Composition of the mitochondrial electron transport chain in acanthamoeba castellanii: structural and evolutionary insights.

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Gawryluk, Ryan M R; Chisholm, Kenneth A; Pinto, Devanand M; Gray, Michael W

2012-11-01

The mitochondrion, derived in evolution from an α-proteobacterial progenitor, plays a key metabolic role in eukaryotes. Mitochondria house the electron transport chain (ETC) that couples oxidation of organic substrates and electron transfer to proton pumping and synthesis of ATP. The ETC comprises several multiprotein enzyme complexes, all of which have counterparts in bacteria. However, mitochondrial ETC assemblies from animals, plants and fungi are generally more complex than their bacterial counterparts, with a number of 'supernumerary' subunits appearing early in eukaryotic evolution. Little is known, however, about the ETC of unicellular eukaryotes (protists), which are key to understanding the evolution of mitochondria and the ETC. We present an analysis of the ETC proteome from Acanthamoeba castellanii, an ecologically, medically and evolutionarily important member of Amoebozoa (sister to Opisthokonta). Data obtained from tandem mass spectrometric (MS/MS) analyses of purified mitochondria as well as ETC complexes isolated via blue native polyacrylamide gel electrophoresis are combined with the results of bioinformatic queries of sequence databases. Our bioinformatic analyses have identified most of the ETC subunits found in other eukaryotes, confirming and extending previous observations. The assignment of proteins as ETC subunits by MS/MS provides important insights into the primary structures of ETC proteins and makes possible, through the use of sensitive profile-based similarity searches, the identification of novel constituents of the ETC along with the annotation of highly divergent but phylogenetically conserved ETC subunits. © 2012 Elsevier B.V. All rights reserved.

Trehalose During Two Stress Responsesin Acanthamoeba: DifferentiationBetween Encystation and Pseudocyst Formation

Czech Academy of Sciences Publication Activity Database

Bínová, E.; Bína, David; Ashford, D.; Thomas-Oates, J.; Nohýnková, E.

2017-01-01

Roč. 168, č. 6 (2017), s. 649-662 ISSN 1434-4610 Institutional support: RVO:60077344 Keywords : Trehalose Acanthamoeba * Acanthamoeba * cyst - pseudocyst Subject RIV: CE - Biochemistry OBOR OECD: Biochemistry and molecular biology Impact factor: 2.794, year: 2016

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

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Nicole S. Ribeiro

2017-08-01

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

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

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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

Chemical composition and anti-Acanthamoeba activity of Melaleuca styphelioides essential oil.

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Albouchi, Ferdaous; Sifaoui, Ines; Reyes-Batlle, Maria; López-Arencibia, Atteneri; Piñero, José E; Lorenzo-Morales, Jacob; Abderrabba, Manef

2017-12-01

Acanthamoeba infections cause serious humans diseases, such as amoebic keratitis and granulomatous amoebic encephalitis. Melaleuca essential oil has been reported to be effective in treating bacterial and fungal infections. However, the anti-parasitic effects of this essential oil are not well studied. In the present study, we first characterized the composition of the essential oil, extracted from the Tunisian Melaleuca styphelioides leaves, and then tested its effect on the Acanthamoeba castellanii Neff. Gas chromatography-mass spectrometry (GC-MS) analysis revealed that the major common compounds were Caryophyllene oxide (23.42%), Spathulenol (20.5%), Isoaromadendrene epoxide (7.45%), Ledol (5.98%), α-Pinene (3.82%), Isopinocarveol (2.18%). Our data also showed that M. styphelioides essential oil inhibited the growth of Acanthamoeba with an IC 50 value of 69.03 ± 9.17 μg/ml. This work suggests M. styphelioides essential oil as a potential anti amoeba drug. Nevertheless, further studies are still needed to further verify the cytotoxicity of the studied oil on human macrophages and check its applicability to treat Acanthamoeba infections in vivo. Copyright © 2017 Elsevier Inc. All rights reserved.

Programmed cell death in Acanthamoeba castellanii Neff induced by several molecules present in olive leaf extracts.

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Ines Sifaoui

Full Text Available Therapy against Acanthamoeba infections such as Granulomatous Amoebic Encephalitis (GAE and Acanthamoeba Keratitis (AK, remains as an issue to be solved due to the existence of a cyst stage which is highly resistant to most chemical and physical agents. Recently, the activity of Olive Leaf Extracts (OLE was demonstrated against Acanthamoeba species. However, the molecules involved in this activity were not identified and/or evaluated. Therefore, the aim of this study was to evaluate the activity of the main molecules which are present in OLE and secondly to study their mechanism of action in Acanthamoeba. Among the tested molecules, the observed activities ranged from an IC50 of 6.59 in the case of apigenine to an IC50 > 100 μg/ml for other molecules. After that, elucidation of the mechanism of action of these molecules was evaluated by the detection of changes in the phosphatidylserine (PS exposure, the permeability of the plasma membrane, the mitochondrial membrane potential and the ATP levels in the treated cells. Vanillic, syringic and ursolic acids induced the higher permeabilization of the plasma membrane. Nevertheless, the mitochondrial membrane was altered by all tested molecules which were also able to decrease the ATP levels to less than 50% in IC90 treated cells after 24 h. Therefore, all the molecules tested in this study could be considered as a future therapeutic alternative against Acanthamoeba spp. Further studies are needed in order to establish the true potential of these molecules against these emerging opportunistic pathogenic protozoa.

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

DEFF Research Database (Denmark)

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

2013-01-01

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, wher......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......-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...

Inefficacy of marketed contact lens disinfection solutions against keratitis-causing Acanthamoeba castellanii belonging to the T4 genotype.

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

2014-06-01

The aim of this study was to assess the anti-amoebic effects of marketed contact lens disinfecting solutions. Using amoebistatic, amoebicidal, and cysticidal assays, nine different contact lens solutions were tested including: ReNu MultiPlus, DuraPlus, Ultimate Plus, OptiFree Replenish, OptiFree Express, Kontex Clean, Kontex Normal, Kontex Multisol extra+, Kontex Soak. In vitro growth inhibition (amoebistatic) assays were performed by incubating Acanthamoeba castellanii with aforementioned contact lens disinfection solutions as per manufacturer's instructions in the growth medium for up to 48h at 30°C. To determine amoebicidal and cysticidal effects, amoebae were incubated with contact lens solutions in phosphate buffered saline for 24h and viability was determined by haemocytometer counting as well as re-inoculating them in the growth medium. For controls, solutions were tested against bacterial corneal pathogen, Pseudomonas aeruginosa, as well as amoebae were incubated with the solvent alone. Of the nine contact lens solutions tested, none of them showed potent amoebicidal effects. Only DuraPlus and OptiFree Replenish exhibited trophozoite lysis of 85.3% and 73.7% respectively. In contrast, all contact lens disinfection solutions except Kontex Clean, Kontex Normal, Kontex Multisol extra+, tested showed amoebistatic effects. Importantly, none of the contact lens disinfection solutions exhibited cysticidal effects using qualitative assays, i.e., cysts treated with aforementioned solutions re-emerged as viable amoebae upon inoculation in the growth medium. However, more than 3-log reduction was observed when ReNu MultiPlus, DuraPlus and OptiFree Express were tested against P. aeruginosa which is in accordance with the ISO Stand-Alone Primary acceptance criteria. These findings are of great concern for contact lens users. Copyright © 2014 Elsevier Inc. All rights reserved.

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

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

2015-01-14

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 cellulose degradation offers a potential therapeutic strategy in the effective targeting of trophozoite encased within the cyst walls. Here, we present a comprehensive report on the structure of cellulose and cellulases, as well as known cellulose degradation mechanisms with an eye to target the Acanthamoeba cyst wall. The disruption of the cyst wall will make amoeba (concealed within) susceptible to chemotherapeutic agents, and at the very least inhibition of the excystment process will impede infection recurrence, as we bring these promising drug targets into focus so that they can be explored to their fullest.

Long-term survival and virulence of Mycobacterium leprae in amoebal cysts.

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Wheat, William H; Casali, Amy L; Thomas, Vincent; Spencer, John S; Lahiri, Ramanuj; Williams, Diana L; McDonnell, Gerald E; Gonzalez-Juarrero, Mercedes; Brennan, Patrick J; Jackson, Mary

2014-12-01

Leprosy is a curable neglected disease of humans caused by Mycobacterium leprae that affects the skin and peripheral nerves and manifests clinically in various forms ranging from self-resolving, tuberculoid leprosy to lepromatous leprosy having significant pathology with ensuing disfiguration disability and social stigma. Despite the global success of multi-drug therapy (MDT), incidences of clinical leprosy have been observed in individuals with no apparent exposure to other cases, suggestive of possible non-human sources of the bacteria. In this study we show that common free-living amoebae (FLA) can phagocytose M. leprae, and allow the bacillus to remain viable for up to 8 months within amoebic cysts. Viable bacilli were extracted from separate encysted cocultures comprising three common Acanthamoeba spp.: A. lenticulata, A. castellanii, and A. polyphaga and two strains of Hartmannella vermiformis. Trophozoites of these common FLA take up M. leprae by phagocytosis. M. leprae from infected trophozoites induced to encyst for long-term storage of the bacilli emerged viable by assessment of membrane integrity. The majority (80%) of mice that were injected with bacilli extracted from 35 day cocultures of encysted/excysted A. castellanii and A. polyphaga showed lesion development that was similar to mice challenged with fresh M. leprae from passage mice albeit at a slower initial rate. Mice challenged with coculture-extracted bacilli showed evidence of acid-fast bacteria and positive PCR signal for M. leprae. These data support the conclusion that M. leprae can remain viable long-term in environmentally ubiquitous FLA and retain virulence as assessed in the nu/nu mouse model. Additionally, this work supports the idea that M. leprae might be sustained in the environment between hosts in FLA and such residence in FLA may provide a macrophage-like niche contributing to the higher-than-expected rate of leprosy transmission despite a significant decrease in human reservoirs

Regulation of carbohydrate metabolism during Giardia encystment

NARCIS (Netherlands)

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

Long-term survival and virulence of Mycobacterium leprae in amoebal cysts.

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William H Wheat

2014-12-01

Full Text Available Leprosy is a curable neglected disease of humans caused by Mycobacterium leprae that affects the skin and peripheral nerves and manifests clinically in various forms ranging from self-resolving, tuberculoid leprosy to lepromatous leprosy having significant pathology with ensuing disfiguration disability and social stigma. Despite the global success of multi-drug therapy (MDT, incidences of clinical leprosy have been observed in individuals with no apparent exposure to other cases, suggestive of possible non-human sources of the bacteria. In this study we show that common free-living amoebae (FLA can phagocytose M. leprae, and allow the bacillus to remain viable for up to 8 months within amoebic cysts. Viable bacilli were extracted from separate encysted cocultures comprising three common Acanthamoeba spp.: A. lenticulata, A. castellanii, and A. polyphaga and two strains of Hartmannella vermiformis. Trophozoites of these common FLA take up M. leprae by phagocytosis. M. leprae from infected trophozoites induced to encyst for long-term storage of the bacilli emerged viable by assessment of membrane integrity. The majority (80% of mice that were injected with bacilli extracted from 35 day cocultures of encysted/excysted A. castellanii and A. polyphaga showed lesion development that was similar to mice challenged with fresh M. leprae from passage mice albeit at a slower initial rate. Mice challenged with coculture-extracted bacilli showed evidence of acid-fast bacteria and positive PCR signal for M. leprae. These data support the conclusion that M. leprae can remain viable long-term in environmentally ubiquitous FLA and retain virulence as assessed in the nu/nu mouse model. Additionally, this work supports the idea that M. leprae might be sustained in the environment between hosts in FLA and such residence in FLA may provide a macrophage-like niche contributing to the higher-than-expected rate of leprosy transmission despite a significant decrease in

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

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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

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

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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.

Conjuntivite presumível por Acanthamoeba Conjunctivitis presumably due to Acanthamoeba

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Ana Cristina de Carvalho Ruthes

2004-12-01

Full Text Available OBJETIVO: Abordar quatro casos de conjuntivite presumível por Acanthamoeba, descrevendo o diagnóstico, considerando sinais e sintomas e o tratamento instituído. MÉTODOS: Foram estudados casos de conjuntivite presumível por Acanthamoeba diagnosticados no Hospital de Olhos do Paraná (HOP, no período de setembro/1998 a janeiro/2002. Todos os olhos estudados foram submetidos a um protocolo de investigação que incluía exame oftalmológico completo, microbiologia e cultura de secreções conjuntivais. RESULTADOS: Os exames laboratoriais de microscopia e cultura do material colhido estes pacientes revelaram o diagnóstico de Acanthamoeba. A maioria dos pacientes referia olhos vermelhos e irritação ocular de longa data. Os autores encontraram correlação entre a cultura e o exame direto, em que se evidenciou a presença de cistos e trofozoítas do protozoário. CONCLUSÃO: Este é o primeiro relato de conjuntivite provavelmente por Acanthamoeba de acordo com a literatura revisada. Pacientes selecionados e refratários ao tratamento habitual de infecção ocular externa devem ser considerados para estudo laboratorial adequado à procura etiológica da doença.PURPOSE: To describe four cases of conjunctivitis presumably due to Acanthamoeba considering diagnosis, signs, symptoms and treatment. METHODS: We reviewed the medical records of all patients who presented a clinical diagnosis of Acanthamoeba conjunctivitis between September/1998 to January/2001 at the "Hospital de Olhos do Paraná (HOP". All eyes were submitted to a protocol of investigation that included ophthalmologic examination, microscopic examination and culture exams of conjunctival smears for adequate treatment. RESULTS: The laboratorial results of conjunctival smears revealed contamination with Acanthamoeba by direct examination and thereafter, confirmed by culture. The authors observed cysts and trophozoites of Acanthamoeba. CONCLUSION: This is the first report of

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

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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.

Membrane Trafficking Modulation during Entamoeba Encystation.

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Herman, Emily; Siegesmund, Maria A; Bottery, Michael J; van Aerle, Ronny; Shather, Maulood Mohammed; Caler, Elisabet; Dacks, Joel B; van der Giezen, Mark

2017-10-09

Entamoeba histolytica is an intestinal parasite that infects 50-100 million people and causes up to 55,000 deaths annually. The transmissive form of E. histolytica is the cyst, with a single infected individual passing up to 45 million cysts per day, making cyst production an attractive target for infection control. Lectins and chitin are secreted to form the cyst wall, although little is known about the underlying membrane trafficking processes supporting encystation. As E. histolytica does not readily form cysts in vitro, we assessed membrane trafficking gene expression during encystation in the closely related model Entamoeba invadens. Genes involved in secretion are up-regulated during cyst formation, as are some trans-Golgi network-to-endosome trafficking genes. Furthermore, endocytic and general trafficking genes are up-regulated in the mature cyst, potentially preserved as mRNA in preparation for excystation. Two divergent dynamin-related proteins found in Entamoeba are predominantly expressed during cyst formation. Phylogenetic analyses indicate that they are paralogous to, but quite distinct from, classical dynamins found in human, suggesting that they may be potential drug targets to block encystation. The membrane-trafficking machinery is clearly regulated during encystation, providing an additional facet to understanding this crucial parasitic process.

Co-isolation of Vahlkampfia and acanthamoeba in acanthamoeba-like keratitis in a Spanish population.

Science.gov (United States)

Arnalich-Montiel, Francisco; Lorenzo-Morales, Jacob; Irigoyen, Cristina; Morcillo-Laiz, Rafael; López-Vélez, Rogelio; Muñoz-Negrete, Francisco; Piñero, Jose E; Valladares, Basilio

2013-05-01

To report the co-isolation incidence of Acanthamoeba and Vahlkampfia in amoebic keratitis from a tertiary care institution in Madrid, Spain. In this retrospective case series, 7 eyes of 7 consecutive patients with culture-proven or polymerase chain reaction-positive Acanthamoeba keratitis were seen at a tertiary care institution from January 2010 to April 2011, and their charts were reviewed. Two of 7 patients showed mixed Acanthamoeba and Vahlkampfia keratitis. Good clinical response to the treatment was strongly correlated with early diagnosis, whereas delayed diagnosis resulted in poor response to the treatment in single or mixed infection. Co-isolation of Vahlkampfia and Acanthamoeba in Acanthamoeba-like keratitis has recently been detected in our population. This finding should raise awareness of the existence of other amoeba different from Acanthamoeba causing keratitis. There are not enough cases yet to determine the impact of mixed amoebic keratitis in the prognosis of this disease.

Acanthamoeba and Dictyostelium as Cellular Models for Legionella Infection

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Swart, A. Leoni; Harrison, Christopher F.; Eichinger, Ludwig; Steinert, Michael; Hilbi, Hubert

2018-01-01

Environmental bacteria of the genus Legionella naturally parasitize free-living amoebae. Upon inhalation of bacteria-laden aerosols, the opportunistic pathogens grow intracellularly in alveolar macrophages and can cause a life-threatening pneumonia termed Legionnaires' disease. Intracellular replication in amoebae and macrophages takes place in a unique membrane-bound compartment, the Legionella-containing vacuole (LCV). LCV formation requires the bacterial Icm/Dot type IV secretion system, which translocates literally hundreds of “effector” proteins into host cells, where they modulate crucial cellular processes for the pathogen's benefit. The mechanism of LCV formation appears to be evolutionarily conserved, and therefore, amoebae are not only ecologically significant niches for Legionella spp., but also useful cellular models for eukaryotic phagocytes. In particular, Acanthamoeba castellanii and Dictyostelium discoideum emerged over the last years as versatile and powerful models. Using genetic, biochemical and cell biological approaches, molecular interactions between amoebae and Legionella pneumophila have recently been investigated in detail with a focus on the role of phosphoinositide lipids, small and large GTPases, autophagy components and the retromer complex, as well as on bacterial effectors targeting these host factors. PMID:29552544

Acanthamoeba and Dictyostelium as Cellular Models for Legionella Infection

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A. Leoni Swart

2018-03-01

Full Text Available Environmental bacteria of the genus Legionella naturally parasitize free-living amoebae. Upon inhalation of bacteria-laden aerosols, the opportunistic pathogens grow intracellularly in alveolar macrophages and can cause a life-threatening pneumonia termed Legionnaires' disease. Intracellular replication in amoebae and macrophages takes place in a unique membrane-bound compartment, the Legionella-containing vacuole (LCV. LCV formation requires the bacterial Icm/Dot type IV secretion system, which translocates literally hundreds of “effector” proteins into host cells, where they modulate crucial cellular processes for the pathogen's benefit. The mechanism of LCV formation appears to be evolutionarily conserved, and therefore, amoebae are not only ecologically significant niches for Legionella spp., but also useful cellular models for eukaryotic phagocytes. In particular, Acanthamoeba castellanii and Dictyostelium discoideum emerged over the last years as versatile and powerful models. Using genetic, biochemical and cell biological approaches, molecular interactions between amoebae and Legionella pneumophila have recently been investigated in detail with a focus on the role of phosphoinositide lipids, small and large GTPases, autophagy components and the retromer complex, as well as on bacterial effectors targeting these host factors.

Biology and pathogenesis of Acanthamoeba

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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.

Acanthamoeba meningoencephalitis

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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.

Biology and pathogenesis of Acanthamoeba

OpenAIRE

Siddiqui, Ruqaiyyah; Khan, Naveed Ahmed

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...

Killing the dead: chemotherapeutic strategies against free-living cyst-forming protists (Acanthamoeba sp. and Balamuthia mandrillaris).

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

2013-01-01

The opportunist free-living protists such as Acanthamoeba spp. and Balamuthia mandrillaris have become a serious threat to human life. As most available drugs target functional aspects of pathogens, the ability of free-living protists to transform into metabolically inactive cyst forms presents a challenge in treatment. It is hoped, that the development of broad spectrum antiprotist agents acting against multiple cyst-forming protists to provide target-directed inhibition will offer a viable drug strategy in the treatment of these rare infections. Here, we present a comprehensive report on upcoming drug targets, with emphasis on cyst wall biosynthesis along with the related biochemistry of encystment pathways, as we strive to bring ourselves a step closer to being able to combat these deadly diseases. © 2013 The Author(s) Journal of Eukaryotic Microbiology © 2013 International Society of Protistologists.

A comparison of regimen methods for the removal and inactivation of bacteria, fungi and Acanthamoeba from two types of silicone hydrogel lenses.

Science.gov (United States)

Kilvington, Simon; Lonnen, James

2009-04-01

To compare the antimicrobial efficacy of commercial contact lens solutions when used according to the manufacturers' recommended regimens with two types of silicone hydrogel lenses. Four multipurpose contact lens care solutions were examined, representing manufacturer recommended regimens of "rub & rinse", "no rub, rinse" or "no rub, no rinse". Test organisms were Pseudomonas aeruginosa, Serratia marcescens, Staphylococcus aureus, Fusarium solani, Candida albicans and Acanthamoeba castellanii (trophozoites and cysts). Organisms, in the presence of organic soil, were inoculated on to Acuvue Oasys or Air Optix lenses and subjected to the solution manufacturer's recommended regimen. The number of surviving organisms on the lenses and in the soak solution was enumerated in accordance with ISO 14729. ISO 14729 dictates that for a given organism the combined average number of surviving microbes from the lenses and disinfectant soaking solution must be bacteria, fungi and Acanthamoeba with both lens types. Solutions employing "no rub, rinse" were less satisfactory but significantly better than "no rub, no rinse". Significant differences were found in organism survival on the lenses with greater numbers remaining on the Air Optix compared to Oasys (plenses. Accordingly, it would seem prudent to recommend that contact lens care systems include a rub step as part of the hygiene regimen.

Protease-activated receptor 2 (PAR2) is upregulated by Acanthamoeba plasminogen activator (aPA) and induces proinflammatory cytokine in human corneal epithelial cells.

Science.gov (United States)

Tripathi, Trivendra; Abdi, Mahshid; Alizadeh, Hassan

2014-05-29

Acanthamoeba plasminogen activator (aPA) is a serine protease elaborated by Acanthamoeba trophozoites that facilitates the invasion of trophozoites to the host and contributes to the pathogenesis of Acanthamoeba keratitis (AK). The aim of this study was to explore if aPA stimulates proinflammatory cytokine in human corneal epithelial (HCE) cells via the protease-activated receptors (PARs) pathway. Acanthamoeba castellanii trophozoites were grown in peptone-yeast extract glucose for 7 days, and the supernatants were collected and centrifuged. The aPA was purified using the fast protein liquid chromatography system, and aPA activity was determined by zymography assays. Human corneal epithelial cells were incubated with or without aPA (100 μg/mL), PAR1 agonists (thrombin, 10 μM; TRAP-6, 10 μM), and PAR2 agonists (SLIGRL-NH2, 100 μM; AC 55541, 10 μM) for 24 and 48 hours. Inhibition of PAR1 and PAR2 involved preincubating the HCE cells for 1 hour with the antagonist of PAR1 (SCH 79797, 60 μM) and PAR2 (FSLLRY-NH2, 100 μM) with or without aPA. Human corneal epithelial cells also were preincubated with PAR1 and PAR2 antagonists and then incubated with or without PAR1 agonists (thrombin and TRAP-6) and PAR2 agonists (SLIGRL-NH2 and AC 55541). Expression of PAR1 and PAR2 was examined by quantitative RT-PCR (qRT-PCR), flow cytometry, and immunocytochemistry. Interleukin-8 expression was quantified by qRT-PCR and ELISA. Human corneal epithelial cells constitutively expressed PAR1 and PAR2 mRNA. Acanthamoeba plasminogen activator and PAR2 agonists significantly upregulated PAR2 mRNA expression (1- and 2-fold, respectively) (P aPA, and PAR2 agonists induced PAR2 mRNA expression in HCE cells (P aPA, significantly upregulated PAR1 mRNA expression, which was significantly inhibited by PAR1 antagonist in HCE cells. Acanthamoeba plasminogen activator and PAR2 agonists stimulated IL-8 mRNA expression and protein production, which is significantly diminished by PAR2 antagonist

Label-free imaging of acanthamoeba using multimodal nonlinear optical microscopy

Science.gov (United States)

Kobayashi, Tsubasa; Cha, Yu-Rok; Kaji, Yuichi; Oshika, Tetsuro; Leproux, Philippe; Couderc, Vincent; Kano, Hideaki

2018-02-01

Acanthamoeba keratitis is a disease in which amoebae named Acanthamoeba invade the cornea of an eye. To diagnose this disease before it becomes serious, it is important to detect the cyst state of Acanthamoeba in the early stage of infection. In the present study, we explored spectroscopic signitures of the cyst state of Acanthamoeba using multimodal nonlinear optical microscopy with the channels of multiplex coherent anti-Stokes Raman scattering (CARS), second harmonic generation (SHG), and third harmonic generation (THG). A sharp band at around 1603 cm-1 in the CARS (Im[χ(3)]) spectrum was found at the cyst state of Acanthamoeba, which possibly originates from ergosterol and/or 7-dehydrostigmasterol. It can be used as a maker band of Acanthamoeba for medical treatment. Keyword: Acanthamoeba keratitis, coherent anti-Stokes Raman scattering, CARS, second harmonic generation, SHG, microspectroscopy, multiphoton microscopy

A case of trauma related Acanthamoeba keratitis.

Science.gov (United States)

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

2004-12-01

Acanthamoeba is an uncommon cause of keratitis but one of the most severe because of the prolonged and painful course of the disease and poor visual outcome. Although contact lens use is the principal risk factor, about 10% of cases occur following trauma and exposure to contaminated soil or water. Two cases of Acanthamoeba keratitis involving women contact lens wearers have previously been reported in Malaysia but this is the first time, a non contact lens related Acanthamoeba keratitis is reported. The case involved a 28 year old Indonesian male construction worker who had a trauma of the right eye during work. His eye was struck by sand and dust particles after which he quickly washed with water from an open tank at the construction site. He experienced pain, redness, glaring and blurring of vision of the right eye three days later. The diagnosis was missed at initial presentation but culture of the corneal scraping had proven Acanthamoeba as the aetiological agent. The history and clinical findings of this trauma related Acanthamoeba keratitis are briefly discussed.

Acanthamoeba keratitis challenges a case report.

Science.gov (United States)

Cristina, Stan; Cristina, Vlăduţiu; Mihaela, Popovici

2016-01-01

Acanthamoeba keratitis is a rare, chronic, mainly contact lens-related infection caused by a free-living amoeba found ubiquitously in water and soil. A case of a 9-year-old child, who presented to our clinic with painful, red left eye, associated with photophobia, and decreased visual acuity, wais reported. The clinical examination revealed a discoid opacity inferiorly bounded by a dense, gray infiltrate. The progressive nature of the corneal infiltrate, the epithelial defect, and the lack of response to treatment was highly suggestive for Acanthamoeba keratitis. The distinctiveness of this case was the presence of Acanthamoeba keratitis in a child without a history of trauma or contact lens usage, the lack of an appropriate diagnosis and management of this vision-threatening infection.

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

International Nuclear Information System (INIS)

Issa, R.M.

2007-01-01

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)

Failure of molecular diagnostics of a keratitis-inducing Acanthamoeba strain.

Science.gov (United States)

Scheid, Patrick L; Balczun, Carsten

2017-12-01

An otherwise healthy 49-year-old female patient presented at the local hospital with severe keratitis in both inflamed eyes. She was a contact lens wearer and had no history of a corneal trauma. In our laboratory for medical parasitology Acanthamoebae were detected microscopically from the cornea scraping and from the fluid of the contact lens storage case after xenical culture and showed the typical cyst morphology of Acanthamoebae group II. The diagnosis of "Acanthamoeba keratitis" was established and successful therapy was provided. While the morphological microscopic method led to the correct diagnosis in this case, an in-house multiplex qPCR and a commercial qPCR showed false negative results regarding Acanthamoeba sp. The subsequent sequencing revealed the Acanthamoeba genotype T4. In the present case report, the inability to detect Acanthamoebae using qPCR only is presented. Therefore, we recommend the utilization of combined different assays for optimal diagnostic purposes. Copyright © 2017 Elsevier Inc. All rights reserved.

First time identification of Acanthamoeba genotypes in the cornea samples of wild birds; Is Acanthamoeba keratitis making the predatory birds a target?

Science.gov (United States)

Karakavuk, Muhammet; Aykur, Mehmet; Şahar, Esra Atalay; Karakuş, Mehmet; Aldemir, Duygu; Döndüren, Ömer; Özdemir, Hüseyin Gökhan; Can, Hüseyin; Gürüz, Adnan Yüksel; Dağcı, Hande; Döşkaya, Mert

2017-12-01

Acanthamoeba is a free-living amoeba which can be isolated from environment and among others well known as an opportunist protozoan parasite causing infections in humans and animals. Eyes are extremely important for the wild birds and losing sight ability due to Acanthamoeba can be dangerous. The studies on Acanthamoeba infection in wild birds is very few in world and Turkey therefore we aimed to screen deceased wild birds found in İzmir and Manisa provinces located in western Turkey using PCR and non-nutrition agar (NNA) plate method. Cornea samples were obtained from 18 deceased wild birds. During the external examination, signs of keratitis were observed in two Eurasian sparrowhawks (Accipiter nisus). All of the corneal samples were analyzed by two PCR methods and NNA plate. According to results, the Acanthamoeba positivity in corneal samples was 16.6% and 5.5% by PCR and plate method, respectively. According to sequencing data, two of isolates belonged to genotype T5 and one was genotype T4. In conclusion, Acanthamoeba infection was detected in wild bird cornea samples with/without keratitis for the first time in the world. The result of this study also show that Acanthamoeba can be a cause of keratitis in wild birds of Turkey and thus these predator birds can be a target of other wild animals due to loss of sight ability. In terms of public health, these results show the importance of wild birds as a source of Acanthamoeba infection in nature. Copyright © 2017 Elsevier Inc. All rights reserved.

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

Directory of Open Access Journals (Sweden)

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.

Defective zoospore encystment and suppressed cyst germination of Phytophthora palmivora caused by transient leaching treatments

NARCIS (Netherlands)

Dijksterhuis, J; Deacon, J W

2003-01-01

The behaviour of encysting zoospores of Phytophthora palmivora during leaching conditions was studied. Zoospores encysted and germinated successfully on polycarbonate membranes after mechanical agitation. Transient (10 min) leaching treatments with nutrient-free buffer underneath the membranes

Caspase-like proteins: Acanthamoeba castellanii metacaspase

Indian Academy of Sciences (India)

2014-10-20

Oct 20, 2014 ... ... cellular death. Several molecular pathways that lead to cell death have been .... cellular stresses such as viral infections and heat shock. (Ivanovska and ..... of how these two essential processes occur and how they may be .... Trypanosoma brucei causes loss of respiration competence and clonal death ...

A case of radial keratoneuritis in non-Acanthamoeba keratitis

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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

Genotyping of clinical isolates of Acanthamoeba genus in Venezuela.

Science.gov (United States)

Wagner, Carolina; Reyes-Batlle, María; Ysea, María Alejandra Vethencourt; Pérez, Mónica V Galindo; de Rondón, Carmen Guzmán; Paduani, Anaibeth J Nessi; Pérez, Angelyseb Dorta; López-Arencibia, Atteneri; Sifaoui, Ines; de Galindo, María Virginia Pérez; de Suárez, Eva Pérez; Martínez-Carretero, Enrique; Valladares, Basilio; Piñero, José E; Lorenzo-Morales, Jacob

2016-12-01

Free-living amoebae of Acanthamoeba genus are opportunistic pathogens distributed worldwide. Strains included in this genus are causative agents of a fatal encephalitis and a sight-threating keratitis in humans and other animals. In this study, 550 clinical samples which were collected between 1984 and 2014 from different patients with suspected infections due to Acanthamoeba were initially screened for the presence of this amoebic genus at the Laboratorio de Amibiasis-Escuela de Bioanálisis at the Universidad Central de Venezuela. Samples were cultured in 2% Non-Nutrient agar plates seeded with a layer of heat killed Escherichia coli. From the 550 clinical samples included in this study, 18 of them were positive for Acanthamoeba genus after culture identification. Moreover, positive samples were confirmed after amplification of the Diagnostic Fragment 3 (DF3) of the Acanthamoeba18S rDNA genus and sequencing was carried out in order to genotype the isolated strains of Acanthamoeba. Furthermore, the pathogenic potential of the strains was checked by performing thermotolerance and osmotolerance assays. Sequencing of the DF3 region resulted in the identification of genotype T4 in all the isolated strains. Moreover, most isolates were thermotolerant or both thermotolerant and osmotolerant and thus were classified as potentially pathogenic strains. To the best of our knowledge, this is the first report on the molecular characterization at the genotype level of Acanthamoeba strains in Venezuela.

Area 51: How do Acanthamoeba invade the central nervous system?

Science.gov (United States)

Siddiqui, Ruqaiyyah; Emes, Richard; Elsheikha, Hany; Khan, Naveed Ahmed

2011-05-01

Acanthamoeba granulomatous encephalitis generally develops as a result of haematogenous spread, but it is unclear how circulating amoebae enter the central nervous system (CNS) and cause inflammation. At present, the mechanisms which Acanthamoeba use to invade this incredibly well-protected area of the CNS and produce infection are not well understood. In this paper, we propose two key virulence factors: mannose-binding protein and extracellular serine proteases as key players in Acanthamoeba traversal of the blood-brain barrier leading to neuronal injury. Both molecules should provide excellent opportunities as potential targets in the rational development of therapeutic interventions against Acanthamoeba encephalitis. Copyright © 2011 Elsevier Ltd. All rights reserved.

AcEST: BP911838 [AcEST

Lifescience Database Archive (English)

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

Prevalence of encysted Toxoplasma gondii in raptors from Alabama.

Science.gov (United States)

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.

The Development of Drugs against Acanthamoeba Infections

OpenAIRE

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

2016-01-01

For the past several decades, there has been little improvement in the morbidity and mortality associated with Acanthamoeba keratitis and Acanthamoeba encephalitis, respectively. The discovery of a plethora of antiacanthamoebic compounds has not yielded effective marketed chemotherapeutics. The rate of development of novel antiacanthamoebic chemotherapies of translational value and the lack of interest of the pharmaceutical industry in developing such chemotherapies have been disappointing. O...

Acanthamoeba Sp. S-11 phagocytotic activity on Mycobacterium ...

African Journals Online (AJOL)

Background: Mycobacterium leprae (M. leprae) is a pathogenic bacterium that causes leprosy. The presence of M. leprae in the environment is supported by microorganisms that act as the new host for M. leprae. Acanthamoeba's potential to be a host of M. leprae in the environment. Acanthamoeba sp. is Free Living ...

Evaluation of immunoperoxidase staining technique in the diagnosis of Acanthamoeba keratitis

Directory of Open Access Journals (Sweden)

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

Heat Shock Protein 90 regulates encystation in Entamoeba

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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.

ACANTHAMOEBA SP.S-11 PHAGOCYTOTIC ACTIVITY ON MYCOBACTERIUM LEPRAE IN DIFFERENT NUTRIENT CONDITIONS.

Science.gov (United States)

Paling, Sepling; Wahyuni, Ratna; Ni'matuzahroh; Winarni, Dwi; Iswahyudi; Astari, Linda; Adriaty, Dinar; Agusni, Indropo; Izumi, Shinzo

2018-01-01

Mycobacterium leprae ( M. leprae ) is a pathogenic bacterium that causes leprosy. The presence of M. leprae in the environment is supported by microorganisms that act as the new host for M. leprae . Acanthamoeba 's potential to be a host of M. leprae in the environment. Acanthamoeba sp. is Free Living Amoeba (FLA) that classified as holozoic, saprophytic, and saprozoic. The existence of nutrients in the environment influence Acanthamoeba ability to phagocytosis or pinocytosis. This study is aimed to determine Acanthamoeba sp.S-11 phagocytic activity to Mycobacterium leprae ( M. leprae ) which cultured in non-nutrient media and riched-nutrient media. This research conducted by culturing Acanthamoeba sp.S-11 and M. leprae on different nutrient media conditions. M. leprae intracellular DNA were isolated and amplified by M. leprae specific primers through Real Time PCR (Q-PCR). The results showed that Acanthamoeba co-cultured on non-nutrient media were more active to phagocyte M. leprae than on rich-nutrient media. The use of non-nutrient media is recommended to optimize Acanthamoeba sp. phagocytic activity to M. leprae .

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

Directory of Open Access Journals (Sweden)

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.

The assembly of GM1 glycolipid- and cholesterol-enriched raft-like membrane microdomains is important for giardial encystation.

Science.gov (United States)

De Chatterjee, Atasi; Mendez, Tavis L; Roychowdhury, Sukla; Das, Siddhartha

2015-05-01

Although encystation (or cyst formation) is an important step of the life cycle of Giardia, the cellular events that trigger encystation are poorly understood. Because membrane microdomains are involved in inducing growth and differentiation in many eukaryotes, we wondered if these raft-like domains are assembled by this parasite and participate in the encystation process. Since the GM1 ganglioside is a major constituent of mammalian lipid rafts (LRs) and known to react with cholera toxin B (CTXB), we used Alexa Fluor-conjugated CTXB and GM1 antibodies to detect giardial LRs. Raft-like structures in trophozoites are located in the plasma membranes and on the periphery of ventral discs. In cysts, however, they are localized in the membranes beneath the cyst wall. Nystatin and filipin III, two cholesterol-binding agents, and oseltamivir (Tamiflu), a viral neuraminidase inhibitor, disassembled the microdomains, as evidenced by reduced staining of trophozoites with CTXB and GM1 antibodies. GM1- and cholesterol-enriched LRs were isolated from Giardia by density gradient centrifugation and found to be sensitive to nystatin and oseltamivir. The involvement of LRs in encystation could be supported by the observation that raft inhibitors interrupted the biogenesis of encystation-specific vesicles and cyst production. Furthermore, culturing of trophozoites in dialyzed medium containing fetal bovine serum (which is low in cholesterol) reduced raft assembly and encystation, which could be rescued by adding cholesterol from the outside. Our results suggest that Giardia is able to form GM1- and cholesterol-enriched lipid rafts and these raft domains are important for encystation. Copyright © 2015, American Society for Microbiology. All Rights Reserved.

Combating Acanthamoeba spp. cysts: what are the options?

OpenAIRE

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

2018-01-01

Acanthamoeba spp. are protist pathogens and causative agents of serious infections including keratitis and granulomatous amoebic encephalitis. Its ability to convert into dormant and highly resistant cysts form limits effectiveness of available therapeutic agents and presents a pivotal challenge for drug development. During the cyst stage, Acanthamoeba is protected by the presence of hardy cyst walls, comprised primarily of carbohydrates and cyst-specific proteins, hence synthesis inhibition ...

Acanthamoeba keratitis: an emerging disease gathering importance worldwide?

Science.gov (United States)

Lorenzo-Morales, Jacob; Martín-Navarro, Carmen María; López-Arencibia, Atteneri; Arnalich-Montiel, Francisco; Piñero, José E; Valladares, Basilio

2013-04-01

Acanthamoeba keratitis (AK) is increasingly being recognized as a severe sight-threatening ocular infection worldwide. Although contact lens wear is the leading risk factor for AK, Acanthamoeba parasites are also an important cause of keratitis in non-contact lens wearers. Diagnosis of AK is challenging, and the available treatments are lengthy and not fully effective against all strains. The pathogenesis of Acanthamoeba is still under study, and the identification of the key factors involved in this process should be useful for the development of fully effective therapies. This review focuses on recent developments on AK pathogenesis and diagnosis as well as novel strategies for the evaluation of anti-amoebic agents that could be applied in the near future against these pathogens. Copyright © 2013 Elsevier Ltd. All rights reserved.

Vectorial role of Acanthamoeba in Legionella propagation in water for human use.

Science.gov (United States)

Magnet, A; Peralta, R H S; Gomes, T S; Izquierdo, F; Fernandez-Vadillo, C; Galvan, A L; Pozuelo, M J; Pelaz, C; Fenoy, S; Del Águila, C

2015-02-01

Legionella spp. is the causative agent of Legionnaires' disease and is transmitted through aerosols emanating from man-made water systems. Legionella resistance to water treatments has been related to its association with environmental amoebae such as Acanthamoeba. Due to the high presence of this protozoon in Spain and the high rate of notification of Legionnaires' disease of this country, the aims of this work were to study the coexistence of these bacteria and protozoa in water as well as their interaction. The usefulness of Acanthamoeba co-culture for the isolation of environmental Legionella was also studied. For this purpose, 70 water samples were collected in 2011 from three Drinking Water Treatment Plants, three Wastewater Treatment Plants and five Natural Pools in Spain. Acanthamoeba was found by PCR in 87.1% (61/70) samples and, by culture in 85.7% (60/70) samples. Legionella was detected by PCR in 58.6% (41/70) of water samples, in 5.7% (4/70) by agar culture and 75.7% (53/70) by Acanthamoeba co-culture. From the 54 Acanthamoeba water isolates, Legionella was detected in 43 of them independently of Acanthamoeba's genotype (T3, T4 and T11). Legionella feeleii, Legionella birminghamiensis, Legionella gresilensis/berliardensis, Legionella fairfieldensis, Legionella drozanski and Legionella falloni were identified. In conclusion, our results showed that environmental Acanthamoeba is infected by Legionella to a high percentage, and due to its ubiquity, high resistance and its pathogenic potential per se, new methods for its elimination should be studied. Also, the high effectivity of Acanthamoeba co-culture for Legionella detection has been shown. Copyright © 2014 Elsevier B.V. All rights reserved.

Pathobiology and Immunobiology of Acanthamoeba Keratitis: Insights from Animal Models
.

Science.gov (United States)

Neelam, Sudha; Niederkorn, Jerry Y

2017-06-01

Acanthamoeba keratitis (AK) is a rare but sight-threatening disease caused by pathogenic species of Acanthamoeba . Despite its ubiquitous nature, the incidence of AK is relatively low compared to other forms of infectious keratitis. Although contact lens wear is a major risk factor, exposure to contaminated water and ocular trauma are also associated with AK. Once a patient develops AK the prognosis is very poor unless an aggressive treatment regimen is initiated early. Some of the intriguing features of AK are the lack of immunological memory, resistance of the dormant cyst form to treatment, differences between the pathogenic strains and soil isolates of Acanthamoeba and the unique role of the innate immune system in controlling this disease. Understanding the series of steps involved in the pathogenesis of the disease and the host immune response against Acanthamoeba antigens is crucial for developing effective therapeutic strategies targeting the disease.

