the Phytophthora infection, comparing to sand filtration and fungicide application. For poinsettia, sand filtration performed better in controlling

  6. Proteomic Study of Entamoeba histolytica Trophozoites, Cysts, and Cyst-Like Structures.

    Luna-Nácar, Milka; Navarrete-Perea, José; Moguel, Bárbara; Bobes, Raúl J; Laclette, Juan P; Carrero, Julio C

    2016-01-01

    The cyst stage of Entamoeba histolytica is a promising therapeutic target against human amoebiasis. Our research team previously reported the production in vitro of Cyst-Like Structures (CLS) sharing structural features with cysts, including rounded shape, size reduction, multinucleation, and the formation of a chitin wall coupled to the overexpression of glucosamine 6-phosphate isomerase, the rate-limiting enzyme of the chitin synthesis pathway. A proteomic study of E. histolytica trophozoites, cysts, and in vitro-produced CLS is reported herein to determine the nature of CLS, widen our knowledge on the cyst stage, and identify possible proteins and pathways involved in the encystment process. Total protein extracts were obtained from E. histolytica trophozoites, CLS, and partially purified cysts recovered from the feces of amoebic human patients; extracts were trypsin-digested and analyzed by LC-MS/MS. In total, 1029 proteins were identified in trophozoites, 550 in CLS, and 411 in cysts, with 539, 299, and 84 proteins unique to each sample, respectively, and only 74 proteins shared by all three stages. About 70% of CLS proteins were shared with trophozoites, even though differences were observed in the relative protein abundance. While trophozoites showed a greater abundance of proteins associated to a metabolically active cell, CLS showed higher expression of proteins related to proteolysis, redox homeostasis, and stress response. In addition, the expression of genes encoding for the cyst wall proteins Jessie and Jacob was detected by RT-PCR and the Jacob protein identified by Western blotting and immunofluorescence in CLS. However, the proteomic profile of cysts as determined by LC-MS/MS was very dissimilar to that of trophozoites and CLS, with almost 40% of hypothetical proteins. Our global results suggest that CLS are more alike to trophozoites than to cysts, and they could be generated as a rapid survival response of trophozoites to a stressful condition

  7. Molecular mimicry by an F-box effector of Legionella pneumophila hijacks a conserved polyubiquitination machinery within macrophages and protozoa.

    Christopher T Price

    2009-12-01

    Full Text Available The ability of Legionella pneumophila to proliferate within various protozoa in the aquatic environment and in macrophages indicates a remarkable evolution and microbial exploitation of evolutionarily conserved eukaryotic processes. Ankyrin B (AnkB of L. pneumophila is a non-canonical F-box-containing protein, and is the only known Dot/Icm-translocated effector of L. pneumophila essential for intra-vacuolar proliferation within both macrophages and protozoan hosts. We show that the F-box domain of AnkB and the (9L(10P conserved residues are essential for intracellular bacterial proliferation and for rapid acquisition of polyubiquitinated proteins by the Legionella-containing vacuole (LCV within macrophages, Dictyostelium discoideum, and Acanthamoeba. Interestingly, translocation of AnkB and recruitment of polyubiquitinated proteins in macrophages and Acanthamoeba is rapidly triggered by extracellular bacteria within 5 min of bacterial attachment. Ectopically expressed AnkB within mammalian cells is localized to the periphery of the cell where it co-localizes with host SKP1 and recruits polyubiquitinated proteins, which results in restoration of intracellular growth to the ankB mutant similar to the parental strain. While an ectopically expressed AnkB-(9L(10P/AA variant is localized to the cell periphery, it does not recruit polyubiquitinated proteins and fails to trans-rescue the ankB mutant intracellular growth defect. Direct in vivo interaction of AnkB but not the AnkB-(9L(10P/AA variant with the host SKP1 is demonstrated. Importantly, RNAi-mediated silencing of expression of SKP1 renders the cells non-permissive for intracellular proliferation of L. pneumophila. The role of AnkB in exploitation of the polyubiquitination machinery is essential for intrapulmonary bacterial proliferation in the mouse model of Legionnaires' disease. Therefore, AnkB exhibits a novel molecular and functional mimicry of eukaryotic F-box proteins that exploits

  8. Neogenesis and maturation of transient Golgi-like cisternae in a simple eukaryote.

    Stefanic, Sasa; Morf, Laura; Kulangara, Caroline; Regös, Attila; Sonda, Sabrina; Schraner, Elisabeth; Spycher, Cornelia; Wild, Peter; Hehl, Adrian B

    2009-08-15

    The highly reduced protozoan parasite Giardia lamblia has minimal machinery for cellular processes such as protein trafficking. Giardia trophozoites maintain diverse and regulated secretory pathways but lack an identifiable Golgi complex. During differentiation to cysts, however, they produce specialized compartments termed encystation-specific vesicles (ESVs). ESVs are hypothesized to be unique developmentally regulated Golgi-like organelles dedicated to maturation and export of pre-sorted cyst wall proteins. Here we present a functional analysis of this unusual compartment by direct interference with the functions of the small GTPases Sar1, Rab1 and Arf1. Conditional expression of dominant-negative variants revealed an essential role of Sar1 in early events of organelle neogenesis, whilst inhibition of Arf1 uncoupled morphological changes and cell cycle progression from extracellular matrix export. The latter led to development of ;naked cysts', which lacked water resistance and thus infectivity. Time-lapse microscopy and photobleaching experiments showed that putative Golgi-like cisternae in Giardia develop into a network capable of exchanging soluble cargo at a high rate via dynamic, tubular connections, presumably to synchronize maturation. The minimized and naturally pulsed trafficking machinery for export of the cyst wall biopolymer in Giardia is a simple model for investigating basic principles of neogenesis and maturation of Golgi compartments. PMID:19622633

  9. Membrane trafficking and organelle biogenesis in Giardia lamblia: use it or lose it.

    Faso, Carmen; Hehl, Adrian B

    2011-04-01

    The secretory transport capacity of Giardia trophozoites is perfectly adapted to the changing environment in the small intestine of the host and is able to deploy essential protective surface coats as well as molecules which act on epithelia. These lumen-dwelling parasites take up nutrients by bulk endocytosis through peripheral vesicles or by receptor-mediated transport. The environmentally-resistant cyst form is quiescent but poised for activation following stomach passage. Its versatility and fidelity notwithstanding, the giardial trafficking systems appear to be the product of a general secondary reduction process geared towards minimization of all components and machineries identified to date. Since membrane transport is directly linked to organelle biogenesis and maintenance, less complexity also means loss of organelle structures and functions. A case in point is the Golgi apparatus which is missing as a steady-state organelle system. Only a few basic Golgi functions have been experimentally demonstrated in trophozoites undergoing encystation. Similarly, mitochondrial remnants have reached a terminally minimized state and appear to be functionally restricted to essential iron-sulfur protein maturation processes. Giardia's minimized organization combined with its genetic tractability provides unique opportunities to study basic principles of secretory transport in an uncluttered cellular environment. Not surprisingly, Giardia is gaining increasing attention as a model for the investigation of gene regulation, organelle biogenesis, and export of simple but highly protective cell wall biopolymers, a hallmark of all perorally transmitted protozoan and metazoan parasites. PMID:21296082

  10. Non-indigenous benthic fishes as new hosts for Bucephalus polymorphus Baer, 1827 (Digenea: Bucephalidae in the Vistula River basin, Poland

    Kvach Y.

    2011-02-01

    Full Text Available Two Ponto-Caspian gobiid species, the racer goby, Babka gymnotrachelus, and the monkey goby, Neogobius fluviatilis, the introduced fishes in the Vistula River, Poland, were found being infected with metacercariae of Bucephalus polymorphus. The encysted larvae were located in muscles and fins of these fish. The adult specimens occurred in the gut of another non-indigenous species, the Chinese sleeper Perccottus glenii. The life cycle of B. polymorphus in the Vistula River drainage includes three non-indigenous components: 1 the zebra mussel Dreissena polymorpha, the source of cercariae; 2 gobiids (the racer and monkey gobies, which are new in this water drainage but could play more important role in the life cycle than habitual cyprinids, thanks to their small size (favorable in transmission through the food-web; 3 the Chinese sleeper, a new accidental definitive host. The presence of these non-indigenous components in the life cycle maintains the distribution and further spread of B. polymorphus in the colonized area.

  11. Ocular toxoplasmosis in immunosuppressed nonhuman primates

    Holland, G.N.; O' Connor, G.R.; Diaz, R.F.; Minasi, P.; Wara, W.M.

    1988-06-01

    To investigate the role of cellular immunodeficiency in recurrent toxoplasmic retinochoroiditis, six Cynomolgus monkeys (Macaca fascicularis) with healed toxoplasmic lesions of the retina were immunosuppressed by total lymphoid irradiation. Three months prior to irradiation 30,000 Toxoplasma gondii organisms of the Beverley strain had been inoculated onto the macula of eye in each monkey via a pars plana approach. Toxoplasmic retinochoroiditis developed in each animal, and lesions were allowed to heal without treatment. During total lymphoid irradiation animals received 2000 centigrays (cGy) over a 7-week period. Irradiation resulted in an immediate drop in total lymphocyte counts and decreased ability to stimulate lymphocytes by phytohemagglutinin. Weekly ophthalmoscopic examinations following irradiation failed to show evidence of recurrent ocular disease despite persistent immunodeficiency. Four months after irradiation live organisms were reinoculated onto the nasal retina of the same eye in each animal. Retinochoroidal lesions identical to those seen in primary disease developed in five of six animals. Toxoplasma organisms therefore were able to proliferate in ocular tissue following the administration of immunosuppressive therapy. This study fails to support the hypothesis that cellular immunodeficiency alone will initiate recurrent toxoplasmic retinochoroiditis. Results suggest that reactivation of disease from encysted organisms involves factors other than suppression of Toxoplasma proliferation. If reactivation occurs by other mechanisms, however, cellular immunodeficiency then may allow development of extensive disease.

  12. Role of the Ubiquitin-Proteasome Systems in the Biology and Virulence of Protozoan Parasites

    Christian Muñoz

    2015-01-01

    Full Text Available In eukaryotic cells, proteasomes perform crucial roles in many cellular pathways by degrading proteins to enforce quality control and regulate many cellular processes such as cell cycle progression, signal transduction, cell death, immune responses, metabolism, protein-quality control, and development. The catalytic heart of these complexes, the 20S proteasome, is highly conserved in bacteria, yeast, and humans. However, until a few years ago, the role of proteasomes in parasite biology was completely unknown. Here, we summarize findings about the role of proteasomes in protozoan parasites biology and virulence. Several reports have confirmed the role of proteasomes in parasite biological processes such as cell differentiation, cell cycle, proliferation, and encystation. Proliferation and cell differentiation are key steps in host colonization. Considering the importance of proteasomes in both processes in many different parasites such as Trypanosoma, Leishmania, Toxoplasma, and Entamoeba, parasite proteasomes might serve as virulence factors. Several pieces of evidence strongly suggest that the ubiquitin-proteasome pathway is also a viable parasitic therapeutic target. Research in recent years has shown that the proteasome is a valid drug target for sleeping sickness and malaria. Then, proteasomes are a key organelle in parasite biology and virulence and appear to be an attractive new chemotherapeutic target.

  13. Ascocotyle (A.) nunezae n. sp. (Digenea: Heterophyidae) from Yucatan, Mexico.

    Scholz, T; Vargas-Vázquez, J; Vidal-Martínez, V M; Aguirre-Macedo, L

    1997-02-01

    A new heterophyid species, Ascocotyle (Ascocotyle) nunezae n. sp., is described from adults found in the intestine of naturally infected heron, Casmerodius albus (type host), from the coastal lagoon of Celestún. Yucatan, Mexico, and a domestic chick (Gallus gallus), experimentally infected with metacercariae from Cichlasoma octofasciatum. The new species is characterized mainly by the number (32-37) and arrangement of circumoral spines, which form I complete row of 25-27 circumoral spines and 6-10 accessory spines on the dorsal side, and by the morphology of the ventrogenital sac with a large gonotyl, consisting of 2 indistinctly separated lobes of vesicular tissue. Ascocotyle (A.) nunezae is placed into the nominotypical subgenus Ascocotyle because of the presence of uterine loops at the pharyngeal region and position of vitelline follicles. However, it differs distinctly from other members of this subgenus by the presence of long intestinal ceca reaching posterior to the ventral sucker. Cichlids of the genus Cichlasoma from cenotes, lakes, and the river Río Hondo in the Yucatan Peninsula were natural second intermediate hosts of A. (A.) nunezae, with metacercariae encysted on their gills. Cichlasoma meeki (Brind) was the most heavily infected fish host (total prevalence 75%; mean intensity 11 +/- 9). PMID:9057711