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

DEFF Research Database (Denmark)

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....

Crosslinking and corneal cryotherapy in acanthamoeba keratitis -- a histological study.

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Hager, Tobias; Hasenfus, A; Stachon, T; Seitz, B; Szentmáry, N

2016-01-01

Acanthamoeba keratitis is rare, but difficult to treat. Penetrating keratoplasty is performed in therapy-resistant cases. Nevertheless, subsequent recurrences occur in 40 % of the cases. In addition to triple-topical therapy (polyhexamid, propamidinisoethionat, neomycin), treatment alternatives are corneal cryotherapy and/or crosslinking (CXL). The aim of our present histological study was to analyze the persistence of acanthamoebatrophozoites and cysts, the persistence of bacteria, and activation of keratocytes in corneas of acanthamoeba keratitis patients following corneal cryotherapy and/or CXL. We analyzed histologically corneal buttons (from penetrating keratoplasties) of nine patients with acanthamoeba keratitis, following corneal cryotherapy (two patients) or a combination of crosslinking and corneal cryotherapy (seven patients), using haematoxilin–eosin, periodic acid Schiff (PAS), Gram and alpha-smooth muscle actin (alpha-SMA) stainings. Acanthamoeba trophozoites persisted in three corneas after cryotherapy and CXL. Cysts persisted in one of two corneas following corneal cryotherapy and in six of seven corneas after a combination of CXL and cryotherapy. One cornea showed positive Gram staining, but there were no alpha-SMA positive keratocytes in any of the corneas. Crosslinking and corneal cryotherapy have only limited impact on killing of acanthamoeba trophozoites, cysts, or bacteria. Corneal cryotherapy and CXL did not stimulate myofibroblastic transformation of keratocytes.

Acanthamoeba keratitis in Scotland: risk factors for contact lens wearers.

Science.gov (United States)

Seal, D V; Kirkness, C M; Bennett, H G; Peterson, M

1999-01-01

To investigate risk factors for Acanthamoeba keratitis amongst contact lens wearers in Scotland. Patients with Acanthamoeba keratitis in the Scottish study, all of whom wore contact lenses, were compared with 46 healthy asymptomatic contact lens-wearing controls. They were all visited at home for contact lens and environmental microbiological sampling. In addition, all 288 optical practices in the West of Scotland were polled for contact lens types and disinfecting solutions sold in 1995, and a sample, each of whom fitted more than 500 contact lenses per year, were polled for a second time. Independently, a poll was commissioned by the Eyecare Information Service in July/August 1995 to estimate the numbers of contact lens wearers in Scotland and the UK. Industry was polled for numbers of each contact lens disinfecting regimen sold in Scotland in 1995. West of Scotland, UK. All contact lens wearers among the 3 million population of the West of Scotland Health Board Areas. Risk factors for Acanthamoeba infection and recommendations for its prevention. When Acanthamoeba infection occurred, patients' home water systems were frequently (54%) found to be colonised by this amoeba. Patients more frequently washed their storage cases in tap water than controls (Pmethods (chlorine tablets or no disinfection). However further investigation showed that these methods were associated with an increased probability of rinsing the storage case in tap water, so that these two behaviours are confounded in the group studied. Failure to disinfect contact lenses, non-compliant use of chlorine tablets and/or introduction of tap water rinsing of storage cases were associated with increased risk of Acanthamoeba infection. New multipurpose solutions and hydrogen peroxide gave the lowest risk of Acanthamoeba infection, with no statistically significant difference between them. Ionic high-water content (FDA group 4) contact lenses were at increased risk of being associated with Acanthamoeba

Prevalence of encysted apicomplexans in muscles of raptors.

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Lindsay, D S; Blagburn, B L

1999-01-28

An acid-pepsin digestion technique was used to examine portions of breast muscle and heart from raptors for encysted protozoans. Apicomplexan zoites were present in 52 (45.6%) of the 114 samples examined: 11 of 12 (91.7%) red-shouldered hawks (Buteo lineatus), 20 of 34 (58.8%) red-tailed hawks (Buteo jamaicensis), two of seven (28.6%) Cooper's hawks (Accipiter cooperi), three of four (75%) sharp-shinned hawks (Accipiter striatus), one (100%) Mississippi kites (Ictinia misisippiensis), one of two (50%) American kestrels (Falco sparverius), one bald eagle (Haliaeetus leucocephalus), one of two (50%) golden eagles (Aquila chrysaetos), one of three (33%) turkey vultures (Cathartes aura), two of three (66.7%) black vultures (Coragyps atratus), three of six (50%) great-horned owls (Bubo virginianus), five of 15 (33.3%) barred owls (Strix varia), and one of 12 (8.3%) screech owls (Asio otus). Encysted protozoans were not observed in digests of tissues from three broad-winged hawks (Buteo platypterus), four ospreys (Pandion haliaetus), and five barn owls (Tyto alba). Apicomplexan cysts resembling Sarcocystis species were observed in tissue sections of muscles from 28 (37.8%) of 74 raptors.

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

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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

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

DEFF Research Database (Denmark)

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 Acanthamoeba......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...

Acanthamoeba Keratitis at the King Khaled Eye Specialist Hospital

International Nuclear Information System (INIS)

Al-Mezaine, H.; Al-Muteb, M.; Al-Motowa, S.; Wagoner, M. D.

2005-01-01

To quantify the number of cases and clinical courses of Acanthamoeba keratitis treated at the King Khaled Eye Specialist Hospital (KKESH) between December 1982 and May 2005, and to review their clinical course and response to medical and surgical therapy. Retrospective review of medical records of all patients with Acanthamoeba keratitis diagnosed by the KKESH Microbiology or Pathology or Pathology Laboratory since the opening of the hospital. During a period of over 22 years, only six cases of confirmed. Acanthamoeba keratitis were diagnosedand treated at KKESH. In two cases the diagnosis was made by corneal biopsy, while four cases required examination of a histopathology specimen obtained at the time of therapeutic keratoplasty to establish the diagnosis. One eye was cured by medical therapy, while five eyes required one (4eyes) or two (1 eye) therapeutic penetrating keratoplasty (PKP) to achieve a clinical cure. Three eyes achieved good visual acuity (20/40, 20/50, 20/60), one eye achieved fair visual acuity (20/160), and two eyes suffered profound visual loss (hand motions). The incidence of Acanthamoeba keratitis may be less in Saudi Arabia than reported in Western countries although the clinical is similarly complicated, with most cases requiring therapeutic keratoplasty for definitive education of the organism. (author)

A multisystemic Acanthamoeba infection in a dog in Tenerife, Canary Islands, Spain.

Science.gov (United States)

Valladares, María; Reyes-Batlle, María; Mora-Peces, Inmaculada; Martín-Navarro, Carmen M; López-Arencibia, Atteneri; Dorta-Gorrín, Alexis; Comyn-Afonso, Estefanía; Martínez-Carretero, Enrique; Maciver, Sutherland K; Piñero, José E; Valladares, Basilio; Lorenzo-Morales, Jacob

2014-10-15

A 22-month-old male Spanish water dog was hospitalized after its physical examination revealed fever and movement difficulty. After 24h, the dog was found to have a high fever (39.5 °C) and was treated empirically with doxycycline/ciprofloxacin. At 48 h, after submission the fever rose to 41 °C and the animal presented with a stiff neck and dehydration. Peripheral blood and cerebrospinal fluid (CSF) were sampled and trophozoites with an Acanthamoeba-like morphology were observed in the CSF. PCR specific for Acanthamoeba, Naegleria fowleri and Balamuthia mandrillaris were performed and the CSF sample found positive for Acanthamoeba. Lungs, kidney, liver and spleen samples were collected post mortem. All collected organ samples were positive for Acanthamoeba by PCR, thus confirming a multisystemic infection. Water samples taken at a suspected site of infection yielded an almost identical PCR fragment to those of the clinical samples, indicating that this was probably where the infection originated. This is the first report of a fatal case of Acanthamoeba disseminated infection in a dog in Spain. Copyright © 2014 Elsevier B.V. All rights reserved.

Granulomatous inflammation in Acanthamoeba sclerokeratitis

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Samrat Chatterjee

2013-01-01

Full Text Available This report describes the histopathological findings in a patient with Acanthamoeba sclerokeratitis (ASK. A 58-year-old patient with ASK underwent enucleation and sections of the cornea and sclera were subjected to histopathology and immunohistochemistry with monoclonal mouse antihuman antibodies against T cell CD3 and B cell CD20 antigens. Hematoxylin and Eosin stained sections of the cornea revealed epithelial ulceration, Bowman′s membrane destruction, stromal vascularization, infiltration with lymphocytes, plasma cells, and granulomatous inflammation with multinucleated giant cells (MNGC. The areas of scleritis showed complete disruption of sclera collagen, necrosis and infiltration with neutrophils, macrophages, lymphocytes, and granulomatous inflammation with MNGC. No cyst or trophozoites of Acanthamoeba were seen in the cornea or sclera. Immunophenotyping revealed that the population of lymphocytes was predominantly of T cells. Granulomatous inflammation in ASK is probably responsible for the continuance and progression of the scleritis and management protocols should include immunosuppressive agents alongside amoebicidal drugs.

[Development of a liquid fermentation system and encystment for a nitrogen-fixing bacterium strain having biofertilizer potential].

Science.gov (United States)

Camelo-Rusinque, Mauricio; Moreno-Galván, Andrés; Romero-Perdomo, Felipe; Bonilla-Buitrago, Ruth

The indiscriminate use of chemical fertilizers has contributed to the deterioration of the biological, physical and chemical properties of the soil, resulting in the loss of its productive capacity. For this reason, the use of biofertilizers has emerged as a technological alternative. The objective of this research was to develop a suitable liquid fermentation system and encystment for the multiplication of Azotobacter chroococcum AC1 strain, a bacterium employed in a biofertilizer formulation produced at present by CARPOICA, Colombia. Sequential statistical designs were used to determine the conditions in the fermentation system. The interaction between agitation, aeration and pH was evaluated on the viable biomass (CFU/ml) of AC1. In addition, the encystment ability of the strain was evaluated using two encystment agents and the potential plant growth-promoting rhizobacteria (PGPR) activity was assessed by different techniques, such as nitrogen fixation by ARA, phosphate solubilization by the phospho-molybdenum-blue reaction and indolic compound production by colorimetric reaction using the Salkowski reagent. Results showed significant effects (p<0.05) on the viable biomass in the three conditions (pH, aeration and agitation) tested individually, in one dual interaction and one tripartite interaction, were demonstrated to have a positive effect on the response variable aeration and agitation. The addition of the two encystment agents evaluated, AE01 and AE02, demonstrated the ability of AC1 to form cysts under stress conditions. Likewise, fermentation and encystment conditions did not affect the biological activities tested. Copyright © 2017 Asociación Argentina de Microbiología. Publicado por Elsevier España, S.L.U. All rights reserved.

Acanthamoeba polyphaga resuscitates viable non-culturable Legionella pneumophila after disinfection.

Science.gov (United States)

García, María Teresa; Jones, Snake; Pelaz, Carmen; Millar, Richard D; Abu Kwaik, Yousef

2007-05-01

Amoebae are the natural hosts for Legionella pneumophila and play essential roles in bacterial ecology and infectivity to humans. When L. pneumophila colonizes an aquatic installation, it can persist for years despite repeated treatments with disinfectants. We hypothesized that freshwater amoebae play an important role in bacterial resistance to disinfectants, and in subsequent resuscitation of viable non-culturable (VNC) L. pneumophila that results in re-emergence of the disease-causing strain in the disinfected water source. Our work showed that in the absence of Acanthamoeba polyphaga, seven L. pneumophila strains became non-culturable after treatment by 256 p.p.m. of sodium hypochlorite (NaOCl). In contrast, intracellular L. pneumophila within A. polyphaga was resistant to 1024 p.p.m. of NaOCl. In addition, L. pneumophila-infected A. polyphaga exhibited increased resistance to NaOCl. When chlorine-sterilized water samples were co-cultured with A. polyphaga, the non-culturable L. pneumophila were resuscitated and proliferated robustly within A. polyphaga. Upon treatment by NaOCl, uninfected amoebae differentiated into cysts within 48 h. In contrast, L. pneumophila-infected A. polyphaga failed to differentiate into cysts, and L. pneumophila was never detected in cysts of A. polyphaga. We conclude that amoebic trophozoites protect intracellular L. pneumophila from eradication by NaOCl, and play an essential role in resuscitation of VNC L. pneumophila in NaOCl-disinfected water sources. Intracellular L. pneumophila within trophozoites of A. polyphaga block encystation of the amoebae, and the resistance of both organisms to NaOCl is enhanced. To ensure long-term eradication and complete loss of the VNC state of L. pneumophila, we recommend that Legionella-protozoa co-culture should be an important tool to ensure complete loss of the VNC state of L. pneumophila.

Interaction between Mycobacterium avium subsp. paratuberculosis and environmental protozoa

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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.

Acanthamoeba sp.: un caso de queratitis no relacionada con el uso de lentes de contacto Acanthamoeba sp.: a case report in a non-contact lens wearer

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Claudia Menghi

2012-12-01

Full Text Available La queratitis por Acanthamoeba sp. generalmente se relaciona con el uso de lentes de contacto y en menor proporción se vincula con el contacto con agua contaminada. Se caracteriza por una disminución de la capacidad visual y la presencia de un fuerte dolor ocular. al examen clínico, puede confundirse con una infección herpética. Si la infección no es diagnosticada y tratada a tiempo en forma adecuada, puede culminar en la perforación de la córnea y, eventualmente, en la pérdida del ojo. en este trabajo se informa uno de los pocos casos registrados de queratitis por Acanthamoeba sp. no relacionada con el uso de lentes de contacto.Acanthamoeba sp. keratitis is generally related to wearing contact lenses, and, to a lesser extent, to contaminated water. it is characterized by reduced visual capacity and the presence of severe ocular pain. Clinically, it can be mistaken for a herpes infection. if it is not diagnosed, and timely and adequately treated, it can result in corneal perforation and, eventually, in vision loss. One of the few registered cases of Acanthamoeba sp. keratitis not related to the use of contact lenses is herein reported.

Patient characteristics, diagnosis, and treatment of non-contact lens related Acanthamoeba keratitis.

Science.gov (United States)

Sharma, S; Garg, P; Rao, G N

2000-10-01

To review the clinical characteristics, diagnosis, and visual outcome in patients with non-contact lens related Acanthamoeba keratitis and compare the findings with reported series of contact lens associated Acanthamoeba keratitis. Medical and microbiology records of 39 consecutive patients with a diagnosis of Acanthamoeba keratitis, at a tertiary eyecare centre in India between January 1996 and June 1998, were analysed retrospectively. A majority of the patients presented with poor visual acuity and large corneal stromal infiltrates (mean size 38.20 (SD 26. 18) mm). A predisposing factor was elicited in 19/39 (48.7%) patients (trauma 15, dirty water splash three, leaf juice one). None of the patients had worn contact lenses. Most patients (26/39 (66. 6%)) came from a low socioeconomic background. Complaint of severe pain was not a significant feature and radial keratoneuritis was seen in 1/39 (2.5%) patients. A ring infiltrate was present in 41.1% of cases. A clinical diagnosis of fungal keratitis was made in 45% of the patients before they were seen by us. However, all patients were diagnosed microbiologically at our institute based on demonstration of Acanthamoeba cysts in corneal scrapings (34/39) and/or culture of Acanthamoeba (34/39). Treatment with biguanides (PHMB, 15/38 (39.4%), PHMB with CHx, 23/38 (60.5%), one patient did not return for treatment) resulted in healing with scar formation in 27 out of 31(87.0%) followed up patients (mean time to healing 106.9 days). Overall visual outcome was poor with no statistical difference between cases diagnosed within 30 days (early) or 30 days after (late) start of symptoms. The visual outcome in cases requiring tissue adhesive (five) and keratoplasty (three) was also poor. This is thought to be the largest series of cases of Acanthamoeba keratitis in non-contact lens wearers. In such cases, the disease is advanced at presentation in most patients, pathognomonic clinical features are often not seen, disease

Broad spectrum antimicrobial activity of melimine covalently bound to contact lenses.

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Dutta, Debarun; Cole, Nerida; Kumar, Naresh; Willcox, Mark D P

2013-01-07

To develop a stable antimicrobial contact lens, which is effective against the International Organization for Standardization (ISO) panel microorganisms, Acanthamoeba castellanii and drug resistant strains of Pseudomonas aeruginosa and Staphylococcus aureus. Melimine was covalently incorporated into etafilcon A lenses. The amount of peptide present on the lens surface was quantified using amino acid analysis. After coating, the heat stability (121°C), lens surface hydrophobicity (by captive bubble), and in vitro cytotoxicity to mouse L929 cells of the lenses were investigated. Antimicrobial activity against the micro-organisms was evaluated by viable plate count and fluorescence microscopy, measuring the proportion of cell death compared with control lenses with no melimine. The most effective concentration was determined to be 152 ± 44 μg lens(-1) melimine on the lens surface. After coating, lenses were relatively hydrophilic and were nontoxic to mammalian cells. The activity remained high after autoclaving (e.g., 3.1, 3.9, 1.2, and 1.0 log inhibition against P. aeruginosa, S. aureus, A. castellanii, and Fusarium solani, respectively). Fluorescence microscopy confirmed significantly reduced (P bacteria to melimine contact lenses. Viable count confirmed that lenses were active against all the bacteria and fungi from the ISO panel, Acanthamoeba and gave at least 2 log inhibition against all the multidrug resistant S. aureus and P. aeruginosa strains. Melimine may offer excellent potential for development as a broad spectrum antimicrobial coating for contact lenses, showing activity against all the bacterial and fungal ISO panel microorganisms, Acanthamoeba, and antibiotic resistant strains of P. aeruginosa and S. aureus.

[A new strategy for enhancing acanthamoebicidal activity with synthesis of nanoflower of Laurocerausus officinalis Roemer (cherry laurel) fruit extracts].

Science.gov (United States)

Baldemir, Ayşe; Karaman, Ülkü; Yusufbeyoğlu, Sadi; Eken, Ayşe; Ildız, Nilay; İlgün, Selen; Çolak, Cemil; Kaçmaz, Gamze; Öçsoy, İsmail; Çankaya, Soner

2018-01-01

Pathogenic Acanthamoeba species often cause infection known as Acanthamoeba keratitis among people who use contact lenses. It is a type of infection that can result in corneal ulceration, visual loss or even blindness, if not treated. There are various therapeutic options available in the treatment of Acanthamoeba infections but they are usually tough treatments with limited efficacy. For instance, hydrogen peroxide (H 2 O 2 ) is a commonly used contact lens disinfectant which is effective against Acanthamoeba but it is toxic to the cornea. For these reasons, new and more efficacious treatment options are required for Acanthamoeba infections. In this context, plants are considered natural resources for the discovery of new drugs. Laurocerasus officinalis Roem. (cherry laurel) (Rosaceae) grows in Black Sea region; and it is known as "Taflan", "Laz kirazı" or "Karayemis". Local people are using the seeds against diabetes, while the fruits are consuming as food, and used fordiuretic and passing kidney stones. It has also been reported that the seeds of the cherry laurel are used as an antiparasitic agent in this area. The aim of the study was to confirm the traditionally use of antiparasitic activity of this fruit and to increase the potential effect by means of organic-inorganic hybrid synthesis. Total phenol contents of methanol extracts prepared from endocarp, mesocarp and seeds of the fruit were calculated. The effects of methanol extracts and nano flower (NFs) plants synthesized from these extracts on the proliferation of Acanthamoeba castellanii were investigated. Thus, for the first time, novel organic-inorganic nanobio-antiparasitic agents called NFs were produced from cherry laurel and the increase in the amoebicidal activity of the NFs was elucidated. The characterization of NFs were determined with Scanning Electron Microscopy (SEM), Fourier Transform Infrared Spectrometer (FT-IR) and Energy-Dispersive X-ray (EDX) techniques. In addition, the catalytic

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

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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.

Isolation and Genotyping of Acanthamoeba Strains from Environmental Sources in Ahvaz City, Khuzestan Province, Southern Iran

Science.gov (United States)

Rahdar, M; Niyyati, M; Salehi, M; Feghhi, M; Makvandi, M; Pourmehdi, M; Farnia, S

2012-01-01

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. Results 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. PMID:23323088

Encystment of parasitic freshwater pearl mussel (Margaritifera margaritifera) larvae coincides with increased metabolic rate and haematocrit in juvenile brown trout (Salmo trutta).

Science.gov (United States)

Filipsson, Karl; Brijs, Jeroen; Näslund, Joacim; Wengström, Niklas; Adamsson, Marie; Závorka, Libor; Österling, E Martin; Höjesjö, Johan

2017-04-01

Gill parasites on fish are likely to negatively influence their host by inhibiting respiration, oxygen transport capacity and overall fitness. The glochidia larvae of the endangered freshwater pearl mussel (FPM, Margaritifera margaritifera (Linnaeus, 1758)) are obligate parasites on the gills of juvenile salmonid fish. We investigated the effects of FPM glochidia encystment on the metabolism and haematology of brown trout (Salmo trutta Linnaeus, 1758). Specifically, we measured whole-animal oxygen uptake rates at rest and following an exhaustive exercise protocol using intermittent flow-through respirometry, as well as haematocrit, in infested and uninfested trout. Glochidia encystment significantly affected whole-animal metabolic rate, as infested trout exhibited higher standard and maximum metabolic rates. Furthermore, glochidia-infested trout also had elevated levels of haematocrit. The combination of an increased metabolism and haematocrit in infested fish indicates that glochidia encystment has a physiological effect on the trout, perhaps as a compensatory response to the potential respiratory stress caused by the glochidia. When relating glochidia load to metabolism and haematocrit, fish with low numbers of encysted glochidia were the ones with particularly elevated metabolism and haematocrit. Standard metabolic rate decreased with substantial glochidia loads towards levels similar to those of uninfested fish. This suggests that initial effects visible at low levels of encystment may be countered by additional physiological effects at high loads, e.g. potential changes in energy utilization, and also that high numbers of glochidia may restrict oxygen uptake by the gills.

Confocal microscopy as an early relapse marker for acanthamoeba keratitis.

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Daas, Loay; Viestenz, Arne; Schnabel, Philipp Albert; Fries, Fabian N; Hager, Tobias; SzentmÁry, Nora; Seitz, Berthold

2018-01-01

Acanthameoba keratitis is a serious ophthalmological condition with a potentially vision-threatening prognosis. Early diagnosis and recognition of relapse, and the detection of persistent Acanthamoeba cysts, are essential for informing the prognosis and managing the condition. We suggest the use of in vivo confocal microscopy not only to identify the early signs of relapse after keratoplasty in patients with Acanthamoeba keratitis, but also as an additional follow-up tool after antimicrobial crosslinking. This study shows that in vivo confocal microscopy is, in experienced hands, a quick and reliable diagnostic tool. Clin. Anat. 31:60-63, 2018. © 2017 Wiley Periodicals, Inc. © 2017 Wiley Periodicals, Inc.

Quick survey for detection, identification and characterization of Acanthamoeba genotypes from some selected soil and water samples in Pakistan

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Tania Tanveer

2015-05-01

Full Text Available Acanthamoeba is an opportunistic protozoan pathogen which is widely distributed in nature and plays a pivotal role in ecosystem. Acanthamoeba species may cause blinding keratitis and fatal granulomatous encephalitis involving central nervous system. In this study, we investigated the presence of Acanthamoeba in soil and water resources of Pakistan. Here, Acanthamoeba were recovered on non-nutrient agar plate lawn with E.coli and identified by morphological characteristics of the cyst. Furthermore PCR was performed with genus-specific primers followed by direct sequencing of the PCR product for molecular identification. Overall our PCR and sequencing results confirmed pathogenic genotypes including T4 and T15 from both soil and water samples. This is our first report of Acanthamoeba isolation from both soil and water resources of Pakistan which may serve as a potential treat to human health across the country.

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

International Nuclear Information System (INIS)

Pan, Hong; Wu, Xinyi

2012-01-01

Highlights: ► Hypoxia attenuates Acanthamoeba-induced the production of IL-8 and IFN-β. ► Hypoxia inhibits TLR4 expression in a time-dependent manner in HCECs. ► Hypoxia inhibits Acanthamoeba-induced the activation of NF-κB and ERK1/2 in HCECs. ► Hypoxia decreases Acanthamoeba-induced inflammatory response via TLR4 signaling. ► 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-β (IFN-β) 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-κB) 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-β. Furthermore, hypoxia attenuated Acanthamoeba-triggered TLR4 expression as well as the activation of NF-κB 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) MyD88 expression and NF-κB activation, confirming that hypoxia suppressed the LPS-induced inflammatory response by affecting TLR4 signaling. In conclusion

Acanthamoeba keratitis in Sudan: outcome of ketoconazole ...

African Journals Online (AJOL)

In an exploration of the acanthamoebic aetiology of chronic keratitis cases (not responding to antibiotics, antiviral or antifungal therapy), 138 Sudanese patients were seen over a 2-year period at two teaching eye hospitals in Khartoum State. Six (4.3%) of these patients were found to be suffering from Acanthamoeba ...

An apparent Acanthamoeba genotype is the product of a chimeric 18S rDNA artifact.

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Corsaro, Daniele; Venditti, Danielle

2018-02-01

Free-living amoebae of the genus Acanthamoeba are potentially pathogenic protozoa widespread in the environment. The detection/diagnosis as well as environmental survey strategies is mainly based on the identification of the 18S rDNA sequences of the strains that allow the recovery of various distinct genotypes/subgenotypes. The accurate recording of such data is important to better know the environmental distribution of distinct genotypes and how they may be preferentially associated with disease. Recently, a putative new acanthamoebal genotype T99 was introduced, which comprises only environmental clones apparently with some anomalous features. Here, we analyze these sequences through partial treeing and BLAST analyses and find that they are actually chimeras. Our results show that the putative T99 genotype is very likely formed by chimeric sequences including a middle fragment from acanthamoebae of genotype T13, while the 5'- and 3'-end fragments came from a nematode and a cercozoan, respectively. Molecular phylogenies of Acanthamoeba including T99 are consequently erroneous as genotype T99 does not exist in nature. Careful identification of Acanthamoeba genotypes is therefore critical for both phylogenetic and diagnostic applications.

Azasterols impair Giardia lamblia proliferation and induces encystation

International Nuclear Information System (INIS)

Maia, Claudia; Attias, Marcia; Urbina, Julio; Gilbert, Ian; Magaraci, Filippo; Souza, Wanderley de

2007-01-01

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 IC 50 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

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

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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.

The Trojan Horse of the microbiological arms race: phage-encoded toxins as a defence against eukaryotic predators.

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Arnold, Jason W; Koudelka, Gerald B

2014-02-01

Phage-encoded Shiga toxin (Stx) acts as a bacterial defence against the eukaryotic predator Tetrahymena. To function as an effective bacterial anti-predator defence, Stx must kill a broad spectrum of predators. Consistent with that assertion, we show here that bacterially encoded Stx efficiently kills the bacteriovore Acanthamoeba castellanii in co-culture. We also show that, in addition to Stx, the phage-encoded exotoxin, diphtheria toxin (Dtx) expressed by Corynebacterium diphtheriae also can function as part of an anti-predator strategy; it kills Acanthamoeba in co-culture. Interestingly, only exotoxins produced by bacteria internalized by the Acanthamoeba predator are cytolethal; the presence of purified Dtx or Stx in culture medium has no effect on predator viability. This finding is consistent with our results indicating that intoxication of Acanthamoeba by these exotoxins does not require a receptor. Thus bacteria, in the disguise of a food source, function as a 'Trojan Horse', carrying genes encoding an exotoxin into target organisms. This 'Trojan Horse' mechanism of exotoxin delivery into predator cells allows intoxication of predators that lack a cell surface receptor for the particular toxin, allowing bacteria-bearing exotoxins to kill a broader spectrum of predators, increasing the fitness of the otherwise 'defenceless' prey bacteria. © 2013 Society for Applied Microbiology and John Wiley & Sons Ltd.

Acanthamoeba keratitis challenges a case report

OpenAIRE

Cristina, Stan; Cristina, Vlăduţiu; Mihaela, Popovici

2016-01-01

Acanthamoeba keratitis is a rare, chronic, mainly contact lens-related infection caused by a free-living amoeba found ubiquitously in water and soil. A case of a 9-year-old child, who presented to our clinic with painful, red left eye, associated with photophobia, and decreased visual acuity, wais reported. The clinical examination revealed a discoid opacity inferiorly bounded by a dense, gray infiltrate. The progressive nature of the corneal infiltrate, the epithelial defect, and the lack of...

Effects of Zataria, Mentha Pulegium, Oregano spp Essential Oil and Hydroalcholic Extract of Hypericum perforatum on Cyst of Acanthamoeba spp In Vitro

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Ali Arjmand Shabestary

2017-11-01

Full Text Available Abstract Background: Resistance of Acanthamoeba cysts causes recurrence of the disease; so, the patient should be monitored regularly ،The aim of the study was to examine the effect of a few herbal materials on Acanthamoeba cysts in vitro. Materials and Methods: Essential oils (EOs of Zataria, Mint, and Oregano were prepared by steam distillation. The EOs and Hypericum perforatum extract were prepared in three concentrations (0.6%, 1% and 10%، Acanthamoeba cysts in various time intervals (30, 60, 120, 180 and 1440 minutes were exposed with plant extracts. Then, the viability of parasite was investigated by eosin 0.1%. Results: Comparison of the parasite mortality rate between control and case groups showed that the mortality of Acanthamoeba cysts was higher in the case groups that exposed to herbal materials. At the equal concentration (10% and time (24 h, the Zataria and mint EOs produced the highest (22% and lowest (4% mortality, respectively. The results showed the mortality rate of Acanthamoeba was time-dependent. Conclusion: Zataria showed the most fatality effect against Acanthamoeba cysts. In this respect, clinical trial studies are suggested.

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

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

2015-03-01

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.

Disinfection capacity of PuriLens contact lens cleaning unit against Acanthamoeba.

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Hwang, Thomas S; Hyon, Joon Young; Song, Jae Kyung; Reviglio, Victor E; Spahr, Harry T; O'Brien, Terrence P

2004-01-01

The PuriLens contact lens system is indicated for cleaning and disinfection of soft (hydrophilic) contact lenses by means of subsonic agitation to remove lens deposits and microorganisms, and ultraviolet irradiation of the storage solution for disinfection. The capacity of the PuriLens system to disinfect storage solutions contaminated with known concentrations of Staphylococcus aureus, Pseudomonas aeruginosa, and Acanthamoeba species was evaluated. An in vitro assessment of the antibacterial and antiparasitic efficacy of the PuriLens system was performed. Separated batches of the storage solution for the cleansing system were contaminated with stock strains of S. aureus and P. aeruginosa. A comparison of the microbiologic content was made between the solution before and after the cycle. The PuriLens system effectively eradicated S. aureus and P. aeruginosa organisms after a 15-minute cycle. However, viable cysts of acanthamoeba were recovered in the solution after the 15-minute cycle. The PuriLens system is highly efficient in protecting against contamination with common bacterial ocular pathogens. Acanthamoeba cysts, however, can survive in the solution or contact lens bath undergoing integrated subsonic debridement and indirect ultraviolet light disinfection. Use of chemical disinfecting solutions that contain agents such as chlorhexidine or other cationic antiseptics may be advisable in conjunction with use of the PuriLens device, especially in high-risk settings.

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

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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

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

OpenAIRE

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.

Acanthamoeba belonging to T3, T4, and T11: genotypes isolated from air-conditioning units in Santiago, Chile.

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Astorga, Berbeli; Lorenzo-Morales, Jacob; Martín-Navarro, Carmen M; Alarcón, Verónica; Moreno, Johanna; González, Ana C; Navarrete, Elizabeth; Piñero, José E; Valladares, Basilio

2011-01-01

Free-living amoebae (FLA) of the genus Acanthamoeba are widely distributed in the environment, in the air, soil, and water, and have also been isolated from air-conditioning units. The objective of this work was to investigate the presence of this genus of FLA in the air-conditioning equipment at the Institute of Public Health of Chile in Santiago, Chile. Water and air samples were collected from air-conditioning systems and were checked for the presence of Acanthamoeba spp. Positive samples were further classified at the genotype level after sequencing the highly variable diagnostic fragment 3 (DF3) region of the 18S rRNA gene. This is the first report of the T3, T4, and T11 genotypes of Acanthamoeba in air-conditioning units from Chile. Overall, the widespread distribution of potentially pathogenic Acanthamoeba strains in the studied source demands more awareness within the public and health professionals in Chile as this pathogen is emerging as a risk for human health worldwide. © 2011 The Author(s) Journal of Eukaryotic Microbiology © 2011 International Society of Protistologists.

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

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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.

The correlation of Acanthamoeba from the ventilation system with other environmental parameters in commercial buildings as possible indicator for indoor air quality.

Science.gov (United States)

Ooi, Soo Shen; Mak, Joon Wah; Chen, Donald K F; Ambu, Stephen

2017-02-07

The free-living protozoan Acanthamoeba is an opportunistic pathogen that is ubiquitous in our environment. However, its role in affecting indoor air quality and ill-health of indoor occupants is relatively unknown. The present study investigated the presence of Acanthamoeba from the ventilation system and its correlation with other indoor air quality parameters, used in the industry code of practice and its potential as an indicator for indoor air quality. Indoor air quality assessments were carried out in nine commercial buildings with approval from the building management, and the parameters assessed were as recommended by the Department of Occupational Safety and Health. The presence of Acanthamoeba was determined through dust swabs from the ventilation system and indoor furniture. Logistic regression was performed to study the correlation between assessed parameters and occupants' complaints. A total of 107 sampling points were assessed and 40.2% of the supplying air diffuser and blowing fan and 15% of the furniture were positive for cysts. There was a significant correlation between Acanthamoeba detected from the ventilation system with ambient total fungus count (r=0.327; p=0.01) and respirable particulates (r=0.276; p=0.01). Occupants' sick building syndrome experience also correlated with the presence of Acanthamoeba in the ventilation system (r=0.361; p=0.01) and those detected on the furniture (r=0.290; p=0.01). Logistic regression showed that there was a five-fold probability of sick building syndrome among occupants when Acanthamoeba was detected in the ventilation system.

Fibrous Catalyst-Enhanced Acanthamoeba Disinfection by Hydrogen Peroxide.

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Kilvington, Simon; Winterton, Lynn

2017-11-01

Hydrogen peroxide (H2O2) disinfection systems are contact-lens-patient problem solvers. The current one-step, criterion-standard version has been widely used since the mid-1980s, without any significant improvement. This work identifies a potential next-generation, one-step H2O2, not based on the solution formulation but rather on a case-based peroxide catalyst. One-step H2O2 systems are widely used for contact lens disinfection. However, antimicrobial efficacy can be limited because of the rapid neutralization of the peroxide from the catalytic component of the systems. We studied whether the addition of an iron-containing catalyst bound to a nonfunctional propylene:polyacryonitrile fabric matrix could enhance the antimicrobial efficacy of these one-step H2O2 systems. Bausch + Lomb PeroxiClear and AOSept Plus (both based on 3% H2O2 with a platinum-neutralizing disc) were the test systems. These were tested with and without the presence of the catalyst fabric using Acanthamoeba cysts as the challenge organism. After 6 hours' disinfection, the number of viable cysts was determined. In other studies, the experiments were also conducted with biofilm formed by Stenotrophomonas maltophilia and Elizabethkingia meningoseptica bacteria. Both control systems gave approximately 1-log10 kill of Acanthamoeba cysts compared with 3.0-log10 kill in the presence of the catalyst (P catalyst compared with ≥3.0-log10 kill when it was omitted. In 30 rounds' recurrent usage, the experiments, in which the AOSept Plus system was subjected to 30 rounds of H2O2 neutralization with or without the presence of catalytic fabric, showed no loss in enhanced biocidal efficacy of the material. The catalytic fabric was also shown to not retard or increase the rate of H2O2 neutralization. We have demonstrated the catalyst significantly increases the efficacy of one-step H2O2 disinfection systems using highly resistant Acanthamoeba cysts and bacterial biofilm. Incorporating the catalyst into the

The potential pathogenicity of chlorhexidine-sensitive Acanthamoeba strains isolated from contact lens cases from asymptomatic individuals in Tenerife, Canary Islands, Spain.

Science.gov (United States)

Martín-Navarro, Carmen M; Lorenzo-Morales, Jacob; Cabrera-Serra, M Gabriela; Rancel, Fernando; Coronado-Alvarez, Nieves M; Piñero, José E; Valladares, Basilio

2008-11-01

Pathogenic strains of the genus Acanthamoeba are causative agents of a serious sight-threatening infection of the eye known as Acanthamoeba keratitis. The prevalence of this infection has risen in the past 20 years, mainly due to the increase in number of contact lens wearers. In this study, the prevalence of Acanthamoeba in a risk group constituted by asymptomatic contact lens wearers from Tenerife, Canary Islands, Spain, was evaluated. Contact lenses and contact lens cases were analysed for the presence of Acanthamoeba isolates. The isolates' genotypes were also determined after rDNA sequencing. The pathogenic potential of the isolated strains was subsequently established using previously described molecular and biochemical assays, which allowed the selection of three strains with high pathogenic potential. Furthermore, the sensitivity of these isolates against two standard drugs, ciprofloxacin and chlorhexidine, was analysed. As the three selected strains were sensitive to chlorhexidine, its activity and IC(50) were evaluated. Chlorhexidine was found to be active against these strains and the obtained IC(50) values were compared to the concentrations of this drug present in contact lens maintenance solutions. It was observed that the measured IC(50) was higher than the concentration found in these maintenance solutions. Therefore, the ineffectiveness of chlorhexidine-containing contact lens maintenance solutions against potentially pathogenic strains of Acanthamoeba is demonstrated in this study.

Phylogeography, genetic variability and structure of Acanthamoeba metapopulations in Iran inferred by 18S ribosomal RNA sequences: A systematic review and meta-analysis.

Science.gov (United States)

Spotin, Adel; Moslemzadeh, Hamid Reza; Mahami-Oskouei, Mahmoud; Ahmadpour, Ehsan; Niyyati, Maryam; Hejazi, Seyed Hossein; Memari, Fatemeh; Noori, Jafar

2017-09-01

To verify phylogeography and genetic structure of Acanthamoeba populations among the Iranian clinical isolates and natural/artificial environments distributed in various regions of the country. We searched electronic databases including Medline, PubMed, Science Direct, Scopus and Google Scholar from 2005 to 2016. To explore the genetic variability of Acanthamoeba sp, 205 sequences were retrieved from keratitis patients, immunosuppressed cases and environmental sources as of various geographies of Iran. T4 genotype was the predominant strain in Iran, and the rare genotypes belonged to T2, T3, T5 (Acanthamoeba lenticulata), T6, T9, T11, T13 and T15 (Acanthamoeba jacobsi). A total of 47 unique haplotypes of T4 were identified. A parsimonious network of the sequence haplotypes demonstrated star-like feature containing haplogroups IR6 (34.1%) and IR7 (31.2%) as the most common haplotypes. In accordance with the analysis of molecular variance, the high value of haplotype diversity (0.612-0.848) of Acanthamoeba T4 represented genetic variability within populations. Neutrality indices of the 18S ribosomal RNA demonstrated negative values in all populations which represented a considerable divergence from neutrality. The majority of genetic diversity belonged to the infected contact lens and dust samples in immunodeficiency and ophthalmology wards, which indicated potential routes for exposure to a pathogenic Acanthamoeba sp. in at-risk individuals. A pairwise fixation index (F ST ) was from low to high values (0.02433-0.41892). The statistically F ST points out that T4 is genetically differentiated between north-west, north-south and central-south metapopulations, but not differentiated between west-central, west-south, central-south, and north-central isolates. An occurrence of IR6 and IR7 displays that possibly a gene flow of Acanthamoeba T4 occurred after the founder effect or bottleneck experience through ecological changes or host mobility. This is the first

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

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M. L. Gertiser

2010-06-01

Full Text Available 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 parasitaria se realizó una biopsia de la zona lesionada, que se destinó al análisis bacteriológico y parasitológico en el laboratorio. También se remitieron los líquidos de lavado y el estuche de las lentes. No se detectó crecimiento bacteriano en las muestras. Los cultivos para la búsqueda de Acanthamoeba resultaron positivos en todos los especímenes analizados. Las amebas aisladas fueron clasificadas morfológica y molecularmente como pertenecientes al género Acanthamoeba. El presente trabajo muestra el primer caso de queratitis por Acanthamoeba de la ciudad de Bahía Blanca identificado por el laboratorio mediante procedimientos de alto valor diagnóstico.Some species of the Acanthamoeba genus cause keratitis, a very painful, most likely unilateral corneal infection , associated with eye and vision impairment. We here present a case of a 31-year-old female patient, a regular user of soft contact lenses without good practices of lens hygiene and handling. The patient attended medical consultation after two months of inflammation and pain in her right eye. After ophthalmological studies, and due to suspicion of a parasitic infection, a biopsy was performed and the sample submitted for bacteriological and parasitological analyses. Moreover, contact lens holders and lens cleaning solutions were studied. The samples yielded negative results for bacterial infection. However, cultivation of all samples showed the presence of amoeboid parasites

Acanthamoeba keratitis: study of the 5-year incidence in Israel.

Science.gov (United States)

Graffi, Shmuel; Peretz, Avi; Jabaly, Haneen; Koiefman, Anna; Naftali, Modi

2013-11-01

Acanthamoeba keratitis (AK) is not a notifiable disease in Israel, so there are no accurate incidence rates for this condition in Israel. The aim of this study was to estimate the incidence of AK in Israel for the years 2008-2012. We distributed a survey questionnaire to laboratory managers in Israel. The laboratories were affiliated to medical institutes that either provided ophthalmology services or served community ophthalmology clinics. Our questionnaire requested survey respondents to provide information regarding the methods used to diagnose AK, and the number of positive and negative cultures for Acanthamoebae species performed for each of the years from 2008 to 2012. Six laboratories used non-nutrient agar with Escherichia coli as the culture medium, one used calcofluor-white staining with fluorescent microscopy, and two used PCR for diagnosing AK. Twenty-three AK cases were identified, to give an estimated incidence of 1/1 668 552. AK is mostly attributable to the use of contact lenses. As contact lenses are popular in Israel, we expected a higher incidence rate. A lower than expected incidence rate may indicate insufficient awareness of AK in Israel.

Deciphering the genomes of 16 Acanthamoeba species does not provide evidence of integration of known giant virus-associated mobile genetic elements.

Science.gov (United States)

Chelkha, Nisrine; Colson, Philippe; Levasseur, Anthony; La Scola, Bernard

2018-06-02

Giant viruses infect protozoa, especially amoebae of the genus Acanthamoeba. These viruses possess genetic elements named Mobilome. So far, this mobilome comprises provirophages which are integrated into the genome of their hosts, transpovirons, and Maverick/Polintons. Virophages replicate inside virus factories within Acanthamoeba and can decrease the infectivity of giant viruses. The virophage infecting CroV was found to be integrated in the host of CroV, Cafeteria roenbergensis, thus protecting C. roenbergensis by reduction of CroV multiplication. Because of this unique property, assessment of the mechanisms of replication of virophages and their relationship with giant viruses is a key element of this investigation. This work aimed at evaluating the presence and the dynamic of these mobile elements in sixteen Acanthamoeba genomes. No significant traces of the integration of genomes or sequences from known virophages were identified in all the available Acanthamoeba genomes. These results brought us to hypothesize that the interactions between mimiviruses and their virophages might occur through different mechanisms, or at low frequency. An additional explanation could be that our knowledge of the diversity of virophages is still very limited. Copyright © 2018 Elsevier B.V. All rights reserved.

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

International Nuclear Information System (INIS)

Maruta, H.; Korn, E.D.

1981-01-01

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 0 0 C. 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

Comparison of Fluorescence Microscopy and Different Growth Media Culture Methods for Acanthamoeba Keratitis Diagnosis.

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Peretz, Avi; Geffen, Yuval; Socea, Soergiu D; Pastukh, Nina; Graffi, Shmuel

2015-08-01

Acanthamoeba keratitis (AK), a potentially blinding infection of the cornea, is caused by a free-living protozoan. Culture and microscopic examination of corneal scraping tissue material is the conventional method for identifying Acanthamoeba. In this article, we compared several methods for AK diagnosis of 32 patients: microscopic examination using fluorescent dye, specific culture on growth media-non-nutrient agar (NNA), culture on liquid growth media-peptone yeast glucose (PYG), and TYI-S-33. AK was found in 14 patients. Thirteen of the specimens were found AK positive by fluorescence microscopic examination, 11 specimens were found AK positive on PYG growth media, and 9 specimens were found AK positive on TYI-S-33 growth media. Only five specimens were found AK positive on NNA growth media. Therefore, we recommend using fluorescence microscopy technique and culture method, especially PYG liquid media. © The American Society of Tropical Medicine and Hygiene.

Possible environmental sources of Acanthamoeba spp in contact lens wearers.

OpenAIRE

Seal, D; Stapleton, F; Dart, J

1992-01-01

The water supply and dust samples from the home environment (bathrooms and kitchens) of 50 wearers of contact lenses (CLs) were cultured for the presence of free-living amoebae. CL cases, solutions, and water taps were cultured for bacteria, which amoebae require for growth. Acanthamoeba spp were isolated from water drawn from six bathroom cold water taps (tank supplied), five in the presence of limescale, and from one kitchen cold water tap (mains supplied). There was an association between ...

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

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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

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

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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.

Evaluation of loop-mediated isothermal amplification assay for rapid diagnosis of Acanthamoeba keratitis

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Abhishek Mewara

2017-01-01

Full Text Available Background: The clinical features of Acanthamoeba keratitis (AK are non-specific and closely resemble bacterial, viral and fungal keratitis. Materials and Methods: We compared loop-mediated isothermal amplification (LAMP with microscopy, non-nutrient agar (NNA culture and polymerase chain reaction (PCR in clinical suspects of AK. Results: Of 52 clinical samples (42 AK suspects and 10 proven bacterial, viral or fungal keratitis, 3 were positive by direct microscopy (sensitivity 60%, confidence interval [CI]: 17%–92.7%, and 5 by NNA culture, 18S rDNA PCR and LAMP (sensitivity 100%, CI: 46.3%–100%. The limit of detection of Acanthamoeba DNA was 1 pg/μl by both LAMP and PCR. Conclusion: PCR and LAMP assays targeting 18S rDNA gene were found particularly suitable for a rapid and accurate diagnosis of AK. LAMP assay takes 2–3 h lesser than PCR, and thus offers a rapid, highly sensitive and specific, simple and affordable diagnostic modality for patients suspected of AK, especially in resource limited settings

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

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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.

Molecular and morphological characterization of Acanthamoeba isolated from corneal scrapes and contact lens wearers in Argentina.

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Casero, Rodolfo D; Mongi, Florencia; Laconte, Laura; Rivero, Fernando; Sastre, Dario; Teherán, Aníbal; Herrera, Giovanny; Ramírez, Juan David

2017-10-01

In this study, we describe the frequency of Acanthamoeba keratitis (AK) in patients that assisted in the Ophthalmology Department and determine the species/genotypes of free living amoebas (FLA) isolates. FLA from Corneal scrapes (CS) and contact lens (CL) wearers were studied by morphological and molecular characterization. A database was constructed with sociodemographic, clinical findings and history of use of CL variables. During January 2000 and September 2016 patients with corneal pathology admitted to the Ophthalmology Service of the University Hospital in Córdoba city, Argentina were included in the study. FLA were detected in 1.5% (11/739) and in 17% (11/65) of CS and CL analyzed respectively. FLA isolates from CL users evidenced an 80.9% of inappropriate lens maintenance, 4.8% (1/21) were not CL users that have been in contact with waters in outdoor environment and 14,3% (3/21) with no data about CL users. Acanthamoeba was confirmed in 100% and 82% of CS and LC respectively. The most frequent symptom associated with AK was red eye and photophobia. FLA from CS belonged to group II but 82% (9/11) and 18% (2/11) from CL belonged to group II and III respectively. T4 genotype and A. polyphaga species were detected in 100% of Acanthamoeba isolates. Poor CL hygiene practices, highlights the need for improved education about the severity of AK and consequences of improper CL hygiene. Genotype T4 detected in 100% of both CS and CL samples, consistently with previous findings indicating that this genotype is by far the most prevalent isolated from ocular infection. Copyright © 2017 Elsevier B.V. All rights reserved.

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

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Anirvan Chatterjee

2016-09-01

Full Text Available 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.

Ultradian metronome: timekeeper for orchestration of cellular coherence.

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Lloyd, David; Murray, Douglas B

2005-07-01

Dynamic intracellular spatial and temporal organization emerges from spontaneous synchronization of a massive array of weakly coupled oscillators; the majority of subcellular processes are implicated in this integrated expression of cellular physiology. Evidence for this view comes mainly from studies of Saccharomyces cerevisiae growing in self-synchronized continuous cultures, in which a temperature-compensated ultradian clock (period of approximately 40 min) couples fermentation with redox state in addition to the transcriptome and cell-division-cycle progression. Functions for ultradian clocks have also been determined in other yeasts (e.g. Schizosaccharomyces pombe and Candida utilis), seven protists (e.g. Acanthamoeba castellanii and Paramecium tetraurelia), as well as cultured mammalian cells. We suggest that ultradian timekeeping is a basic universal necessity for coordinated intracellular coherence.

Molecular characterization of Acanthamoeba strains isolated from domestic dogs in Tenerife, Canary Islands, Spain.

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Valladares, María; Reyes-Batlle, María; Martín-Navarro, Carmen M; López-Arencibia, Atteneri; Dorta-Gorrín, Alexis; Wagner, Carolina; Martínez-Carretero, Enrique; Piñero, José E; Valladares, Basilio; Lorenzo-Morales, Jacob

2015-06-01

The present study describes two cases of Acanthamoeba infections (keratitis and ascites/peritonitis) in small breed domestic dogs in Tenerife, Canary Islands, Spain. In both cases, amoebic trophozoites were observed under the inverted microscope and isolated from the infected tissues and/or fluids, without detecting the presence of other viral, fungal or bacterial pathogens. Amoebae were isolated using 2 % non-nutrient agar plates and axenified for further biochemical and molecular analyses. Osmotolerance and thermotolerance assays revealed that both isolates were able to grow up to 37 °C and 1 M of mannitol and were thus considered as potentially pathogenic. Moreover, the strains were classified as highly cytotoxic as they cause more than 75 % of toxicity when incubated with two eukaryotic cell lines. In order to classify the strains at the molecular level, the diagnostic fragment 3 (DF3) region of the 18S rDNA of Acanthamoeba was amplified and sequenced, revealing that both isolates belonged to genotype T4. In both cases, owners of the animals did not allow any further studies or follow-up and therefore the current status of these animals is unknown. Furthermore, the isolation of these pathogenic amoebae should raise awareness with the veterinary community locally and worldwide.

Revisiting the Acanthamoeba species that form star-shaped cysts (genotypes T7, T8, T9, and T17): characterization of seven new Brazilian environmental isolates and phylogenetic inferences.

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Magliano, Ana C M; Teixeira, Marta M G; Alfieri, Silvia C

2012-01-01

Free-living amoebae of the genus Acanthamoeba are the agents of both opportunistic and non-opportunistic infections and are frequently isolated from the environment. Of the 17 genotypes (T1-T17) identified thus far, 4 (T7, T8, T9, and T17) accommodate the rarely investigated species of morphological group I, those that form large, star-shaped cysts. We report the isolation and characterization of 7 new Brazilian environmental Acanthamoeba isolates, all assigned to group I. Phylogenetic analyses based on partial (~1200 bp) SSU rRNA gene sequences placed the new isolates in the robustly supported clade composed of the species of morphological group I. One of the Brazilian isolates is closely related to A. comandoni (genotype T9), while the other 6, together with 2 isolates recently assigned to genotype T17, form a homogeneous, well-supported group (2·0% sequence divergence) that likely represents a new Acanthamoeba species. Thermotolerance, osmotolerance, and cytophatic effects, features often associated with pathogenic potential, were also examined. The results indicated that all 7 Brazilian isolates grow at temperatures up to 40°C, and resist under hyperosmotic conditions. Additionally, media conditioned by each of the new Acanthamoeba isolates induced the disruption of SIRC and HeLa cell monolayers.

Resistance of Acanthamoeba Cysts to Disinfection Treatments Used in Health Care Settings▿

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Coulon, Céline; Collignon, Anne; McDonnell, Gerald; Thomas, Vincent

2010-01-01

Free-living amoebae that belong to the genus Acanthamoeba are widespread in the environment, including water. They are responsible for human infections and can host pathogenic microorganisms. Under unfavorable conditions, they form cysts with high levels of resistance to disinfection methods, thus potentially representing a threat to public health. In the present study we evaluated the efficacies of various biocides against trophozoites and cysts of several Acanthamoeba strains. We demonstrated that disinfectant efficacy varied depending on the strains tested, with environmental strains demonstrating greater resistance than collection strains. Trophozoites were inactivated by all treatments except those using glutaraldehyde as an active compound: for these treatments, we observed resistance even after 30 min exposure. Cysts resisted many treatments, including certain conditions with glutaraldehyde and other biocides. Moist heat at 55°C was not efficient against cysts, whereas exposure at 65°C was. Several chemical formulations containing peracetic acid, hydrogen peroxide, or ortho-phthalaldehyde presented greater efficacy than glutaraldehyde, as did ethanol and sodium hypochlorite; however, some of these treatments required relatively long incubation times to achieve cyst inactivation. Amoebal cysts can be highly resistant to some high-level disinfectants, which has implications for clinical practice. These results highlight the need to consider the effective disinfection of protozoa in their vegetative and resistant forms due to their intrinsic resistance. This is important not only to prevent the transmission of protozoa themselves but also due to the risks associated with a range of microbial pathogens that are found to be associated intracellularly with these microorganisms. PMID:20519477

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

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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.

Morphological characteristics of developmental stages of Acanthamoeba and Naegleria species before and after staining by various techniques.

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Ithoi, Init; Ahmad, Arine-Fadzlun; Mak, J W; Nissapatorn, Veeranoot; Lau, Yee-Ling; Mahmud, Rohela

2011-11-01

Seven stains were studied to determine the best color and contrast for staining the developmental stages of free living pathogenic Acanthamoeba and Naegleria species. The acid-fast bacilli stain (AFB) produced a blue color without contrast; trichrome-eosin and modified Field's showed various color contrasts; Giemsa, iron-hematoxylin, modified AFB and Gram produced only one color which distinguished the nucleus, nucleolus, cytoplasm, food- and water-vacuoles. The motile organs (acanthopodia, pseudopodia, lobopodia and flagella) were also clearly differentiated but produced a similar color as the cytoplasm. These motile organelles were first induced by incubating at 37 degrees C for at least 15 minutes and then fixing with methanol in order to preserve the protruding morphology prior to staining. The trichrome-eosin and iron-hematoxylin stains showed good color contrast for detecting all three stages, the trophozoite, cyst and flagellate; Giemsa and Gram stained the trophozoite and flagellate stages; the modified Field's and modified AFB stains stained only the trophozoite stage. Depending on the purpose, all these stains (except the AFB stain) can be used to identify the developmental stages of Acanthamoeba and Naegleria for clinical, epidemiological or public health use.

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

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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...

UV inactivation of pathogenic and indicator microorganisms

International Nuclear Information System (INIS)

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

1985-01-01

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

UV inactivation of pathogenic and indicator microorganisms

Energy Technology Data Exchange (ETDEWEB)

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.

Visual outcome in Japanese patients with Acanthamoeba keratitis.

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Yamazoe, K; Yamamoto, Y; Shimazaki-Den, S; Shimazaki, J

2012-04-01

To identify prognostic factors affecting visual outcome in Acanthamoeba keratitis (AK) treated with topical chlorhexidine gluconate (CHG). A total of 35 eyes in 34 patients with AK were treated with 0.02% topical CHG. Patients were divided into two groups according to the final visual outcome: Group 1, final visual acuity (VA) of 20/25 or greater (22 eyes); Group 2, less than 20/25 (13 eyes). We compared these groups and evaluated the effectiveness of topical CHG compared with outcomes in previous reports. Ring infiltrate was observed more often in Group 2 (4.5% vs 61.5%, OR 33.6, 95% confidence interval (CI) 3.4-333.9, PAK was significantly longer (24.9 days vs 48.4 days, OR 1.03, 95% CI 1.00-1.06, P = 0.04) and VA at initial examination (log MAR) significantly lower (0.47 vs 1.59, OR 25.5, 95% CI 3.4-186.7, PAK. Topical CHG was comparably effective to other treatments, including polyhexamethyl biguanide and propamidine isethionate.

Free-living Acanthamoeba and Naegleria spp. amebae in water sources of León, Nicaragua

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Byron Leiva

2008-06-01

Full Text Available free-living amebae (FLA are known to occur worldwide in water-related biotopes, but only limited information is available on these organisms in developing countries and so far no information on their presence is available from Nicaragua. The aims of this study were to evaluate the prevalence of potentially pathogenic Acanthamoeba spp. and Naegleria spp. in different water sources to which the population of León municipality is exposed. Since pathogenic amebae are thermotolerant, we were especially interested in the occurrence of FLA in geothermal areas. Water samples were collected from León area in Nicaragua: 88 samples were from rivers and springs, 111 from wells, 74 from water taps and 21 from water tanks in urban and suburban León and from three nearby geothermal areas of San Jacinto, Posoltega and Tipitapa. Amebae were identified using morphological and physiological criteria, immunohistochemical staining procedures and molecular methods. Indirect immunofluorescent test was performed on cysts and trophozoites fixed on microscopical slides and incubated for 30 min at room temperature in separate experiments with the following antibodies: rabbit-anti N. fowleri/N. lovanensis (Nf-Pab, mouse monoclonal antibody anti N. fowleri (Nf-5D12u, rabbit antibodies against Acanthamoeba spp. And fluorescent in situ hybridization (FISH was performed using 18S rRNA-targeted fluorescent oligonucleotide probes. Probes: GSP for the detection of Acanthamoeba and NAEG1088 for the detection of Naegleria. Free-living amebae were recovered from approximately 43 % of the samples. Acanthamoeba spp was found in 21 % of samples from León municipality and in 2 % of samples from geothermal areas. Amoeboflagellates were found in 10 % of samples from León and in 19 % in geothermal areas. Fifty three percent of tested wells in the geothermal area contained thermotolerant amoeboflagellates. Naegleria spp. was identified in 24 out of 39 (61.5 % of isolated

[Survey of the number of Acanthamoeba keratitis cases in Japan].

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Toriyama, Koji; Suzuki, Takashi; Ohashi, Yuichi

2014-01-01

To investigate the trend in the number of Acanthamoeba keratitis (AK) cases in Japan. A survey was conducted in 48 university hospitals. Patients who were diagnosed with AK from January 2007 to December 2011 were enrolled. The trend in the number of cases and the type of contact lenses (CLs) that patients used were studied. A total of 524 patients was studied. The numbers of AK cases in each year, from 2007 to 2011, were 105, 152, 155, 65, and 47. The number dropped markedly after 2009. The percentage of conventional soft CLs and frequent replacement soft CL users that needed daily care such as rubbing-washing also dropped after 2008. The number of AK cases in Japan has been decreasing in recent years. The cause is uncertain, but one possibility is that information about proper CL care promulgated by ophthalmic societies in recent years is producing results.

Prognostic factors in Acanthamoeba keratitis.

Science.gov (United States)

Kaiserman, Igor; Bahar, Irit; McAllum, Penny; Srinivasan, Sathish; Elbaz, Uri; Slomovic, Allan R; Rootman, David S

2012-06-01

To assess the prognostic factors influencing visual prognosis and length of treatment after acanthamoeba keratitis (AK). Forty-two AK eyes of 41 patients treated between 1999 and 2006 were included. A diagnosis of AK was made on the basis of culture results with a corresponding clinical presentation. We calculated the prognostic effect of the various factors on final visual acuity and the length of treatment. Multivariate regression analysis was used to adjust for the simultaneous effects of the various prognostic factors. Mean follow-up was 19.7 ± 21.0 months. Sixty-four percent of cases had > 1 identified risk factor for AK, the most common risk factor being contact lens wear (92.9% of eyes). At presentation, median best spectacle corrected visual acuity (BCVA) was 20/200 (20/30 to Hand Motion [HM]) that improved after treatment to 20/50 (20/20 to Counting Fingers [CF]). Infection acquired by swimming or related to contact lenses had significantly better final BCVA (p = 0.03 and p = 0.007, respectively). Neuritis and pseudodendrites were also associated with better final BCVA (p = 0.04 and p = 0.05, respectively). Having had an epithelial defect on presentation and having been treated with topical steroid were associated with worse final best spectacle corrected visual acuity (BSCVA) (p = 0.0006 and p = 0.04). Multivariate regression analysis found a good initial visual acuity (p = 0.002), infections related to swimming (p = 0.01), the absence of an epithelial defect (p = 0.03), having been treated with chlorhexidine (p = 0.05), and not having receive steroids (p = 0.003) to significantly forecast a good final BCVA. We identified several prognostic factors that can help clinicians evaluate the expected visual damage of the AK infection and thus tailor treatment accordingly. Copyright © 2012 Canadian Ophthalmological Society. All rights reserved.

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

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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.

Protozoa Drive the Dynamics of Culturable Biocontrol Bacterial Communities.

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Müller, Maren Stella; Scheu, Stefan; Jousset, Alexandre

2013-01-01

Some soil bacteria protect plants against soil-borne diseases by producing toxic secondary metabolites. Such beneficial biocontrol bacteria can be used in agricultural systems as alternative to agrochemicals. The broad spectrum toxins responsible for plant protection also inhibit predation by protozoa and nematodes, the main consumers of bacteria in soil. Therefore, predation pressure may favour biocontrol bacteria and contribute to plant health. We analyzed the effect of Acanthamoeba castellanii on semi-natural soil bacterial communities in a microcosm experiment. We determined the frequency of culturable bacteria carrying genes responsible for the production of the antifungal compounds 2,4-diacetylphloroglucinol (DAPG), pyrrolnitrin (PRN) and hydrogen cyanide (HCN) in presence and absence of A. castellanii. We then measured if amoebae affected soil suppressiveness in a bioassay with sugar beet seedlings confronted to the fungal pathogen Rhizoctonia solani. Amoebae increased the frequency of both DAPG and HCN positive bacteria in later plant growth phases (2 and 3 weeks), as well as the average number of biocontrol genes per bacterium. The abundance of DAPG positive bacteria correlated with disease suppression, suggesting that their promotion by amoebae may enhance soil health. However, the net effect of amoebae on soil suppressiveness was neutral to slightly negative, possibly because amoebae slow down the establishment of biocontrol bacteria on the recently emerged seedlings used in the assay. The results indicate that microfaunal predators foster biocontrol bacterial communities. Understanding interactions between biocontrol bacteria and their predators may thus help developing environmentally friendly management practices of agricultural systems.

Protozoa Drive the Dynamics of Culturable Biocontrol Bacterial Communities.

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Maren Stella Müller

Full Text Available Some soil bacteria protect plants against soil-borne diseases by producing toxic secondary metabolites. Such beneficial biocontrol bacteria can be used in agricultural systems as alternative to agrochemicals. The broad spectrum toxins responsible for plant protection also inhibit predation by protozoa and nematodes, the main consumers of bacteria in soil. Therefore, predation pressure may favour biocontrol bacteria and contribute to plant health. We analyzed the effect of Acanthamoeba castellanii on semi-natural soil bacterial communities in a microcosm experiment. We determined the frequency of culturable bacteria carrying genes responsible for the production of the antifungal compounds 2,4-diacetylphloroglucinol (DAPG, pyrrolnitrin (PRN and hydrogen cyanide (HCN in presence and absence of A. castellanii. We then measured if amoebae affected soil suppressiveness in a bioassay with sugar beet seedlings confronted to the fungal pathogen Rhizoctonia solani. Amoebae increased the frequency of both DAPG and HCN positive bacteria in later plant growth phases (2 and 3 weeks, as well as the average number of biocontrol genes per bacterium. The abundance of DAPG positive bacteria correlated with disease suppression, suggesting that their promotion by amoebae may enhance soil health. However, the net effect of amoebae on soil suppressiveness was neutral to slightly negative, possibly because amoebae slow down the establishment of biocontrol bacteria on the recently emerged seedlings used in the assay. The results indicate that microfaunal predators foster biocontrol bacterial communities. Understanding interactions between biocontrol bacteria and their predators may thus help developing environmentally friendly management practices of agricultural systems.

Non-contact lens related Acanthamoeba keratitis

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Prashant Garg

2017-01-01

Full Text Available The purpose of the study is to describe epidemiology, clinical features, diagnosis, and treatment of Acanthamoeba keratitis (AK with special focus on the disease in nonusers of contact lenses (CLs. This study was a perspective based on authors' experience and review of published literature. AK accounts for 2% of microbiology-proven cases of keratitis. Trauma and exposure to contaminated water are the main predisposing factors for the disease. Association with CLs is seen only in small fraction of cases. Contrary to classical description experience in India suggests that out of proportion pain, ring infiltrate, and radial keratoneuritis are seen in less than a third of cases. Majority of cases present with diffuse infiltrate, mimicking herpes simplex or fungal keratitis. The diagnosis can be confirmed by microscopic examination of corneal scraping material and culture on nonnutrient agar with an overlay of Escherichia coli. Confocal microscopy can help diagnosis in patients with deep infiltrate; however, experience with technique and interpretation of images influences its true value. Primary treatment of the infection is biguanides with or without diamidines. Most patients respond to medical treatment. Corticosteroids play an important role in the management and can be used when indicated after due consideration to established protocols. Surgery is rarely needed in patients where definitive management is initiated within 3 weeks of onset of symptoms. Lamellar keratoplasty has been shown to have good outcome in cases needing surgery. Since the clinical features of AK in nonusers of CL are different, it will be important for ophthalmologists to be aware of the scenario wherein to suspect this infection. Medical treatment is successful if the disease is diagnosed early and management is initiated soon.

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

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Yanet Romero

Full Text Available 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.

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

DEFF Research Database (Denmark)

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

2012-01-01

-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......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......, heat treated or osmotically stressed bacteria than with control bacteria. Also, while similar to 1.5 x 10(3) colony forming unit/ml internalized bacteria could typically be recovered 24 h post-gentamicin treatment with control bacteria, no starved, heat treated or osmotically stressed bacteria could...

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

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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

Legionella pneumophila transcriptome during intracellular multiplication in human macrophages

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Sebastien P Faucher

2011-04-01

Full Text Available 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 (TFBSS, which is used to translocate more than 150 effector proteins to host cells. While the transcriptional response of Legionella to the intracellular environment of A. castellanii has been investigated, much less is known about the Legionella transcriptional response inside human macrophages. In this study, the transcriptome of L. pneumophila was monitored during exponential and post-exponential phase in rich AYE broth as well as during infection of human cultured macrophages. This was accomplished with microarrays and an RNA amplification procedure called SCOTS to detect small amounts of mRNA from low numbers of intracellular bacteria. Among the genes induced intracellularly are those involved in amino acid biosynthetic pathways leading to L-arginine, L-histidine and L-proline as well as many transport systems involved in amino acid and iron uptake. Gene involved in catabolism of glycerol is also induced during intracellular growth and could be used as a carbon source. The genes encoding the Icm/Dot system are not differentially expressed inside cells compared to control bacteria grown in rich broth, but the genes encoding several translocated effectors are strongly induced. Moreover, we used the transcriptome data to predict previously unrecognized Icm/Dot effector genes based on their expression pattern and confirmed translocation for three candidates. This study provides a comprehensive view of how L. pneumophila responds to the human macrophage intracellular environment.

One Year Genome Evolution of Lausannevirus in Allopatric versus Sympatric Conditions.

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Mueller, Linda; Bertelli, Claire; Pillonel, Trestan; Salamin, Nicolas; Greub, Gilbert

2017-06-01

Amoeba-resisting microorganisms raised a great interest during the last decade. Among them, some large DNA viruses present huge genomes up to 2.5 Mb long, exceeding the size of small bacterial genomes. The rate of genome evolution in terms of mutation, deletion, and gene acquisition in these genomes is yet unknown. Given the suspected high plasticity of viral genomes, the microevolution of the 346 kb genome of Lausannevirus, a member of Megavirales, was studied. Hence, Lausannevirus was co-cultured within the amoeba Acanthamoeba castellanii over one year. Despite a low number of mutations, the virus showed a genome reduction of 3.7% after 12 months. Lausannevirus genome evolution in sympatric conditions was investigated by its co-culture with Estrella lausannensis, an obligate intracellular bacterium, in the amoeba A. castellanii during one year. Cultures were split every 3 months. Genome sequencing revealed that in these conditions both, Lausannevirus and E. lausannensis, show stable genome, presenting no major rearrangement. In fact, after one year they acquired from 2 to 7 and from 4 to 10 mutations per culture for Lausannevirus and E. lausannensis, respectively. Interestingly, different mutations in the endonuclease encoding genes of Lausannevirus were observed in different subcultures, highlighting the importance of this gene product in the replication of Lausannevirus. Conversely, mutations in E. lausannensis were mainly located in a gene encoding for a phosphoenolpyruvate-protein phosphotransferase (PtsI), implicated in sugar metabolism. Moreover, in our conditions and with our analyses we detected no horizontal gene transfer during one year of co-culture. © The Author(s) 2017. Published by Oxford University Press on behalf of the Society for Molecular Biology and Evolution.

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

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

2015-05-01

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

Studio del comportamento di Acanthamoeba. polyphaga in presenza di Legionella pneumophila e di altri batteri ad habitat acquatico

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M. Bondi

2003-05-01

Full Text Available Le amebe a vita libera sono state oggetto di diversi studi negli ultimi anni, non solo per le loro potenzialità patogene nei confronti dell’uomo, ma anche per l’importante ruolo che svolgono in natura, dove agiscono come predatori in grado di controllare le popolazioni batteriche. Alcuni degli organismi fagocitati però possono evitare la lisi fagosomiale e mantenere la loro condizione vitale a livello intracellulare, divenendo endosimbionti. Le amebe fungono così da riserva per questi batteri, proteggendoli da difficili condizioni extracellulari e provvedendo ad un ambiente consono alla loro replicazione. Tale tipo di interazione è particolarmente studiata in Legionella pneumophila, dal momento che l’ampia diffusione di questo germe, nonché la sua virulenza, pare siano fortemente influenzate dalla capacità di parassitare protozoi appartenenti ai generi Acanthamoeba, Naegleria e Balamuthia. Al fine di ottenere maggiori informazioni sui fattori favorenti o inibenti lo sviluppo di questi protozoi, è stato studiato il comportamento di un ceppo di Acanthamoeba polyphaga coltivato, in solido e in liquido, in associazione con L. pneumophila ed altri batteri ad habitat acquatico (Pseudomonas, Aeromonas, Achromobacter, Burkholderia. Su tappeti di cellule batteriche allestiti in Non Nutrient Agar (NNA, A.polyphaga si è mostrata in grado di moltiplicarsi utilizzando come nutrimento tutti i ceppi testati, nonostante alcuni, come Burkholderia cepacia SSV6 e Achromobacter xylox SS28, risultino più idonei al suo sviluppo. In piastre a pozzetti addizionate di acqua condottata autoclavata, il protozoo ha mostrato una buona capacità di sopravvivenza, non risultando inoltre influenzato dalla presenza di legionella o dei batteri acquatici testati. Dal momento che, fra i batteri descritti come capaci di vita intra-amebica, sono inclusi patogeni quali Chlamydia, Legionella, Listeria e Rickettsiae, risulta necessario riconsiderare la rilevanza clinica

Interstrain polymorphisms of isoenzyme profiles and mitochondrial DNA fingerprints among seven strains assigned to Acanthamoeba polyphaga.

Science.gov (United States)

Kong, H H; Park, J H; Chung, D I

1995-12-01

Interstrain polymorphisms of isoenzyme profiles and mitochondrial (Mt) DNA fingerprints were observed among seven strains of Acanthamoeba isolated from different sources and morphologically assigned to A. polyphaga. Mt DNA fingerprints by eight restriction endonucleases (Bgl II, Sca I, Cla I, EcoR I, Xba I, Kpn I, Sal I, and Sst I) revealed considerable interstrain polymorphisms. Isoenzyme profiles revealed considerable interstrain polymorphisms for acid phosphatase, lactate dehydrogenase, and glucose-6-phosphate dehydrogenase while those for glucose phosphate isomerase, leucine aminopeptidase, and malate dehydrogenase showed similarity. Despite of the interstrain polymorphisms, the isoenzyme profiles and Mt DNA fingerprints of the strain Ap were found to be identical with those of the strain Jones. Mt DNA fingerprinting was found to be highly applicable for the strain identification, characterization, and differentiation.

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

LENUS (Irish Health Repository)

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.

Survey on medicinal plants traditionally used in Senegal for the treatment of tuberculosis (TB) and assessment of their antimycobacterial activity.

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Diop, ElHadji Assane; Queiroz, Emerson Ferreira; Kicka, Sébastien; Rudaz, Serge; Diop, Tahir; Soldati, Thierry; Wolfender, Jean-Luc

2018-04-24

In West Africa, populations are used to taking traditional medicine as a first aid against common health problems. In this aspect, many plants are claimed to be effective in the treatment of Tuberculosis (TB), which according to the World Health Organization (WHO) remains one of the world's deadliest communicable diseases. The main aim of this study was to identify plants used to treat TB-symptoms by the population of Senegal and to evaluate their possible concomitant use with clinically approved TB-drugs. This approach allowed the selection of plants effectively used in traditional medicine. In order to verify if the usage of some of these plants can be rationalized, the activity of their traditional preparations was assessed with both an intracellular and extracellular antimycobacterial host-pathogen assays. An ethnopharmacological survey conducted on 117 TB-patients and 30 healers in Senegal from March to May 2014. The questionnaires were focused on the use of medicinal plants to treat common TB -symptoms (cough longer than 2 weeks, fever, night sweats, weight loss and bloody sputum). Local plant names, utilized organs (herbal drugs) and traditional formulations of the plants were recorded. Extracts were prepared by mimicking the traditional decoction in boiling water and screened for their antimycobacterial activity using Mycobacterium marinum, as a validated TB surrogate, and an Acanthamoeba castellanii - M. marinum whole-cell based host-pathogen assay, to detect anti-infective activities. By the end of the survey, nearly 30 plants were cited and the 12 most cited herbal drugs were collected and their usage documented by extensive literature search. Extracts of the chosen herbs were screened with the described assays; with a main focus on traditional formulas (mainly herbal decoctions). Two of the water extracts from Combretum aculeatum and Guiera senegalensis showed significant antimycobacterial activities when compared to the positive control drug (rifampin

Distinct DNA exit and packaging portals in the virus Acanthamoeba polyphaga mimivirus.

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Zauberman, Nathan; Mutsafi, Yael; Halevy, Daniel Ben; Shimoni, Eyal; Klein, Eugenia; Xiao, Chuan; Sun, Siyang; Minsky, Abraham

2008-05-13

Icosahedral double-stranded DNA viruses use a single portal for genome delivery and packaging. The extensive structural similarity revealed by such portals in diverse viruses, as well as their invariable positioning at a unique icosahedral vertex, led to the consensus that a particular, highly conserved vertex-portal architecture is essential for viral DNA translocations. Here we present an exception to this paradigm by demonstrating that genome delivery and packaging in the virus Acanthamoeba polyphaga mimivirus occur through two distinct portals. By using high-resolution techniques, including electron tomography and cryo-scanning electron microscopy, we show that Mimivirus genome delivery entails a large-scale conformational change of the capsid, whereby five icosahedral faces open up. This opening, which occurs at a unique vertex of the capsid that we coined the "stargate", allows for the formation of a massive membrane conduit through which the viral DNA is released. A transient aperture centered at an icosahedral face distal to the DNA delivery site acts as a non-vertex DNA packaging portal. In conjunction with comparative genomic studies, our observations imply a viral packaging pathway akin to bacterial DNA segregation, which might be shared by diverse internal membrane-containing viruses.

Distinct DNA exit and packaging portals in the virus Acanthamoeba polyphaga mimivirus.

Directory of Open Access Journals (Sweden)

Nathan Zauberman

2008-05-01

Full Text Available Icosahedral double-stranded DNA viruses use a single portal for genome delivery and packaging. The extensive structural similarity revealed by such portals in diverse viruses, as well as their invariable positioning at a unique icosahedral vertex, led to the consensus that a particular, highly conserved vertex-portal architecture is essential for viral DNA translocations. Here we present an exception to this paradigm by demonstrating that genome delivery and packaging in the virus Acanthamoeba polyphaga mimivirus occur through two distinct portals. By using high-resolution techniques, including electron tomography and cryo-scanning electron microscopy, we show that Mimivirus genome delivery entails a large-scale conformational change of the capsid, whereby five icosahedral faces open up. This opening, which occurs at a unique vertex of the capsid that we coined the "stargate", allows for the formation of a massive membrane conduit through which the viral DNA is released. A transient aperture centered at an icosahedral face distal to the DNA delivery site acts as a non-vertex DNA packaging portal. In conjunction with comparative genomic studies, our observations imply a viral packaging pathway akin to bacterial DNA segregation, which might be shared by diverse internal membrane-containing viruses.

The rising tide of Acanthamoeba keratitis in Auckland, New Zealand: a 7-year review of presentation, diagnosis and outcomes (2009-2016).

Science.gov (United States)

McKelvie, James; Alshiakhi, Moaz; Ziaei, Mohammed; Patel, Dipika V; McGhee, Charles Nj

2018-02-07

Acanthamoeba is an increasingly prevalent cause of vision-threatening microbial keratitis. To assess the incidence, clinical presentation, diagnosis and outcomes of patients with Acanthamoeba keratitis (AK) in Auckland, New Zealand over a 7-year period. Retrospective observational consecutive case series. Fifty-eight eyes of 52 patients diagnosed with AK. All cases of AK were identified using a cross-referenced search of clinical, laboratory and pharmacy records from March 2009 to May 2016. Demographic and clinical data were collected including age, gender, risk factors, clinical manifestations, initial diagnosis, diagnostic investigations, treatment, presenting and final visual acuity and surgical interventions. Contact lens (CL) use was noted in 96% of unilateral and 100% of bilateral cases. The mean duration of symptoms at presentation was 21 days and the mean duration from presentation to definitive diagnosis was 14 days. Initial diagnosis was recorded as CL-related keratitis in 70.6%, viral keratitis in 15.5% and AK in 12.0%. The diagnosis was confirmed with In vivo confocal microscopy (IVCM) in 67.2%, corneal scrape in 22.4%, corneal biopsy in 1.7% and clinically in 8.6%. IVCM sensitivity was 83.0%. Surgical intervention was required in four patients, all with delayed diagnosis (range 63-125 days). The incidence of AK has more than doubled when compared with the preceding 7-year period. AK is a rare vision-threatening protozoal infection with rapidly-increasing incidence in New Zealand, predominantly affecting CL users. Diagnosis is often challenging and when delayed is associated with worse outcomes. IVCM offers rapid diagnosis with high sensitivity. © 2018 Royal Australian and New Zealand College of Ophthalmologists.

Multiple identification of most important waterborne protozoa in surface water used for irrigation purposes by 18S rRNA amplicon-based metagenomics.

Science.gov (United States)

Moreno, Y; Moreno-Mesonero, L; Amorós, I; Pérez, R; Morillo, J A; Alonso, J L

2018-01-01

Understanding waterborne protozoan parasites (WPPs) diversity has important implications in public health. In this study, we evaluated a NGS-based method as a detection approach to identify simultaneously most important WPPs using 18S rRNA high-throughput sequencing. A set of primers to target the V4 18S rRNA region of WPPs such as Cryptosporidium spp., Giardia sp., Blastocystis sp., Entamoeba spp, Toxoplasma sp. and free-living amoebae (FLA) was designed. In order to optimize PCR conditions before sequencing, both a mock community with a defined composition of representative WPPs and a real water sample inoculated with specific WPPs DNA were prepared. Using the method proposed in this study, we have detected the presence of Giardia intestinalis, Acanthamoeba castellanii, Toxoplasma gondii, Entamoeba histolytica and Blastocystis sp. at species level in real irrigation water samples. Our results showed that untreated surface irrigation water in open fields can provide an important source of WPPs. Therefore, the methodology proposed in this study can establish a basis for an accurate and effective diagnostic of WPPs to provide a better understanding of the risk associated to irrigation water. Copyright © 2017 The Authors. Published by Elsevier GmbH.. All rights reserved.

An investigation of virulence factors of Legionella pneumophila environmental isolates

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Elif Özlem Arslan-Aydoğdu

Full Text Available ABSTRACT Nine Legionella pneumophila strains isolated from cooling towers and a standard strain (L. pneumophila serogroup 1, ATCC 33152, Philadelphia 1 were analyzed and compared in terms of motility, flagella structure, ability to form biofilms, enzymatic activities (hemolysin, nucleases, protease, phospholipase A, phospholipase C, acid phosphatase, alkaline phosphatase and lipase, hemagglutination capabilities, and pathogenicity in various host cells (Acanthamoeba castellanii ATCC 30234, mouse peritoneal macrophages and human peripheral monocytes. All the isolates of bacteria appeared to be motile and polar-flagellated and possessed the type-IV fimbria. Upon the evaluation of virulence factors, isolate 4 was found to be the most pathogenic strain, while 6 out of the 9 isolates (the isolates 1, 2, 3, 4, 5, and 7 were more virulent than the ATCC 33152 strain. The different bacterial strains exhibited differences in properties such as adhesion, penetration and reproduction in the hosts, and preferred host type. To our knowledge, this is the first study to compare the virulence of environmental L. pneumophila strains isolated in Turkey, and it provides important information relevant for understanding the epidemiology of L. pneumophila.

Characterization of the efficiency and uncertainty of skimmed milk flocculation for the simultaneous concentration and quantification of water-borne viruses, bacteria and protozoa.

Science.gov (United States)

Gonzales-Gustavson, Eloy; Cárdenas-Youngs, Yexenia; Calvo, Miquel; da Silva, Marcelle Figueira Marques; Hundesa, Ayalkibet; Amorós, Inmaculada; Moreno, Yolanda; Moreno-Mesonero, Laura; Rosell, Rosa; Ganges, Llilianne; Araujo, Rosa; Girones, Rosina

2017-03-01

In this study, the use of skimmed milk flocculation (SMF) to simultaneously concentrate viruses, bacteria and protozoa was evaluated. We selected strains of faecal indicator bacteria and pathogens, such as Escherichia coli and Helicobacter pylori. The viruses selected were adenovirus (HAdV 35), rotavirus (RoV SA-11), the bacteriophage MS2 and bovine viral diarrhoea virus (BVDV). The protozoa tested were Acanthamoeba, Giardia and Cryptosporidium. The mean recoveries with q(RT)PCR were 66% (HAdV 35), 24% (MS2), 28% (RoV SA-11), 15% (BVDV), 60% (E. coli), 30% (H. pylori) and 21% (Acanthamoeba castellanii). When testing the infectivity, the mean recoveries were 59% (HAdV 35), 12% (MS2), 26% (RoV SA-11) and 0.7% (BVDV). The protozoa Giardia lamblia and Cryptosporidium parvum were studied by immunofluorescence with recoveries of 18% and 13%, respectively. Although q(RT)PCR consistently showed higher quantification values (as expected), q(RT)PCR and the infectivity assays showed similar recoveries for HAdV 35 and RoV SA-11. Additionally, we investigated modelling the variability and uncertainty of the recovery with this method to extrapolate the quantification obtained by q(RT)PCR and estimate the real concentration. The 95% prediction intervals of the real concentration of the microorganisms inoculated were calculated using a general non-parametric bootstrap procedure adapted in our context to estimate the technical error of the measurements. SMF shows recoveries with a low variability that permits the use of a mathematical approximation to predict the concentration of the pathogen and indicator with acceptable low intervals. The values of uncertainty may be used for a quantitative microbial risk analysis or diagnostic purposes. Copyright © 2017 The Author(s). Published by Elsevier B.V. All rights reserved.