  14. Resveratrol Induces Apoptosis-Like Death and Prevents In Vitro and In Vivo Virulence of Entamoeba histolytica

    Pais-Morales, Jonnatan; Betanzos, Abigail; García-Rivera, Guillermina; Chávez-Munguía, Bibiana; Shibayama, Mineko; Orozco, Esther

    2016-01-01

    Entamoeba histolytica causes amoebiasis, an infection that kills 100,000 individuals each year. Metronidazole and its derivatives are currently used against this protozoan, but these drugs present adverse effects on human health. Here, we investigated the effect of resveratrol (a natural compound) on E. histolytica trophozoites viability, as well as its influence on the parasite virulence. Trophozoites growth was arrested by 72 μM resveratrol and the IC50 was determined as 220 μM at 48 h. Cells appeared smaller, rounded and in clusters, with debris-containing vacuoles and with abnormally condensed chromatin. Resveratrol triggered reactive oxygen species production. It caused lipid peroxidation and produced phosphatidylserine externalization and DNA fragmentation this latter evidenced by TUNEL assays. It also provoked an increase of intracellular Ca2+ concentration, activated calpain and decreased superoxide dismutase activity, indicating that an apoptosis-like event occurred; however, autophagy was not detected. Cytopathic activity, phagocytosis, encystment and in vivo virulence were diminished dramatically by pre-incubation of trophozoites with resveratrol, evidencing that resveratrol attenuated the trophozoite virulence in vitro. Interestingly, after the inoculation of virulent trophozoites, animals treated with the drug did not develop or developed very small abscesses. Our findings propose that resveratrol could be an alternative to contend amoebiasis. PMID:26731663

  15. A Combined Morphometric and Molecular Approach to Identifying Metacercariae of Euclinostomum heterostomum (Digenea: Clinostomidae).

    Caffara, Monica; Locke, Sean A; Cristanini, Cecilia; Davidovich, Nadav; Markovich, Michal Perry; Fioravanti, Maria L

    2016-04-01

    Metacercariae of species of Euclinostomum have been found encysted in kidney, liver, and muscles of several fish species, while adults occur in the mouth, pharynx, larynx, trachea, and upper esophagus of fish-eating birds. The aim of this work was to gather molecular and morphological data from the type species, Euclinostomum heterostomum, as a starting point for needed revisions of the genus. Metacercariae were collected from cichlids in Lake Kinneret, Israel, and all were identified as E. heterostomum based on morphology. This identification was further confirmed by principal components analyses that revealed no partitions in morphometric resemblance, indicating that the material represents a single species, and that the specimens were morphometrically similar to other descriptions of E. heterostomum. In phylogenetic analysis of concatenated partial internal transcribed spacer 1 and 2, 5.8S rDNA, and partial cytochrome c oxidase I barcode sequences, the isolates of Euclinostomum obtained in this study form a single, monophyletic group separate from recently published data from Euclinostomum sp. from Thailand and from Clinostomum species, which are also monophyletic. The morphological and molecular data reported in the present work can provide a useful point of reference for future studies. PMID:26762958

  16. Draft genome sequencing of giardia intestinalis assemblage B isolate GS: is human giardiasis caused by two different species?

    Oscar Franzén

    2009-08-01

    Full Text Available Giardia intestinalis is a major cause of diarrheal disease worldwide and two major Giardia genotypes, assemblages A and B, infect humans. The genome of assemblage A parasite WB was recently sequenced, and the structurally compact 11.7 Mbp genome contains simplified basic cellular machineries and metabolism. We here performed 454 sequencing to 16x coverage of the assemblage B isolate GS, the only Giardia isolate successfully used to experimentally infect animals and humans. The two genomes show 77% nucleotide and 78% amino-acid identity in protein coding regions. Comparative analysis identified 28 unique GS and 3 unique WB protein coding genes, and the variable surface protein (VSP repertoires of the two isolates are completely different. The promoters of several enzymes involved in the synthesis of the cyst-wall lack binding sites for encystation-specific transcription factors in GS. Several synteny-breaks were detected and verified. The tetraploid GS genome shows higher levels of overall allelic sequence polymorphism (0.5 versus <0.01% in WB. The genomic differences between WB and GS may explain some of the observed biological and clinical differences between the two isolates, and it suggests that assemblage A and B Giardia can be two different species.

  17. Drepanotrema kermatoides (Orbigny, 1835 (Mollusca, Planorbidae, hospedeiro de um paranfistomídeo (Trematoda, no Rio Grande do Sul, Brasil Drepanotrema kermatoides (Orbigny, 1835 (Mollusca, Planorbidae, host of a paramphistomid (Trematoda, in Rio Grande do Sul, Brazil

    Inga Ludmila Veitenheimer-Mendes

    1989-03-01

    Full Text Available São descritas e ilustrradas rédia, cercária imatura e cercária madura e metacercária de um paranfistomídeo, provavelmente do gênero Paramphistomum (Fischoeder, 1901. Estas formas evolutivas foram obtidas de um exemplar de Drepanotrema kermatoides (Orbigny, 1835 coletado, em maio de 1983, no arroio Chico Lumã (29º56'03"S e 50º35'59"W, município de Santo Antônio da Patrulha, Rio Grande do Sul, Brasil.Morphological descriptions and illustrations of the redia, the imature cercaria and mature cercaria, and encysted metacercaria, found in a infected specimen of Drepanotrema kermatoides (Orbigny, 1835 are presented. The infected mollusk was found in Chico Lumã creek (29º56'03"S and 50º35'59"W, Santo Antônio County, Rio Grande do Sul State, in May of 1983. These larval stages probably are part of a cycle of a rumen parasite of the genus Paramphistomum (Fischoeder, 1901.

  18. The Neurotropic Parasite Toxoplasma Gondii Increases Dopamine Metabolism

    Prandovszky, Emese; Gaskell, Elizabeth; Martin, Heather; Dubey, J. P.; Webster, Joanne P.; McConkey, Glenn A.

    2011-01-01

    The highly prevalent parasite Toxoplasma gondii manipulates its host's behavior. In infected rodents, the behavioral changes increase the likelihood that the parasite will be transmitted back to its definitive cat host, an essential step in completion of the parasite's life cycle. The mechanism(s) responsible for behavioral changes in the host is unknown but two lines of published evidence suggest that the parasite alters neurotransmitter signal transduction: the disruption of the parasite-induced behavioral changes with medications used to treat psychiatric disease (specifically dopamine antagonists) and identification of a tyrosine hydroxylase encoded in the parasite genome. In this study, infection of mammalian dopaminergic cells with T. gondii enhanced the levels of K+-induced release of dopamine several-fold, with a direct correlation between the number of infected cells and the quantity of dopamine released. Immunostaining brain sections of infected mice with dopamine antibody showed intense staining of encysted parasites. Based on these analyses, T. gondii orchestrates a significant increase in dopamine metabolism in neural cells. Tyrosine hydroxylase, the rate-limiting enzyme for dopamine synthesis, was also found in intracellular tissue cysts in brain tissue with antibodies specific for the parasite-encoded tyrosine hydroxylase. These observations provide a mechanism for parasite-induced behavioral changes. The observed effects on dopamine metabolism could also be relevant in interpreting reports of psychobehavioral changes in toxoplasmosis-infected humans. PMID:21957440

  19. The neurotropic parasite Toxoplasma gondii increases dopamine metabolism.

    Emese Prandovszky

    Full Text Available The highly prevalent parasite Toxoplasma gondii manipulates its host's behavior. In infected rodents, the behavioral changes increase the likelihood that the parasite will be transmitted back to its definitive cat host, an essential step in completion of the parasite's life cycle. The mechanism(s responsible for behavioral changes in the host is unknown but two lines of published evidence suggest that the parasite alters neurotransmitter signal transduction: the disruption of the parasite-induced behavioral changes with medications used to treat psychiatric disease (specifically dopamine antagonists and identification of a tyrosine hydroxylase encoded in the parasite genome. In this study, infection of mammalian dopaminergic cells with T. gondii enhanced the levels of K+-induced release of dopamine several-fold, with a direct correlation between the number of infected cells and the quantity of dopamine released. Immunostaining brain sections of infected mice with dopamine antibody showed intense staining of encysted parasites. Based on these analyses, T. gondii orchestrates a significant increase in dopamine metabolism in neural cells. Tyrosine hydroxylase, the rate-limiting enzyme for dopamine synthesis, was also found in intracellular tissue cysts in brain tissue with antibodies specific for the parasite-encoded tyrosine hydroxylase. These observations provide a mechanism for parasite-induced behavioral changes. The observed effects on dopamine metabolism could also be relevant in interpreting reports of psychobehavioral changes in toxoplasmosis-infected humans.

  20. Human Extraintestinal Sarcocystosis: What We Know, and What We don't Know.

    Harris, V C; van Vugt, M; Aronica, E; de Bree, G J; Stijnis, C; Goorhuis, A; Grobusch, M P

    2015-08-01

    There are over 150 known Sarcocystis species, and at least one is capable of infecting and causing disease in man. Extraintestinal (muscular) sarcocystosis and intestinal sarcocystosis are the two known manifestations of disease in humans. In this series of six cases and review, we focus on the invasive extraintestinal ("muscular") form of sarcocystosis in humans. This disease, which until recently was rarely described, has become relevant particularly as an imported condition in travelers due to a recent series of outbreaks reported from Malaysia. Human intestinal sarcocystosis is ubiquitous across the globe. However, absolute numbers of probable and particularly confirmed cases are few, with only several hundred described to date. Characteristically, patients exhibit signs and symptoms either 1-2 weeks after exposure, or after 4-8 weeks. Whether people remain asymptomatic or develop disease apparently depends on the infecting species, host factors, and the inoculum size. The definitive host(s) remain uncertain, and identification of the animal reservoir(s) requires further research. A better understanding of the epidemiology of the disease, as well as its immunological determinants, is hampered by the lack of reliable serological diagnostic methods. Additionally, DNA seems to be contained very effectively within the encysted parasite, thereby rendering PCR detection unreliable. Physicians should suspect the condition in patients with suggestive symptoms and a possible history of exposure. Surveillance networks for imported infectious diseases are formidable tools to help detect and localize outbreaks. PMID:26115699

  1. Pimply gut condition in mithun (Bos frontalis) calves.

    Chamuah, J K; Pegu, S R; Raina, O K; Siju, S J; Sakhrie, A; Deka, Anil; Rajkhowa, C

    2016-06-01

    The present observation was recorded at National Research Centre on Mithun, Jharnapani from May 2010 to September 2012. A total of 15 mithun calves, which died in and around Jharnapani, were attended and detailed post-mortem examination was carried out. Out of these, five calves (33.33 %) aging between 1 and 1.5 years exhibiting the condition of chronic wasting and diarrhoea were found positive for pimply gut condition based on gross and microscopic examination. Post-mortem examination revealed extensive nodule formation on the wall of the rectum; however, the entire lumen did not reveal any of adult parasites. In all the cases, there were congestion in the mucous layer and thickening of the intestinal wall. Histopathological examination revealed chronic enteritis with mononuclear cell infiltration comprising mostly of macrophages, lymphocytes and eosinophils. In the muscularis mucosae, encysted larvae were found along with fibrous tissue proliferation. These lesions gave the intestine a nodular appearance as they thickened the wall and projected from the serosal surface. These extensive numbers of nodules in the intestine might have interfered with peristalsis and intestinal absorption which led to chronic wasting and diarrhoea in the calves. PMID:27413288

  2. Group 1 LEA proteins contribute to the desiccation and freeze tolerance of Artemia franciscana embryos during diapause.