Detection limits of Legionella pneumophila in environmental samples after co-culture with Acanthamoeba polyphaga

Science.gov (United States)

2013-01-01

Background The efficiency of recovery and the detection limit of Legionella after co-culture with Acanthamoeba polyphaga are not known and so far no investigations have been carried out to determine the efficiency of the recovery of Legionella spp. by co-culture and compare it with that of conventional culturing methods. This study aimed to assess the detection limits of co-culture compared to culture for Legionella pneumophila in compost and air samples. Compost and air samples were spiked with known concentrations of L. pneumophila. Direct culturing and co-culture with amoebae were used in parallel to isolate L. pneumophila and recovery standard curves for both methods were produced for each sample. Results The co-culture proved to be more sensitive than the reference method, detecting 102-103 L. pneumophila cells in 1 g of spiked compost or 1 m3 of spiked air, as compared to 105-106 cells in 1 g of spiked compost and 1 m3 of spiked air. Conclusions Co-culture with amoebae is a useful, sensitive and reliable technique to enrich L. pneumophila in environmental samples that contain only low amounts of bacterial cells. PMID:23442526

Protozoan Cysts Act as a Survival Niche and Protective Shelter for Foodborne Pathogenic Bacteria

Science.gov (United States)

Lambrecht, Ellen; Baré, Julie; Chavatte, Natascha; Bert, Wim; Sabbe, Koen

2015-01-01

The production of cysts, an integral part of the life cycle of many free-living protozoa, allows these organisms to survive adverse environmental conditions. Given the prevalence of free-living protozoa in food-related environments, it is hypothesized that these organisms play an important yet currently underinvestigated role in the epidemiology of foodborne pathogenic bacteria. Intracystic bacterial survival is highly relevant, as this would allow bacteria to survive the stringent cleaning and disinfection measures applied in food-related environments. The present study shows that strains of widespread and important foodborne bacteria (Salmonella enterica, Escherichia coli, Yersinia enterocolitica, and Listeria monocytogenes) survive inside cysts of the ubiquitous amoeba Acanthamoeba castellanii, even when exposed to either antibiotic treatment (100 μg/ml gentamicin) or highly acidic conditions (pH 0.2) and resume active growth in broth media following excystment. Strain- and species-specific differences in survival periods were observed, with Salmonella enterica surviving up to 3 weeks inside amoebal cysts. Up to 53% of the cysts were infected with pathogenic bacteria, which were located in the cyst cytosol. Our study suggests that the role of free-living protozoa and especially their cysts in the persistence and epidemiology of foodborne bacterial pathogens in food-related environments may be much more important than hitherto assumed. PMID:26070667

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

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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.

The Legionella pneumophila Collagen-Like Protein Mediates Sedimentation, Autoaggregation, and Pathogen-Phagocyte Interactions

Science.gov (United States)

Abdel-Nour, Mena; Duncan, Carla; Prashar, Akriti; Rao, Chitong; Ginevra, Christophe; Jarraud, Sophie; Low, Donald E.; Ensminger, Alexander W.; Terebiznik, Mauricio R.

2014-01-01

Although only partially understood, multicellular behavior is relatively common in bacterial pathogens. Bacterial aggregates can resist various host defenses and colonize their environment more efficiently than planktonic cells. For the waterborne pathogen Legionella pneumophila, little is known about the roles of autoaggregation or the parameters which allow cell-cell interactions to occur. Here, we determined the endogenous and exogenous factors sufficient to allow autoaggregation to take place in L. pneumophila. We show that isolates from Legionella species which do not produce the Legionella collagen-like protein (Lcl) are deficient in autoaggregation. Targeted deletion of the Lcl-encoding gene (lpg2644) and the addition of Lcl ligands impair the autoaggregation of L. pneumophila. In addition, Lcl-induced autoaggregation requires divalent cations. Escherichia coli producing surface-exposed Lcl is able to autoaggregate and shows increased biofilm production. We also demonstrate that L. pneumophila infection of Acanthamoeba castellanii and Hartmanella vermiformis is potentiated under conditions which promote Lcl dependent autoaggregation. Overall, this study shows that L. pneumophila is capable of autoaggregating in a process that is mediated by Lcl in a divalent-cation-dependent manner. It also reveals that Lcl potentiates the ability of L. pneumophila to come in contact, attach, and infect amoebae. PMID:24334670

Intermediary metabolism in protists: a sequence-based view of facultative anaerobic metabolism in evolutionarily diverse eukaryotes.

Science.gov (United States)

Ginger, Michael L; Fritz-Laylin, Lillian K; Fulton, Chandler; Cande, W Zacheus; Dawson, Scott C

2010-12-01

Protists account for the bulk of eukaryotic diversity. Through studies of gene and especially genome sequences the molecular basis for this diversity can be determined. Evident from genome sequencing are examples of versatile metabolism that go far beyond the canonical pathways described for eukaryotes in textbooks. In the last 2-3 years, genome sequencing and transcript profiling has unveiled several examples of heterotrophic and phototrophic protists that are unexpectedly well-equipped for ATP production using a facultative anaerobic metabolism, including some protists that can (Chlamydomonas reinhardtii) or are predicted (Naegleria gruberi, Acanthamoeba castellanii, Amoebidium parasiticum) to produce H(2) in their metabolism. It is possible that some enzymes of anaerobic metabolism were acquired and distributed among eukaryotes by lateral transfer, but it is also likely that the common ancestor of eukaryotes already had far more metabolic versatility than was widely thought a few years ago. The discussion of core energy metabolism in unicellular eukaryotes is the subject of this review. Since genomic sequencing has so far only touched the surface of protist diversity, it is anticipated that sequences of additional protists may reveal an even wider range of metabolic capabilities, while simultaneously enriching our understanding of the early evolution of eukaryotes. Copyright © 2010 Elsevier GmbH. All rights reserved.

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

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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

Molecular identification of t4 and t5 genotypes in isolates from acanthamoeba keratitis patients.

Science.gov (United States)

Ledee, D R; Iovieno, A; Miller, D; Mandal, N; Diaz, M; Fell, J; Fini, M E; Alfonso, E C

2009-05-01

Acanthamoeba keratitis (AK) is a rare but sight-threatening ocular infection. Outbreaks have been associated with contaminated water and contact lens wear. The epidemiology and pathology may be associated with unique genotypes. We determined the Rns genotype for 37 clinical isolates from 23 patients presenting at the University of Miami Bascom Palmer Eye Institute with confirmed AK infections in 2006 to 2008. The genus-specific ASA.S1 amplicon allowed for rapid genotyping of the nonaxenic cultures. Of the 37 isolates, 36 were of the T4 genotype. Within this group, 13 unique diagnostic fragment 3 sequences were identified, 3 of which were not in GenBank. The 37th isolate was a T5, the first in the United States and second worldwide to be found in AK. For five patients with isolates from the cornea and contact lens/case, identical sequences within each patient cluster were observed, confirming the link between contact lens contamination and AK infection. Genotyping is an important tool in the epidemiological study of AK. In this study, it allowed for the detection of new strains and provided an etiological link between source and infection. Additionally, it can allow for accurate categorizing of physiological differences, such as strain virulence, between isolates and clades.

Resurgence of Acanthamoeba keratitis in Auckland, New Zealand: a 7-year review of presentation and outcomes.

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Patel, Dipika V; Rayner, Sandra; McGhee, Charles N J

2010-01-01

To investigate the presentation, clinical characteristics and outcomes of Acanthamoeba keratitis (AK) in Auckland, New Zealand over a 7-year period. Retrospective analysis of all cases of AK treated by the tertiary corneal service at Auckland City Hospital/ University of Auckland Department of Ophthalmology (August 2001 to May 2008). Data were collected regarding age, gender, contact lens history, presenting signs and symptoms, diagnosis at first presentation, time to final diagnosis, identifiable risk factors, presenting and final visual acuity, results of microbiological testing, medical treatment, surgical interventions, recurrence of disease and length of follow up. All photographs and in vivo confocal microscopy images were reviewed. Twenty-five eyes of 25 patients were identified with a diagnosis of AK (mean age 40 +/- 13 years). Ninety-six per cent were contact lens wearers. Mean time to diagnosis was 41 +/- 49 days (range 0-181 days, median 21 days). Fourteen patients (56%) had been treated with topical corticosteroids prior to the diagnosis. Early diagnosis of AK (New Zealand in the current decade. This study highlights the fundamental importance of early diagnosis and appropriate management in ensuring favourable outcomes. Practitioners should maintain a clinical suspicion of AK, especially as 96% of the subjects in this study were contact lens wearers.

Gold Nanoparticle Conjugation Enhances the Antiacanthamoebic Effects of Chlorhexidine

Science.gov (United States)

Aqeel, Yousuf; Siddiqui, Ruqaiyyah; Anwar, Ayaz; Shah, Muhammad Raza

2015-01-01

Acanthamoeba keratitis is a serious infection with blinding consequences and often associated with contact lens wear. Early diagnosis, followed by aggressive topical application of drugs, is a prerequisite in successful treatment, but even then prognosis remains poor. Several drugs have shown promise, including chlorhexidine gluconate; however, host cell toxicity at physiologically relevant concentrations remains a challenge. Nanoparticles, subcolloidal structures ranging in size from 10 to 100 nm, are effective drug carriers for enhancing drug potency. The overall aim of the present study was to determine whether conjugation with gold nanoparticles enhances the antiacanthamoebic potential of chlorhexidine. Gold-conjugated chlorhexidine nanoparticles were synthesized. Briefly, gold solution was mixed with chlorhexidine and reduced by adding sodium borohydride, resulting in an intense deep red color, indicative of colloidal gold-conjugated chlorhexidine nanoparticles. The synthesis was confirmed using UV-visible spectrophotometry that shows a plasmon resonance peak of 500 to 550 nm, indicative of gold nanoparticles. Further characterization using matrix-assisted laser desorption ionization-mass spectrometry showed a gold-conjugated chlorhexidine complex at m/z 699 ranging in size from 20 to 100 nm, as determined using atomic force microscopy. To determine the amoebicidal and amoebistatic effects, amoebae were incubated with gold-conjugated chlorhexidine nanoparticles. For controls, amoebae also were incubated with gold and silver nanoparticles alone, chlorhexidine alone, neomycin-conjugated nanoparticles, and neomycin alone. The findings showed that gold-conjugated chlorhexidine nanoparticles exhibited significant amoebicidal and amoebistatic effects at 5 μM. Amoebicidal effects were observed by parasite viability testing using a Trypan blue exclusion assay and flow-cytometric analysis using propidium iodide, while amoebistatic effects were observed using growth

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

DEFF Research Database (Denmark)

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

2014-01-01

) in 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...

Acanthamoeba keratitis in 194 patients: risk factors for bad outcomes and severe inflammatory complications.

Science.gov (United States)

Carnt, Nicole; Robaei, Dana; Minassian, Darwin C; Dart, John K G

2018-01-03

To determine demographic and clinical features of patients with Acanthamoeba keratitis (AK) that are independent risk factors both for bad outcomes and for severe inflammatory complications (SIC). A retrospective audit of medical records of AK cases at Moorfields Eye Hospital from July 2000 to April 2012, including 12 earlier surgical cases. Cases with a bad outcome were defined as those having one or more of the following: corneal perforation, keratoplasty, other surgery (except biopsy), duration of antiamoebic therapy (AAT) ≥10.5 months (the 75th percentile of the whole cohort) and final visual acuity ≤20/80. SICs were defined as having scleritis and/or a stromal ring infiltrate. Multivariable analysis was used to identify independent risk factors for both bad outcomes and SICs. Records of 194 eyes (194 patients) were included, having bad outcomes in 93 (48%). Bad outcomes were associated with the presence of SIC, aged >34 years, corticosteroids used before giving AAT and symptom duration >37 days before AAT. The development of SIC was independently associated with aged >34 years, corticosteroids used before giving AAT and herpes simplex virus (HSV) keratitis treatment before AAT. The prompt diagnosis of AK, avoidance of a misdiagnosis of HSV keratitis and corticosteroid use before the exclusion of AK as a potential cause of keratitis are essential to the provision of a good outcome for patients and for the avoidance of SIC. Older age is an unmodifiable risk factor that may reflect differences in the immune response to AK in this patient subset. © Article author(s) (or their employer(s) unless otherwise stated in the text of the article) 2018. All rights reserved. No commercial use is permitted unless otherwise expressly granted.

Exploring Anti-Bacterial Compounds against Intracellular Legionella

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Harrison, Christopher F.; Kicka, Sébastien; Trofimov, Valentin; Berschl, Kathrin; Ouertatani-Sakouhi, Hajer; Ackermann, Nikolaus; Hedberg, Christian; Cosson, Pierre; Soldati, Thierry; Hilbi, Hubert

2013-01-01

Legionella pneumophila is a ubiquitous fresh-water bacterium which reproduces within its erstwhile predators, environmental amoeba, by subverting the normal pathway of phagocytosis and degradation. The molecular mechanisms which confer resistance to amoeba are apparently conserved and also allow replication within macrophages. Thus, L. pneumophila can act as an ‘accidental’ human pathogen and cause a severe pneumonia known as Legionnaires’ disease. The intracellular localisation of L. pneumophila protects it from some antibiotics, and this fact must be taken into account to develop new anti-bacterial compounds. In addition, the intracellular lifestyle of L. pneumophila may render the bacteria susceptible to compounds diminishing bacterial virulence and decreasing intracellular survival and replication of this pathogen. The development of a single infection cycle intracellular replication assay using GFP-producing L. pneumophila and Acanthamoeba castellanii amoeba is reported here. This fluorescence-based assay allows for continuous monitoring of intracellular replication rates, revealing the effect of bacterial gene deletions or drug treatment. To examine how perturbations of the host cell affect L. pneumophila replication, several known host-targeting compounds were tested, including modulators of cytoskeletal dynamics, vesicle scission and Ras GTPase localisation. Our results reveal a hitherto unrealized potential antibiotic property of the β-lactone-based Ras depalmitoylation inhibitor palmostatin M, but not the closely related inhibitor palmostatin B. Further characterisation indicated that this compound caused specific growth inhibition of Legionella and Mycobacterium species, suggesting that it may act on a common bacterial target. PMID:24058631

An atypical proprotein convertase in Giardia lamblia differentiation.

Science.gov (United States)

Davids, B J; Gilbert, M A; Liu, Q; Reiner, D S; Smith, A J; Lauwaet, T; Lee, C; McArthur, A G; Gillin, F D

2011-02-01

Proteolytic activity is important in the lifecycles of parasites and their interactions with hosts. Cysteine proteases have been best studied in Giardia, but other protease classes have been implicated in growth and/or differentiation. In this study, we employed bioinformatics to reveal the complete set of putative proteases in the Giardia genome. We identified 73 peptidase homologs distributed over 5 catalytic classes in the genome. Serial analysis of gene expression of the G. lamblia lifecycle found thirteen protease genes with significant transcriptional variation over the lifecycle, with only one serine protease transcript upregulated late in encystation. The translated gene sequence of this encystation-specific transcript was most similar to eukaryotic subtilisin-like proprotein convertases (SPC), although the typical catalytic triad was not identified. Epitope-tagged gSPC protein expressed in Giardia under its own promoter was upregulated during encystation with highest expression in cysts and it localized to encystation-specific secretory vesicles (ESV). Total gSPC from encysting cells produced proteolysis in gelatin gels that co-migrated with the epitope-tagged protease in immunoblots. Immuno-purified gSPC also had gelatinase activity. To test whether endogenous gSPC activity is involved in differentiation, trophozoites and cysts were exposed to the specific serine proteinase inhibitor 4-(2-aminoethyl)-benzenesulfonyl fluoride hydrochloride (AEBSF). After 21 h encystation, a significant decrease in ESV was observed with 1mM AEBSF and by 42 h the number of cysts was significantly reduced, but trophozoite growth was not inhibited. Concurrently, levels of cyst wall proteins 1 and 2, and AU1-tagged gSPC protein itself were decreased. Excystation of G. muris cysts was also significantly reduced in the presence of AEBSF. These results support the idea that serine protease activity is essential for Giardia encystation and excystation. Copyright © 2010 Elsevier B

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

DEFF Research Database (Denmark)

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

2002-01-01

of the 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...... microcosms, the active and total populations of ciliates, their potential prey (bacteria and small protozoa), their potential competitors (amoebae, flagellates, and nematodes), and their potential predators (nematodes). We sampled with short time intervals (2 to 6 days) and generated a data set, suitable...

Genome Segregation and Packaging Machinery in Acanthamoeba polyphaga Mimivirus Is Reminiscent of Bacterial Apparatus

Science.gov (United States)

Chelikani, Venkata; Ranjan, Tushar; Zade, Amrutraj; Shukla, Avi

2014-01-01

ABSTRACT Genome packaging is a critical step in the virion assembly process. The putative ATP-driven genome packaging motor of Acanthamoeba polyphaga mimivirus (APMV) and other nucleocytoplasmic large DNA viruses (NCLDVs) is a distant ortholog of prokaryotic chromosome segregation motors, such as FtsK and HerA, rather than other viral packaging motors, such as large terminase. Intriguingly, APMV also encodes other components, i.e., three putative serine recombinases and a putative type II topoisomerase, all of which are essential for chromosome segregation in prokaryotes. Based on our analyses of these components and taking the limited available literature into account, here we propose for the first time a model for genome segregation and packaging in APMV that can possibly be extended to NCLDV subfamilies, except perhaps Poxviridae and Ascoviridae. This model might represent a unique variation of the prokaryotic system acquired and contrived by the large DNA viruses of eukaryotes. It is also consistent with previous observations that unicellular eukaryotes, such as amoebae, are melting pots for the advent of chimeric organisms with novel mechanisms. IMPORTANCE Extremely large viruses with DNA genomes infect a wide range of eukaryotes, from human beings to amoebae and from crocodiles to algae. These large DNA viruses, unlike their much smaller cousins, have the capability of making most of the protein components required for their multiplication. Once they infect the cell, these viruses set up viral replication centers, known as viral factories, to carry out their multiplication with very little help from the host. Our sequence analyses show that there is remarkable similarity between prokaryotes (bacteria and archaea) and large DNA viruses, such as mimivirus, vaccinia virus, and pandoravirus, in the way that they process their newly synthesized genetic material to make sure that only one copy of the complete genome is generated and is meticulously placed inside

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

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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.

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

Science.gov (United States)

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

2013-04-01

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

Disseminated Acanthamoeba Infection Presenting With Cutaneous Lesions in an Immunocompromised Patient: A Case Report, Review of Histomorphologic Findings, and Potential Diagnostic Pitfalls.

Science.gov (United States)

Morrison, Annie O; Morris, Robert; Shannon, Amie; Lauer, Scott R; Guarner, Jeannette; Kraft, Colleen S

2016-02-01

Free-living amoebas are exceedingly rare causes of cutaneous infections and present unique diagnostic and therapeutic challenges. We describe a case of disseminated acanthamoebiasis with cutaneous manifestations and summarize additional diagnostic, prognostic, and therapeutic highlights. A 58-year-old man with relapsed chronic lymphocytic leukemia had several weeks of progressive, painful ulcerations on the forehead, arms, abdomen, and thighs. A biopsy was performed for histopathologic evaluation. The biopsy specimen showed inflammatory infiltrate with abscess formation involving the epidermis, dermis, and subcutis. Scattered cells showed nuclei with a prominent central karyosome, dispersed chromatin, and either abundant foamy basophilic cytoplasm or two well-demarcated cytoplasmic walls. Acanthamoeba species was confirmed by polymerase chain reaction from the formalin-fixed, paraffin-embedded tissue. Cutaneous lesions from acanthamoebiasis are exceptionally rare but should be included in the differential diagnosis of necrotic cutaneous lesions in immunocompromised patients. Although infrequently encountered, pathologists need to be aware of the morphologic features of free-living amoebas. Immunohistochemical and molecular studies can confirm the diagnosis. Multiagent treatment regimens, when initiated empirically, have been more successful than single-agent regimens, but infections involving the central nervous system are almost universally fatal. © American Society for Clinical Pathology, 2016. All rights reserved. For permissions, please e-mail: journals.permissions@oup.com.

The membrane protein LasM Promotes the Culturability of Legionella pneumophila in Water

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Laam Li

2016-09-01

Full Text Available The water-borne pathogen Legionella pneumophila (Lp strongly expresses the lpg1659 gene in water. This gene encodes a hypothetical protein predicted to be a membrane protein using in silico analysis. While no conserved domains were identified in Lpg1659, similar proteins are found in many Legionella species and other aquatic bacteria. RT-qPCR showed that lpg1659 is positively regulated by the alternative sigma factor RpoS, which is essential for Lp to survive in water. These observations suggest an important role of this novel protein in the survival of Lp in water. Deletion of lpg1659 did not affect cell morphology, membrane integrity or tolerance to high temperature. Moreover, lpg1659 was dispensable for growth of Lp in rich medium, and during infection of the amoeba Acanthamoeba castellanii and of THP-1 human macrophages. However, deletion of lpg1659 resulted in an early loss of culturability in water, while over-expression of this gene promoted the culturability of Lp. Therefore, these results suggest that lpg1659 is required for Lp to maintain culturability, and possibly long-term survival, in water. Since the loss of culturability observed in the absence of Lpg1659 was complemented by the addition of trace metals into water, this membrane protein is likely a transporter for acquiring essential trace metal for maintaining culturability in water and potentially in other metal-deprived conditions. Given its role in the survival of Lp in water, Lpg1659 was named LasM for Legionella aquatic survival membrane protein.

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

Directory of Open Access Journals (Sweden)

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

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

International Nuclear Information System (INIS)

Torian, B.E.; Stroeher, V.L.; Stamm, W.E.; Flores, B.M.; Hagen, F.S.

1990-01-01

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 35 S-labeled live trophozoites. The surface location of the antigen was corroborated by mAb immunoprecipitation of a 29-kDa protein from surface- 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

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

Energy Technology Data Exchange (ETDEWEB)

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.

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

Science.gov (United States)

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

2012-04-01

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

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?

Science.gov (United States)

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

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, barely controlled by topical medication. Actually, the physical features and the wearing time characteristics of the disposable soft contact lenses created unprecedented difficulties to the medical surveillance of their wearers. Without the right assistance they tend to become careless regarding routine cleaning. They also feel free to buy less expensive lenses, to use saline instead of lens solutions, to violate the limits of wearing time and to extend the use over the sleeping period. Additionally, the aggressive marketing and the wide distribution of these lenses increase the chances that economically or educationally unprepared people will acquire them. The question that remains is: Is the present case an accidental event or an example of what is likely to happen in the future if the indiscriminate selling of disposable soft contact lenses continues to evolve?

Acanthamoebicidal activity of periglaucine A and betulinic acid from Pericampylus glaucus (Lam.) Merr. in vitro.

Science.gov (United States)

Mahboob, Tooba; Azlan, Abdul-Majid; Shipton, Fiona Natalia; Boonroumkaew, Patcharaporn; Nor Azman, Nadiah Syafiqah; Sekaran, Shamala Devi; Ithoi, Init; Tan, Tian-Chye; Samudi, Chandramathi; Wiart, Christophe; Nissapatorn, Veeranoot

2017-12-01

Acanthamoeba species are pathogenic protozoa which account for amoebic keratitis, conjunctivitis and granulomatous amoebic encephalitis. These amoebae form cysts which resist drugs and more effective acanthamoebicidal agents are needed. Medicinal plants could be useful in improving the current treatment strategies for Acanthamoeba infections. In the present study, we examined the amoebicidal effects of Pericampylus glaucus (Lam.) Merr., a medicinal plant used for the treatment of conjunctivitis in Malaysia. Pathogenic Acanthamoeba triangularis were isolated from environmental water samples and treated with different concentrations of fractions obtained from Pericampylus glaucus (Lam.) Merr. as well as main constituents for 24-72 h. Chlorhexidine was used as a reference drug. Ethanol fraction of stem showed significant (p < 0.05) inhibition of trophozoites survival. Betulinic acid and periglaucine A from this plant at 100 μg/mL inhibited more than 70% survival of both cysts and trophozoites. The calculated therapeutic index for betulinic acid and periglaucine A was 170 and 1.5 for trophozoites stage and 3.75 and 8.5 for cysts stage. The observed amoebicidal efficacies indicate the beneficial aspects of this plant in the treatment of Acanthamoeba infection. Periglaucine A could also be of value for the treatment of Acanthamoeba infection. Copyright © 2017 Elsevier Inc. All rights reserved.

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

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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

In the case of transmission of Mycobacterium ulcerans in Buruli ulcer ...

African Journals Online (AJOL)

Also, Acanthamoeba-pathogen association has a long evolutionary history because the same set of bacterial genes and gene products e.g. in L. pneumophila are required for survival in both mammalian and protozoan host cells. We suggest that the involvement of Acanthamoeba in the transmission of M. ulcerans to ...

Production and characterization of monoclonal antibodies against cathepsin B and cathepsin B-Like proteins of Naegleria fowleri.

Science.gov (United States)

Seong, Gi-Sang; Sohn, Hae-Jin; Kang, Heekyoung; Seo, Ga-Eun; Kim, Jong-Hyun; Shin, Ho-Joon

2017-12-01

Naegleria fowleri causes fatal primary amoebic meningoencephalitis (PAM) in humans and experimental animals. In previous studies, cathepsin B (nfcpb) and cathepsin B-like (nfcpb-L) genes of N. fowleri were cloned, and it was suggested that refolding rNfCPB and rNfCPB-L proteins could play important roles in host tissue invasion, immune response evasion and nutrient uptake. In this study, we produced anti-NfCPB and anti-NfCPB-L monoclonal antibodies (McAb) using a cell fusion technique, and observed their immunological characteristics. Seven hybridoma cells secreting rNfCPB McAbs and three hybridoma cells secreting rNfCPB-L McAbs were produced. Among these, 2C9 (monoclone for rNfCPB) and 1C8 (monoclone for rNfCPB-L) McAb showed high antibody titres and were finally selected for use. As determined by western blotting, 2C9 McAb bound to N. fowleri lysates, specifically the rNfCPB protein, which had bands of 28 kDa and 38.4 kDa. 1C8 McAb reacted with N. fowleri lysates, specifically the rNfCPB-L protein, which had bands of 24 kDa and 34 kDa. 2C9 and 1C8 monoclonal antibodies did not bind to lysates of other amoebae, such as N. gruberi, Acanthamoeba castellanii and A. polyphaga in western blot analyses. Immuno-cytochemistry analysis detected NfCPB and NfCPB-L proteins in the cytoplasm of N. fowleri trophozoites, particularly in the pseudopodia and food-cup. These results suggest that monoclonal antibodies produced against rNfCPB and rNfCPB-L proteins may be useful for further immunological study of PAM. Copyright © 2017 Elsevier Inc. All rights reserved.

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

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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

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

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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

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

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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.

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?

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Sidney Júlio de Faria e Sousa

2008-06-01

Full Text Available 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, barely controlled by topical medication. Actually, the physical features and the wearing time characteristics of the disposable soft contact lenses created unprecedented difficulties to the medical surveillance of their wearers. Without the right assistance they tend to become careless regarding routine cleaning. They also feel free to buy less expensive lenses, to use saline instead of lens solutions, to violate the limits of wearing time and to extend the use over the sleeping period. Additionally, the aggressive marketing and the wide distribution of these lenses increase the chances that economically or educationally unprepared people will acquire them. The question that remains is: Is the present case an accidental event or an example of what is likely to happen in the future if the indiscriminate selling of disposable soft contact lenses continues to evolve?Este é o relato de uma paciente do sexo feminino, de 19 anos, com ceratite bilateral por Acanthamoeba, devido ao uso de lentes de contato descartáveis, adquiridas em uma butique. Fez uso do mesmo par de lentes durante três meses, sem os cuidados adequados. Foi submetida a transplante penetrante de córnea e facectomia bilateral, além de trabeculectomia no olho direito. Quando vista pela última vez, os enxertos estavam transparentes, mas a visão de ambos os olhos não era satisfatória. Ela também apresentava glaucoma secundário bilateral, controlado precariamente com medicação tópica. Ocorre

Dicty_cDB: Contig-U06541-1 [Dicty_cDB

Lifescience Database Archive (English)

Full Text Available 1 ( CW927554 ) EDCBM45TF A. castellanii, 6-8 kb library from tot... 44 7.5 1 ( CL534328 ) trib011xa13.g1 T. reesei HindIII BAC librar...y Hypo... 44 7.5 1 ( CL531533 ) trib005xd13.b1 T. reesei HindIII BAC library...CCNP644_b1 Cowpea UCR 779 Mixed Tissue an... 44 7.5 1 ( DQ121376 ) Uncultured bacterium clone YC01G10, parti..., ge... 44 7.5 1 ( CW927722 ) EDCBN39TR A. castellanii, 6-8 kb library from tot... 44 7.5 ...4 7.5 1 ( DW476283 ) GH_RMIRS_002_B08_F Cotton Normalized Library rand... 44 7.5 1 ( DR951133 ) EST1142672 Aquilegia cDNA library

Effects of X-ray on the metacestodes of Echinococcus granulosus in vitro.

Science.gov (United States)

Mao, Rui; Wu, Ge; Wang, Hui; Lu, Pengfei; Li, Jun; Li, Haitao; Ainiwaer, Aimudula; Bai, Yiwei; Shu, Mingyang; Bao, Yongxing; Zhang, Wenbao

2017-09-21

Radiotherapy may represent an alternative treatment modality for cystic echinococcosis (CE), but there is no adequate evidence for it up to now. In this study, we aim to investigate the parasiticidal effects of X-ray on the metacestodes of Echinococcus granulosus in vitro. Protoscoleces obtained from sheep naturally infected with CE were cultivated in RPMI 1640 medium containing 10% fetal bovine serum (FBS) at 37 °C in 5% CO 2 . Upon encystation on day 14, the metacestodes were subjected to various intensities of X-ray. Metacestode structures were observed using light microscope and transmission electron microscopy (TEM), and Real-Time PCR was carried out to determine the expression of EgTPX, EgHSP70, EgEPC1 and Caspase-3. On day 14, encystation was noticed in the majority of protoscoleces in the control group. In the X-ray groups, the encystation rate showed significant decrease compared with that of the control group (P granulosus. Irradiation triggered increased expression of EgTPX, EgHSP70, EgEPC1 and Caspase-3.

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

Science.gov (United States)

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

2013-12-01

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

Interactions of Bacteria and Amoebae with Ocular Biomaterials

OpenAIRE

John, Thomas

1991-01-01

The use of biomaterials in periocular and intraocular sites has resulted in some ocular inflammations and infections which can result in vision-threatening ocular disease. This review addresses bacterial interactions with, and adherence to ocular biomaterials such as soft contact lenses, surgical suture materials, and intraocular lenses. In addition, adherence of Acanthamoeba to soft contact lenses is described, and the role of these lenses in the development of Acanthamoeba keratitis is disc...

Nye diagnostiske tiltag ved akantamøbekeratitis

DEFF Research Database (Denmark)

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....

Survival and Virulence of Campylobacter spp. in the Environment

DEFF Research Database (Denmark)

Bui, Thanh Xuan

of environmental stress factors, namely heat shock, starvation, osmosis, and oxidation, on the expression of three virulence genes (ciaB, dnaJ, and htrA) of C. jejuni and its uptake by and intracellular survival within A. castellanii. I also investigated the mechanism(s) involved in phagocytosis and killing of C....... jejuni by A. castellanii. I observed that heat and osmotic stresses reduced the survival of C. jejuni significantly, whereas oxidative stress had no effect. The results of qRT-PCR experiments showed that the transcription of virulence genes of C. jejuni was slightly up-regulated under heat and oxidative...... soil flagellates may play a role for the survival of these food-borne pathogens on plant surfaces and in soil. It would be very interesting to further investigate the impacts of this soil flagellate on the survival of different food-borne pathogens in soil and in plant surface that may explain...

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

Directory of Open Access Journals (Sweden)

Chenyu Zhang

2009-05-01

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

Primary amebic meningoencephalitis due to Naegleria fowleri in a South American tapir.

Science.gov (United States)

Lozano-Alarcón, F; Bradley, G A; Houser, B S; Visvesvara, G S

1997-05-01

Naegleria fowleri, Acanthamoeba spp., and Balamuthia mandrillaris are known to cause fatal central nervous system (CNS) disease in human beings. N. fowleri causes acute, fulminating primary amebic meningoencephalitis (PAM), which generally leads to death within 10 days. Acanthamoeba spp. and B. mandrillaris cause chronic granulomatous amebic encephalitis, which may last for 8 weeks. Acanthamoeba spp. and B. mandrillaris also cause CNS disease in animals. N. fowleri, however, has been described only in human beings. This report is the first of PAM in an animal, a South American tapir. Dry cough, lethargy, and coma developed in the animal, and its condition progressed to death. At necropsy, lesions were seen in the cerebrum, cerebellum, and lungs. The CNS had severe, suppurative meningoencephalitis with many neutrophils, fibrin, plasma cells, and amebas. Amebas were 6.5 microns to 9 microns in diameter and had a nucleus containing a large nucleolus. Amebas in the sections reacted with a monoclonal antibody specific for N. fowleri in the immunofluorescent assay and appeared bright green.

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

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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.

Parasitic amoebae found in water bodies of Ukraine.

Science.gov (United States)

Patsyuk, Marina

2017-12-01

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

Dendritiform Keratopathy Associated with Exposure to Polyquarternium-1, a Common Ophthalmic Preservative.

Science.gov (United States)

Matoba, Alice Y; Peterson, Jeff R; Wilhelmus, Kirk R

2016-03-01

To describe dendritiform keratopathy associated with exposure to polyquaternium-1, a common preservative found in contact lens solutions and tear replacement products. Case series. Sixteen patients who demonstrated dendritiform keratopathy during topical ophthalmic exposure to polyquaternium-1. Records were reviewed of all patients diagnosed with dendritiform keratopathy between 1999 and 2014 who had documented exposure to contact lens care disinfecting solutions or artificial tear solutions containing polyquaternium-1. Patients were excluded who had coexisting potential causes for dendritiform keratopathy, such as prior herpes simplex keratitis, varicella-zoster viral keratitis, the linear form of Thygeson's superficial keratitis, epithelial regeneration line, Acanthamoeba keratitis, mucus plaque keratopathy, medication-related keratopathy, or limbal stem cell deficiency characterized by conjunctivalization of the corneal epithelium. Effect of discontinuation of exposure to polyquaternium-1 on the dendritiform keratopathy. Sixteen patients demonstrated dendritiform keratopathy after exposure to the preservative polyquaternium-1. Thirteen patients had a history of recent exposure to contact lens disinfecting solutions (Opti-Free, Equate) containing polyquaternium-1. Three patients used a tear replacement product (Systane) containing a polyquaternium-1 preservative. Four patients were treated with antiviral medications for presumed herpes simplex keratitis; 4 patients underwent diagnostic testing for Acanthamoeba keratitis. Two additional patients were diagnosed sequentially with herpes simplex keratitis, then Acanthamoeba keratitis before referral. All dendritiform lesions resolved within 2 to 6 weeks after elimination of exposure to polyquaternium-1. Ophthalmic products containing polyquaternium-1 may cause dendritiform keratopathy that may be confused with infections of the superficial cornea, such as herpes simplex virus keratitis or Acanthamoeba keratitis

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

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Parumon Noinarin

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

Experimental Analysis of Mimivirus Translation Initiation Factor 4a Reveals Its Importance in Viral Protein Translation during Infection of Acanthamoeba polyphaga.

Science.gov (United States)

Bekliz, Meriem; Azza, Said; Seligmann, Hervé; Decloquement, Philippe; Raoult, Didier; La Scola, Bernard

2018-05-15

The Acanthamoeba polyphaga mimivirus is the first giant virus ever described, with a 1.2-Mb genome which encodes 979 proteins, including central components of the translation apparatus. One of these proteins, R458, was predicted to initiate translation, although its specific role remains unknown. We silenced the R458 gene using small interfering RNA (siRNA) and compared levels of viral fitness and protein expression in silenced versus wild-type mimivirus. Silencing decreased the growth rate, but viral particle production at the end of the viral cycle was unaffected. A comparative proteomic approach using two-dimensional difference-in-gel electrophoresis (2D-DIGE) revealed deregulation of the expression of 32 proteins in silenced mimivirus, which were defined as up- or downregulated. Besides revealing proteins with unknown functions, silencing R458 also revealed deregulation in proteins associated with viral particle structures, transcriptional machinery, oxidative pathways, modification of proteins/lipids, and DNA topology/repair. Most of these proteins belong to genes transcribed at the end of the viral cycle. Overall, our data suggest that the R458 protein regulates the expression of mimivirus proteins and, thus, that mimivirus translational proteins may not be strictly redundant in relation to those from the amoeba host. As is the case for eukaryotic initiation factor 4a (eIF4a), the R458 protein is the prototypical member of the ATP-dependent DEAD box RNA helicase mechanism. We suggest that the R458 protein is required to unwind the secondary structures at the 5' ends of mRNAs and to bind the mRNA to the ribosome, making it possible to scan for the start codon. These data are the first experimental evidence of mimivirus translation-related genes, predicted to initiate protein biosynthesis. IMPORTANCE The presence in the genome of a mimivirus of genes coding for many translational processes, with the exception of ribosome constituents, has been the subject of

[Evaluation of chemokines in tears of patients with infectious keratitis].

Science.gov (United States)

Hori, Shinsuke; Shoji, Jun; Inada, Noriko; Sawa, Mitsuru

2013-02-01

To investigate the chemokine profile in tears of patients with infectious keratitis. Subjects were 32 eyes of 16 patients with infectious keratitis and 5 eyes of 5 healthy volunteers as a control. The patients with infectious keratitis were classified into two groups of eyes: 10 with bacterial keratitis and 6 with Acanthamoeba keratitis. Tear fluid was obtained from both eyes of the patients with infectious keratitis and from the right eyes of the control subjects using filter paper. Chemokine concentration (unit: Odu/mm2) and its profile in tears was analyzed using an antibody-array. In terms of chemokine profile in the bacterial keratitis group, the expression volume of interleukin-8 (IL-8) and monocyte chemoattractant protein-1 (MCP-1) in the diseased eyes was significantly higher than in the healthy eyes (p tears of the Acanthamoeba keratitis group. Regarding the chemokine ratio, the IL-8/MEC ratio in the diseased eyes of the Pseudomonas keratitis group and the MCP-1/IL-8 in the diseased eyes of the Acanthamoeba keratitis group showed a significantly high level (p tears of infectious keratitis patients is useful as a clinical tear laboratory test to interpret the pathologic condition of infectious keratitis

Comparison of bioindicator eukaryotes of activated sludge biocenoses on two water-treatment plants: a case study

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Achmadulina Farida Y.

2017-06-01

Full Text Available Activated sludge biocenoses were compared on waste-water treatment plants in the city of Kazan, Russian Federation and the city of Teplice, Czech Republic. Based on Palia-Kovnatski index, Acanthamoeba in Kazan, Epistylis in Teplice, and Acanthamoeba and Centropyxis were dominant genera in both plants. The major subdominant generas identified were Arcella, Opercularia and Aspidisca. This indicates high nitrification ability, high water purification potential and matured activated sludge. Chemical composition of the waste-water was identified as the main factor determining the sludge biocenoses diversity. Higher sludge biodiversity (Shannon, Margalef, and Sorensen indexes was found in Kazan corresponding to more concentrated inflow water.

Filling Knowledge Gaps for Mimivirus Entry, Uncoating, and Morphogenesis.

Science.gov (United States)

Andrade, Ana Cláudia Dos Santos Pereira; Rodrigues, Rodrigo Araújo Lima; Oliveira, Graziele Pereira; Andrade, Kétyllen Reis; Bonjardim, Cláudio Antônio; La Scola, Bernard; Kroon, Erna Geessien; Abrahão, Jônatas Santos

2017-11-15

Since the discovery of mimivirus, its unusual structural and genomic features have raised great interest in the study of its biology; however, many aspects concerning its replication cycle remain uncertain. In this study, extensive analyses of electron microscope images, as well as biological assay results, shed light on unclear points concerning the mimivirus replication cycle. We found that treatment with cytochalasin, a phagocytosis inhibitor, negatively impacted the incorporation of mimivirus particles by Acanthamoeba castellanii , causing a negative effect on viral growth in amoeba monolayers. Treatment of amoebas with bafilomicin significantly impacted mimivirus uncoating and replication. In conjunction with microscopic analyses, these data suggest that mimiviruses indeed depend on phagocytosis for entry into amoebas, and particle uncoating (and stargate opening) appears to be dependent on phagosome acidification. In-depth analyses of particle morphogenesis suggest that the mimivirus capsids are assembled from growing lamellar structures. Despite proposals from previous studies that genome acquisition occurs before the acquisition of fibrils, our results clearly demonstrate that the genome and fibrils can be acquired simultaneously. Our data suggest the existence of a specific area surrounding the core of the viral factory where particles acquire the surface fibrils. Furthermore, we reinforce the concept that defective particles can be formed even in the absence of virophages. Our work provides new information about unexplored steps in the life cycle of mimivirus. IMPORTANCE Investigating the viral life cycle is essential to a better understanding of virus biology. The combination of biological assays and microscopic images allows a clear view of the biological features of viruses. Since the discovery of mimivirus, many studies have been conducted to characterize its replication cycle, but many knowledge gaps remain to be filled. In this study, we conducted a

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

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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...

The neurotropic parasite Toxoplasma gondii increases dopamine metabolism

Science.gov (United States)

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...

Biofumigation for control of pale potato cyst nematodes: activity of brassica leaf extracts and green manures on Globodera pallida in vitro and in soil.

Science.gov (United States)

Lord, James S; Lazzeri, Luca; Atkinson, Howard J; Urwin, Peter E

2011-07-27

The effects of brassica green manures on Globodera pallida were assessed in vitro and in soil microcosms. Twelve of 22 brassica accessions significantly inhibited the motility of G. pallida infective juveniles in vitro. Green manures of selected brassicas were then incorporated into soil containing encysted eggs of G. pallida. Their effect on egg viability was estimated by quantifying nematode actin 1 mRNA by RT-qPCR. The leaf glucosinolate profiles of the plants were determined by high-performance liquid chromatography. Three Brassica juncea lines (Nemfix, Fumus, and ISCI99) containing high concentrations of 2-propenyl glucosinolate were the most effective, causing over 95% mortality of encysted eggs of G. pallida in polyethylene-covered soil. The toxic effects of green manures were greater in polyethylene-covered than in open soil. Toxicity in soil correlated with the concentration of isothiocyanate-producing glucosinolate but not total glucosinolate in green manures.