    Toxopeus, Jantina; Warner, Alden H; MacRae, Thomas H

    2014-11-01

    Water loss either by desiccation or freezing causes multiple forms of cellular damage. The encysted embryos (cysts) of the crustacean Artemia franciscana have several molecular mechanisms to enable anhydrobiosis-life without water-during diapause. To better understand how cysts survive reduced hydration, group 1 late embryogenesis abundant (LEA) proteins, hydrophilic unstructured proteins that accumulate in the stress-tolerant cysts of A. franciscana, were knocked down using RNA interference (RNAi). Embryos lacking group 1 LEA proteins showed significantly lower survival than control embryos after desiccation and freezing, or freezing alone, demonstrating a role for group 1 LEA proteins in A. franciscana tolerance of low water conditions. In contrast, regardless of group 1 LEA protein presence, cysts responded similarly to hydrogen peroxide (H2O2) exposure, indicating little to no function for these proteins in diapause termination. This is the first in vivo study of group 1 LEA proteins in an animal and it contributes to the fundamental understanding of these proteins. Knowing how LEA proteins protect A. franciscana cysts from desiccation and freezing may have applied significance in aquaculture, where Artemia is an important feed source, and in the cryopreservation of cells for therapeutic applications. PMID:24846336

  3. Establishment of a continuous culture system for Entamoeba muris and analysis of the small subunit rRNA gene

    Kobayashi S.

    2009-06-01

    Full Text Available We established a culture system for Entamoeba muris (MG-EM-01 strain isolated from a Mongolian gerbil using a modified Balamuth’s egg yolk infusion medium supplemented with 4% adult bovine serum and Bacteroides fragilis cocultured with Escherichia coli. Further, encystation was observed in the culture medium. The morphological characteristics of E. muris are similar to those of Entamoeba coli (E. coli; moreover, the malic isoenzyme electrophoretic band, which shows species-specific electrophoretic mobility, of E. muris had almost the same mobility as that observed with the malic isoenzyme electrophorectic band of E. coli (UZG-EC-01 strain isolated from a gorilla. We determined the small subunit rRNA (SSU-rRNA gene sequence of the MG-EM-01 strain, and this sequence was observed to show 82.7% homology with that of the UZG-EC-01 strain. Further, the resultant phylogenetic tree for molecular taxonomy based on the SSU-rRNA genes of the 21 strains of the intestinal parasitic amoeba species indicated that the MG-EM-01 strain was most closely related to E. coli.

  4. Molecular phylogeny and ultrastructure of Aphelidium aff. melosirae (Aphelida, Opisthosporidia).

    Karpov, Sergey A; Mamkaeva, Maria A; Benzerara, Karim; Moreira, David; López-García, Purificación

    2014-08-01

    Aphelids are a poorly known group of parasitoids of algae that have raised considerable interest due to their pivotal phylogenetic position. Together with Cryptomycota and the highly derived Microsporidia, they have been recently re-classified as Opisthosporidia, being the sister group to fungi. Despite their huge diversity, as revealed by molecular environmental studies, and their phylogenetic interest, only three genera have been described (Aphelidium, Amoeboaphelidium, and Pseudaphelidium), from which 18S rRNA gene sequences exist only for Amoeboaphelidium species. Here, we describe the life cycle and ultrastructure of Aphelidium aff. melosirae, and provide the first 18S rRNA gene sequence obtained for this genus. Molecular phylogeny analysis indicates that Aphelidium is very distantly related to Amoebaphelidium, highlighting the wide genetic diversity of the aphelids. The parasitoid encysts and penetrates the host alga, Tribonema gayanum through an infection tube. Cyst germination leads to a young trophont that phagocytes the algal cell content and progressively develops a plasmodium, which becomes a zoospore-producing sporangium. Aphelidium aff. melosirae has amoeboflagellate zoospores, tubular/lamellar mitochondrial cristae, a metazoan type of centrosome, and closed orthomitosis with an intranuclear spindle. These features together with trophont phagocytosis distinguish Aphelidium from fungi and support the erection of the new superphylum Opisthosporidia as sister to fungi. PMID:24995586

  5. Use of the vital stain FUN-1 indicates viability of Phytophthora capsici propagules and can be used to predict maximum zoospore production.

    Lewis Ivey, Melanie L; Miller, Sally A

    2014-01-01

    The fluorescent vital dye FUN®-1 (2-chloro-4-[2,3-dihydro-3-methyl-{benzo-1,3-thiazol-2-yl}-methylidene]-1-phenylquinolinium iodide) was evaluated as a tool to assess Phytophthora capsici sporangia and zoospore metabolic activity and viability. Under aerobic conditions, mycelia, sporangia and zoospores cultured on agar medium and stained with FUN-1 exhibited red fluorescent cylindrical intravacuolar structures (CIVS) that were clearly visible at 100× magnification. Encysted zoospores did not exhibit CIVS after exposure to FUN-1 dye. Over 7 d there was a significant reduction in the percent of sporangia containing CIVS, which corresponded with a significant increase in zoospore formation and release. The decline in the percentage of metabolically active sporangia and increase in the number of zoospores fit both a linear and log regression model. The FUN-1 dye was suitable for distinguishing between live and dead sporangia and effective in monitoring the change in metabolic activity of sporangia over time. It will be useful in determining parameters, including P. capsici culture age, that maximize production of zoospores in vitro. PMID:24782503

  6. Inhibition of Phytophthora parasitica and P. capsici by Silver Nanoparticles Synthesized Using Aqueous Extract of Artemisia absinthium.

    Ali, Mohammad; Kim, Bosung; Belfield, Kevin D; Norman, David; Brennan, Mary; Ali, Gul Shad

    2015-09-01

    Application of nanoparticles for controlling plant pathogens is a rapidly emerging area in plant disease management, and nanoparticles synthesis methods that are economical and ecofriendly are extensively investigated. In this project, we investigated the potential of silver nanoparticles (AgNPs) synthesized with aqueous extract of Artemisia absinthium against several Phytophthora spp., which cause many economically important crop diseases. In in vitro dose-response tests conducted in microtiter plates, 10 µg ml⁻¹ of AgNPs inhibited mycelial growth of P. parasitica, P. infestans, P. palmivora, P. cinnamomi, P. tropicalis, P. capsici, and P. katsurae. Detailed in vitro dose-response analyses conducted with P. parasitica and P. capsici revealed that AgNPs synthesized with A. absinthium extract were highly potent (IC50: 2.1 to 8.3 µg ml⁻¹) and efficacious (100%) in inhibiting mycelial growth, zoospore germination, germ tube elongation, and zoospore production. Interestingly, AgNP treatment accelerated encystment of zoospores. Consistent with in vitro results, in planta experiments conducted in a greenhouse revealed that AgNP treatments prevented Phytophthora infection and improved plant survival. Moreover, AgNP in in planta experiments did not produce any adverse effects on plant growth. These investigations provide a simple and economical method for controlling Phytophthora with AgNP without affecting normal plant physiology. PMID:25871856

  7. Bold, Sedentary Fathead Minnows Have More Parasites.

    Pan, Tiffany; Gladen, Kelsey; Duncan, Elizabeth C; Cotner, Sehoya; Cotner, James B; McEwen, Daniel C; Wisenden, Brian D

    2016-08-01

    Parasites that rely on trophic transmission can manipulate the behavior of an intermediate host to compromise the host's antipredator competence and increase the probability of reaching the next host. Selection for parasite manipulation is diminished when there is significant risk of host death to causes other than consumption by a suitable definitive host for the parasite. Consequently, behavioral manipulation by parasites can be expected to be subtle. Ornithodiplostomum ptychocheilus (Op) is a trematode parasite that has a bird-snail-fish host life cycle. Fathead minnows are a common intermediate host of Op, where metacercariae encyst in the minnow brain. In this study, we report a link between metacercarial intensity and behavior in fathead minnows. In the field, we found that roaming distance by free-living minnows over 24 h was negatively correlated with parasite intensity. In the laboratory, we found that boldness in an open field test was positively correlated with parasite intensity. These parasite-induced behavioral changes may render infected minnows more susceptible to predators, which would serve to facilitate trophic transmission of parasites to the bird host. PMID:27093037

  8. Transient fusion and selective secretion of vesicle proteins in Phytophthora nicotianae zoospores

    Weiwei Zhang

    2013-12-01

    Full Text Available Secretion of pathogen proteins is crucial for the establishment of disease in animals and plants. Typically, early interactions between host and pathogen trigger regulated secretion of pathogenicity factors that function in pathogen adhesion and host penetration. During the onset of plant infection by spores of the Oomycete, Phytophthora nicotianae, proteins are secreted from three types of cortical vesicles. Following induction of spore encystment, two vesicle types undergo full fusion, releasing their entire contents onto the cell surface. However, the third vesicle type, so-called large peripheral vesicles, selectively secretes a small Sushi domain-containing protein, PnCcp, while retaining a large glycoprotein, PnLpv, before moving away from the plasma membrane. Selective secretion of PnCcp is associated with its compartmentalization within the vesicle periphery. Pharmacological inhibition of dynamin function, purportedly in vesicle fission, by dynasore treatment provides evidence that selective secretion of PnCcp requires transient fusion of the large peripheral vesicles. This is the first report of selective protein secretion via transient fusion outside mammalian cells. Selective secretion is likely to be an important aspect of plant infection by this destructive pathogen.

  9. Transient fusion and selective secretion of vesicle proteins in Phytophthora nicotianae zoospores.

    Zhang, Weiwei; Blackman, Leila M; Hardham, Adrienne R

    2013-01-01

    Secretion of pathogen proteins is crucial for the establishment of disease in animals and plants. Typically, early interactions between host and pathogen trigger regulated secretion of pathogenicity factors that function in pathogen adhesion and host penetration. During the onset of plant infection by spores of the Oomycete, Phytophthora nicotianae, proteins are secreted from three types of cortical vesicles. Following induction of spore encystment, two vesicle types undergo full fusion, releasing their entire contents onto the cell surface. However, the third vesicle type, so-called large peripheral vesicles, selectively secretes a small Sushi domain-containing protein, PnCcp, while retaining a large glycoprotein, PnLpv, before moving away from the plasma membrane. Selective secretion of PnCcp is associated with its compartmentalization within the vesicle periphery. Pharmacological inhibition of dynamin function, purportedly in vesicle fission, by dynasore treatment provides evidence that selective secretion of PnCcp requires transient fusion of the large peripheral vesicles. This is the first report of selective protein secretion via transient fusion outside mammalian cells. Selective secretion is likely to be an important aspect of plant infection by this destructive pathogen. PMID:24392285

  10. Self-Fertilization and the Role of Males in Populations of Tadpole Shrimp (Branchiopoda: Notostraca: Triops).

    Horn, Rebekah L; Cowley, David E

    2016-11-01

    Self-fertilization has both negative and positive fitness effects on species evolution. Selfing can increase inbreeding depression, thereby decreasing genetic diversity. In contrast, self-fertilization can preserve beneficial gene combinations and facilitate colonization success. Within the class of crustaceans Branchiopoda, selfing is a primary reproductive mode. Some species of Triops, in the family Notostraca, are a few of the animal species thought to have a mixed mating system between hermaphrodites and males termed androdioecy. The objective of this study is to validate the reproductive mode utilized by Triops newberryi in southern New Mexico by the use of progeny arrays and population simulations. Individuals were reared in the lab from dried soil collected from temporary ponds inhabited by T. newberryi The adults reared and the encysted embryos contained within their brood pouches were genotyped using 7 T. newberryi specific microsatellite markers to determine the relatedness between parent and offspring. Overall microsatellite diversity was low with few heterozygous individuals and limited polymorphisms. Simulated populations and allele segregation analysis suggest hermaphroditism is the primary reproductive mode for T. newberryi In addition, based on the offspring's alleles, there was no direct evidence that a male (ovisacless) T. newberryi outcrossed with a female. Population simulations further suggest that the rate of successful outcrossing events must be low and could explain why outcrossing was not observed in the laboratory rearing trials. PMID:27358476

  11. Use of gamma radiation to prevent infectivity of meat-borne parasites and effect of irradiation on the biochemical and its properties of meats

    The pre-liminary study performed indicated that the least effective irradiation dose to control the infectivity of Trichinela spirales was 0.2 kGy, by which neither worms nor encysted larvae were found in the laboratory animals on the 3rd, 7th and 21st day post infection. The effect of this effective irradiation dose on the biochemical constituents and acceptability of beef and pork meat was evaluated. The results showed that there was no significant differences in the biochemical constituents and organoleptic properties between unirradiated and irradiated meats. By GC-MS analysis no new fatty acids were found in beef and pork lipids by using 0.2 kGy irradiation dose. By amino acids analysis, 17 amino acids were detected on all unirradiated and irradiated samples. 0.2 kGy irradiation dose almost had no effect on the amino acids profiles. However, some of the mino acids slightly increased and others slightly decreased. It could be concluded that low irradiation dose used approved to be efficient in controlling infectivity of meat-borne parasites with minimal changes in organoleptic properties and biochemical constitution of meat

  12. Experimental Infections of Bluegill with the Trematode Ribeiroia ondatrae (Digenea: Cathaemasiidae): Histopathology and Hematological Response.