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

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

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

Science.gov (United States)

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

2017-01-01

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

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

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Farzana Abubakar Yousuf

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

Experiences with dormancy in tardigrades

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Deborah BOSCHINI

2004-09-01

Full Text Available Tardigrades often colonise extreme habitats, in which they survive using both types of dormancy: quiescence and diapause. Together with nematodes and bdelloid rotifers, tardigrades are known to enter quiescence (with several forms of cryptobiosis: anhydrobiosis, cryobiosis, anoxybiosis, osmobiosis at any stage of their life cycle, from egg to adult. Entering anhydrobiosis, tardigrades contract their body into a so-called tun, loosing most of their free and bound water (>95%, synthesizing cell protectants (e.g., trehalose, glycerol, heat shock proteins and strongly reducing or suspending their metabolism. Our research on cryptobiosis focused on some ecological and evolutionary aspects. We evaluated: i the long-term anhydrobiotic survival by comparing quantitative data on recovery from naturally induced desiccation in several species of tardigrades; ii differences in survival patterns between species and populations by experimentally inducing anhydrobiosis and cryobiosis; iii phenotypic factors affecting anhydrobiotic survival. As regards diapause, we considered encystment and eggs. Encystment involves at least the synthesis of new cuticular structures. Morphological changes during cyst formation are more complex than those involved in tun formation. We analyzed more in detail encystment processes, comparing a semiterrestrial with a limnic species. Several inter-specific differences have been identified, other than the production of two types of cysts in the semiterrestrial species. Our analysis of life history traits of a laboratory reared strain of a soil tardigrade revealed a particular hatching phenology that involved the production of both subitaneous and resting eggs. The latter need a cue to hatch (dehydration followed by re-hydration. In addition, the evolutionary meaning of dormancy in tardigrades is discussed.

Gene expression profiling during asexual development of the late blight pathogen Phytophthora infestans reveals a highly dynamic transcriptome

NARCIS (Netherlands)

Judelson, H.S.; Ah-Fong, A.M.V.; Aux, G.; Avrova, A.O.; Bruce, C.; Cakir, C.; Cunha, da L.; Grenville-Briggs, L.; Latijnhouwers, M.; Ligterink, W.; Meijer, H.J.G.; Roberts, S.; Thurber, C.S.; Whisson, S.C.; Birch, P.R.J.; Govers, F.; Kamoun, S.; West, van P.; Windass, J.

2008-01-01

Much of the pathogenic success of Phytophthora infestans, the potato and tomato late blight agent, relies on its ability to generate from mycelia large amounts of sporangia, which release zoospores that encyst and form infection structures. To better understand these stages, Affymetrix GeneChips

Induction of cell death in the testis of Heteracris littoralis by gamma rays

International Nuclear Information System (INIS)

Al-Taweel, A.A.; Shawkit, M.A.

1983-01-01

Cell killing by gamma rays was studied in Heteracris littoralis. Primary spermatocytes, which are encysted and have cytoplasmic connection, show unusual dose-response kinetics with no shoulder at low doses. Also, the spermatocytes do not die independently but in groups, usually with whole cysts degenerating synchronously. (author)

High Prevalence of Porocephalus crotali Infection on a Barrier Island (Cumberland Island) off the Coast of Georgia, with Identification of Novel Intermediate Hosts.

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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.

Developmental changes in the adhesive disk during Giardia differentiation.

Science.gov (United States)

Palm, Daniel; Weiland, Malin; McArthur, Andrew G; Winiecka-Krusnell, Jadwiga; Cipriano, Michael J; Birkeland, Shanda R; Pacocha, Sarah E; Davids, Barbara; Gillin, Frances; Linder, Ewert; Svärd, Staffan

2005-06-01

Giardia lamblia is a protozoan parasite infecting the upper mammalian small intestine. Infection relies upon the ability of the parasite to attach to the intestine via a unique cytoskeletal organelle, the ventral disk. We determined the composition and structure of the disk throughout the life cycle of the parasite and identified a new disk protein, SALP-1. SALP-1 is an immunodominant protein related to striated fiber-assemblin (SFA). The disk is disassembled during encystation and stored as four fragments in the immobile cyst. Serial Analysis of Gene Expression (SAGE) showed that the mRNA levels of the disk proteins decreased in encystation but two-dimensional protein gels showed that the protein levels were more constant. The parasite emerges without a functional disk but the four disk fragments are quickly reassembled into two new disks on the dividing, early excysting form. Thus, disk proteins are stored within the cyst, ready to be used in the rapid steps of excystation.

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

Science.gov (United States)

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.

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

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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.

Journal of Biosciences | Indian Academy of Sciences

Indian Academy of Sciences (India)

2004-10-27

Oct 27, 2004 ... 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, ...

Journal of Biosciences | Indian Academy of Sciences

Indian Academy of Sciences (India)

Clipboard: Order in stress – Lessons from the inanimate world .... indica A. Juss and their role in a multicomponent system against lepidopteran larvae .... Heat stress response in plants: a complex game with chaperones and more than twenty heat ... Encysted embryos (cysts) of the brine shrimp, Artemia, provide excellent ...

Monitoring of Waterborne Parasites in Two Drinking Water Treatment Plants: A Study in Sarawak, Malaysia.

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Richard, Reena Leeba; Ithoi, Init; Abd Majid, Mohamad Azlan; Wan Sulaiman, Wan Yusoff; Tan, Tian Chye; Nissapatorn, Veeranoot; Lim, Yvonne Ai Lian

2016-06-28

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.

Formulation and stability of an extemporaneous 0.02% chlorhexidine digluconate ophthalmic solution.

Science.gov (United States)

Lin, Shu-Chiao; Huang, Chih-Fen; Shen, Li-Jiuan; Wang, Hsueh-Ju; Lin, Chia-Yu; Wu, Fe-Lin Lin

2015-12-01

Acanthamoeba keratitis is difficult to treat because Acanthamoeba cysts are resistant to the majority of antimicrobial agents. Despite the efficacy of 0.02% chlorhexidine in treating Acanthamoeba keratitis, a lack of data in the literature regarding the formulation's stability limits its clinical use. The objective of this study was to develop an optimal extemporaneous 0.02% chlorhexidine digluconate ophthalmic formulation for patients in need. With available active pharmaceutical ingredients, 0.02% chlorhexidine digluconate sample solutions were prepared by diluting with BSS Plus Solution or acetate buffer. Influences of the buffer, type of container, and temperature under daily-open condition were assessed based on the changes of pH values and chlorhexidine concentrations of the test samples weekly. To determine the beyond-use date, the optimal samples were stored at 2-8°C or room temperature, and analyzed at time 0 and at Week 1, Week 2, Week 3, Week 4, Week 5, Week 8, Week 12, and Week 24. Despite chlorhexidine exhibiting better stability in acetate buffer than in BSS solution, its shelf-life was stability at 2-25°C for 6 months after being sealed and for 1 month after opening. This finding will enable us to prepare 0.02% chlorhexidine digluconate ophthalmic solutions based on a doctor's prescription. Copyright © 2014. Published by Elsevier B.V.

Paling et al., Afr., J. Infect. Dis. (2018) 12(S): 44-48 https://doi.org ...

African Journals Online (AJOL)

Riset SINAS

2017-10-22

Oct 22, 2017 ... after Brazil and India in 2014. ... though the World Health Organization (WHO) Multidrug Therapy (MDT) program has been implemented (Ministry of .... always moves to occupy the entire space in the media, so Acanthamoeba.

Efecto de la temperatura sobre la viabilidad de larvas de Trichinella spiralis Effect of temperature on the viability of Trichinella spiralis larvae

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Viviana R Randazzo

2011-12-01

Full Text Available El objetivo de este estudio fue determinar el efecto de diferentes temperaturas sobre la viabilidad de larvas libres y enquistadas de Trichinella spiralis aisladas en el sudoeste de la provincia de Buenos Aires, Argentina. Se trataron larvas libres y enquistadas a diferentes temperaturas (-30 °C, -20 °C, 4 °C, 20 °C, calentamiento gradual entre 0-100 °C. Se determinó el tiempo necesario para matar el 100 % de las larvas. Durante los primeros días, la mortalidad larvaria en todos los tratamientos con frío aumentó signifcativamente en función del tiempo. En todos los casos, las larvas libres sobrevivieron menor cantidad de días que las enquistadas. A -30 °C, -20 °C y 20 °C no se observaron diferencias signifcativas entre las curvas de mortalidad de cada estadio larvario, pero a 4 °C la mortalidad fue menos intensa entre las larvas enquistadas. El calentamiento disminuyó la viabilidad, sin observarse diferencias entre estadios larvarios. La totalidad de las larvas libres y enquistadas había muerto a los 61 y 95 días (-30 °C, a los 160 y 180 días (-20 °C, a los 280 y 330 días (4° C, y a los 460 y 590 días (20 °C, respectivamente. Fue necesaria una cocción durante 15 minutos a 90 °C para matar al 100 % de las larvas libres y a 100 °C para lograr igual mortalidad de las enquistadas. Nuestros resultados indican que la temperatura y los tiempos tradicionalmente utilizados para tratar productos cárnicos con potencial de transmisión de T. spiralis no serían los más efectivos para lograr la inactivación de la totalidad de larvas vivas de este parásito.The aim of this work was to study the effect of temperature on the viability of free and encysted larvae of Trichinella spiralis from southwest Buenos Aires province, Argentina. Larvae were treated at variable temperatures (-30 °C, -20 °C, 4 °C, 20 °C, gradual heating between 0-100 °C. The time necessary to kill 100 % of larvae was calculated. During the frst days of

Self-mating in the definitive host potentiates clonal outbreaks of the apicomplexan parasites Sarcocystis neurona and Toxoplasma gondii.

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Jered M Wendte

2010-12-01

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

Self-mating in the definitive host potentiates clonal outbreaks of the apicomplexan parasites Sarcocystis neurona and Toxoplasma gondii.

Science.gov (United States)

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

2010-12-23

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

Physiologic and systemic acute phase inflammatory responses in young horses repeatedly infected with cyathostomins and Strongylus vulgaris.

Science.gov (United States)

Andersen, U V; Reinemeyer, C R; Toft, N; Olsen, S N; Jacobsen, S; Nielsen, M K

2014-03-17

Migrating Strongylus vulgaris and encysted cyathostomin larvae cause a localized inflammatory response in horses. It is unknown whether these larvae elicit a systemic acute phase response (APR), evidenced by changes in serum amyloid A (SAA), haptoglobin (Hp), iron (Fe), albumin, or albumin/globulin (A/G) ratio. In this study, 28 horses were randomly allocated to receive either pyrantel tartrate or a pelleted placebo formulation in their daily feed. Concurrent with treatment, all the horses were administered 5000 pyrantel-susceptible cyathostomin infective larvae once daily, 5 days a week, for 24 weeks. Beginning in the fifth week, the horses also received 25 S. vulgaris larvae once weekly for the remainder of the study. At regular biweekly intervals, fecal samples were collected for quantitative egg counts, and whole blood and serum samples were collected for measurement of packed cell volume, total protein, albumin, globulin, A/G ratio, SAA, Hp, and Fe. On days 161-164, all the horses were euthanatized and necropsied. Samples were collected for enumeration of total luminal worm burdens, encysted cyathostomin larval populations, and migrating S. vulgaris larvae. Concentrations of Hp, Fe, and A/G ratio were associated significantly with strongyle burdens. Only treated male horses had significant increases in serum albumin. Larval S. vulgaris did not associate with Fe, whereas Fe was associated negatively with both total cyathostomin burdens and encysted L4s. The A/G ratios differed significantly between the two treatment groups. Significant differences between groups and individual time points were also observed for Hp and Fe, whereas SAA concentrations remained low throughout the study. In general, this study illustrated that experimental inoculations with S. vulgaris and cyathostomins may be associated with changes in Hp, Fe, and serum proteins, but not with SAA. Overall, these changes suggest that mixed strongyle infections elicit a mild acute phase reaction

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

Science.gov (United States)

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

2017-01-01

Data on the distribution of free-living amoebae is still lacking especially in Southeast Asian region. The aquatic environment revealed a high occurrence of free-living amoebae (FLA) due to its suitable condition and availability of food source, which subsequently causes infection to humans. A total of 94 water samples consisted of both treated and untreated from Laos (31), Myanmar (42), and Singapore (21) were investigated for the presence of pathogenic FLA. Each water sample was filtered and cultured onto non-nutrient agar seeded with live suspension of Escherichia coli and incubated at room temperature. Morphological identification was conducted for both trophozoites and cysts via microscopic stains (Giemsa and immunofluorescence). The presence of Naegleria-like structures was the most frequently encountered in both treated and untreated water samples, followed by Acanthamoeba-like and Vermamoeba-like features. To identify the pathogenic isolates, species-specific primer sets were applied for molecular identification of Acanthamoeba, Naegleria, and Vermamoeba. The pathogenic species of Acanthamoeba lenticulata and A. triangularis were detected from untreated water samples, while Vermamoeba vermiformis was found in both treated and untreated water samples. Our results suggested that poor water quality as well as inadequate maintenance and treatment might be the cause of this alarming problem since chlorine disinfection is ineffective in eradicating these amoebas in treated water samples. Regular monitoring and examination of water qualities are necessary in order to control the growth, hence, further preventing the widespread of FLA infections among the public. PMID:28212409

ASPECTOS BIOLÓGICOS DAS PRINCIPAIS AMEBAS DE VIDA-LIVRE DE IMPORTÂNCIA MÉDICA

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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

Habitat diversity and adaptation to environmental stress in encysted ...

Indian Academy of Sciences (India)

Unknown

intense study (for example, Morimoto et al 1994; Fiege et al 1996; Feder and ... Current research on the stress response is aimed chiefly at the cellular and .... removed (1600 g, 5 min) and the supernatant applied to a column of Sepharose ...

Counting Legionella cells within single amoeba host cells

Science.gov (United States)

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

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

Science.gov (United States)

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

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

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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.

Fimbrolide-coated antimicrobial lenses: their in vitro and in vivo effects.

Science.gov (United States)

Zhu, Hua; Kumar, Ajay; Ozkan, Jerome; Bandara, Rani; Ding, Aidong; Perera, Indrani; Steinberg, Peter; Kumar, Naresh; Lao, William; Griesser, Stefani S; Britcher, Leanne; Griesser, Hans J; Willcox, Mark D P

2008-05-01

To examine the ability of contact lenses coated with fimbrolides, inhibitors of bacterial quorum sensing, to prevent microbial adhesion and their safety during short-term clinical assessment. A fimbrolide was covalently attached to commercially available high Dk contact lenses. Subsequently Pseudomonas aeruginosa, Staphylococcus aureus, Serratia marcescens, or Acanthamoeba sp. were added to the lenses and control uncoated contact lenses. Lenses plus microbes were incubated for 24 h, then washed thoroughly to remove non-adherent microbes. Lenses were macerated and resulting slurry plated onto agar plates. After appropriate incubation, the numbers of colony forming units of bacteria (or numbers of Acanthamoeba trophozoites measured using a hemocytometer) from fimbrolide-coated and uncoated lenses were examined. A Guinea Pig model of lens wear was used to assess the safety of lenses worn on a continuous basis for 1 month. In a separate study, 10 subjects wore fimbrolide-coated lenses for 24 h. The responses of the Guinea Pigs and human volunteers to the lenses were assessed by slit lamp examination. The fimbrolides-coated lenses reduced the adhesion of all bacterial strains tested, with reductions occurring of between 67 and 92%. For Acanthamoeba a reduction of 70% was seen. There were no significant differences in ocular responses to fimbrolide-coated lenses compared with controls in either the 1 month animal model or overnight human trial. Fimbrolide-coated lenses show promise as an antibacterial and anti-acanthamoebal coating on contact lenses and appear to be safe when worn on the eye in an animal model.

Sudan Journal of Medical Sciences - Vol 1, No 1 (2006)

African Journals Online (AJOL)

Acanthamoeba keratitis in Sudan: outcome of ketoconazole treatment in six patients · EMAIL FULL TEXT EMAIL FULL TEXT DOWNLOAD FULL TEXT DOWNLOAD FULL TEXT. Abdel Mageed M Imam, El Sheikh Mahgoub, 34-36. http://dx.doi.org/10.4314/sjms.v1i1.38437 ...

Potentially pathogenic free-living amoebae in some flood-affected areas during 2011 Chiang Mai flood.

Science.gov (United States)

Wannasan, Anchalee; Uparanukraw, Pichart; Songsangchun, Apichart; Morakote, Nimit

2013-01-01

The survey was carried out to investigate the presence of potentially pathogenic free-living amoebae (FLA) during flood in Chiang Mai, Thailand in 2011. From different crisis flood areas, seven water samples were collected and tested for the presence of amoebae using culture and molecular methods. By monoxenic culture, FLA were detected from all samples at 37 °C incubation. The FLA growing at 37 °C were morphologically identified as Acanthamoeba spp., Naegleria spp. and some unidentified amoebae. Only three samples (42.8%), defined as thermotolerant FLA, continued to grow at 42 °C. By molecular methods, two non-thermotolerant FlA were shown to have 99% identity to Acanthamoeba sp. and 98% identity to Hartmannella vermiformis while the two thermotolerant FLA were identified as Echinamoeba exundans (100% identity) and Hartmannella sp. (99% identity). This first report of the occurrence of FLA in water during the flood disaster will provide information to the public to be aware of potentially pathogenic FLA.

Marine mimivirus relatives are probably large algal viruses

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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.

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

Science.gov (United States)

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

2014-12-15

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

Clinical care of acanthamoeba keratitis patients

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Yelena V. Skryabina

2017-12-01

Full Text Available Recently, akanthamoeba keratitis (AK is seen more and more often in ophthalmological practice. However, today there are no standard guidelines concerning diagnosis and treatment of patients with AK. In the article, the experience in care for such patients is presented. Purpose: to estimate the efficiency of diagnosis and treatment of patients with AK. Materials and methods. Case histories of patients, who received treatment for akanthamoeba keratitis in the Eye Microsurgery Department No. 4, City Ophthalmologic Center of the City Hospital No. 2, from 2011 to 2016, were analyzed. Under observation, there were 25 patients (26 eyes with akanthamoeba keratitis aged from 18 to 77 years; there were 15 men and 10 women. Patients were observed during 1 year. Full ophthalmologic examination was conducted in all patients. Additional diagnostic methods included microbiological investigation of corneal scrapes and washings, culturing them on innutritious agar (with E. сoli covering, confocal corneal microscopy (HRT 3 with cornea module, Heidelberg Retina Tomograph Rostock Cornea Module. A superficial punctate keratits (AK stage 2 was found in one patient. All other patients were divided into two groups. Stromal ring-shaped keratitis was diagnosed in patients of the first group (7 patients, AK stage 3. The 2nd group consisted of 17 patients with corneal ulcer (AK stage 4. All patients received medicamentous treatment. However patients of the 2nd group required different kinds of surgical treatment. Results. In AK diagnosis, corneal confocal microscopy is the most informative method. In patients with AK stages 2 and 3, there was an improvement in visual functions as a result of medicamentous therapy. As a result of treatment at the discharge from the hospital, the best corrected visual acuity was 0.5-1.0 for most patients. In the 2nd group patients, who were subjects to different types of surgical treatment visual functions stabilized. However non-compliance with recommendations led to disease recurrences with worse outcomes in four cases. Conclusion. It is possible to stop the inflammatory process preserving at the same time high visual functions only when patients are addressed in time, and when appropriate AK therapy is prescribed and patients are compliant with it for a long time.

CLINICAL FEATURES, DIAGNOSIS, THE RESULTS OF THERAPEUTIC AND SURGICAL TREATMENT OF ACANTHAMOEBIC KERATITIS

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Evg. A. Kasparova

2017-01-01

Full Text Available Purpose: to describe our experience in Acanthamoeba keratitis diagnostics and treatment in the FGBNU Research Institute of eye diseases, Moscow.Patients and Methods. We observed 24 patients (25 eyes with the Acanthamoeba keratitis (AK. The age ranged from 18 to 47 years. All patients, except one, were contact lenses wearers. Clinical signs included superficial epithelial-stromal lesions in 8 patients (8 eyes, stromal forms of AK- in 16 patients (17 eyes, and mixed keratitis in 9 (9 eyes. We used confocal microscopy, conjunctival smearing and blood immunofluorescent analysis for HSV types I and II. 8 patients (8 eyes underwent penetrating keratoplasty (PKP and their corneal buttons were morphologically examined. AK treatment included 2 biguanid antiseptics — a PHMB ("Comfort-drops" — solution for contact lenses care and 0.025% solution of a chlorhexidini bigluconati, or "Vitabact" in frequent instillations. We also used Diflucan solution 0.2% instillations — 6–8 times a day in, and Orungal or Diflucan per os (200 mg once a day. Eye drops of aminoglycozide or fluorhinolon groups were added to the treatment as well. In the cases of mixed Acanthamoeba and HSV keratitis we used anti-herpetic medications (Poludan, Acyclovir.Results. Cysts were found with confocal microscopy in 66% examined patients, and in 75% of the corneal buttons after keratoplasty. 15 cases (60% healed with various intensity opacities. We removed corneal epithelium in 2 patients with poor effect of the medication treatment. 8 patients (8 eyes underwent PKP with transparent (2 patients and a semi-transparent engraftment (2 patients, in 4 cases (4 eyes AK recurrences had occurred, what required repeated surgery. Long persistence of Acanthamoeba cysts in the cornea after clinical recovery caused the admission of 2 antiseptic biguanids eye drops for 6–20 months in all patients. There were no recurrences in the group of patients during the observation period (range 1 to 6

Parachlamydia spp. and Related Chlamydia-like Organisms and Bovine Abortion

Science.gov (United States)

Borel, Nicole; Ruhl, Silke; Casson, Nicola; Kaiser, Carmen; Pospischil, Andreas

2007-01-01

Chlamydophila abortus and Waddlia chondrophila cause abortion in ruminants. We investigated the role of Parachlamydia acanthamoebae in bovine abortion. Results of immunohistochemical analyses were positive in 30 (70%) of 43 placentas from which Chlamydia-like DNA was amplified, which supports the role of Parachlamydia spp. in bovine abortion. PMID:18258043

Microbial keratitis in Gujarat, Western India: findings from 200 cases ...

African Journals Online (AJOL)

Results: Of the 200 ulcers 55% were culture positive, 26.5% were bacterial ulcers of which 47% were due to Staphylococcus spp. Pure fungal growth was seen in 22% while 6% were mixed ulcers. Fusarium spp. (30%) was the most common fungus followed by Aspergillus spp. (21%). Only one case of Acanthamoeba ...

Small-sized euryhaline fish as intermediate hosts of the digenetic trematode Cryptocotyle concavum

Science.gov (United States)

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.

The life cycle of Ascocotyle (Phagicola) longa (Digenea: Heterophyidae), a causative agent of fish-borne trematodosis.

Science.gov (United States)

Simões, Susana Balmant Emerique; Barbosa, Helene Santos; Santos, Cláudia Portes

2010-03-01

The complete life cycle of the trematode Ascocotyle (Phagicola) longa (Digenea: Heterophyidae) is elucidated by natural observation validated by experimental infections. The natural first intermediate host of A. (P.) longa, an agent of human heterophyiasis in Brazil, is the cochliopid snail Heleobia australis (new first intermediate host). Metacercariae were found encysted in the body musculature, heart, stomach, liver, kidney, spleen, gonads and mesentery of mullets Mugil liza. Hamsters Mesocricetus auratus were experimentally infected with metacercariae of A. (P.) longa obtained from the mullets, and the adults recovered were used to infect the snails H. australis. Rediae and cercariae of A. (P.) longa are described for the first time. The ultrastructure of the tegument of A. (P.) longa shows a change in spination pattern from the cercaria with single-pointed spines to the metacercaria and adult with multipointed, brush-shaped spines. The life cycle of A. (P.) longa is related to estuaries and coastal lagoons where the recruitment of mugilid juveniles occurs. The high prevalence (100%) of A. (P.) longa encysted in the mullets examined within the urban area of Rio de Janeiro indicates the potentially great public health impact of the consumption of raw mullets. Copyright 2009 Elsevier B.V. All rights reserved.

Contribution of computed tomography on chest drainage guidance

International Nuclear Information System (INIS)

Douvlou, E.; Tzortzis, D.; Vlachou, I.; Petrocheilou, G.; Safarika, V.; Fragopoulou, L.; Stathopoulou, S.; Kokkinis, C.

2012-01-01

Full text: Introduction: Thoracic collections (encysted pleural, endopulmonary, mediastinal) are common findings in major trauma hospitals mainly in need of further treatment as drainage. Objectives and tasks: To evaluate the CT-guidance, as a method of choice for thoracic drainages. Material and methods: 35 CT-guided chest drainages were performed for diagnostic and therapeutic purpose in 33 patients, with a mean age of 62 years. Of the 35 drainages, 31 were encysted at the pleura, 3 of them were endopulmonary and 1 was in the mediastinum. During the procedure we used needles of 15cm long and 18-22 G diameter for small collections while for 'large' collections drainage catheters of 10-16 F were placed. Results: All the CT-guided drainages of the chest were successful. Of them, 7 were pleural effusion collections while 24 were exudate collections (18 inflammatory and 6 neoplastic) and all were developed in the lungs or the mediastinum. In all the cases that a catheter was placed, full removal of the collections was achieved leading to a remarkable improvement of the patient's condition. Non-significant pneumothorax and tiny endopulmonary bleeding were the complications that occurred. Conclusion: CT-guided drainage of thoracic collections is an accurate and secure procedure and achieves high diagnostic and therapeutic results

Establishment of a continuous culture system for Entamoeba muris and analysis of the small subunit rRNA gene

OpenAIRE

Kobayashi S.; Suzuki J.; Takeuchi T.

2009-01-01

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 electrop...

A PRACTICAL METHOD FOR QUANTIFICATION OF PLEURAL EFFUSION BY USG

OpenAIRE

Swish Kumar; Dinesh Kumar; Suganita; Singh; Vijay Shankar; Rajeev; Ajay; Anjali

2016-01-01

OBJECTIVE The aim of this study is to find a correlation between pleural separation and amount of aspirated effusion. METHODS Total 20 adult patients with 25 effusions were taken into the study with chest x-ray showing homogeneous opacity in either one or both of the lung field, which was confirmed on USG. Only uncomplicated pleural effusion were taken into study. Effusion with septations or encysted effusion or pyothorax were excluded from the study. RESULTS...

An ultrastructural and ecological study of Ceratium hirundinella (Dinophyceae)

OpenAIRE

Chapman, Deborah Vivien

1981-01-01

Vegetative cells of Ceratium hirundinella taken from Esthwaite Water, a small productive lake in the English Lake District, were studied each month by scanning and transmission electron microscopy. Changes in size and abundance of chloroplasts and their lamellar spacing was shown to be related to their exposure to light within the lake. Changes in the quantities of starch and lipid present were mainly related to cyst formation or recent excystment. Excystment and encystment were studied by li...

Incidence of adult brain cancers is higher in countries where the protozoan parasite Toxoplasma gondii is common

OpenAIRE

Thomas, Frédéric; Lafferty, Kevin D.; Brodeur, Jacques; Elguero, Eric; Gauthier-Clerc, Michel; Missé, Dorothée

2011-01-01

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 b...

Alternative mounting media for preservation of some protozoa.

Science.gov (United States)

Criado-Fornelio, A; Heredero-Bermejo, I; Pérez-Serrano, J

2014-10-01

Protozoa resistant stages are disintegrated when mounted in toluene-based media. To overcome such problem, three toluene-free mountants were tested on preserve Acanthamoeba spp and gregarines. Two commercial glues based on cyanoacrylate or trimethoxysilane were suitable for preserving both cysts and trophozoites. Hoyer's medium showed good results for mounting gregarine oocysts. Copyright © 2014 Elsevier B.V. All rights reserved.

Consequence of temperature changes on cercarial shedding from Galba truncatula infected with Fasciola hepatica or Paramphistomum daubneyi

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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.

Wild Rodents as Experimental Intermediate Hosts of Lagochilascaris minor Leiper, 1909

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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.

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

OpenAIRE

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...

Distribution and Characterization of Antigens Found in Subcellular Fractions of African Trypanosomes.

Science.gov (United States)

1979-08-01

34 • . . .. . -. 11. (i.e., Trypanosoma cruzi, Pereira et al 1978; Entamoeba histolytica, McLaughlin and Muller in preparation...Partial purification and some properties of a neutral sulfhydryl and an acid proteinase from Entamoeba histolytica. Can. J. Microbiol. 23 420-425...in membrane fragments isolated from Acanthamoeba sp . Biochem. Biophys. Acta 193 203-211. Voorheis, H.P. Gale, J.S., Owen, M.J. and Edwards W. (1979

[Contribution to the study of Microphallidae Travassos 1920 (trematoda). XXXII. Microphallus breviatus n. sp., a species with an abbreviated evolutive cycle from a Mediterranean pond in the Languedoc].

Science.gov (United States)

Deblock, S; Maillard, C

1975-01-01

Contribution to the study of Microphallidae Travassos, 1920 (Trematoda). XXXII. - Microphallus breviatus n. sp., a short life-cycle species of a mediterranean pond of Languedoc. The whole larval life-cycle of M. breviatus takes place in one host, Hydrobia ventrosa (Montagu), Mollusc Hydrobiidae. Hepato-pancreatic sporocysts produce morphologically altered xiphidio-cercariae which become encysted metacercariae in the sporocyts themselves. This species is defined in the genus by the anatomic characteristics of its cercariae and metacercariae, allied with its uncommon biology including two hosts only.

Local and systemic inflammatory and immunologic reactions to cyathostomin larvicidal therapy in horses.

Science.gov (United States)

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

Comparison of the larvicidal efficacies of moxidectin or a five-day regimen of fenbendazole in horses harboring cyathostomin populations resistant to the adulticidal dosage of fenbendazole.

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Reinemeyer, C R; Prado, J C; Nielsen, M K

2015-11-30

Despite widespread acknowledgement of cyathostomin resistance to adult icidal dosages of benzimidazole (BZD) anthelmintics, many strongyle control programs continue to feature regularly scheduled larvicidal treatment with fenbendazole (FBZ). However, no studies have been conducted to evaluate the efficacy of larvicidal regimens against encysted cyathostomins in a BZD-resistant (BZD-R) population. A masked, randomized, controlled clinical study was conducted with 18 juvenile horses harboring populations of cyathostomins that were considered BZD-R on the basis of fecal egg count reduction (FECR). Horses were blocked by prior history, ranked by egg counts, and allocated randomly to one of three treatment groups: 1--control, 2--FBZ >10mg/kg once daily for five consecutive days, or 3--moxidectin (MOX) >0.4 mg/kg once. Fecal samples were collected prior to treatment and seven and 14 days after the final dose of anthelmintic. On Days 18-20, complete replicates of horses were euthanatized and necropsied, and 1% aliquots of large intestinal contents were recovered for determination of complete worm counts. The cecum and ventral colon were weighed, and measured proportions of the respective organ walls were processed for quantitation and characterization of encysted cyathostomin populations. The five-day regimen of FBZ achieved 44.6% fecal egg count reduction, had 56.4% activity against luminal adults and larvae, and was 38.6% and 71.2% effective against encysted early third stage (EL3) and late third stage/ fourth stage (LL3/L4) cyathostomin larvae, respectively. In contrast, MOX provided 99.9% FECR, removed 99.8% of luminal stages, and exhibited 63.6% and 85.2% efficacy against EL3 and LL3/L4 mucosal cyathostomins, respectively. Although BZD-R was the most feasible explanation for the lower larvicidal efficacies of FBZ, mean larval counts of moxidectin-treated horses were not significantly different from controls or those treated with FBZ. The lack of significant

Oral immunization of BALB/c mice with Giardia duodenalis recombinant cyst wall protein inhibits shedding of cysts.

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

2003-10-01

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 immunogenicity of recombinant Giardia CWP is unknown, we have investigated the immunogenicity of recombinant CWP2 (rCWP2) and its efficacy in interfering with the phenomenon of encystation taking place in the small bowels of BALB/c mice vaccinated with the recombinant protein. Here we report that the immunization of BALB/c mice with rCWP2 stimulated the immune system in a manner comparable to that for a live infection with Giardia muris cysts. Fecal and serum anti-rCWP2 immunoglobulin A (IgA) antibodies were detected in the immunized mice. In addition, anti-rCWP2 IgG1 and IgG2a antibodies were detected in the serum. mRNAs coding for Th1 and Th2 types of cytokines were detected in spleen and Peyer's patch cells from immunized mice. When the vaccinated mice were challenged with live cysts, the animals shed fewer cysts. We conclude that rCWP2 is a possible candidate antigen for the development of a transmission-blocking vaccine.

A case of ectopic intraabdominal fascioliasis presented with acute abdomen.

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Tanir, Gönül; Karaman, Ayşe; Tüfekçı, Sehra Birgül; Erdoğan, Duygu; Tuygun, Nilden; Ozkan, Ayşegül Taylan

2011-06-01

Human fascioliasis with Fasciola species occurs worldwide and is most common among rural people who tend sheep and eat uncooked water vegetables, particularly watercress. The natural history of the acute phase begins with ingestion of metacercariae encysted on various kinds of aquatic vegetation such as watercress. Fascioliasis primarily involves the liver, bile ducts, gallbladder, and occasionally ectopic sites. We describe herein a case of ectopic fascioliasis. This uncommon form of disease was peritonitis; both visceral and parietal peritoneal layers were affected with the formation of multiple nodules and ascites.

Real-time PCR systems targeting giant viruses of amoebae and their virophages.

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Ngounga, Tatsiana; Pagnier, Isabelle; Reteno, Dorine-Gaelle Ikanga; Raoult, Didier; La Scola, Bernard; Colson, Philippe

2013-01-01

Giant viruses that infect amoebae, including mimiviruses and marseilleviruses, were first described in 2003. Virophages were subsequently described that infect mimiviruses. Culture isolation with Acanthamoeba spp. and metagenomic studies have shown that these giant viruses are common inhabitants of our biosphere and have enabled the recent detection of these viruses in human samples. However, the genomes of these viruses display substantial genetic diversity, making it a challenge to examine their presence in environmental and clinical samples using conventional and real-time PCR. We designed and evaluated the performance of PCR systems capable of detecting all currently isolated mimiviruses, marseilleviruses and virophages to assess their prevalence in various samples. Our real-time PCR assays accurately detected all or most of the members of the currently delineated lineages of giant viruses infecting acanthamoebae as well as the mimivirus virophages, and enabled accurate classification of the mimiviruses of amoebae in lineages A, B or C. We were able to detect four new mimiviruses directly from environmental samples and correctly classified these viruses within mimivirus lineage C. This was subsequently confirmed by culture on amoebae followed by partial Sanger sequencing. PCR systems such as those implemented here may contribute to an improved understanding of the prevalence of mimiviruses, their virophages and marseilleviruses in humans.

Cell Imaging by Spontaneous and Amplified Raman Spectroscopies

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Giulia Rusciano

2017-01-01

Full Text Available Raman spectroscopy (RS is a powerful, noninvasive optical technique able to detect vibrational modes of chemical bonds. The high chemical specificity due to its fingerprinting character and the minimal requests for sample preparation have rendered it nowadays very popular in the analysis of biosystems for diagnostic purposes. In this paper, we first discuss the main advantages of spontaneous RS by describing the study of a single protozoan (Acanthamoeba, which plays an important role in a severe ophthalmological disease (Acanthamoeba keratitis. Later on, we point out that the weak signals that originated from Raman scattering do not allow probing optically thin samples, such as cellular membrane. Experimental approaches able to overcome this drawback are based on the use of metallic nanostructures, which lead to a huge amplification of the Raman yields thanks to the excitation of localized surface plasmon resonances. Surface-enhanced Raman scattering (SERS and tip-enhanced Raman scattering (TERS are examples of such innovative techniques, in which metallic nanostructures are assembled on a flat surface or on the tip of a scanning probe microscope, respectively. Herein, we provide a couple of examples (red blood cells and bacterial spores aimed at studying cell membranes with these techniques.

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

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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.

Operation, Maintenance and Performance Evaluation of the Potomac Estuary Experimental Water Treatment Plant. Main Volume.

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1983-09-01

was stained with Lugol’s or D’Antonlo’s iodine stain and then examined microscopically for Giardia cysts, Entamoeba histolytica, Acanthamoeba...five-tube ..ON procedure was used for quantitating Salmonella sp . in water. The enrichmentp isolation and primary biochemical test methods are found in...8217 . . ..*- * .* . • .* .- .,,.. - ... - . .. .-. * . . . .- . ....... .. .. . . .i. . -. ’.. IO. Sampling Began I I , ta . __ _ _ _ __ _ _ _ a Grab Samples taken at 4 PM 88. I -MAR I -JUN I-SEP IDEC I-PAR I t- SP

Brain-Eating Amoebae: Predilection Sites in the Brain and Disease Outcome

OpenAIRE

Ong, Timothy Yu Yee; Khan, Naveed Ahmed; Siddiqui, Ruqaiyyah

2017-01-01

Acanthamoeba spp. and Balamuthia mandrillaris are causative agents of granulomatous amoebic encephalitis (GAE), while Naegleria fowleri causes primary amoebic meningoencephalitis (PAM). PAM is an acute infection that lasts a few days, while GAE is a chronic to subacute infection that can last up to several months. Here, we present a literature review of 86 case reports from 1968 to 2016, in order to explore the affinity of these amoebae for particular sites of the brain, diagnostic modalities...

Taeniasis: A possible cause of ileal bleeding.

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Settesoldi, Alessia; Tozzi, Alessandro; Tarantino, Ottaviano

2017-12-16

Taenia spp. are flatworms of the class Cestoda, whose definitive hosts are humans and primates. Human infestation (taeniasis) results from the ingestion of raw meat contaminated with encysted larval tapeworms and is considered relatively harmless and mostly asymptomatic. Anemia is not recognized as a possible sign of taeniasis and taeniasis-induced hemorrhage is not described in medical books. Its therapy is based on anthelmintics such praziquantel, niclosamide or albendazole. Here we describe a case of acute ileal bleeding in an Italian man affected with both Taenia spp. infestation resistant to albendazole and Helicobacter pylori -associated duodenal ulcers.

Recent advances in diagnosing pathogenic equine gastrointestinal helminths

DEFF Research Database (Denmark)

Andersen, Ulla Vestergaard; Howe, D. K.; Olsen, Susanne Nautrup

2013-01-01

that the pathogenic larval stages of cyathostomins and large strongyles cannot be detected by any of the available diagnostics. With the lengthy prepatency periods characterizing these parasites, there is a huge need for developing such assays. The recent identification of a possible diagnostic marker for encysted...... for validated and useful assays in the relative near future....... the needs for reliable and practical diagnostic tools for detection of major parasites infecting equines. The current, widely used coprological techniques are important and useful, but they do have considerable limitations as they are incapable of diagnosing the pathogenic migrating stages. Species...

Waterborne parasites and physico-chemical assessment of selected lakes in Malaysia.

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Onichandran, Subashini; Kumar, Thulasi; Lim, Yvonne A L; Sawangjaroen, Nongyao; Andiappan, Hemah; Salibay, Cristina C; Chye, Tan Tian; Ithoi, Init; Dungca, Julieta Z; Sulaiman, Wan Y W; Ling, Lau Yee; Nissapatorn, Veeranoot

2013-12-01

The objective of this study was to assess the physico-chemical parameters and waterborne parasites in selected recreational lakes from Malaysia. Samples were collected from seven stations of Recreational Lake A (RL-A) and six stations of Recreational Lake B (RL-B). The samples were processed to detect the presence of Giardia spp. and Cryptosporidium spp. using immunomagnetic separation kit, helminth eggs or ova by bright field microscopy and Acanthamoeba spp. by cultivation in non-nutrient agar. Chemical parameters such as ammonia, chlorine, fluoride, nitrate and nitrite and physical parameters such as dissolved oxygen, electrical conductivity, pH, salinity, temperature and total dissolved solid were also measured. Both lakes were freshwater with salinity ranging from 0.05 to 0.09 ppt. Most stations of these lakes were contaminated with Cryptosporidium spp., Giardia spp., Ascaris spp. and hookworm. Schistosoma spp. was found in RL-B only, while Acanthamoeba spp. was found in all stations. Of all sampling sites, station 5 of RL-B is the most contaminated. Linear regression and correlation analysis revealed that Giardia spp. and Schistosoma spp. showed a significant negative correlation with turbidity (p Malaysia. Future work on heavy metals (chromium, copper, mercury and zinc) is recommended to enhance the overall water quality monitoring and to take appropriate safety measures to ensure maintenance of good water standards.

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

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

2017-10-01

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

Artyfechinostomum malayanum: Metacercariae Encysted in Pila sp. Snails Purchased from Phnom Penh, Cambodia.

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Sohn, Woon-Mok; Yong, Tai-Soon; Eom, Keeseon S; Sinuon, Muth; Jeoung, Hoo-Gn; Chai, Jong-Yil

2017-06-01

The metacercariae of Artyfechinostomum malayanum (Leiper, 1911) Mendheim, 1943 were discovered in Pila sp. snails purchased from a market in Phnom Penh, Cambodia. They were isolated from the snails using the artificial digestion technique and were orally fed to 2 hamsters, 1 rat, and 2 mice to obtain the adult flukes. The metacercariae were round, 145-165 μm in diameter, having a cyst wall of 6-10 μm in thickness, a head collar and collar spines, and characteristic features of excretory granules. Adult flukes were recovered in the small intestines of the animals at days 14 and 32 post infection and were morphologically observed using a light microscope and a scanning electron microscope. They were plump or elongated, ventrally curved, 6.0-8.1 × 1.6-2.0 mm in size, and characterized by the head collar bearing 43 collar spines, including 5 end group ones on each side, a long cirrus sac extending beyond the posterior margin of the ventral sucker, a submedian ovary, and 2 deeply lobed testes. Eggs in uteri were operculate, ovoid to ellipsoid, and 120-135 × 68-75 μm in size. In scanning electron microscopy, the head collar was prominent with collar spines looking like horns. Scale-like tegumental spines were densely distributed on the ventral surface between the head collar and ventral sucker. Sensory papillae were distributed mainly on the tegument around suckers. By this study, it has been first confirmed that the life cycle of A. malayanum exists in Cambodia.

Experimental life history and biological characteristics of Fasciola gigantica (Digenea: Fasciolidae).

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Phalee, Anawat; Wongsawad, Chalobol; Rojanapaibul, Amnat; Chai, Jong-Yil

2015-02-01

This study was conducted to investigate the life history, morphology, and maturation of larval stages and adult worms of Fasciola gigantica in experimental mice. Lymnaea auricularia rubiginosa was used as the intermediate host, and Oryza sativa was used for encystment of the metacercariae, while Mus musculus was used as the definitive host for maturation study. Fresh eggs from the gall bladder of water buffaloes fully developed into embryonated ones and hatched out at days 11-12 after incubation at about 29ºC. Free-swimming miracidia rapidly penetrated into the snail host, and gradually developed into the next larval stages; sporocyst, redia, and daughter redia with cercariae. Fully-developed cercariae were separated from the redia and shed from the snails on day 39 post-infection (PI). Free-swimming cercariae were immediately allowed to adhere to rice plants, and capsules were constructed to protect metacercariae on rice plants. Juvenile worms were detected in intestines of mice at days 3 and 6 PI, but they were found in the bile duct from day 9 PI. Juvenile and adult flukes were recovered from 16 mice experimentally infected with metacercariae, with the average recovery rate of 35.8%. Sexually mature adult flukes were recovered from day 42 PI. It could be confirmed that experimentally encysted metacercariae could infect and develop to maturity in the experimental host. The present study reports for the first time the complete life history of F. gigantica by an experimental study in Thailand. The obtained information can be used as a guide for prevention, elimination, and treatment of F. gigantica at environment and in other hosts.

Use of cross-flow membrane filtration in a recirculating hydroponic system to suppress root disease in pepper caused by Pythium myriotylum.

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Schuerger, Andrew C; Hammer, William

2009-05-01

Zoosporic pathogens in the genera Pythium and Phytophthora cause extensive root disease epiphytotics in recirculating hydroponic vegetable-production greenhouses. Zoospore cysts of Pythium myriotylum Drechsler were used to evaluate the effectiveness of cross-flow membrane filters to control pythiaceous pathogens in recirculating hydroponic systems. Four membrane filter brands (Honeycomb, Polypure, Polymate, and Absolife) were tested alone or in combination to determine which filters would effectively remove infective propagules of P. myriotylum from solutions and reduce disease incidence and severity. Zoospore cysts of P. myriotylum generally measured 8 to 10 microm, and it was hypothesized that filters with pore-sizespepper plants from root infection. Single-filter assays with Honeycomb and Polypure brands removed 85 to 95% of zoospore cysts when pore sizes were rated at 1, 5, 10, 20, or 30 microm. Single-filter assays of Polymate and Absolife brands were more effective, exhibiting apparently 100% removal of zoospore cysts from nutrient solutions on filters rated at 1 to 10 microm. However, plant bioassays with Honeycomb and Polymate single filters failed to give long-term protection of pepper plants. Double-filter assays with 1- and 0.5-microm Polymate filters significantly increased the protection of pepper plants grown in nutrient film technique systems but, eventually, root disease and plant wilt could be observed. Insect transmissions by shore flies were not factors in disease development. Scanning electron microscopy images of zoospore cysts entrapped on Polymate filters revealed zoospore cysts that were either fully encysted, partially encysted, or of unusually small size (3 microm in diameter). It was concluded that either the atypically small or pliable pleomorphic zoospore cysts were able to penetrate filter membranes that theoretically should have captured them.

The First Record for the Americas of Loxodes rex, a Flagship Ciliate with an Alleged Restricted Biogeography.

Science.gov (United States)

Hines, Hunter N; McCarthy, Peter J; Esteban, Genoveva F

2016-01-01

As the foundations of food webs, protozoa are essential to the success of an ecological system. These organisms are often overlooked, and research in the Americas is sparse. Recent samplings conducted in freshwater canals and ponds in Florida, USA, have revealed Loxodes rex, an alleged endemic ciliate species. Originally described as endemic to tropical Africa, L. rex has been considered a prime candidate for proof of microbial endemism. Our studies have shown this giant, non-encysting ciliate to be thriving in subtropical Florida. Our observations are novel and include both the first record of occurrence for the Americas and the first high-quality in vivo images for this charismatic species.

Location on chitin in the cyst wall of Entamoeba invadens with colloidal gold tracers.

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Arroyo-Begovich, A; Cárabez-Trejo, A

1982-04-01

Chitin was located in the cyst wall of Entamoeba invadens with colloidal gold-linked wheat germ agglutinin. Cysts stained differentially from trophozoites when encysting cultures were treated with the gold tracer; cysts acquired a wine-red coloration while, in general trophozoites remained unstained. Observation of cells with the electron microscope revealed that the tracer particles were bound specifically to the walls of the surface of the cyst when cells were exposed in suspension, and to the cyst wall cross-section, when cells were exposed to the tracer in thin section, indicating that chitin fibers were distributed on the surface as well as throughout the matrix of the cyst wall.

Comparison between Two Methods for Diagnosis of Trichinellosis: Trichinoscopy and Artificial Digestion

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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

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

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Bischoff, Paul J; Wetmore, Scott

2009-01-01

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

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.

Science.gov (United States)

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. Copyright © 2016 Elsevier B.V. All rights reserved.

Pseudomonas aeruginosa adaptation to lungs of cystic fibrosis patients leads to lowered resistance to phage and protist enemies

DEFF Research Database (Denmark)

Friman, Ville-Petri; Ghoul, Melanie; Molin, Søren

2013-01-01

) patients affects its survival in the presence of natural phage (14/1, ΦKZ, PNM and PT7) and protist (Tetrahymena thermophila and Acanthamoebae polyphaga) enemies. We found that most of the bacteria isolated from relatively recently intermittently colonised patients (1-25 months), were innately phage......-resistant and highly toxic for protists. In contrast, bacteria isolated from long time chronically infected patients (2-23 years), were less efficient in both resisting phages and killing protists. Moreover, chronic isolates showed reduced killing of wax moth larvae (Galleria mellonella) probably due to weaker...

Brain-Eating Amoebae: Predilection Sites in the Brain and Disease Outcome

Science.gov (United States)

Ong, Timothy Yu Yee; Siddiqui, Ruqaiyyah

2017-01-01

ABSTRACT Acanthamoeba spp. and Balamuthia mandrillaris are causative agents of granulomatous amoebic encephalitis (GAE), while Naegleria fowleri causes primary amoebic meningoencephalitis (PAM). PAM is an acute infection that lasts a few days, while GAE is a chronic to subacute infection that can last up to several months. Here, we present a literature review of 86 case reports from 1968 to 2016, in order to explore the affinity of these amoebae for particular sites of the brain, diagnostic modalities, treatment options, and disease outcomes in a comparative manner. PMID:28404683

EVALUATION OF THE THERAPEUTIC EFFICACY OF LEVAMISOLE HYDROCHLORIDE ON THIRD-STAGE LARVAE OF Lagochilascaris minor IN EXPERIMENTALLY INFECTED MICE

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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.

Characterization of a Rab11-like GTPase, EhRab11, of Entamoeba histolytica.

Science.gov (United States)

McGugan, Glen C; Temesvari, Lesly A

2003-07-01

The Entamoeba histolytica Rab11 family of small molecular weight GTPases consists of three members, EhRab11, EhRab11B, and EhRab11C. The functions of these Rabs in Entamoeba have not been determined. Therefore, as an approach to elucidate the role of the Rab11 family of GTPases in Entamoeba, immunofluorescence microscopy was undertaken to define the subcellular localization of one member of this family, EhRab11. Under conditions of growth, EhRab11 displayed a punctate pattern in the cytoplasm of trophozoites. EhRab11 did not colocalize with markers for the Golgi apparatus, endoplasmic reticulum, pinosomes, phagosomes, or compartments formed by receptor-mediated endocytosis, suggesting that this Rab may not play a role in vesicle trafficking between these organelles. Under conditions of iron and serum starvation, EhRab11 was translocated to the periphery of the cell. The altered cellular localization was accompanied by multinucleation of the cells as well as the acquisition of detergent resistance by the cells, features that are characteristic of Entamoeba cysts. The translocation of EhRab11 to the periphery of the cell during iron and serum starvation was specific as the subcellular localizations of two other Rab GTPases, EhRab7 and EhRabA, were not altered under the same conditions. In addition, the formation of multinucleated cells by inhibition of cytokinesis was not sufficient to induce the translocation of EhRab11 to the cell periphery. Taken together, the data suggest that iron and serum starvation may induce encystation in E. histolytica and that EhRab11 may play a role in this process. Moreover, these studies are the first to describe a putative role for a Rab GTPase in encystation in Entamoeba sp.

Selective condensation drives partitioning and sequential secretion of cyst wall proteins in differentiating Giardia lamblia.

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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

Testicular Descend, How and Why: A Review Article

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Sujan Narayan Agrawal

2017-08-01

Full Text Available Background: The testis develops in the dorsal abdominal wall, and then descends to scrotum. The development begins as early as 6th week of intrauterine life and is completed by fifth month of intrauterine life. The testis may get arrested during its descent from dorsal abdominal wall to scrotum. The anomalies of descent includes cryptorchism (and its variant like anarchism, monarchism or partially descended testis, ectopic testis, persistent processus vaginalis and encysted hydrocoel of spermatic cord etc. Cryptorchism is usually diagnosed during the new born examination. The recognition of this condition, identification of associated syndromes, proper diagnostic evaluation and timely treatment by surgical urologist is important to prevent adverse consequences like sterility, congenital hernia & hydrocoel, testicular carcinoma etc. Objectives: the objective of this review is to study the role of gubernaculum in the testicular migration process. Material & Method: We performed a descriptive review of the literature about the role of the gubernaculum in testicular migration during the human fetal life. This article provides an overview of role of gubernaculum and other factors responsible for gonadal migration. Results: In the first phase of testicular migration the gubernaculum enlarges to hold the testis near groin and in the second phase the gubernaculum migrates across the pubic region to reach the scrotum. The proximal end of gubernaculum is attached to the testis and epididymis. The lower end reaches to bottom of scrotum. A failure in the proper functioning of gubernaculum causes cryptorchism. Rarely male gonads may deviate from main pathway due to presence of many tails of distal gubernaculum, and it may give rise to ectopic testis. The processus vaginalis usually closes by birth. If it remains patent, it leads to congenital hernia, hydrocoel, encysted hydrocoel etc. Conclusion: the gubernaculum presents a significant structure during

Disseminated cysticercosis in a child: whole-body MR diagnosis with the use of parallel imaging

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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.)

The Histopathology of the Infection of Tilapia rendalli and Hypostomus regani (Osteichthyes by Lasidium Larvae of Anodontites trapesialis (Mollusca, Bivalvia

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Ângela Teresa Silva-Souza

2002-04-01

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.

Comparison of ultrasound and computer tomography in the diagnosis of pleural shadowing

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Kurtz, B.; Schmitt, W.G.H.

1983-05-01

Sonography was performed in 40 patients with pleural shadowing of uncertain origin in order to complement conventional X-ray examination. Where the findings were not clear-cut, computer tomography was performed subsequently and the diagnosis ascertained by biopsy or operation. Fluid-containing pleural lesions, such as encysted effusions and empyemas, pericardial cysts and one echinococcus cyst were identified by sonography from their appearance and biopsy was performed; computer tomography adds little additional information in these conditions. Solid pleural masses, on the other hand, can be more accurately defined by computer tomography. This shows the entire pleura, including small areas of calcification. Density measurements are able to differentiate various tissues, and particularly fat.

A comparison of ultrasound and computer tomography in the diagnosis of pleural shadowing

International Nuclear Information System (INIS)

Kurtz, B.; Schmitt, W.G.H.

1983-01-01

Sonography was performed in 40 patients with pleural shadowing of uncertain origin in order to complement conventional X-ray examination. Where the findings were not clear-cut, computer tomography was performed subsequently and the diagnosis ascertained by biopsy or operation. Fluid-containing pleural lesions, such as encysted effusions and empyemas, pericardial cysts and one echinococcus cyst were identified by sonography from their appearance and biopsy was performed; computer tomography adds little additional information in these conditions. Solid pleural masses, on the other hand, can be more accurately defined by computer tomography. This shows the entire pleura, including small areas of calcification. Density measurements are able to differentiate various tissues, and particularly fat. (orig.) [de

Ectopic fascioliasis mimicking a colon tumor

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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 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. PMID:17552017

Revised Mimivirus major capsid protein sequence reveals intron-containing gene structure and extra domain

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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.

Lamarckian evolution of the giant Mimivirus in allopatric laboratory culture on amoebae.

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Colson, Philippe; Raoult, Didier

2012-01-01

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

[IMPACT OF CASPIAN SEA LEVEL FLUCTUATIONS ON THE EPIZOOTIC ACTIVITY OF THE CASPIAN SANDY NATURAL PLAGUE FOCUS].

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Popov, N V; Udovikov, A I; Eroshenko, G A; Karavaeva, T B; Yakovlev, S A; Porshakov, A M; Zenkevich, E S; Kutyrev, V V

2016-01-01

There is evidence that in 1923-2014 the sharp aggravations of the epizootic situation of plague in the area of its Caspian sandy natural focus after long interepizootic periods are in time with the ups of the Caspian Sea in the extrema of 11-year solar cycles. There were cases of multiple manifestations of plague in the same areas in the epizootic cycles of 1946-1954, 1979-1996, 2001, and 2013-2014. The paper considers the possible role of amebae of the genus Acanthamoeba and nematodes, the representatives of the orders Rhabditida and Tylenchida in the microfocal pattern of plague manifestations.

Dilated cardiomyopathy and amebic gastritis in a giant anteater (Myrmecophaga tridactyla).

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Coke, Rob L; Carpenter, James W; Aboellail, Tawfk; Armbrust, Laura; Isaza, Ramiro

2002-09-01

An approximately 11-mo-old female giant anteater (Myrmecophaga tridactyla) exhibited anorexia, lethargy, hypothermia, depression, and minimal response to external stimuli. Radiography and ultrasonography revealed an enlarged heart, with free gas and fluid in the abdomen. Abdominocentesis produced a clear brown fluid with an acute to subacute septic suppurative exudate. Cardiac ultrasonography revealed a dilated, thin-walled left ventricle with a comparatively low fractional shortening. Despite intensive supportive care, the anteater died. Postmortem findings included gastric ulceration with perforation near the pylorus. Entameba spp. and Acanthamoeba spp. were both identified in large numbers at the site of the gastric ulceration and perforation.

Exposure to synthetic greywater inhibits amoebae encystation and alters expression of Legionella pneumophila virulence genes

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Water conservation efforts have focused on greywater (GW) usage, especially for applications that do not require potable water quality. However, there is a need to better understand environmental pathogens and their free-living amoebae (FLA) hosts within GW. Using synthetic gre...

Morphological and molecular identification of the fish-borne metacercaria of Ascocotyle (Phagicola) longa Ransom, 1920 in Mugil liza from Argentina.

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Martorelli, S R; Lino, A; Marcotegui, P; Montes, M M; Alda, P; Panei, C J

2012-12-21

This is the first report of Ascocotyle (Phagicola) longa Ransom, 1920 (Digenea: Heterophyidae) in Argentina confirmed by morphological and molecular studies. The metacercaria was found encysted in myotomal musculature, heart and mesentery of the mullet Mugil liza (Pisces: Mugilidae) from Samborombon bay. We provide a morphological description of the metacercaria which we identified using species-specific primers for A. (Phagicola) longa and nucleotid sequence. This worldwide parasite has been reported as one of the causative agents of heterophyiosis, an emerging fish-borne disease of humans, contracted by the consumption of raw mullet. The discovery of A. (Phagicola) longa in Argentina represents a warning of the potentially great impact of this parasite on public health. Copyright © 2012 Elsevier B.V. All rights reserved.

Third-stage larvae of the enoplid nematode Dioctophyme renale (Goeze, 1782) in the freshwater turtle Trachemys dorbigni from southern Brazil.

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Mascarenhas, C S; Müller, G

2015-09-01

The giant kidney worm Dioctophyme renale is normally found in wild carnivores and domestic dogs, with aquatic oligochaetes acting as intermediate hosts. In the present study a prevalence of 50% of third-stage larvae of D. renale was recorded in 60 specimens of the freshwater turtle Trachemys dorbigni from southern Brazil. Larvae were encysted in muscles, the coelomic cavity and mesentery, the serous lining of the stomach and on the surfaces of the lung, heart, liver, pancreas, spleen and intestines. There are no previous records of reptiles being part of the life cycle of D. renale, although fish and amphibians normally act as paratenic hosts. This is the first report of third-stage D. renale larvae in the freshwater turtle, T. dorbigni.

Are soil and waterborne parasitic infections health risk for worker populations in southeast Turkey?

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Sibel Ak

2017-06-01

Full Text Available Objective: The soil and waterborne parasitic infections rate is high degree in developed and developing countries. Migratory workers have greater exposure to these parasitic infections and a lot of morbidity due to these infections in workers. For this reason, we aimed to investigate the presence of soil and waterborne parasites in the Gaziantep Organized Industrial Zone of southeast Turkey. Methods: A total of 25 environmental samples (18 soil samples and 7 water samples were taken from The Gaziantep Organized Industrial Zone, in two different seasons (summer and winter. All of the samples were screened for parasites using microscopic examination and culture methods. The parasites were genotyped with polymerase chain reaction and DNA sequencing analysis. Results: The prevalence of soil and water transmitted parasites was found to be positive 52% (13/25 in summer while there is no any parasites in winter. It was found 22.3% (4/18 Acanthamoeba (genotype4, 16.6% (3/18 Ascaris lumbricoides, 11.1% (2/18 Strongoides stercoralis in soil samples and 14.3% (1/7 Acanthamoeba (genotype 4, 42.9% (3/7 Blastocystis (subtype3 in all of water samples. Conclusion: The migratory worker waves have always shaped the ethnic composition and public health problem of the province of Gaziantep. Climate change has the potential to influence prevalence of parasite and our study has shown that increased prevalence of parasite in summer. The global target for the coming years should be to remove the deaths from earth and waterborne parasitic infections in the worker populations. Thus, we prevent the distribution of parasitic infections in our country.

Twelve-year analysis of microbial keratitis trends at a UK tertiary hospital.

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Tan, S Z; Walkden, A; Au, L; Fullwood, C; Hamilton, A; Qamruddin, A; Armstrong, M; Brahma, A K; Carley, F

2017-08-01

PurposeTo investigate the frequencies, trends, and in vitro drug susceptibilities of the causative pathogens in microbial keratitis in Manchester Royal Eye Hospital.Patients and methodsCorneal scrape results recorded by the microbiology service between 2004 and 2015 were extracted from an established database. A total of 4229 corneal scrape specimens were identified from an established database. First-line antibiotic treatment in our centre during the study period was ofloxacin and second line was cefuroxime and gentamicin.ResultsMean age was 45.9±21.0. A total of 1379 samples (32.6%) were culture positive. One hundred forty-eight (10.7%) specimens cultured multiple organisms. Of the 1539 organisms identified, 63.3% were Gram-positive bacteria, 27.3% Gram-negative bacteria, 7.1% fungi, and 2.3% Acanthamoebae. A decreasing trend in Gram-positive isolates was found together with a stable trend in Gram negatives and an increasing trend in Acanthamoeba and fungi. There appeared to be a significant increasing trend of Moraxella infection (P=0.001). In all, 83.1 and 90.8% of Gram-positive and -negative isolates tested were susceptible to ofloxacin, respectively. Cefuroxime covered 86.6% of Gram-positive and 61.4% of Gram-negative isolates, whereas gentamicin covered 88.8 and 96.5% of Gram-positive and -negative isolates, respectively.ConclusionWe found a change in the type of Gram-negative organisms isolated over time, with the Moraxella species on the rise. Reassuringly, no significant increase in resistance was observed in vitro for any of the commonly used antibiotics. Ofloxacin remains a good first-line antibiotic treatment but duo-therapy does have broader coverage and should be considered in non-responsive cases.

High prevalence of Angiostrongylus cantonensis (rat lungworm on eastern Hawai'i Island: A closer look at life cycle traits and patterns of infection in wild rats (Rattus spp..

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Susan I Jarvi

Full Text Available The nematode Angiostrongylus cantonensis is a zoonotic pathogen and the etiological agent of human angiostrongyliasis or rat lungworm disease. Hawai'i, particularly east Hawai'i Island, is the epicenter for angiostrongyliasis in the USA. Rats (Rattus spp. are the definitive hosts while gastropods are intermediate hosts. The main objective of this study was to collect adult A. cantonensis from wild rats to isolate protein for the development of a blood-based diagnostic, in the process we evaluated the prevalence of infection in wild rats. A total of 545 wild rats were sampled from multiple sites in the South Hilo District of east Hawai'i Island. Adult male and female A. cantonensis (3,148 were collected from the hearts and lungs of humanely euthanized Rattus rattus, and R. exulans. Photomicrography and documentation of multiple stages of this parasitic nematode in situ were recorded. A total of 45.5% (197/433 of rats inspected had lung lobe(s (mostly upper right which appeared granular indicating this lobe may serve as a filter for worm passage to the rest of the lung. Across Rattus spp., 72.7% (396/545 were infected with adult worms, but 93.9% (512/545 of the rats were positive for A. cantonensis infection based on presence of live adult worms, encysted adult worms, L3 larvae and/or by PCR analysis of brain tissue. In R. rattus we observed an inverse correlation with increased body mass and infection level of adult worms, and a direct correlation between body mass and encysted adult worms in the lung tissue, indicating that larger (older rats may have developed a means of clearing infections or regulating the worm burden upon reinfection. The exceptionally high prevalence of A. cantonensis infection in Rattus spp. in east Hawai'i Island is cause for concern and indicates the potential for human infection with this emerging zoonosis is greater than previously thought.

Pathogen filtration to control plant disease outbreak in greenhouse production

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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

An unusual case of cycticercosis of the tendon sheath of the tendoachilles.

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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.

Histological identification of muscular sarcocystis: A report of two cases

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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.

The effect of toxic loads on effluent purification systems

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Cillie, G.C.; Hart, O.O.; Davies, T.R.; Hassett, A.J.

1977-04-01

Deterioration of the performance of biological wastewater treatment plants at Daspoort Sewage Works, Pretoria, South Africa, during 1975 was attributed to toxic material in the sewage. Unusually high concentrations of mercury, phenol, chromium, and an organophosphorus compound were detected. These substances singly or in combination could have been responsible for the deterioration. Data from the biological plants and research units at Daspoort are collected and reviewed to evaluate the water quality effects of the period of low performance. Encystation by peritrichous ciliates caused high bacteria counts in biofilter effluents. Nitrification declined rapidly and recovery was delayed, while carbon oxidation was affected to a limited extent. The water product was of the same quality throughout, even when the influent was little better than settled sewage. (12 graphs, 4 references, 3 tables)

An unusual case of intracystic papillary carcinoma of breast with invasive component

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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.

Effect of infection by irradiated Trichinella Spirals larvae on mice and assessment the role of Al bendazole in treating them

International Nuclear Information System (INIS)

Moawad, M.A.F.; Amin, M.M.

2005-01-01

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

Opportunistic Pathogens and Microbial Communities and Their Associations with Sediment Physical Parameters in Drinking Water Storage Tank Sediments

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Qin, Ke; Struewing, Ian; Domingo, Jorge Santo; Lytle, Darren

2017-01-01

The occurrence and densities of opportunistic pathogens (OPs), the microbial community structure, and their associations with sediment elements from eight water storage tanks in Ohio, West Virginia, and Texas were investigated. The elemental composition of sediments was measured through X-ray fluorescence (XRF) spectra. The occurrence and densities of OPs and amoeba hosts (i.e., Legionella spp. and L. pneumophila, Mycobacterium spp., P. aeruginosa, V. vermiformis, Acanthamoeba spp.) were determined using genus- or species-specific qPCR assays. Microbial community analysis was performed using next generation sequencing on the Illumina Miseq platform. Mycobacterium spp. were most frequently detected in the sediments and water samples (88% and 88%), followed by Legionella spp. (50% and 50%), Acanthamoeba spp. (63% and 13%), V. vermiformis (50% and 25%), and P. aeruginosa (0 and 50%) by qPCR method. Comamonadaceae (22.8%), Sphingomonadaceae (10.3%), and Oxalobacteraceae (10.1%) were the most dominant families by sequencing method. Microbial communities in water samples were mostly separated with those in sediment samples, suggesting differences of communities between two matrices even in the same location. There were associations of OPs with microbial communities. Both OPs and microbial community structures were positively associated with some elements (Al and K) in sediments mainly from pipe material corrosions. Opportunistic pathogens presented in both water and sediments, and the latter could act as a reservoir of microbial contamination. There appears to be an association between potential opportunistic pathogens and microbial community structures. These microbial communities may be influenced by constituents within storage tank sediments. The results imply that compositions of microbial community and elements may influence and indicate microbial water quality and pipeline corrosion, and that these constituents may be important for optimal storage tank management

Molecular survey of occurrence and quantity of Legionella spp., Mycobacterium spp., Pseudomonas aeruginosa and amoeba hosts in municipal drinking water storage tank sediments.

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Lu, J; Struewing, I; Yelton, S; Ashbolt, N

2015-07-01

To examine the occurrence and quantity of potential pathogens and an indicator of microbial contamination in the sediments of municipal drinking water storage tanks (MDWSTs), given the absence of such data across the United States. Sediment samples (87 MDWST) from eighteen locations across ten states of the United States were collected and assayed by qPCR for a range of potential enteric and opportunistic microbial pathogens and a sewage-associated Bacteroides marker. Potential opportunistic pathogens dominated, with the highest detection of occurrence (per cent positive detection; average cell equivalence (CE)) being Mycobacterium spp. (88·9%; 6·7 ± 8·5 × 10(4) CE g(-1) ), followed by Legionella spp. (66·7%; 5·2 ± 5·9 × 10(3) CE g(-1) ), Pseudomonas aeruginosa (22·2%; 250 ± 880 CE g(-1) ) and Acanthamoeba spp. (38·9%; 53 ± 70 CE g(-1) ), with no detected Naegleria fowleri. Most enteric pathogens (Campylobacter jejuni, Escherichia coli 0157:H7, Salmonella enterica, Cryptosporidium parvum and Giardia duodenalis) were not detected, except for a trace signal for Campylobacter spp. There was significant correlation between the qPCR signals of Legionella spp. and Acanthamoeba spp. (R(2) = 0·61, n = 87, P = 0·0001). Diverse Legionella spp. including Leg. pneumophila, Leg. pneumophila sg1 and Leg. anisa were identified, each of which might cause legionellosis. These results imply that potential opportunistic pathogens are common within MDWST sediments and could act as a source of microbial contamination, but need downstream growth to be of potential concern. The results imply that opportunistic pathogen risks may need to be managed by regular tank cleaning or other management practices. 2015 The Society for Applied Microbiology.

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

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

2016-02-01

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

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

Science.gov (United States)

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

2016-08-03

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

Contact lens disinfection by ultraviolet light

International Nuclear Information System (INIS)

Dolman, P.J.; Dobrogowski, M.J.

1989-01-01

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

Dicty_cDB: Contig-U04013-1 [Dicty_cDB

Lifescience Database Archive (English)

Full Text Available pid:none) Flavobacterium psychrophilum JIP... 98 3e-19 EU678894_1( EU678894 |pid:none) Bacillus intermedius ...46 3e-04 2 ( EH672780 ) CCIL10412.b1_G11.ab1 CCI(LMS) chicory Cichorium i... 46 3e-04 2 ( EJ368820 ) 1092963715450 Global-Ocean-Sampl...41 |pid:none) Sulfolobus solfataricus P2, com... 89 2e-16 AH1770( AH1770 ) conserved hypothetical protein lin2710 [imported...AY653733 ) Acanthamoeba polyphaga mimivirus, complete genome. 62 3e-05 1 ( EH678404 ) CCIL3410.b1_D14.ab1 CC...ngs S... 44 6.8 1 >( AC116963 ) Dictyostelium discoideum chromosome 2 map 4657875-4914984 strain AX4, compl

Occurrence and Control of Legionella in Recycled Water Systems

Science.gov (United States)

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

Presence of pathogenic microorganisms in power-plant cooling waters. Report for October 1, 1979-September 30, 1981

Energy Technology Data Exchange (ETDEWEB)

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.

Helminths Parasite Larvae collected from Arabian Gulf Fish. 4. Description of four Larvae including two Metacercarae, one Didymozoid and one Acanthocephalan from Emirati Coasts

International Nuclear Information System (INIS)

Kardousha, M.M.

2005-01-01

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)

Parachordodes tegonotus n. sp. (Gordioidea: Nematomorpha), a hairworm parasite of ground beetles (Carabidae: Coleoptera), with a summary of gordiid parasites of carabids.

Science.gov (United States)

Poinar, George; Rykken, Jessica; LaBonte, Jim

2004-06-01

A new species of hairworm, Parachordodes tegonotus n. sp. (Gordioidea: Nematomorpha) is described from three species of ground beetles (Carabidae: Coleoptera) from the state of Oregon. This is the first record of Parachordodes Camerano parasitising carabid beetles in North America. Diagnostic characters for the new species include size, colour, the nature and arrangement of the areoles, and the character, shape and extant of ornamentation on the ventral surface of the male tail. Encysted hairworm larvae found in the internal tissues of mayfly and caddisfly larvae at the type-locality were presumed to be those of P. tegonotus, indicating an indirect life-cycle involving paratenic hosts. A worldwide host list shows that some 70 species of ground beetles have been documented as developmental hosts to hairworms belonging to at least five genera, namely Gordius, Parachordodes, Dacochordodes, Gordionus and Paragordionus.

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

Czech Academy of Sciences Publication Activity Database

Veverková, Marie; Dyková, Iva; Pecková, Hana

2002-01-01

Roč. 49, č. 3 (2002), s. 243-245 ISSN 0015-5683 R&D Projects: GA AV ČR KSK6005114 Institutional research plan: CEZ:AV0Z6022909 Keywords : amoeba * fish * pathology Subject RIV: EA - Cell Biology Impact factor: 0.515, year: 2002

cDNA-AFLP analysis of differential gene expression related to cell chemotactic and encystment of Azospirillum brasilense.

Science.gov (United States)

Li, Huamin; Cui, Yanhua; Wu, Lixian; Tu, Ran; Chen, Sanfeng

2011-12-20

Our previous study indicated org35 was involved in chemotaxis and interacted with nitrogen fixation transcriptional activator NifA via PAS domain. In order to reveal the role of org35 in nitrogen regulation, the downstream target genes of org35 were identified. We here report differentially expressed genes in org35 mutants comparing with wild type Sp7 by means of cDNA-AFLP. Four up-regulated transcript-derived fragments (TDFs) homologues of chemotaxis transduction proteins were found, including CheW, methyl-accepting chemotaxis protein and response regulator CheY-like receiver. Three distinct TDFs (AB46, AB58 and AB63) were similar to PHB de-polymerase C-terminus, cell shape-determining protein and flagellin domain protein. And 11 TDFs showed similarities with signal transduction proteins, including homologous protein of the nitrogen regulation protein NtrY and nitrate/nitrite response regulator protein NarL. These data suggested that the Azospirillum brasilense org35 was a multi-effecter and involved in chemotaxis, cyst development and regulation of nitrogen fixation. Copyright © 2010 Elsevier GmbH. All rights reserved.

Monitoring of Legionella pneumophila viability after chlorine dioxide treatment using flow cytometry.

Science.gov (United States)

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. Copyright © 2015 Institut Pasteur. Published by Elsevier Masson SAS. All rights reserved.

Nematode eel parasite found inside acanthocephalan cysts--a "Trojan horse" strategy?

Science.gov (United States)

Emde, Sebastian; Rueckert, Sonja; Kochmann, Judith; Knopf, Klaus; Sures, Bernd; Klimpel, Sven

2014-11-18

The invasive eel parasite Anguillicoloides crassus (syn. Anguillicola crassus) is considered one of the major causes for the decline of the European eel (Anguilla anguilla) panmictic population. It impairs the swim bladder function and reduces swimming performance of its host. The life cycle of this parasite involves different intermediate and paratenic hosts. Despite an efficient immune system of the paratenic fish hosts acting against infections with A. crassus, levels of parasitized eels remain high in European river systems. Recently, the round goby Neogobius melanostomus (Gobiidae) has become dominant in many rivers in Europe and is still spreading at a rapid pace. This highly invasive species might potentially act as an important, so far neglected paratenic fish host for A. crassus. Based on own observations and earlier single sightings of A. crassus in N. melanostomus, 60 fresh individuals of N. melanostomus were caught in the Rhine River and examined to assess the infection levels with metazoan parasites, especially A. crassus. Glycerin preparations were used for parasite identification. The parasite most frequently found in N. melanostomus was the acanthocephalan Pomphorhynchus sp. (subadult stage) which occurred mainly encysted in the mesenteries and liver. Every third gobiid (P = 31.7%) was infected by A. crassus larvae (L3) which exclusively occurred inside the acanthocephalan cysts. No intact or degenerated larvae of A. crassus were detected elsewhere in the goby, neither in the body cavity and mesenteries nor in other organs. Affected cysts contained the acanthocephalan larvae and 1-12 (mI =3) living A. crassus larvae. Additionally, encysted larvae of the nematode Raphidascaris acus were detected in the gobies, but only in the body cavity and not inside the acanthocephalan cysts. Based on our observations, we suggest that A. crassus might actively bypass the immune response of N. melanostomus by invading the cysts of acanthocephalan parasites of the

Pathology of AIDS-Study from a Neuropsychiatric Centre from South India

Directory of Open Access Journals (Sweden)

Vani Santosh

1998-01-01

Full Text Available The pathomorphological features noted in 47 cases of HIV/AIDS studied on autopsy (39 cases and surgical biopsy material (8 cases over a period of 8.5 years are described here. The serum samples of all cases and 26/29 CSF samples tested were positive for anti HIV antibodies. Majority of patients were young adult males in the age group 25-35 years. History of high risk behaviour was available in 59.6% cases. 38/39 autopsied cases succumbed to various opportunistic infections. Infection by a single pathogen was noted in 76.3% cases and due to multiple infectious agents in 23.7%. The commonest opportunistic infection was cryptococcosis (59% followed by toxoplasma encephalities (28.2% and tuberculous meningitis (25.6%. The others included meningococcal meningitis, herpes simplex, cytomegalovirus and acanthamoeba encephalitis noted in one case each. Features meningococcal meningitis, herpes simplex, cytomegalovirus and acanthamoeba encephalitis noted in one case each. Features of HIV leucoencephalitis was noted in one autopsy and tow brain biopsy specimens. Complete body necropsy carried out in 11/39 cases revealed disseminated cryptococcosis in 5, tuberculosis in 4 and features of HIV related changes of lymphoreticular organs in 3 cases. Pneumocystis carinil pneumonia was noted in 2 patients along with cryptococcosis. Diagnostic lymph node biopsy in 3 patients revealed tuberculous lymphadenitis in 2 and infection by non-tuberculous mycobacteria in one. Evidence of IIIV associated neuropathy was noted in 3 cases. In the present series, the incidence of cryptococcal meningitis was very high compared to neurotuberculosis as reported from other clinical and one autopsy series from other parts of India. Similarly, the incidence of toxoplasma encephalitis was also higher. The type of laboratory support and the experience of the pathologist play a crucial role in accurate identification of the pathogens. Therefore, a high index of clinical suspicion and a

A Student's Guide to Giant Viruses Infecting Small Eukaryotes: From Acanthamoeba to Zooxanthellae.

Science.gov (United States)

Wilhelm, Steven W; Bird, Jordan T; Bonifer, Kyle S; Calfee, Benjamin C; Chen, Tian; Coy, Samantha R; Gainer, P Jackson; Gann, Eric R; Heatherly, Huston T; Lee, Jasper; Liang, Xiaolong; Liu, Jiang; Armes, April C; Moniruzzaman, Mohammad; Rice, J Hunter; Stough, Joshua M A; Tams, Robert N; Williams, Evan P; LeCleir, Gary R

2017-03-17

The discovery of infectious particles that challenge conventional thoughts concerning "what is a virus" has led to the evolution a new field of study in the past decade. Here, we review knowledge and information concerning "giant viruses", with a focus not only on some of the best studied systems, but also provide an effort to illuminate systems yet to be better resolved. We conclude by demonstrating that there is an abundance of new host-virus systems that fall into this "giant" category, demonstrating that this field of inquiry presents great opportunities for future research.

Transmission of some species of internal parasites in horse foals born in 2013 in the same pasture on a farm in Central Kentucky

Directory of Open Access Journals (Sweden)

Tolliver S. C.

2015-09-01

Full Text Available The present research is a continuation of studies conducted periodically over 40 years on transmission of natural infections of internal parasites in the same horse herd on pasture (Field 10 on a farm in Central Kentucky. It included 12 mixed light horse foals born in 2013 and euthanatized between July, 2013 and April, 2014 for collection of internal parasites. Parasites found: Gasterophilus intestinalis, Strongyloides westeri, Parascaris equorum, Anoplocephala perfoliata, small strongyles (cyathostomes, Strongylus vulgaris, Strongylus edentatus and Thelazia lacrymalis. Prevalence generally was related to age of the foals. Overall prevalence and number of specimens were lower than in earlier studies except for P. equorum. There were 15 species (much fewer than previously of small strongyles found and recorded by location in the large intestines. All stages of small strongyles encysted in the mucosa of the large intestine were recovered by artificial digestion and in significantly lower numbers in older foals

Hydrocele of the canal of Nuck

Directory of Open Access Journals (Sweden)

Nourah ALSaleh

2018-02-01

Full Text Available Hydrocele of the canal of Nuck is a rare condition in female children caused by a failure of complete obliteration of the canal of Nuck. The canal of Nuck is an abnormal patent pouch of the peritoneum extending anterior to the round ligament of the uterus into the labia majora. Incomplete obliteration of this canal (patent processus vaginalis can result in either an inguinal hernia or a hydrocele. Here, we report two cases of hydrocele of the canal of Nuck presented within a two-month period. In the first case, incarcerated right-sided inguinal hernia was suspected while the second case was clinically diagnosed as encysted hydrocele of canal of Nuck. Both patients underwent surgical exploration. Hydrocelectomy with high ligation was also performed. One month after the surgery, the first patient showed signs of hydrocele on contralateral side, while the other patient showed no sign of contralateral hydrocele or hernia.

Modeling effective transmission pathways and control of the world's most successful parasite.

Science.gov (United States)

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. Copyright © 2013 Elsevier Inc. All rights reserved.

Rapid Identification of Seven Waterborne Exophiala Species by RCA DNA Padlock Probes.

Science.gov (United States)

Najafzadeh, M J; Vicente, V A; Feng, Peiying; Naseri, A; Sun, Jiufeng; Rezaei-Matehkolaei, A; de Hoog, G S

2018-03-05

The black yeast genus Exophiala includes numerous potential opportunistic species that potentially cause systematic and disseminated infections in immunocompetent individuals. Species causing systemic disease have ability to grow at 37-40 °C, while others consistently lack thermotolerance and are involved in diseases of cold-blooded, waterborne vertebrates and occasionally invertebrates. We explain a fast and sensitive assay for recognition and identification of waterborne Exophiala species without sequencing. The ITS rDNA region of seven Exophiala species (E. equina, E. salmonis, E. opportunistica, E. pisciphila, E. aquamarina, E. angulospora and E. castellanii) along with the close relative Veronaea botryosa was sequenced and aligned for the design of specific padlock probes for the detection of characteristic single-nucleotide polymorphisms. The assay demonstrated to successfully amplify DNA of target fungi, allowing detection at the species level. Amplification products were visualized on 1% agarose gels to confirm specificity of probe-template binding. Amounts of reagents were reduced to prevent the generation of false positive results. The simplicity, tenderness, robustness and low expenses provide padlock probe assay (RCA) a definite place as a very practical method among isothermal approaches for DNA diagnostics.

Occurrence of Infected Free-Living Amoebae in Cooling Towers of Southern Brazil.

Science.gov (United States)

Soares, Scheila S; Souza, Thamires K; Berté, Francisco K; Cantarelli, Vlademir V; Rott, Marilise B

2017-12-01

This study determined the occurrence of potentially pathogenic free-living amoebae (FLA) and bacteria associated with amoebae in air-conditioning cooling towers in southern Brazil. Water samples were collected from 36 cooling systems from air-conditioning in the state of Rio Grande do Sul, Brazil. The organisms were identified using polymerase chain reaction (PCR) and sequencing automated. The results showed that these aquatic environments, with variable temperature, are potential "hot spots" for emerging human pathogens like free-living amoebae and bacteria associated. In total, 92% of the cooling-tower samples analyzed were positive for FLA, and Acanthamoeba was the dominant genus by culture and PCR. Amoebal isolates revealed intracellular bacteria in 39.3% of them and all were confirmed as members of the genus Pseudomonas. The results obtained show the important role of cooling towers as a source of amoebae-associated pathogens.

Aerobic mitochondria of parasitic protists: Diverse genomes and complex functions.

Science.gov (United States)

Zíková, Alena; Hampl, Vladimír; Paris, Zdeněk; Týč, Jiří; Lukeš, Julius

In this review the main features of the mitochondria of aerobic parasitic protists are discussed. While the best characterized organelles are by far those of kinetoplastid flagellates and Plasmodium, we also consider amoebae Naegleria and Acanthamoeba, a ciliate Ichthyophthirius and related lineages. The simplistic view of the mitochondrion as just a power house of the cell has already been abandoned in multicellular organisms and available data indicate that this also does not apply for protists. We discuss in more details the following mitochondrial features: genomes, post-transcriptional processing, translation, biogenesis of iron-sulfur complexes, heme metabolism and the electron transport chain. Substantial differences in all these core mitochondrial features between lineages are compatible with the view that aerobic protists harbor organelles that are more complex and flexible than previously appreciated. Copyright © 2016 Elsevier B.V. All rights reserved.