    Calhoun, Dana M; Schaffer, Paula A; Gregory, Jacklyn R; Hardy, Katherine M; Johnson, Pieter T J

    2015-12-01

    Infections by the digenetic trematode, Ribeiroia ondatrae, cause severe limb malformations in many North American amphibians. Ribeiroia ondatrae also infects fishes as second intermediate hosts, but less is known about the pathology and immune responses initiated in infected fish, even though reports of infected fish date back to early 1900s. To this end, we experimentally exposed juvenile Bluegills Lepomis macrochirus to three doses of R. ondatrae cercariae and monitored the pathology, parasite infection success, and humoral responses over 648 h. All exposed fish became infected with metacercariae, and the average infection load increased with exposure dose. Histologically, infection was associated with acute hemorrhages in the lateral line and local dermis at 36 h, followed by progressive granulomatous inflammation that led to the destruction of encysted metacercariae. Correspondingly, over the course of 648 h we observed an 85% decline in average infection load among hosts, reflecting the host's clearance of the parasite. Infection was not associated with changes in fish growth or survival, but did correlate with leukocytosis and neutrophilia in circulating host blood. Understanding the physiological responses of R. ondatrae in Bluegill will help to clarify the ecological effects of this parasite and provide a foundation for subsequent comparisons into its effects on behavior, individual health, and population dynamics of Bluegill. PMID:26587684

  13. Protection against heterologous infection by using cross antigenicity between schistosoma mansoni and fasciola hepatica

    Fasciola hepatica is the causative agent of fasciolosis in many areas in America, Europe, Africa, Asia and Australia. There is an urgent need for improved methods to control the parasite's transmission. The present study is parasitological, immunological (interleukine-1β and interleukine-6) and histopathological investigations on the immunizing effect of cross antigenicity between S. mansoni and F. hepatica against schistosomiasis and fasciolosis in mice. Parasitological study showed that vaccination with irradiated cercariae of S. mansoni or vaccination by F. hepatica whole worm extract (FhWWE) before challenged with encysted metacercariae of F. hepatica or cercariae of S. mansoni played a significant control on the parasitic infection manifested by a remarkable reduction in the means of worm count. Assessment of IL-1β and IL-6 in sera of the experimental groups showed that there are cross reactivity between fasciola / schistosoma and its relation to cross protection. Histopathological examination of vaccinated mice livers showed protection against parasite maturation and liver damage after challenged, as compared to mice infected only without vaccination

  14. Pediatric neurocysticercosis: current challenges and future prospects

    Singhi P

    2016-03-01

    Full Text Available Pratibha Singhi, Arushi Gahlot SainiDepartment of Pediatrics, Pediatric Neurology and Neurodevelopment Unit, Advanced Pediatrics Centre, Post Graduate Institute of Medical Education and Research, Chandigarh, IndiaAbstract: Neurocysticercosis (NCC is an acquired infection of the nervous system caused by encysted larvae of Taenia solium. It is a major cause of epilepsy in the tropics and the commonest cause of focal seizures in North Indian children. T. solium teniasis-cysticercosis is considered a parasitic “Neglected Tropical Diseases” endemic throughout Southeast Asia. NCC in children has pleomorphic manifestations depending on the location, number and viability of the cysts, and host response. Even with advancing knowledge of the disease manifestations, many aspects related to diagnosis and treatment, particularly in children, still remain controversial and pose challenges to clinical practice. There is no gold standard test to diagnose NCC and the management recommendations are still emerging. This review provides an overview of diagnosis of NCC in children and its management with special focus on current challenges and future prospects.Keywords: neurocysticercosis, children, epilepsy, ring enhancing lesions, pigs

  15. Experimental infection of selected arthropods with spirurid nematodes Spirocerca lupi Railliet & Henry, 1911 and Gongylonema ingluvicola Molin, 1857.

    Mukaratirwa, S; Pillay, E; Munsammy, K

    2010-12-01

    Gongylonema ingluvicola and Spirocerca lupi are spirurid nematodes that require arthropod intermediate hosts in order to complete their life cycle. Beetles of the family Scarabaeidae are reported to serve as intermediate hosts for both these parasites. In this study selected species of beetles of the family Scarabaeidae as well as other groups of arthropods were screened for susceptibility to infection with S. lupi and G. ingluvicola. Arthropods were exposed to infective eggs of both parasites for a determined period of time and dissected/digested to determine the presence or absence of pre-infective and infective larvae. All the five species of dung beetles exposed to infection with S. lupi, namely, Pachylomerus femoralis, Scarabaeus rugosus, Gymnopleurus humanus, Kheper nigroaeneus and Anachalcos convexus were susceptible and, of the two species exposed to G. ingluvicola, only Gy. humanus was susceptible. Spirocerca lupi eggs developed in millipede species, Daratoagonus cristulatus, and remained as encysted larvae, while in Orthoporoides kyrhocephalus no development was observed. Spirocerca lupi larvae were not detected in the cricket species Gryllus assimilis, or the cockroach species Periplaneta americana, and, similarly, G. ingluvicola larvae were not detected in the millipede species O. kyrhocephalus. The difference in the susceptibility of the arthropods to the two parasite species may depend on their feeding biology. PMID:20132587

  16. A nucleotide sugar transporter involved in glycosylation of the Toxoplasma tissue cyst wall is required for efficient persistence of bradyzoites.