A Student’s Guide to Giant Viruses Infecting Small Eukaryotes: From Acanthamoeba to Zooxanthellae

Directory of Open Access Journals (Sweden)

Steven W. Wilhelm

2017-03-01

Full Text Available The discovery of infectious particles that challenge conventional thoughts concerning “what is a virus” has led to the evolution a new field of study in the past decade. Here, we review knowledge and information concerning “giant viruses”, with a focus not only on some of the best studied systems, but also provide an effort to illuminate systems yet to be better resolved. We conclude by demonstrating that there is an abundance of new host–virus systems that fall into this “giant” category, demonstrating that this field of inquiry presents great opportunities for future research.

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

Energy Technology Data Exchange (ETDEWEB)

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.

Dioctophyma renale (Goeze, 1782) in the abdominal cavity of a capuchin monkey (Cebus apella), Brazil.

Science.gov (United States)

Ishizaki, Mirian Naomi; Imbeloni, Aline Amaral; Muniz, José Augusto Pereira Carneiro; Scalercio, Sarah Raphaella Rocha de Azevedo; Benigno, Raimundo Nonato Moraes; Pereira, Washington Luiz Assunção; Cunha Lacreta Junior, Antonio Carlos

2010-10-29

This study reports a case of parasitism by Dioctophyma renale (Goeze, 1762) encysted in the abdominal cavity of a capuchin monkey (Cebus apella) coming from the Centro Nacional de Primatas, Brazil. The animal was sent to the Veterinary Clinic sector with an increase in abdominal volume and no occurrence of any other clinical signs. Upon palpation, a movable circular mass with a diameter of approximately 10 cm was found. Urinalysis, complete blood count and serum biochemistry were performed without revealing any alterations. The animal was then submitted to an abdominal ultrasound exam. The cyst was punctured and a surgical removal procedure was performed, revealing a brownish-colored cylindrical structure that was already deteriorated, making it impossible to perform morphological analysis and classification. In the sediment of the liquid found, eggs were encountered that had morphological characteristics compatible with D. renale. The objective of this paper is to report the first case of parasitism by D. renale in C. apella (Linnaeus, 1758). Copyright © 2010 Elsevier B.V. All rights reserved.

Molecular characterization of enolase gene from Taenia multiceps.

Science.gov (United States)

Li, W H; Qu, Z G; Zhang, N Z; Yue, L; Jia, W Z; Luo, J X; Yin, H; Fu, B Q

2015-10-01

Taenia multiceps is a cestode parasite with its larval stage, known as Coenurus cerebralis, mainly encysts in the central nervous system of sheep and other livestocks. Enolase is a key glycolytic enzyme and represents multifunction in most organisms. In the present study, a 1617bp full-length cDNA encoding enolase was cloned from T. multiceps and designated as TmENO. A putative encoded protein of 433 amino acid residues that exhibited high similarity to helminth parasites. The recombinant TmENO protein (rTmENO) showed the catalytic and plasminogen-binding characteristics after the TmENO was subcloned and expressed in the pET30a(+) vector. The TmENO gene was transcribed during the adult and larval stages and was also identified in both cyst fluid and as a component of the adult worms and the metacestode by western blot analysis. Taken together, our results will facilitate further structural characterization for TmENO and new potential control strategies for T. multiceps. Copyright © 2015 Elsevier Ltd. All rights reserved.

Establishment of a continuous culture system for Entamoeba muris and analysis of the small subunit rRNA gene

Directory of Open Access Journals (Sweden)

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.

Protection against heterologous infection by using cross antigenicity between schistosoma mansoni and fasciola hepatica

International Nuclear Information System (INIS)

Moawad, M.A.; Amin, M.M.

2004-01-01

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

Liver histopathology in the cane toad, Rhinella marina (Amphibia: Bufonidae), induced by Ortleppascaris sp. larvae (Nematoda: Ascarididae).

Science.gov (United States)

Silva, Jefferson P E; da Silva, Djane C B; Melo, Francisco T V; Giese, Elane G; Furtado, Adriano P; Santos, Jeannie N

2013-04-01

Exposure to parasites is considered to be an important factor in the development of many diseases and histopathologies which are the result of the parasite-host interaction. The present study evaluated the impact of natural infection by larvae of Ortleppascaris sp. (Nematoda: Ascaridida) in the liver of the cane toad Rhinella marina (Linnaeus, 1758). Larvae were encysted in nodules delimited by collagenous fibers and fibroblasts or freely within the hepatic parenchyma, provoking a clear response from the host. The histological examination of the liver revealed viable larvae in a number of different developmental stages, as well as cysts filled with amorphous material and cell residues and surrounded by dense fibrotic tissue. The infection of the liver by these larvae induces a significant increase in the area occupied by melanomacrophages and a reduction or deficit in the vascularization of the liver, hypertrophy of the hepatocytes, vacuolar bodies, and cytoplasmatic granules. Focal concentrations of inflammatory infiltrates were observed enclosing the unencapsulated early-stage larvae. These results indicate that infection by Ortleppascaris sp. induces severe physiological problems and histopathological lesions in the liver of R. marina .

Heterophysiasis, an intestinal fluke infection of man and vertebrates transmitted by euryhaline gastropods and fish

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Taraschewski, H.

1984-03-01

Heterophyes heterophyes, agent of human heterophyiasis in the Near East, is transmitted in marine lagoons and saline inland waters, where the euryhaline intermediate hosts are abundant. In Egypt, mullets, the predominant second intermediate hosts, are customarily consumed raw; thus man becomes infected easily. Symptoms of human infections are usually considered mild. Mullets do not seem to be affected by the metacercariae encysted in the muscles, whereas the growth of the snail host Pirenella conica was found to be enhanced due to the infestation by the trematodes. In laboratory experiments, the flukes were found to be well developed in dogs, foxes and cats, but failed to reach sexual maturity in several other potentially piscivorous mammals and birds. In nature, dogs probably serve as the major reservoir hosts. Heterophyiasis is most prevalent in the Nile Delta, a huge brackish water area which is densely populated by humans and, consequently, also by dogs and cats. In the Far East, besides Heterophyes nocens, several other heterophysids with marine or fresh-water life-cycles are known to infect humans.

Pediatric neurocysticercosis: current challenges and future prospects

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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

High-resolution electron microscopic evidence for the filamentous structure of the cyst wall in Giardia muris and Giardia duodenalis.

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Erlandsen, S L; Bemrick, W J; Pawley, J

1989-10-01

High-resolution morphological studies of the cyst wall of Giardia spp. were performed using low-voltage scanning electron microscopy (LVSEM) and transmission electron microscopy (TEM). The cyst wall was composed of membranous and filamentous layers. The membranous layer consisted of an inner and an outer cyst membrane separated by a thin layer of cytoplasm. The filamentous layer contained individual filaments that ranged from 7 to 20 nm in diameter when measured by LVSEM, formed a dense meshwork with branches or interconnections, and were occasionally arranged on the surface in whorled patterns. Cysts of Giardia muris from mice, Giardia duodenalis from dogs, pigs, voles, beavers, muskrats, and humans, and Giardia psittaci from a bird (parakeet), possessed an essentially identical wall composed of filaments. Inducement of excystation in viable Giardia cysts produced a dramatic increase in the interfilament spacing over an entire cyst, but none was observed in heat-killed or chemically fixed control cysts. These results demonstrated that the cyst wall of Giardia spp. was composed of a complex arrangement of filaments, presumably formed during the process of encystment.

Common variable immune deficiency syndrome

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Ahmed A. El-Masry

2014-07-01

Full Text Available A male patient 40 years old, non smoker, presented with fever, cough and expectoration of greenish sputum and diarrhea of 1 week duration. The condition started 3 years ago, by cough and expectoration of about ¼ cup/day of greenish sputum, not related to posture, along with fever up to 39 °C with loss of weight about 12 kg in one month and associated with diarrhea and mucus shedding. The patient sought medical advice and received empirical antibiotics and symptomatic treatments with partial clinical improvement. The patient showed multiple relapses of same respiratory and gastro-enterology symptoms every 3–4 weeks with 4 hospital admissions. CT-chest was done and revealed emphysematous changes with basal inflammatory reaction (Fig. 1; sputum workup showed no acid fast bacilli and growth of normal flora, fasting blood glucose was 102 mg/dl. A second CT-chest was done after one year and showed right sided pneumonic consolidation, bilateral pneumonic reaction with multiple mediastinal lymphadenopathy, bilateral pleural thickening and right encysted pleural effusion (Fig. 2.

Minor Lagochilascaris Otomastoiditis in a Child: Case Report

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Guimaraes, Valeriana de Castro

2010-09-01

Full Text Available Introduction: The lagochilascariosis is an emerging zoonosis determined by the presence of helminth Lagochilascaris minor in human tissues. Infection occurs through ingestion of larvae encysted in the subcutaneous tissue, muscles and viscera of wild animals consumed by man so raw or undercooked. The disease is rare, insidious, chronic condition, characterized by the appearance of lesions, especially in the neck, mastoid, ear, nose and oropharynx. A serious, sometimes fatal, depend on the location of the parasite. Objective: To describe a case of otomastoiditis by L. minor in child, attended at the Hospital das Clinicas, Federal University of Goias. Case Report: Patient 10 years old female, complaining of otalgia and purulent otorrhea right. Otoscopy left ear was normal, while the right ear had retroauricular edema, polyps in the external ear canal and fistula with drainage of pus. The tympanic membrane was intact.The stages of treatment were described since the initial consultation to the recovery of the patient. Final Comments: The clinician should be alert to the importance in considering the differential diagnosis of this disease on individuals residing in rural areas.

Use of the vital stain FUN-1 indicates viability of Phytophthora capsici propagules and can be used to predict maximum zoospore production.

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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.

Incidence of adult brain cancers is higher in countries where the protozoan parasite Toxoplasma gondii is common

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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.

Systemic acanthamoebiasis associated with canine distemper in dogs in the semiarid region of Paraíba, Brazil

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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

Repertoire of free-living protozoa in contact lens solutions.

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Bouchoucha, Ibtissem; Aziz, Aurore; Hoffart, Louis; Drancourt, Michel

2016-10-29

The repertoire of free-living protozoa in contact lens solutions is poorly known despite the fact that such protozoa may act as direct pathogens and may harbor intra-cellular pathogens. Between 2009 and 2014, the contact lens solutions collected from patients presenting at our Ophthalmology Department for clinically suspected keratitis, were cultured on non-nutrient agar examined by microscope for the presence of free-living protozoa. All protozoa were identified by 18S rRNA gene sequencing. A total of 20 of 233 (8.6 %) contact lens solution specimens collected from 16 patients were cultured. Acanthamoeba amoeba in 16 solutions (80 %) collected from 12 patients and Colpoda steini, Cercozoa sp., Protostelium sp. and a eukaryotic more closely related to Vermamoeba sp., were each isolated in one solution. Cercozoa sp., Colpoda sp., Protostelium sp. and Vermamoeba sp. are reported for the first time as contaminating contact lens solutions. The repertoire of protozoa in contact lens solutions is larger than previously known.

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

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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.

Parasite manipulation of brain monoamines in California killifish (Fundulus parvipinnis) by the trematode Euhaplorchis californiensis

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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.

Relationship between thermal loading and parasitism in the mosquitofish

International Nuclear Information System (INIS)

Aho, J.M.; Gibbons, J.W.; Esch, G.W.

1976-01-01

The relationship between thermal loading and parasitism was examined in 980 mosquitofish, Gambusia affinis, taken from areas of varying thermal conditions at the Savannah River Plant (SRP) near Aiken, S. C. Collections were made at 2-week intervals from May to August 1974 and again from January to February 1975. The metacercaria of two strigeid trematodes, Ornithodiplostomum ptychocheilus and Diplostomum scheuringi were the only species of parasites recovered. Ornithodiplostomum ptychocheilus was always found encysted in the brain and eyes, whereas D. scheuringi was restricted exclusively to the body cavity. The density of the body-cavity parasite was highest in fish from areas of ambient temperatures and declined in fish from areas with higher water temperatures. The density of the brain parasite, on the other hand, was higher in fish from areas directly receiving thermal effluent than in fish from ambient-temperature areas. The body-cavity parasite was absent from mosquitofish from Pond C, but the infection percentages were relatively consistent in fish from other areas in the Par Pond system regardless of water temperature. The brain metacercaria were recovered from 95 percent of fish from Pond C

Anthelminthic and antiallergic activities of Mangifera indica L. stem bark components Vimang and mangiferin.

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García, D; Escalante, M; Delgado, R; Ubeira, F M; Leiro, J

2003-12-01

This study investigated the antiallergic and anthelmintic properties of Vimang (an aqueous extract of Mangifera indica family stem bark) and mangiferin (the major polyphenol present in Vimang) administered orally to mice experimentally infected with the nematode, Trichinella spiralis. Treatment with Vimang or mangiferin (500 or 50 mg per kg body weight per day, respectively) throughout the parasite life cycle led to a significant decline in the number of parasite larvae encysted in the musculature; however, neither treatment was effective against adults in the gut. Treatment with Vimang or mangiferin likewise led to a significant decline in serum levels of specific anti-Trichinella IgE, throughout the parasite life cycle. Finally, oral treatment of rats with Vimang or mangiferin, daily for 50 days, inhibited mast cell degranulation as evaluated by the passive cutaneous anaphylaxis test (sensitization with infected mouse serum with a high IgE titre, then stimulation with the cytosolic fraction of T. spiralis muscle larvae). Since IgE plays a key role in the pathogenesis of allergic diseases, these results suggest that Vimang and mangiferin may be useful in the treatment of diseases of this type. Copyright 2003 John Wiley & Sons, Ltd.

Use of gamma radiation to prevent infectivity of meat-borne parasites and effect of irradiation on the biochemical and its properties of meats

International Nuclear Information System (INIS)

Zahran, D.A.H.

2002-01-01

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

Zoonotic ectopic fascioliasis: review and discussion.

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Rashed, Amr A; Khalil, Hazem H M; Morsy, Ayman T A

2010-12-01

Ectopic fascioliasis (EF) has direct and indirect effects on both humans and animals. The phenomenon of EF was individual cases in the period from 1950 up to the end of last century. From the period of 2000 up to 2006, plenty of researches were on EF in the developed and undeveloped countries. Nineteen EF cases infected with the immature and few with the mature worms were 13 females and 6 males. Three cases of lymphatic, pleural and breast fascioliasis reached the adults and laid their eggs in a lymph node in the cervical region pleural cavity and breast tissues. Until recent, knowledge about the ectopic fascioliasis pathway is little. Fasciola hepatica was the commonest species in most cases. The effect of fascioliasis might be direct to liver as ectopic foci or indirect on other organs due to the metabolites and secretory excretory products. All ages and both sexes were EF infected. Watercress topped the list of water plants born encysted metacercariae followed by lettuce, mint, and alfalfa. Nearly 24 million Egyptians at risk and about 800,000 were infected. On the global scale, about 180 million are at risk of infection.

Persistence of host response against glochidia larvae in Micropterus salmoides.

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Dodd, Benjamin J; Barnhart, M Christopher; Rogers-Lowery, Constance L; Fobian, Todd B; Dimock, Ronald V

2006-11-01

Host fish acquire resistance to the parasitic larvae (glochidia) of freshwater mussels (Unionidae). Glochidia metamorphose into juvenile mussels while encysted on host fish. We investigated the duration of acquired resistance of largemouth bass, Micropterus salmoides (Lacepède, 1802) to glochidia of the broken rays mussel, Lampsilis reeveiana (Call, 1887). Fish received three successive priming infections with glochidia to induce an immune response. Primed fish were held at 22-23 degrees C and were challenged (re-infected) at intervals after priming. Metamorphosis success was quantified as the percent of attached glochidia that metamorphosed to the juvenile stage and were recovered alive. Metamorphosis success at 3, 7, and 12 months after priming was significantly lower on primed fish (26%, 40%, and 68% respectively) than on control fish (85%, 93%, and 92% respectively). A second group of largemouth bass was similarly primed and blood was extracted. Immunoblotting was used to detect host serum antibodies to L. reeveiana glochidia proteins. Serum antibodies were evident in primed fish, but not in naive control fish. Acquired resistance of host fish potentially affects natural reproduction and artificial propagation of unionids, many of which are of conservation concern.

Characterization of the local immune response to cyst antigens during the acute and elimination phases of primary murine giardiasis.

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Abdul-Wahid, Aws; Faubert, Gaétan

2008-05-01

During the course of a giardial infection, the host's immune system is presented with a variety of Giardia antigens as trophozoites differentiate, through encysting cells, to form the infective cysts. Previous studies examining the host's immune response during giardial infections have focused on trophozoite-derived antigens (Ags). In this study, we were interested to determine if the host's immune system reacts to cyst Ags during the acute and elimination phases, when there is cyst shedding. For this purpose, we used antigenic extracts from trophozoites (Troph), encysting cells (ENC), and purified giardial cyst walls (PCW), as well as purified recombinant cyst wall protein 2 (rCWP2). Comparative analysis of the parasite extracts using SDS-PAGE analysis and surface-enhanced laser desorption/ionization time of flight mass spectrometry resulted in the detection of 175 protein entities, of which 26 were Troph-specific proteins, 17 ENC-specific proteins, and 31 were PCW-specific proteins. On the other hand, we detected 34 proteins shared between Troph and ENC, 19 proteins that were shared between ENC and PCW, and 29 proteins that were common to Troph and PCW. Finally, we detected 19 proteins that were shared by all three extract samples. BALB/c mice were infected with 10(5)Giardia muris cysts and sacrificed either at the acute or elimination phases of infection (days 12 and 40, respectively), and lymphocytes were isolated from the Peyer's patches (PP). Using flow cytometry, we detected significant increases in the number of PP-derived CD4(+) and CD19(+), but not CD8(+) lymphocytes. Quantification of the number of mucosal IL-4 and IFN-gamma secreting T-lymphocytes by enzyme-linked immunosorbent spot assay showed that these cells reacted by secreting similar levels of IL-4 and IFN-gamma, regardless of the Ag or the phase of infection. Analysis of intestinal humoral immune responses by ELISA resulted in the detection of Ag-specific IgA and IgG intestinal antibodies

Infection and Proliferation of Giant Viruses in Amoeba Cells.

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Takemura, Masaharu

2016-01-01

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

Metacommunity analysis of amoeboid protists in grassland soils.

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Fiore-Donno, Anna Maria; Weinert, Jan; Wubet, Tesfaye; Bonkowski, Michael

2016-01-11

This study reveals the diversity and distribution of two major ubiquitous groups of soil amoebae, the genus Acanthamoeba and the Myxomycetes (plasmodial slime-moulds) that are rarely, if ever, recovered in environmental sampling studies. We analyzed 150 grassland soil samples from three Biodiversity Exploratories study regions in Germany. We developed specific primers targeting the V2 variable region in the first part of the small subunit of the ribosomal RNA gene for high-throughput pyrotag sequencing. From ca. 1 million reads, applying very stringent filtering and clustering parameters to avoid overestimation of the diversity, we obtained 273 acanthamoebal and 338 myxomycete operational taxonomic units (OTUs, 96% similarity threshold). This number is consistent with the genetic diversity known in the two investigated lineages, but unequalled to date by any environmental sampling study. Only very few OTUs were identical to already known sequences. Strikingly different OTUs assemblages were found between the three German regions (PerMANOVA p.value = 0.001) and even between sites of the same region (multiple-site Simpson-based similarity indices <0.4), showing steep biogeographical gradients.

Mimivirus: leading the way in the discovery of giant viruses of amoebae.

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Colson, Philippe; La Scola, Bernard; Levasseur, Anthony; Caetano-Anollés, Gustavo; Raoult, Didier

2017-04-01

The accidental discovery of the giant virus of amoeba - Acanthamoeba polyphaga mimivirus (APMV; more commonly known as mimivirus) - in 2003 changed the field of virology. Viruses were previously defined by their submicroscopic size, which probably prevented the search for giant viruses, which are visible by light microscopy. Extended studies of giant viruses of amoebae revealed that they have genetic, proteomic and structural complexities that were not thought to exist among viruses and that are comparable to those of bacteria, archaea and small eukaryotes. The giant virus particles contain mRNA and more than 100 proteins, they have gene repertoires that are broader than those of other viruses and, notably, some encode translation components. The infection cycles of giant viruses of amoebae involve virus entry by amoebal phagocytosis and replication in viral factories. In addition, mimiviruses are infected by virophages, defend against them through the mimivirus virophage resistance element (MIMIVIRE) system and have a unique mobilome. Overall, giant viruses of amoebae, including mimiviruses, marseilleviruses, pandoraviruses, pithoviruses, faustoviruses and molliviruses, challenge the definition and classification of viruses, and have increasingly been detected in humans.

Corneal ring infiltration in contact lens wearers

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Seyed Ali Tabatabaei

2017-01-01

Full Text Available To report a case of atypical sterile ring infiltrates during wearing soft silicone hydrogel contact lens due to poor lens care. A 29-year-old woman presented with complaints of pain, redness, and morning discharge. She was wearing soft silicone hydrogel contact lens previously; her current symptoms began 1 week before presentation. On examination, best-corrected visual acuity was 20/40 in that eye. Slit-lamp examination revealed dense, ring-shaped infiltrate involving both the superficial and deep stromal layers with lucid interval to the limbus, edema of the epithelium, epithelial defect, and vascularization of the superior limbus. Cornea-specific in vivo laser confocal microscopy (Heidelberg Retina Tomograph 2 Rostock Cornea Module, HRT 2-RCM, Heidelberg Engineering GmbH, Dossenheim, Germany revealed Langerhans cells and no sign of Acanthamoeba or fungal features, using lid scraping and anti-inflammatory drops; her vision completely recovered. We reported an atypical case of a sterile corneal ring infiltrate associated with soft contact lens wearing; smear, culture, and confocal microscopy confirmed a sterile inflammatory reaction.

Comparative Genomics of Chrysochromulina Ericina Virus and Other Microalga-Infecting Large DNA Viruses Highlights Their Intricate Evolutionary Relationship with the Established Mimiviridae Family.

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Gallot-Lavallée, Lucie; Blanc, Guillaume; Claverie, Jean-Michel

2017-07-15

Chrysochromulina ericina virus CeV-01B (CeV) was isolated from Norwegian coastal waters in 1998. Its icosahedral particle is 160 nm in diameter and encloses a 474-kb double-stranded DNA (dsDNA) genome. This virus, although infecting a microalga (the haptophyceae Haptolina ericina , formerly Chrysochromulina ericina ), is phylogenetically related to members of the Mimiviridae family, initially established with the acanthamoeba-infecting mimivirus and megavirus as prototypes. This family was later split into two genera ( Mimivirus and Cafeteriavirus ) following the characterization of a virus infecting the heterotrophic stramenopile Cafeteria roenbergensis (CroV). CeV, as well as two of its close relatives, which infect the unicellular photosynthetic eukaryotes Phaeocystis globosa (Phaeocystis globosa virus [PgV]) and Aureococcus anophagefferens (Aureococcus anophagefferens virus [AaV]), are currently unclassified by the International Committee on Viral Taxonomy (ICTV). The detailed comparative analysis of the CeV genome presented here confirms the phylogenetic affinity of this emerging group of microalga-infecting viruses with the Mimiviridae but argues in favor of their classification inside a distinct clade within the family. Although CeV, PgV, and AaV share more common features among them than with the larger Mimiviridae , they also exhibit a large complement of unique genes, attesting to their complex evolutionary history. We identified several gene fusion events and cases of convergent evolution involving independent lateral gene acquisitions. Finally, CeV possesses an unusual number of inteins, some of which are closely related despite being inserted in nonhomologous genes. This appears to contradict the paradigm of allele-specific inteins and suggests that the Mimiviridae are especially efficient in spreading inteins while enlarging their repertoire of homing genes. IMPORTANCE Although it infects the microalga Chrysochromulina ericina , CeV is more closely

Antimycobacterial drug discovery using Mycobacteria-infected amoebae identifies anti-infectives and new molecular targets.

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Trofimov, Valentin; Kicka, Sébastien; Mucaria, Sabrina; Hanna, Nabil; Ramon-Olayo, Fernando; Del Peral, Laura Vela-Gonzalez; Lelièvre, Joël; Ballell, Lluís; Scapozza, Leonardo; Besra, Gurdyal S; Cox, Jonathan A G; Soldati, Thierry

2018-03-02

Tuberculosis remains a serious threat to human health world-wide, and improved efficiency of medical treatment requires a better understanding of the pathogenesis and the discovery of new drugs. In the present study, we performed a whole-cell based screen in order to complete the characterization of 168 compounds from the GlaxoSmithKline TB-set. We have established and utilized novel previously unexplored host-model systems to characterize the GSK compounds, i.e. the amoeboid organisms D. discoideum and A. castellanii, as well as a microglial phagocytic cell line, BV2. We infected these host cells with Mycobacterium marinum to monitor and characterize the anti-infective activity of the compounds with quantitative fluorescence measurements and high-content microscopy. In summary, 88.1% of the compounds were confirmed as antibiotics against M. marinum, 11.3% and 4.8% displayed strong anti-infective activity in, respectively, the mammalian and protozoan infection models. Additionally, in the two systems, 13-14% of the compounds displayed pro-infective activity. Our studies underline the relevance of using evolutionarily distant pathogen and host models in order to reveal conserved mechanisms of virulence and defence, respectively, which are potential "universal" targets for intervention. Subsequent mechanism of action studies based on generation of over-expresser M. bovis BCG strains, generation of spontaneous resistant mutants and whole genome sequencing revealed four new molecular targets, including FbpA, MurC, MmpL3 and GlpK.

Risk Assessment for the Spread of Serratia marcescens Within Dental-Unit Waterline Systems Using Vermamoeba vermiformis.

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Lal, Sham; Singhrao, Sim K; Achilles-Day, Undine E M; Morton, L H Glyn; Pearce, Mark; Crean, StJohn

2015-10-01

Vermamoeba vermiformis is associated with the biofilm ecology of dental-unit waterlines (DUWLs). This study investigated whether V. vermiformis is able to act as a vector for potentially pathogenic bacteria and so aid their dispersal within DUWL systems. Clinical dental water was initially examined for Legionella species by inoculating it onto Legionella selective-medium plates. The molecular identity/profile of the glassy colonies obtained indicated none of these isolates were Legionella species. During this work bacterial colonies were identified as a non-pigmented Serratia marcescens. As the water was from a clinical DUWL which had been treated with Alpron™, this prompted the question as to whether S. marcescens had developed resistance to the biocide. Exposure to Alpron™ indicated that this dental biocide was effective, under laboratory conditions, against S. marcescens at up to 1 × 10(8) colony forming units/millilitre (cfu/ml). V. vermiformis was cultured for 8 weeks on cells of S. marcescens and Escherichia coli. Subsequent electron microscopy showed that V. vermiformis grew equally well on S. marcescens and E. coli (P = 0.0001). Failure to detect the presence of S. marcescens within the encysted amoebae suggests that V. vermiformis is unlikely to act as a vector supporting the growth of this newly isolated, nosocomial bacterium.

Gene expression profiling during asexual development of the late blight pathogen Phytophthora infestans reveals a highly dynamic transcriptome.

Science.gov (United States)

Judelson, Howard S; Ah-Fong, Audrey M V; Aux, George; Avrova, Anna O; Bruce, Catherine; Cakir, Cahid; da Cunha, Luis; Grenville-Briggs, Laura; Latijnhouwers, Maita; Ligterink, Wilco; Meijer, Harold J G; Roberts, Samuel; Thurber, Carrie S; Whisson, Stephen C; Birch, Paul R J; Govers, Francine; Kamoun, Sophien; van West, Pieter; Windass, John

2008-04-01

Much of the pathogenic success of Phytophthora infestans, the potato and tomato late blight agent, relies on its ability to generate from mycelia large amounts of sporangia, which release zoospores that encyst and form infection structures. To better understand these stages, Affymetrix GeneChips based on 15,650 unigenes were designed and used to profile the life cycle. Approximately half of P. infestans genes were found to exhibit significant differential expression between developmental transitions, with approximately (1)/(10) being stage-specific and most changes occurring during zoosporogenesis. Quantitative reverse-transcription polymerase chain reaction assays confirmed the robustness of the array results and showed that similar patterns of differential expression were obtained regardless of whether hyphae were from laboratory media or infected tomato. Differentially expressed genes encode potential cellular regulators, especially protein kinases; metabolic enzymes such as those involved in glycolysis, gluconeogenesis, or the biosynthesis of amino acids or lipids; regulators of DNA synthesis; structural proteins, including predicted flagellar proteins; and pathogenicity factors, including cell-wall-degrading enzymes, RXLR effector proteins, and enzymes protecting against plant defense responses. Curiously, some stage-specific transcripts do not appear to encode functional proteins. These findings reveal many new aspects of oomycete biology, as well as potential targets for crop protection chemicals.

Inhibition of Phytophthora parasitica and P. capsici by Silver Nanoparticles Synthesized Using Aqueous Extract of Artemisia absinthium.

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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.

Differences in pigmentation between life cycle stages in Scrippsiella lachrymosa (dinophyceae).

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Persson, Agneta; Smith, Barry C; Cyronak, Tyler; Cooper, Emily; DiTullio, Giacomo R

2016-02-01

Various life cycle stages of cyst-producing dinoflagellates often appear differently colored under the microscope; gametes appear paler while zygotes are darker in comparison to vegetative cells. To compare physiological and photochemical competency, the pigment composition of discrete life cycle stages was determined for the common resting cyst-producing dinoflagellate Scrippsiella lachrymosa. Vegetative cells had the highest cellular pigment content (25.2 ± 0.5 pg · cell(-1) ), whereas gamete pigment content was 22% lower. The pigment content of zygotes was 82% lower than vegetative cells, even though they appeared darker under the microscope. Zygotes of S. lachrymosa contained significantly higher cellular concentrations of β-carotene (0.65 ± 0.15 pg · cell(-1) ) than all other life stages. Photoprotective pigments and the de-epoxidation ratio of xanthophylls-cycle pigments in S. lachrymosa were significantly elevated in zygotes and cysts compared to other stages. This suggests a role for accessory pigments in combating intracellular oxidative stress during sexual reproduction or encystment. Resting cysts contained some pigments even though chloroplasts were not visible, suggesting that the brightly colored accumulation body contained photosynthetic pigments. The differences in pigmentation between life stages have implications for interpretation of pigment data from field samples when sampled during dinoflagellate blooms. © 2015 Phycological Society of America.

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.

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Martorelli, Sergio R; Fredensborg, Brian L; Mouritsen, Kim N; Poulin, Robert

2004-04-01

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 other related species mainly in the shape, size, and patterns of distributions of the spines on the cirrus, the shape of the metraterm, the presence of an unlobed ovary, and the complete ring of the vitelline follicles. Based on morphometric features of metacercariae and adult specimens, the trophic relationships among invertebrate and vertebrate hosts, experimental infections, and previous reports of species of Maritrema with similar transmission patterns, the life cycle of M. novaezealandensis n. sp. is described. A 3-host life cycle is proposed for this parasite. The first intermediate host is the mud snail, Zeacumantus subcarinatus, in which the cercarial stage is produced in sporocysts located within the gonad of the snail. At least 3 crab species (Hemigrapsus crenulatus, Macrophtalmus hirtipes, and Halicarcinus whitei) and several species of amphipods act as second intermediate hosts, with 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.

Studies on vertical transmission of Trichinella spiralis in experimentally infected guinea pigs (Cavia porcellus).

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Riva, Eliana; Fiel, Cesar; Bernat, Gisele; Muchiut, Sebastián; Steffan, Pedro

2017-08-01

An experimental study to enhance knowledge on the capability of Trichenella spiralis to pass from guinea pigs to progeny at different periods of pregnancy or lactation was performed. For this purpose, 18 female adult guinea pigs were inoculated with 100 or 1000 T. spiralis muscle larvae (ML) during early, late gestation and during lactation period. The presence of T. spiralis (ML) in mothers and newborns was studied through enzymatic digestion from muscle samples. ML were observed in 9 of 42 newborn guinea pigs and levels of infection were significantly higher when infections of mothers were done during late gestation (p = 0.0046) with the high infective dose (p = 0.0043). T. spiralis ML were not recovered from any of the newborns from mothers infected in the lactation period. Ten out of 18 infected mothers presented larvae 1 in their mammary glands. Muscle samples from the tongue and the masseter showed the highest larval burdens. These observations confirm previous reports on that ML of T. spiralis are capable to pass through placental tissues to reach and encyst in striated muscle groups of newborn guinea pigs. This study may also reinforce the importance of preventive programs to control trichinellosis in those endemic areas where pregnant women would have high risk of infection.

Resveratrol Induces Apoptosis-Like Death and Prevents In Vitro and In Vivo Virulence of Entamoeba histolytica

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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

Draft genome sequencing of giardia intestinalis assemblage B isolate GS: is human giardiasis caused by two different species?

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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.

An incidental case of biliary fascioliasis mimicking cholangiocellular carcinoma.

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Şenateş, Ebubekir; Doğan, Abdullah; Şenates, Banu Erkalma; Bodakçi, Erdal; Bekçibasi, Muhammet

2014-12-01

Fascioliasis is a zoonotic infestation caused by Fasciola hepatica that usually attacks mammals, such as goats, sheep and cattle. The parasites can infect humans via freshwater plants contaminated with encysted metacercariae. In the acute phase, which involves hepatic invasion, the disease may present with abdominal pain, mild fever and hepatomegaly. In the chronic phase, the parasites settle into the biliary tracts, and then cause cholangitis and cholestasis. Sometimes, the disease may mimic malignancies, creating a mass appearance. Endoscopic retrograde cholangiopancreatography (ERCP) is an important diagnostic and treatment method because it allows simultaneous diagnosis and treatment. Here, we present a 44-year-old female patient who presented to our hospital with complaints of abdominal pain, nausea, vomiting, anorexia and weight loss. After diagnostic investigations with laboratory and imaging methods, she was initially hospitalized with a preliminary diagnosis of cholangiocellular carcinoma (CCC). However, after a full work-up, the patient was diagnosed with Fasciola hepatica via ERCP and parasites were extracted with ERCP at the same time and then treated with a single dose of triclabendazole 10 mg/kg. Two months later, the clinical status of the patient had improved markedly, with resolution of all symptoms and all laboratory and imaging tests returning to within normal limits.

Effects of altered temperature and precipitation on desert protozoa associated with biological soil crusts.

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Darby, Brian J; Housman, David C; Zaki, Amr M; Shamout, Yassein; Adl, Sina M; Belnap, Jayne; Neher, Deborah 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 degrees C. Cysts survived the upper end of daily temperatures (37-55 degrees C), and could be stimulated to excyst if temperatures were reduced to 15 degrees 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.

[Morphological features of utilization intraperitoneal double-sided prostheses in inguinoplasty in dogs].

Science.gov (United States)

de Andrade, Luiz Carlos; Ceneviva, Reginaldo; Coutinho-Netto, Joaquim; Silva Júnior, Orlando de Castro e; dos Santos, José Sebastião; Sukeda, Daniel Hirochi

2009-10-01

To asses the morphological features of the behavior of a double-sided prostheses using inguinoplasty laparotomy in dogs with latex side turned to the visceras. Twenty dogs were divided into two groups of 10 and submitted into infraumbilical laparotomy with double-sided prostheses fixed in an inguinal area and in the other side area a control prostheses of polipropilene (PPL). Macroscopics itens were studied on the 14th and 28th day post-operatory, and they were related to obstruction and intestinal fistulas, encystation, fusion and especially sticker. The microscopic analysis covered the inflammatory process in its acute, chronic and restored phase Infectious process, obstruction or intestinal fistula did not happen. The prostheses presented good accommodation and incorporation. The stickers happened with more prevalent and intensity with the PPL (p0,05). The double-sided prostheses in its parietal side adds the advantages of the incorporation's potential to the noticed material with PPL to the biocompatibility from the latex in its visceral side. The little distance between the PPL disc and the edge of the double-sided prostheses (2 cm) allied to its sticking with just five staples is not enough to avoid gaps, through which the epíploon migrated towards to the inflammatory process provoked by PPL in the parietal side.

Dynamics of initial colonization of nonconserved perennial ryegrass by anaerobic fungi in the bovine rumen.

Science.gov (United States)

Edwards, Joan E; Kingston-Smith, Alison H; Jimenez, Hugo R; Huws, Sharon A; Skøt, Kirsten P; Griffith, Gareth W; McEwan, Neil R; Theodorou, Michael K

2008-12-01

Anaerobic fungi (Neocallimastigales) are active degraders of fibrous plant material in the rumen. However, only limited information is available relating to how quickly they colonize ingested feed particles. The aim of this study was to determine the dynamics of initial colonization of forage by anaerobic fungi in the rumen and the impact of different postsampling wash procedures used to remove loosely associated microorganisms. Neocallimastigales-specific molecular techniques were optimized to ensure maximal coverage before application to assess the population size (quantitative PCR) and composition (automated ribosomal intergenic spacer analysis) of the colonizing anaerobic fungi. Colonization of perennial ryegrass (PRG) was evident within 5 min, with no consistent effect of time or wash procedure on fungal population composition. Wash procedure had no effect on population size unlike time, which had a significant effect. Colonizing fungal population size continued to increase over the incubation period after an initial lag of c. 4 min. This dynamic differs from that reported previously for rumen bacteria, where substantial colonization of PRG occurred within 5 min. The observed delay in colonization of plant material by anaerobic fungi is suggested to be primarily mediated by the time taken for fungal zoospores to locate, attach and encyst on plant material.

Role of the Ubiquitin-Proteasome Systems in the Biology and Virulence of Protozoan Parasites

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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.

Co-infection of visceral leishmaniasis and pulmonary tuberculosis: a case study

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Shweta

2014-02-01

Full Text Available Co-infection of visceral leishmaniasis and pulmonary tuberculosis are increasing public health problem in eastern region of country. A large number of clinical cases of leishmaniasis and tuberculosis have been reported in Sudan. Such type of co-infections lead to decreased host ’s immune system. This is a case report of 48 years old male with visceral leishmaniasis and pulmonary tuberculosis. He arrived at hospital with complaints of fever with rigor, abdominal pain, weakness, loss of appetite, yellowish discoloration of urine and sclerosis at lower back. Bone marrow aspiration cytology revealed the presence of Leishmania donovani bodies (2+. His treatment was initiated with amphotericin B deoxycholate (inj. Fungizone 15 infusions on alternate days with 5% dextrose. He had 20 years past history of pulmonary tuberculosis. His chest X-ray showed increased bronchovascular marking encysted pleural effusion on lower segment of right lung. Ultrasonography guided fine needle aspiration cytology of pleural fluid for protein, sugar, lactate dehydrogenase, adenosine deaminase, cell type and cell count. Cytological reports confirmed pulmonary tuberculosis. Antitubercular therapy (four drug regimen: rifampicin, isoniazid, ethambutal, and pyrazinamide was started. Co-infection of visceral leishmaniasis and pulmonary tuberculosis is a real threat in developing countries. There is a need of cost effective diagnostic and therapeutic facilities for these co-infections.

Morphologic evaluation of the use of a latex prosthesis in videolaparoscopic inguinoplasty: an experimental study in dogs Avaliação morfológica da utilização de prótese de latex na inguinoplastia videolaparoscópica: estudo experimental em cães

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Luiz Henrique de Sousa

2011-01-01

Full Text Available PURPOSE: To evaluate the morphological aspects of the behavior of 4 types of latex biomembranes implanted in preperitoneal videolaparoscopic inguinoplasty. METHODS: Sixteen inguinoplasties were performed in 12 dogs: group 1 received an impermeable latex biomembrane in the right inguinal region and a prolene prosthesis, as control, in the contralateral inguinal region; groups 2, 3 and 4 received latex biomembranes respectively containing impermeable polyamide, 1-mm thick porous polyamide and 0.5-mm thick porous polyamide. Macro- and microscopic evaluations of the inguinal region and of the removed implants were made on the 7th, 14th, 21st and 28th days in group 1 and on the 28th postoperative day in the other groups. RESULTS: We observed absence of hematoma, seroma and infection; presence of tortuosities; induction of vascular neoformation, inflammatory reaction and collagen deposition, and full encystment of the latex biomembranes, except that with fine porous polyamide, which was partially incorporated, with the formation of microcysts. No latex biomembrane induced fibrosis as observed in the prolene control group. CONCLUSIONS: The biomembranes maintain induction of the healing process without fibrosis, are fully encysted and, except for the one with fine porous polyamide, are not incorporated into adjacent tissues. The latex biomembrane, with or without polyamide, is not recommended as a separate material for preperitoneal inguinoplasty.OBJETIVO: Avaliar aspectos morfológicos do comportamento de 4 tipos de biomembranas de latex, colocadas pré-peritonealmente em cães, por inguinoplastia videolaparoscópica. MÉTODOS: Dezesseis inguinoplastias em 12 cães: grupo 1, com biomembrana de latex impermeável inguinal direita em quatro cães e prótese de prolene, como controle, contra-lateral; grupos 2, 3 e 4, com biomembrana de latex respectivamente de poliamida impermeável, poliamida porosa com 1mm de espessura e poliamida porosa com 0,5mm de

Phosphorylated peptides occur in a non-helical portion of the tail of a catch muscle myosin

International Nuclear Information System (INIS)

Castellani, L.; Elliott, B.W. Jr.; Cohen, C.

1987-01-01

Myosin from a molluscan catch muscle (the Anterior Byssus Retractor (ABRM) of Mytilus edulis) is unusual in being phosphorylated in the rod by an endogenous heavy-chain kinase. This phosphorylation enhances myosin solubility at low ionic strength and induces molecular folding of the myosin tail. Papain and chymotryptic cleavage of this myosin, phosphorylated with [γ- 32 P]ATP, indicates that the phosphorylated residues are associated with the carboxy-terminal end of the light meromyosin. Ion-exchange and reverse-phase HPLC of radiolabeled chymotryptic peptides allow the isolation of two different peptides with high specific activity. One of these peptides is rich in lysine and arginine residues, a finding consistent with the observation that basic residues often determine the substrate specificity of protein kinases. The second peptide contains proline residues. Taken together, these results suggest that, as in the case of Acanthamoeba myosin, phosphorylation occurs in a nonhelical portion of the rod that may also control solubility. Identification of the residues that are phosphorylated and their location in the rod may reveal how the phosphorylation-dependent changes observed in the myosin in vitro are related to changes in intermolecular interactions in the thick filaments in vivo

Methodological approaches for monitoring opportunistic pathogens in premise plumbing: A review.