    Carolina E Caffaro

    Full Text Available Toxoplasma gondii is an intracellular parasite that transitions from acute infection to a chronic infective state in its intermediate host via encystation, which enables the parasite to evade immune detection and clearance. It is widely accepted that the tissue cyst perimeter is highly and specifically decorated with glycan modifications; however, the role of these modifications in the establishment and persistence of chronic infection has not been investigated. Here we identify and biochemically and biologically characterize a Toxoplasma nucleotide-sugar transporter (TgNST1 that is required for cyst wall glycosylation. Toxoplasma strains deleted for the TgNST1 gene (Δnst1 form cyst-like structures in vitro but no longer interact with lectins, suggesting that Δnst1 strains are deficient in the transport and use of sugars for the biosynthesis of cyst-wall structures. In vivo infection experiments demonstrate that the lack of TgNST1 activity does not detectably impact the acute (tachyzoite stages of an infection or tropism of the parasite for the brain but that Δnst1 parasites are severely defective in persistence during the chronic stages of the infection. These results demonstrate for the first time the critical role of parasite glycoconjugates in the persistence of Toxoplasma tissue cysts.

  17. Effects of altered temperature and precipitation on desert protozoa associated with biological soil crusts

    Darby, B.J.; Housman, D.C.; Zaki, A.M.; Shamout, Y.; Adl, S.M.; Belnap, J.; Neher, D.A.

    2006-01-01

    Biological soil crusts are diverse assemblages of bacteria, cyanobacteria, algae, fungi, lichens, and mosses that cover much of arid land soils. The objective of this study was to quantify protozoa associated with biological soil crusts and test the response of protozoa to increased temperature and precipitation as is predicted by some global climate models. Protozoa were more abundant when associated with cyanobacteria/lichen crusts than with cyanobacteria crusts alone. Amoebae, flagellates, and ciliates originating from the Colorado Plateau desert (cool desert, primarily winter precipitation) declined 50-, 10-, and 100-fold, respectively, when moved in field mesocosms to the Chihuahuan Desert (hot desert, primarily summer rain). However, this was not observed in protozoa collected from the Chihuahuan Desert and moved to the Sonoran desert (hot desert, also summer rain, but warmer than Chihuahuan Desert). Protozoa in culture began to encyst at 37??C. Cysts survived the upper end of daily temperatures (37-55??C), and could be stimulated to excyst if temperatures were reduced to 15??C or lower. Results from this study suggest that cool desert protozoa are influenced negatively by increased summer precipitation during excessive summer temperatures, and that desert protozoa may be adapted to a specific desert's temperature and precipitation regime. ?? 2006 by the International Society of Protistologists.

  18. Incidence of adult brain cancers is higher in countries where the protozoan parasite Toxoplasma gondii is common

    Thomas, Frédéric; Lafferty, Kevin D.; Brodeur, Jacques; Elguero, Eric; Gauthier-Clerc, Michel; Missé, Dorothée

    2012-01-01

    We explored associations between the common protozoan parasite Toxoplasma gondii and brain cancers in human populations. We predicted that T. gondii could increase the risk of brain cancer because it is a long-lived parasite that encysts in the brain, where it provokes inflammation and inhibits apoptosis. We used a medical geography approach based on the national incidence of brain cancers and seroprevalence of T. gondii. We corrected reports of incidence for national gross domestic product because wealth probably increases the ability to detect cancer. We also included gender, cell phone use and latitude as variables in our initial models. Prevalence of T. gondii explained 19 per cent of the residual variance in brain cancer incidence after controlling for the positive effects of gross domestic product and latitude among nations. Infection with T. gondii was associated with a 1.8-fold increase in the risk of brain cancers across the range of T. gondii prevalence in our dataset (4–67%). These results, though correlational, suggest that T. gondii should be investigated further as a possible oncogenic pathogen of humans.

  19. Parasite manipulation of brain monoamines in California killifish (Fundulus parvipinnis) by the trematode Euhaplorchis californiensis

    Shaw, J.C.; Korzan, W.J.; Carpenter, R.E.; Kuris, A.M.; Lafferty, K.D.; Summers, C.H.; Overli, O.

    2009-01-01

    California killifish (Fundulus parvipinnis) infected with the brain-encysting trematode Euhaplorchis californiensis display conspicuous swimming behaviours rendering them more susceptible to predation by avian final hosts. Heavily infected killifish grow and reproduce normally, despite having thousands of cysts inside their braincases. This suggests that E. californiensis affects only specific locomotory behaviours. We hypothesised that changes in the serotonin and dopamine metabolism, essential for controlling locomotion and arousal may underlie this behaviour modification. We employed micropunch dissection and HPLC to analyse monoamine and monoamine metabolite concentrations in the brain regions of uninfected and experimentally infected fish. The parasites exerted density-dependent changes in monoaminergic activity distinct from those exhibited by fish subjected to stress. Specifically, E. californiensis inhibited a normally occurring, stress-induced elevation of serotonergic metabolism in the raphae nuclei. This effect was particularly evident in the experimentally infected fish, whose low-density infections were concentrated on the brainstem. Furthermore, high E. californiensis density was associated with increased dopaminergic activity in the hypothalamus and decreased serotonergic activity in the hippocampus. In conclusion, the altered monoaminergic metabolism may explain behavioural differences leading to increased predation of the infected killifish by their final host predators. ?? 2008 The Royal Society.

  20. Efficacy of a 2 per cent moxidectin gel against gastrointestinal parasites of ponies.

    Bauer, C; Cirak, V Y; Hermosilla, C; Okoro, H

    1998-11-14

    The efficacy of moxidectin was evaluated in young ponies naturally infected with gastrointestinal parasites. Eight animals were treated orally with moxidectin at 0.4 mg/kg bodyweight and eight received only the vehicle. They were all necropsied two weeks later. Faecal samples were examined daily for egg counts and larval cultures. Parasites were recovered from total faecal samples collected daily and from the gastrointestinal tracts at necropsy. Moxidectin reduced the strongyle egg counts by > 99 per cent from three days after treatment but some individuals remained positive for 10 days. The drug had little or no ovicidal activity. As evaluated in the critical-controlled test, moxidectin was 99 to 100 per cent effective (P Oxyuris equi. Adult S edentatus were also completely removed. Its efficacy against third stage larvae of Gasterophilus intestinalis was 95 per cent (P < 0.05). Luminal nematode stages were removed within a few days, and bots continued to be eliminated for at least two weeks after treatment. No activity was observed against Anoplocephala perforliata. As evaluated in the controlled test, moxidectin was 100 per cent effective against Habronema muscae (P < 0.05) and had a 76 per cent but not significant efficacy against encysted small strongyle larvae. PMID:9854319