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Wang, Hong; Bédard, Emilie; Prévost, Michèle; Camper, Anne K; Hill, Vincent R; Pruden, Amy

2017-06-15

Opportunistic premise (i.e., building) plumbing pathogens (OPPPs, e.g., Legionella pneumophila, Mycobacterium avium complex, Pseudomonas aeruginosa, Acanthamoeba, and Naegleria fowleri) are a significant and growing source of disease. Because OPPPs establish and grow as part of the native drinking water microbiota, they do not correspond to fecal indicators, presenting a major challenge to standard drinking water monitoring practices. Further, different OPPPs present distinct requirements for sampling, preservation, and analysis, creating an impediment to their parallel detection. The aim of this critical review is to evaluate the state of the science of monitoring OPPPs and identify a path forward for their parallel detection and quantification in a manner commensurate with the need for reliable data that is informative to risk assessment and mitigation. Water and biofilm sampling procedures, as well as factors influencing sample representativeness and detection sensitivity, are critically evaluated with respect to the five representative bacterial and amoebal OPPPs noted above. Available culturing and molecular approaches are discussed in terms of their advantages, limitations, and applicability. Knowledge gaps and research needs towards standardized approaches are identified. Copyright © 2017 Elsevier Ltd. All rights reserved.

Ring Keratitis Associated With Topical Abuse of a Dilute Anesthetic After Refractive Surgery

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Yu-Chih Hou

2009-12-01

Full Text Available Anesthetic toxic keratitis is rare and presents as a ring keratitis, which is often misdiagnosed as Acanthamoeba keratitis. Here, we report an unusual case of toxic keratitis caused by topical abuse of a dilute anesthetic. A 26-year-old woman presented with bilateral corneal edema, ring infiltrates, pigmented keratic precipitate, Descemet's membrane folding, and strong anterior chamber reactions 2 weeks after laser subepithelial keratomileusis surgery. Tracing back her medical history, topical dilute 0.1% proparacaine was prescribed and frequently used for 1 month. Toxic keratitis was suspected. After discontinuation of the topical anesthetic and initiation of treatment with topical 20% autologous serum, complete corneal epithelialization was achieved within 1 week. Corneal infiltrates and anterior chamber reaction gradually subsided. Vision improved from finger counting to 20/20 in the right eye and 20/25 in the left eye, but confocal microscopy showed decreased corneal endothelial cells. Topical abuse of a dilute topical anesthetic can cause severe toxic keratitis and endothelial cell loss. The physician must be aware of the signs of topical anesthetic abuse and should not prescribe even a dilute anesthetic for long-term use. Autologous serum can help in the recovery of toxic keratitis.

Microbiological Spectrum of Brain Abscess at a Tertiary Care Hospital in South India: 24-Year Data and Review

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V. Lakshmi

2011-01-01

Full Text Available Intracranial abscesses are life-threatening infections that pose a diagnostic challenge not only to the neurosurgeon but also to the microbiologists. Detailed studies documenting the spectrum of infecting agents involved in brain abscesses are limited from India. Materials and Methods. This is a retrospective analysis of 352 samples from 1987 to 2010 analyzed at a tertiary care hospital in South India from 1987 to 2010, to document the changing trends with time. Results. The age of the patients ranged from 2 to 80 years, a larger number of males being affected. Otogenic infections were the most common cause while cryptogenic abscesses were 20%. Gram stain and culture positivity were 78% each. Gram-positive and negative facultative aerobes and obligate anaerobes were also on the rise. Unusual organisms, like Burkholderia pseudomallei, Salmonella typhi, Nocardia species, Cladosporium bantiana, Fonsecaea pedrosoi, Entamoeba histolytica, and Acanthamoeba were also isolated and/or detected from the brain abscesses aspirate or resected tissue. Summary. New and emerging pathogens associated with brain abscess, especially in immunosuppressed individuals, have renewed the necessity of an early detection, and it will be of great value in appropriate management of patients with brain abscess.

Current and future molecular diagnostics for ocular infectious diseases.

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Doan, Thuy; Pinsky, Benjamin A

2016-11-01

Confirmation of ocular infections can pose great challenges to the clinician. A fundamental limitation is the small amounts of specimen that can be obtained from the eye. Molecular diagnostics can circumvent this limitation and have been shown to be more sensitive than conventional culture. The purpose of this review is to describe new molecular methods and to discuss the applications of next-generation sequencing-based approaches in the diagnosis of ocular infections. Efforts have focused on improving the sensitivity of pathogen detection using molecular methods. This review describes a new molecular target for Toxoplasma gondii-directed polymerase chain reaction assays. Molecular diagnostics for Chlamydia trachomatis and Acanthamoeba species are also discussed. Finally, we describe a hypothesis-free approach, metagenomic deep sequencing, which can detect DNA and RNA pathogens from a single specimen in one test. In some cases, this method can provide the geographic location and timing of the infection. Pathogen-directed PCRs have been powerful tools in the diagnosis of ocular infections for over 20 years. The use of next-generation sequencing-based approaches, when available, will further improve sensitivity of detection with the potential to improve patient care.

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

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

2015-01-01

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

Molecular Survey of the Occurrence of Legionella spp., Mycobacterium spp., Pseudomonas aeruginosa, and Amoeba Hosts in Two Chloraminated Drinking Water Distribution Systems

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Wang, Hong; Edwards, Marc; Falkinham, Joseph O.

2012-01-01

The spread of opportunistic pathogens via public water systems is of growing concern. The purpose of this study was to identify patterns of occurrence among three opportunistic pathogens (Legionella pneumophila, Mycobacterium avium, and Pseudomonas aeruginosa) relative to biotic and abiotic factors in two representative chloraminated drinking water distribution systems using culture-independent methods. Generally, a high occurrence of Legionella (≥69.0%) and mycobacteria (100%), lower occurrence of L. pneumophila (≤20%) and M. avium (≤33.3%), and rare detection of Pseudomonas aeruginosa (≤13.3%) were observed in both systems according to quantitative PCR. Also, Hartmanella vermiformis was more prevalent than Acanthamoeba, both of which are known hosts for opportunistic pathogen amplification, the latter itself containing pathogenic members. Three-minute flushing served to distinguish distribution system water from plumbing in buildings (i.e., premise plumbing water) and resulted in reduced numbers of copies of Legionella, mycobacteria, H. vermiformis, and 16S rRNA genes (P Legionella and H. vermiformis, were noted, emphasizing potential microbial ecological relationships. Overall, the results provide insight into factors that may aid in controlling opportunistic pathogen proliferation in real-world water systems. PMID:22752174

Giant virus Megavirus chilensis encodes the biosynthetic pathway for uncommon acetamido sugars.

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Piacente, Francesco; De Castro, Cristina; Jeudy, Sandra; Molinaro, Antonio; Salis, Annalisa; Damonte, Gianluca; Bernardi, Cinzia; Abergel, Chantal; Tonetti, Michela G

2014-08-29

Giant viruses mimicking microbes, by the sizes of their particles and the heavily glycosylated fibrils surrounding their capsids, infect Acanthamoeba sp., which are ubiquitous unicellular eukaryotes. The glycans on fibrils are produced by virally encoded enzymes, organized in gene clusters. Like Mimivirus, Megavirus glycans are mainly composed of virally synthesized N-acetylglucosamine (GlcNAc). They also contain N-acetylrhamnosamine (RhaNAc), a rare sugar; the enzymes involved in its synthesis are encoded by a gene cluster specific to Megavirus close relatives. We combined activity assays on two enzymes of the pathway with mass spectrometry and NMR studies to characterize their specificities. Mg534 is a 4,6-dehydratase 5-epimerase; its three-dimensional structure suggests that it belongs to a third subfamily of inverting dehydratases. Mg535, next in the pathway, is a bifunctional 3-epimerase 4-reductase. The sequential activity of the two enzymes leads to the formation of UDP-l-RhaNAc. This study is another example of giant viruses performing their glycan synthesis using enzymes different from their cellular counterparts, raising again the question of the origin of these pathways. © 2014 by The American Society for Biochemistry and Molecular Biology, Inc.

Chlamydiaceae family, Parachlamydia spp., and Waddlia spp. in porcine abortion.

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Koschwanez, Maria; Meli, Marina; Vögtlin, Andrea; Greub, Gilbert; Sidler, Xaver; Handke, Martin; Sydler, Titus; Kaiser, Carmen; Pospischil, Andreas; Borel, Nicole

2012-09-01

At present, despite extensive laboratory investigations, most cases of porcine abortion remain without an etiological diagnosis. Due to a lack of recent data on the abortigenic effect of order Chlamydiales, 286 fetuses and their placentae of 113 abortion cases (1-5 fetuses per abortion case) were investigated by polymerase chain reaction (PCR) methods for family Chlamydiaceae and selected Chlamydia-like organisms such as Parachlamydia acanthamoebae and Waddlia chondrophila. In 0.35% of the cases (1/286 fetuses), the Chlamydiaceae real-time PCR was positive. In the Chlamydiaceae-positive fetus, Chlamydia abortus was detected by a commercial microarray and 16S ribosomal RNA PCR followed by sequencing. The positive fetus had a Porcine circovirus-2 coinfection. By the Parachlamydia real-time PCR, 3.5% (10/286 fetuses of 9 abortion cases) were questionable positive (threshold cycle values: 35.0-45.0). In 2 of these 10 cases, a confirmation by Chlamydiales-specific real-time PCR was possible. All samples tested negative by the Waddlia real-time PCR. It seems unlikely that Chlamydiaceae, Parachlamydia, and Waddlia play an important role as abortigenic agents in Swiss sows.

Emerging role of Chlamydia and Chlamydia-like organisms in adverse pregnancy outcomes.

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Baud, David; Regan, Lesley; Greub, Gilbert

2008-02-01

This review considers the roles of Chlamydia spp. and newly identified Chlamydia-like organisms in miscarriage, stillbirths and preterm labour in both animals and humans. The cause of miscarriage, stillbirth and preterm labour often remains unexplained. Intracellular bacteria that grow either poorly or not at all on media used routinely to detect human pathogens could be the aetiological agents of these obstetrical conditions. There is growing evidence that Chlamydia trachomatis, Chlamydophila abortus, Chlamydophila psittaci and Chlamydophila pneumoniae infections may result in adverse pregnancy outcomes in humans and/or animals. Waddlia, a Chlamydia-like organism first isolated from an aborted bovine, has emerged as an agent of abortion in cattle. Recently, Waddlia was also implicated in human foetal death. Moreover, Parachlamydia acanthamoebae is also abortigenic in ruminants. Whether additional novel Chlamydia-like organisms, such as Protochlamydia amoebophila, Neochlamydia hartmanellae, Criblamydia sequanensis, Rhabdochlamydia crassificans and Simkania negevensis, are involved in foetal loss or premature delivery remains to be determined. This review provides an update on the consequences of chlamydial infection during pregnancy and summarizes current evidence suggesting that some Chlamydia-related organisms are probably emerging obstetrical pathogens.

Diversity of protists and bacteria determines predation performance and stability.

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Saleem, Muhammad; Fetzer, Ingo; Harms, Hauke; Chatzinotas, Antonis

2013-10-01

Predation influences prey diversity and productivity while it effectuates the flux and reallocation of organic nutrients into biomass at higher trophic levels. However, it is unknown how bacterivorous protists are influenced by the diversity of their bacterial prey. Using 456 microcosms, in which different bacterial mixtures with equal initial cell numbers were exposed to single or multiple predators (Tetrahymena sp., Poterioochromonas sp. and Acanthamoeba sp.), we showed that increasing prey richness enhanced production of single predators. The extent of the response depended, however, on predator identity. Bacterial prey richness had a stabilizing effect on predator performance in that it reduced variability in predator production. Further, prey richness tended to enhance predator evenness in the predation experiment including all three protists predators (multiple predation experiment). However, we also observed a negative relationship between prey richness and predator production in multiple predation experiments. Mathematical analysis of potential ecological mechanisms of positive predator diversity-functioning relationships revealed predator complementarity as a factor responsible for both enhanced predator production and prey reduction. We suggest that the diversity at both trophic levels interactively determines protistan performance and might have implications in microbial ecosystem processes and services.

Relationships between free-living protozoa, cultivable Legionella spp., and water quality characteristics in three drinking water supplies in the Caribbean.

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Valster, Rinske M; Wullings, Bart A; van den Berg, Riemsdijk; van der Kooij, Dick

2011-10-01

The study whose results are presented here aimed at identifying free-living protozoa (FLP) and conditions favoring the growth of these organisms and cultivable Legionella spp. in drinking water supplies in a tropical region. Treated and distributed water (±30°C) of the water supplies of three Caribbean islands were sampled and investigated with molecular techniques, based on the 18S rRNA gene. The protozoan host Hartmannella vermiformis and cultivable Legionella pneumophila were observed in all three supplies. Operational taxonomic units (OTUs) with the highest similarity to the potential or candidate hosts Acanthamoeba spp., Echinamoeba exundans, E. thermarum, and an Neoparamoeba sp. were detected as well. In total, 59 OTUs of FLP were identified. The estimated protozoan richness did not differ significantly between the three supplies. In supply CA-1, the concentration of H. vermiformis correlated with the concentration of Legionella spp. and clones related to Amoebozoa predominated (82%) in the protozoan community. These observations, the low turbidity (water. The absence of H. vermiformis in most samples from supply CA-3 suggests that growth of this protozoan is limited at ATP concentrations of <1 ng liter(-1).

Relationships between Free-Living Protozoa, Cultivable Legionella spp., and Water Quality Characteristics in Three Drinking Water Supplies in the Caribbean▿†

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Valster, Rinske M.; Wullings, Bart A.; van den Berg, Riemsdijk; van der Kooij, Dick

2011-01-01

The study whose results are presented here aimed at identifying free-living protozoa (FLP) and conditions favoring the growth of these organisms and cultivable Legionella spp. in drinking water supplies in a tropical region. Treated and distributed water (±30°C) of the water supplies of three Caribbean islands were sampled and investigated with molecular techniques, based on the 18S rRNA gene. The protozoan host Hartmannella vermiformis and cultivable Legionella pneumophila were observed in all three supplies. Operational taxonomic units (OTUs) with the highest similarity to the potential or candidate hosts Acanthamoeba spp., Echinamoeba exundans, E. thermarum, and an Neoparamoeba sp. were detected as well. In total, 59 OTUs of FLP were identified. The estimated protozoan richness did not differ significantly between the three supplies. In supply CA-1, the concentration of H. vermiformis correlated with the concentration of Legionella spp. and clones related to Amoebozoa predominated (82%) in the protozoan community. These observations, the low turbidity (water. The absence of H. vermiformis in most samples from supply CA-3 suggests that growth of this protozoan is limited at ATP concentrations of <1 ng liter−1. PMID:21873489

Mitochondrial uncoupling proteins in unicellular eukaryotes.

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Jarmuszkiewicz, Wieslawa; Woyda-Ploszczyca, Andrzej; Antos-Krzeminska, Nina; Sluse, Francis E

2010-01-01

Uncoupling proteins (UCPs) are members of the mitochondrial anion carrier protein family that are present in the mitochondrial inner membrane and mediate free fatty acid (FFA)-activated, purine nucleotide (PN)-inhibited proton conductance. Since 1999, the presence of UCPs has been demonstrated in some non-photosynthesising unicellular eukaryotes, including amoeboid and parasite protists, as well as in non-fermentative yeast and filamentous fungi. In the mitochondria of these organisms, UCP activity is revealed upon FFA-induced, PN-inhibited stimulation of resting respiration and a decrease in membrane potential, which are accompanied by a decrease in membranous ubiquinone (Q) reduction level. UCPs in unicellular eukaryotes are able to divert energy from oxidative phosphorylation and thus compete for a proton electrochemical gradient with ATP synthase. Our recent work indicates that membranous Q is a metabolic sensor that might utilise its redox state to release the PN inhibition of UCP-mediated mitochondrial uncoupling under conditions of phosphorylation and resting respiration. The action of reduced Q (QH2) could allow higher or complete activation of UCP. As this regulatory feature was demonstrated for microorganism UCPs (A. castellanii UCP), plant and mammalian UCP1 analogues, and UCP1 in brown adipose tissue, the process could involve all UCPs. Here, we discuss the functional connection and physiological role of UCP and alternative oxidase, two main energy-dissipating systems in the plant-type mitochondrial respiratory chain of unicellular eukaryotes, including the control of cellular energy balance as well as preventive action against the production of reactive oxygen species. Copyright © 2009 Elsevier B.V. All rights reserved.

Prevalence of Fasciola species in ruminants under extensive management system in Ibadan southwestern Nigeria.

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Adediran, O A; Adebiyi, A I; Uwalaka, E C

2014-12-01

Fasciolosis is a zoonotic trematode of ruminants and a common parasite in cattle in Nigeria. With the increasing numbers of small household cattle herds kept under extensive management system as well as the more usual small ruminants, there is a possibility therefore that parasites common to these ruminants but better adapted to cattle such as Fasciola may also be on the increase within the small ruminant population as they graze freely over the same wide area and sometimes freely mix. The aim of the study is to determine the prevalence of Fasciola spp in sheep, goats and cattle in Ibadan during the short wet season, (August-October 2011). Faecal samples were collected per rectum from 440 each of goats and sheep and 291 cattle. Helminth ova were recovered using sedimentation technique for faecal examination. A total of 40 goats (9.1%), 53 sheep (12.0%) and 158 cattle (54.3%) were positive for Fasciola ova which reflects an increase in prevalence in small ruminants. This implies that the entire Ibadan is endemic for Fasciola spp and small ruminants may serve as an important link in the epidemiology of Fasciola spp infection. More so, this has implications for the scavenging household animals which receive little or no medical attention and man that may acquire this trematode by inadvertently eating vegetables with encysted Fasciola metacercariae.

The neurotropic parasite Toxoplasma gondii increases dopamine metabolism.

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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.

Ocular toxoplasmosis in immunosuppressed nonhuman primates

International Nuclear Information System (INIS)

Holland, G.N.; O'Connor, G.R.; Diaz, R.F.; Minasi, P.; Wara, W.M.

1988-01-01

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

Ocular toxoplasmosis in immunosuppressed nonhuman primates

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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.

Morphological variability, spatial distribution and abundance of Helicostomella species (Ciliophora: Tintinnina in relation to environmental factors (Argentine shelf; 40-55°S

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Luciana F. Santoferrara

2009-12-01

Full Text Available For the first time a taxonomic and ecological study of species belonging to Helicostomella was performed over a mesoscale spatial distribution by examining ca. 3000 loricae collected in Argentine shelf waters during the austral autumn. Microscopic and statistical analysis revealed that the general shape and the oral diameter remained practically constant in the whole area surveyed, despite a continuous length fluctuation of ca. 300 µm, which includes the entire range present in eight previously reported species. Consequently, we consider that the genus may be represented only by H. subulata, whose strong fluctuations in length (mostly attributed to an increase in the collar length, density and biomass seem to respond to temperature, food availability and front-related processes. In Buenos Aires coastal waters associated with a quasi permanent estuarine front (40-40.5°S, mixed conditions would favour moderate abundances of long loricae, whereas in northern (42.5-45.5°S and southern (46-54.5°S Patagonian waters, high (103 ind. L-1 and low (< 5 ind. L-1 densities of short loricae seem to be a consequence of stratification and encystment, respectively. The non-occurrence of the species at 41-42°S, together with a three-fold reduction in length-related parameters between specimens from Buenos Aires and Patagonian waters, suggests a disjunct distribution.

Studies on the epidemiology and histopathology of Euclinostomum heterostomum (Trematoda; Digenea infection in Channa punctata from North India

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Shareef P. A. Ahammed

2015-09-01

Full Text Available A survey on the occurrence and epidemiology of the encysted progenetic metacercariae of Euclinostomum heterostomum infection in Channa punctata in the Aligarh region of North India revealed a mean prevalence, intensity, and abundance of 18.61, 1.52, and 0.38%, respectively, during the period from April 2011 to March 2012. Liver, kidney, peritoneum, muscle, and ovary were found to be infected with this parasite, and the later three are reported for the first time in this fish species. The histopathology of the infected tissues indicated the following at the host-parasite interface: tissue damage, infiltration of immune cells into the cyst wall, chronic inflammatory responses, and granulomatous lesions. The infected liver showed degeneration of hepatocytes, cytoplasmic vacuolation, nuclear alterations, mallory body formation, fibrosis, and necrosis. The pathology of the infected kidney included distortion and dilation of renal tubules, vacuolar degeneration, hypertrophy and hyperplasia of tubular epithelial cells, occlusion of tubules, fibrosis, hemorrhage, and congestion of glomeruli. The infected muscle demonstrated comparatively fewer pathological changes confined only to the circumference of the cyst wall. The ovary displayed the least changes. The conclusions drawn from the study are that the large metacercarial cysts formed by E. heterostomum in the vital organs of the economically important fish C. punctata could result in the impairment of fish physiology and health, thereby affecting their productivity and quality for human consumption.

Increasing Risk Factors for Imported and Domestic Gnathostomiasis in the United States.

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Diaz, James H

2015-01-01

Gnathostomiasis is a foodborne zoonotic helminthic infection, commonly described in Asia and Latin America, which may follow the consumption of raw fish, eels, amphibians, and reptiles infected with muscle-encysted larvae of Gnathostoma species nematodes. After an inoculum of as little as one infective larva and an incubation period of months to years, most infections are characterized by intermittent migratory swellings due to subdermal larval migration. Less commonly, larval migration to the central nervous system may result in radiculomyelopathy or eosinophilic meningoencephalitis with high fatality rates; or larval migration to the eye with resulting blindness in untreated cases. Since the US now supports a zoonosis of Gnathostoma species with infective larvae encysted in imported and domestic fish and eels that may be consumed raw as exotic ethnic dishes, the objectives of this review were to describe the biology and life cycle of Gnathostoma nematodes and the behavioral risk factors for gnathostomiasis; and to describe the clinical manifestations, diagnosis, management, and prevention of human gnathostomiasis. Since the eradication of gnathostomiasis is very unlikely given the global distribution of Gnathostoma nematodes and the increasingly exotic culinary tastes of US residents and travelers to endemic regions, the only effective strategies for gnathostomiasis include: (1) educating citizens in the US and travelers abroad in endemic areas that fish, eels, frogs, snakes, and chicken must be cooked thoroughly first and not eaten raw or marinated; and (2) seeking medical care immediately for evaluation of migratory subcutaneous swellings. The combination of international travel and increased immigration from Asia and Latin America to the US has resulted in greater popularity of exotic ethnic cuisine, especially raw seafood dishes. The ethnic cuisine industry is supported by domestic aquaculture that produces fish-farmed tilapia and trout, and by increased

Ultraviolet radiation for the sterilization of contact lenses

International Nuclear Information System (INIS)

Gritz, D.C.; Lee, T.Y.; McDonnell, P.J.; Shih, K.; Baron, N.

1990-01-01

Two sources of ultraviolet (UV) radiation with peak wavelengths in the UV-C or UV-B ranges were compared for their ability to sterilize contact lenses infected with Pseudomonas aeruginosa, Streptococcus pneumoniae, Acanthamoeba castellani, Candida albicans, and Aspergillus niger. Also examined was the effect of prolonged UV light exposure on soft and rigid gas permeable (RGP) contact lenses. The UV-C lamp (253.7 nm, 250 mW/cm2 at 1 cm) was germicidal for all organisms within 20 minutes but caused destruction of the soft lens polymers within 6 hours of cumulative exposure. UV-C caused damage to RGP lenses in less than 100 hours. The UV-B lamp (290-310 nm, 500 mW/cm2 at 1 cm) was germicidal for all organisms tested (except Aspergillus) with a 180-minute exposure and caused less severe changes in the soft lens polymers than did the UV-C lamp, although cumulative exposure of 300 hours did substantially weaken the soft lens material. RGP materials were minimally affected by exposure to 300 hours of UV-B. Ultraviolet light is an effective germicidal agent but is injurious to soft lens polymers; its possible utility in the sterilization of RGP lenses and lens cases deserves further study

High-resolution, label-free two-photon imaging of diseased human corneas

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Batista, Ana; Breunig, Hans Georg; König, Aisada; Schindele, Andreas; Hager, Tobias; Seitz, Berthold; König, Karsten

2018-03-01

The diagnosis of corneal diseases may be improved by monitoring the metabolism of cells and the structural organization of the stroma using two-photon imaging (TPI). We used TPI to assess the differences between nonpathological (NP) human corneas and corneas diagnosed with either keratoconus, Acanthamoeba keratitis, or stromal corneal scars. Images were acquired using a custom-built five-dimensional laser-scanning microscope with a broadband sub-15 femtosecond near-infrared pulsed excitation laser and a 16-channel photomultiplier tube detector in combination with a time-correlated single photon counting module. Morphological alterations of epithelial cells were observed for all pathologies. Moreover, diseased corneas showed alterations to the cells' metabolism that were revealed using the NAD(P)H free to protein-bound ratios. The mean autofluorescence lifetime of the stroma and the organization of the collagen fibers were also significantly altered due to the pathologies. We demonstrate that TPI can be used to distinguish between NP and diseased human corneas, based not only on alterations of the cells' morphology, which can also be evaluated using current clinical devices, but on additional morphological and functional features such as the organization of the stroma and the cells' metabolism. Therefore, TPI could become an efficient tool for diagnosing corneal diseases and better understanding the biological processes of the diseases.

Mimivirus relatives in the Sargasso sea

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Claverie Jean-Michel

2005-08-01

Full Text Available 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 individualization of cellular organisms from the three established domains, triggering a debate that can only be resolved by generating and analyzing more data. The next step is then to seek some evidence that Mimivirus is not the only representative of its kind and determine where to look for new Mimiviridae. An exhaustive similarity search of all Mimivirus predicted proteins against all publicly available sequences identified many of their closest homologues among the Sargasso Sea environmental sequences. Subsequent phylogenetic analyses suggested that unknown large viruses evolutionarily closer to Mimivirus than to any presently characterized species exist in abundance in the Sargasso Sea. Their isolation and genome sequencing could prove invaluable in understanding the origin and diversity of large DNA viruses, and shed some light on the role they eventually played in the emergence of eukaryotes.

A divergent Cardinium found in daddy long-legs (Arachnida: Opiliones).

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Chang, Jin; Masters, Amber; Avery, Amanda; Werren, John H

2010-11-01

Recent studies indicate that a newly described bacterial endosymbiont, Cardinium, is widespread in arthropods and induces different reproductive manipulations in hosts. In this study, we used a portion of the 16S rRNA gene of the Cardinium to screen 16 Opilionid species from the suborder Palptores. We found the incidence of Cardinium in these Opiliones was significantly higher than in other pooled arthropods (31.2% versus 7.2%, P=0.007). Phylogenetic analyses using maximum parsimony (MP) and Bayesian analysis revealed two distinct clades in Opiliones. One is a divergent monophyletic clade with strong support that has so far not been found in other arthropods, and a second one contains Cardinium both from Opiliones and other arthropods. There is not complete concordance of the Cardinium strains with host phylogeny, suggesting some horizontal movement of the bacteria among Opiliones. Although the divergence in the sequenced 16S rRNA region between the Cardinium infecting Opiliones and Cardinium from other arthropods is greater than among Cardinium found in other arthropods, all are monophyletic with respect to the outgroup bacteria (endosymbionts of Acanthamoeba). Based on high pairwise genetic distances, deep branch, and a distinct phylogenetic grouping, we conclude that some Opiliones harbor a newly discovered Cardinium clade. Copyright © 2010 Elsevier Inc. All rights reserved.

Preliminary crystallographic analysis of a possible transcription factor encoded by the mimivirus L544 gene

International Nuclear Information System (INIS)

Ciaccafava, Alexandre; Lartigue, Audrey; Mansuelle, Pascal; Jeudy, Sandra; Abergel, Chantal

2011-01-01

The mimivirus L544 gene product was expressed in E. coli and crystallized; preliminary phasing of a MAD data set was performed using the selenium signal present in a crystal of recombinant selenomethionine-substituted protein. Mimivirus is the prototype of a new family (the Mimiviridae) of nucleocytoplasmic large DNA viruses (NCLDVs), which already include the Poxviridae, Iridoviridae, Phycodnaviridae and Asfarviridae. Mimivirus specifically replicates in cells from the genus Acanthamoeba. Proteomic analysis of purified mimivirus particles revealed the presence of many subunits of the DNA-directed RNA polymerase II complex. A fully functional pre-transcriptional complex appears to be loaded in the virions, allowing mimivirus to initiate transcription within the host cytoplasm immediately upon infection independently of the host nuclear apparatus. To fully understand this process, a systematic study of mimivirus proteins that are predicted (by bioinformatics) or suspected (by proteomic analysis) to be involved in transcription was initiated by cloning and expressing them in Escherichia coli in order to determine their three-dimensional structures. Here, preliminary crystallographic analysis of the recombinant L544 protein is reported. The crystals belonged to the orthorhombic space group C222 1 with one monomer per asymmetric unit. A MAD data set was used for preliminary phasing using the selenium signal present in a selenomethionine-substituted protein crystal

Phylogenomic analysis of Odyssella thessalonicensis fortifies the common origin of Rickettsiales, Pelagibacter ubique and Reclimonas americana mitochondrion.

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Kalliopi Georgiades

Full Text Available BACKGROUND: The evolution of the Alphaproteobacteria and origin of the mitochondria are topics of considerable debate. Most studies have placed the mitochondria ancestor within the Rickettsiales order. Ten years ago, the bacterium Odyssella thessalonicensis was isolated from Acanthamoeba spp., and the 16S rDNA phylogeny placed it within the Rickettsiales. Recently, the whole genome of O. thessalonicensis has been sequenced, and 16S rDNA phylogeny and more robust and accurate phylogenomic analyses have been performed with 65 highly conserved proteins. METHODOLOGY/PRINCIPAL FINDINGS: The results suggested that the O. thessalonicensis emerged between the Rickettsiales and other Alphaproteobacteria. The mitochondrial proteins of the Reclinomonas americana have been used to locate the phylogenetic position of the mitochondrion ancestor within the Alphaproteobacteria tree. Using the K tree score method, nine mitochondrion-encoded proteins, whose phylogenies were congruent with the Alphaproteobacteria phylogenomic tree, have been selected and concatenated for Bayesian and Maximum Likelihood phylogenies. The Reclinomonas americana mitochondrion is a sister taxon to the free-living bacteria Candidatus Pelagibacter ubique, and together, they form a clade that is deeply rooted in the Rickettsiales clade. CONCLUSIONS/SIGNIFICANCE: The Reclinomonas americana mitochondrion phylogenomic study confirmed that mitochondria emerged deeply in the Rickettsiales clade and that they are closely related to Candidatus Pelagibacter ubique.

Colony types and virulence traits of Legionella feeleii determined by exopolysaccharide materials.

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Wang, Changle; Saito, Mitsumasa; Ogawa, Midori; Yoshida, Shin-Ichi

2016-05-01

Legionella feeleii is a Gram-negative pathogenic bacterium that causes Pontiac fever and pneumonia in humans. When L. feeleii serogroup 1 (ATCC 35072) was cultured on BCYE agar plates, two types of colonies were observed and exhibited differences in color, opacity and morphology. Since the two colony types are white rugose and brown translucent, they were termed as white rugose L. feeleii (WRLf) and brown translucent L. feeleii (BTLf), respectively. They exhibited different growth capacities in BYE broth in vitro, and it was found that WRLf could transform to BTLf. Under the electron microscope, it was observed that WRLf secreted materials which could be stained with ruthenium red, which was absent in BTLf. When U937 macrophages and HeLa cells were infected with the bacteria, WRLf manifested stronger internalization ability than BTLf. Intracellular growth in murine macrophages and Acanthamoeba cells was affected by the level of initial phagocytosis. WRLf was more resistant to human serum bactericidal action than BTLf. After being inoculated to guinea pigs, both organisms caused fever in the animals. These results suggest that ruthenium red-stained materials secreted in the surroundings may play a crucial role in determining L. feeleii colony morphology and virulence traits. © FEMS 2016. All rights reserved. For permissions, please e-mail: journals.permissions@oup.com.

High-throughput isolation of giant viruses in liquid medium using automated flow cytometry and fluorescence staining.

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Jacques Yaacoub Bou Khalil

2016-01-01

Full Text Available The isolation of giant viruses using amoeba co-culture is tedious and fastidious. Recently, the procedure was successfully associated with a method that detects amoebal lysis on agar plates. However, the procedure remains time-consuming and is limited to protozoa growing on agar. We present here advances for the isolation of giant viruses. A high-throughput automated method based on flow cytometry and fluorescent staining was used to detect the presence of giant viruses in liquid medium. Development was carried out with the Acanthamoeba polyphaga strain widely used in past and current co-culture experiments. The proof of concept was validated with virus suspensions: artificially contaminated samples but also environmental samples from which viruses were previously isolated. After validating the technique, and fortuitously isolating a new Mimivirus, we automated the technique on 96-well plates and tested it on clinical and environmental samples using other protozoa. This allowed us to detect more than ten strains of previously known species of giant viruses and 7 new strains of a new virus lineage. This automated high-throughput method demonstrated significant time saving, and higher sensitivity than older techniques. It thus creates the means to isolate giant viruses at high speed.

Abundance, diversity and community composition of free-living protozoa on vegetable sprouts.

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Chavatte, N; Lambrecht, E; Van Damme, I; Sabbe, K; Houf, K

2016-05-01

Interactions with free-living protozoa (FLP) have been implicated in the persistence of pathogenic bacteria on food products. In order to assess the potential involvement of FLP in this contamination, detailed knowledge on their occurrence, abundance and diversity on food products is required. In the present study, enrichment and cultivation methods were used to inventory and quantify FLP on eight types of commercial vegetable sprouts (alfalfa, beetroot, cress, green pea, leek, mung bean, red cabbage and rosabi). In parallel, total aerobic bacteria and Escherichia coli counts were performed. The vegetable sprouts harbored diverse communities of FLP, with Tetrahymena (ciliate), Bodo saltans and cercomonads (flagellates), and Acanthamoeba and Vannella (amoebae) as the dominant taxa. Protozoan community composition and abundance significantly differed between the sprout types. Beetroot harbored the most abundant and diverse FLP communities, with many unique species such as Korotnevella sp., Vannella sp., Chilodonella sp., Podophrya sp. and Sphaerophrya sp. In contrast, mung bean sprouts were species-poor and had low FLP numbers. Sampling month and company had no significant influence, suggesting that seasonal and local factors are of minor importance. Likewise, no significant relationship between protozoan community composition and bacterial load was observed. Copyright © 2015 Elsevier Ltd. All rights reserved.

Variability in prevalence of Cymatocarpus solearis (Trematoda, Brachycoeliidae) in Caribbean spiny lobsters Panulirus argus (Decapoda: Palinuridae) from Bahía de la Ascensión (Mexico).

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Briones-Fourzán, Patricia; Muñoz de Cote-Hernández, Rubén; Lozano-Álvarez, Enrique

2016-06-01

Cymatocarpus solearis, a brachycoeliid trematode that parasitizes sea turtles, uses spiny lobsters Panulirus argus as second intermediate hosts. We examined variability in infection by C. solearis in Bahía de la Ascensión, Mexico, a tropical bay with a highly productive fishery for P. argus that is based on the extensive use of casitas (large artificial shelters), which can harbor multiple juveniles and adults of this gregarious lobster species. We sampled 3956 lobsters from 466 casitas distributed over three bay zones within two closed seasons and one fishing season. In these lobsters (9.5-115.2mm carapace length, CL), the average infection prevalence was 11.7% but the probability of infection increased significantly with size. Prevalence varied with season but was consistently higher in one zone than in the other two zones. Infection with C. solearis was negatively related with clinical infection with Panulirus argus Virus 1 (PaV1). We also sampled 405 lobsters from the commercial catch obtained throughout the bay at the onset of the fishing season. In these relatively larger lobsters (70.3-168.0mm CL), average prevalence of C. solearis was higher (23.5%) but was not affected by lobster size or sex. Encysted metacercariae occurred in both abdominal and cephalothoracic muscles. The effects of C. solearis on phenotypic traits of P. argus that may potentially impact the host population dynamics and fisheries remain to be investigated. Copyright © 2016 Elsevier Inc. All rights reserved.

Grazers and phytoplankton growth in the oceans: an experimental and evolutionary perspective.

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Ratti, Simona; Knoll, Andrew H; Giordano, Mario

2013-01-01

The taxonomic composition of phytoplankton responsible for primary production on continental shelves has changed episodically through Earth history. Geological correlations suggest that major changes in phytoplankton composition correspond in time to changes in grazing and seawater chemistry. Testing hypotheses that arise from these correlations requires experimentation, and so we carried out a series of experiments in which selected phytoplankton species were grown in treatments that differed with respect to the presence or absence of grazers as well as seawater chemistry. Both protistan (Euplotes sp.) and microarthropod (Acartia tonsa) grazers changed the growth dynamics and biochemical composition of the green alga Tetraselmis suecica, the diatom Thalassiosira weissflogii, and the cyanobacterium Synechococcus sp., increasing the specific growth rate and palatability of the eukaryotic algae, while decreasing or leaving unchanged both parameters in the cyanobacteria. Synechococcus (especially) and Thalassiosira produced toxins effective against the copepod, but ciliate growth was unaffected. Acartia induced a 4-6 fold increase of Si cell quota in the diatom, but Euplotes had no similar effect. The differential growth responses of the eukaryotic algae and cyanobacteria to ciliate grazing may help to explain the apparently coeval radiation of eukaryophagic protists and rise of eukaryotes to ecological prominence as primary producers in Neoproterozoic oceans. The experimental results suggest that phytoplankton responses to the later radiation of microarthropod grazers were clade-specific, and included changes in growth dynamics, toxin synthesis, encystment, and (in diatoms) enhanced Si uptake.

An unusual cause of intra-abdominal calcification: A lithopedion

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Ramos-Andrade, Daniel; Ruivo, Catarina; Portilha, M. Antónia; Brito, Jorge B.; Caseiro-Alves, Filipe; Curvo-Semedo, Luís

2014-01-01

We report a case of a 77-year-old female who was admitted to the emergency department complaining of diffuse abdominal pain for five days, associated with nausea, vomiting and constipation. Physical examination disclosed a large incarcerated umbilical hernia, which was readily apparent on supine abdominal plain films. These also showed a calcified heterogeneous mass in the mid-abdominal region, which was further characterized by CT as a lithopedion (calcified ectopic pregnancy). This is one of the few cases studied on a MDCT equipment, and it clearly enhances the post-processing abilities of this imaging method which allows diagnostic high-quality MIP images. Lithopedion is a rare entity, with less than 300 cases previously described in the medical literature. However, many reported cases corresponded to cases of skeletonization or collections of fetal bone fragments discovered encysted in the pelvic region at surgery or autopsy. It is thus estimated that true lithopedion is a much rarer entity. The diagnosis may be reached by a suggestive clinical history and a palpable mass on physical examination, while the value of modern cross-sectional techniques is still virtually unknown. Ultrasonography may depict an empty uterine cavity and a calcified abdominal mass of non-specific characteristics, and computed tomography or magnetic resonance imaging are able to reach a conclusive diagnosis and may additionally define the involvement of adjacent structures. The differential diagnosis includes other calcified pathologic situations, including ovarian tumors, uterine fibroids, urinary tract neoplasms, inflammatory masses or epiploic calcifications

Reduction in fecal excretion of Giardia cysts: effect of cholestasis and diet.

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Erlandsen, Stanley

2005-12-01

Bile is a major growth factor for the proliferation of Giardia spp. trophozoites in the small intestine and, at high concentrations, stimulates encystment of trophozoites. This report demonstrates that surgical cholestasis to interrupt the flow of bile from liver to intestine or the use of bile-binding resins in the diet can both dramatically decrease the fecal excretion of Giardia muris cysts. Cholestasis produced a 3 log reduction in excretion of G. muris cysts within 24 hr of surgery and a 4 log reduction after 3 days. Sham controls showed no difference in cyst excretion from presurgical control values. Two isocaloric diets were studied: a control diet (N) of Purina mouse chow containing 5% celufil and an experimental diet (CR) containing 5% cholestyramine, a resin that binds bile. Compared with the N diet, the CR diet was associated with reductions in cyst excretion of 3 logs within 1 day. Despite lowered excretion of G. muris cysts in mice fed the cholestyramine diet, the trophozoite recovery from the duodenum was similar with both diets. Cyclic feeding of the CR diet and the N diet at 3-day intervals produced significant oscillations (changes of 3-4 logs) in fecal cyst shedding. The significant reductions in fecal excretion of cysts observed with agents that bind bile suggests that diets capable of binding bile might be a therapeutic means to minimize the fecal excretion of cysts and thereby may help to reduce the risk of spreading giardiasis through fecal-oral contamination.

Occurrence and effect of trematode metacercariae in two endangered killifishes from Greece.

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Kalogianni, Eleni; Kmentová, Nikol; Harris, Eileen; Zimmerman, Brian; Giakoumi, Sofia; Chatzinikolaou, Yorgos; Vanhove, Maarten P M

2017-11-01

We report digeneans (Diplostomidae, Crassiphialinae) in the endangered freshwater fishes Valencia letourneuxi and Valencia robertae, endemics of Western Greece. Digenean metacercariae occurred in two forms in the abdominal cavity, excysted and encysted, the latter attached to the gonads, liver and alimentary tract. Parasites were, using morphological and molecular techniques, identified as two representatives of Crassiphialinae, specifically part of the Posthodiplostomum-Ornithodiplostomum clade. The spatial, seasonal, and age class variation in parasite prevalence was examined. Autumn parasite prevalence varied between the six populations sampled (18.2 to 100%). Seasonal prevalence at the two sites sampled quadannually peaked in autumn and reached its lowest value in spring; prevalence increased with size to 100% in young adult fish. We did not find a correlation between prevalence and host sex. Overall parasites' weight averaged 0.64% of the host's, while parasite weight increased with host weight. A comparison of relative condition and hepatosomatic and gonadosomatic indices of infected and metacercariae-free specimens showed that infection did not have a significant effect on host body condition and reproduction. Regarding the parasite's life cycle, planorbid gastropods are proposed as potential first intermediate hosts in view of the host's diet and occurrence data of molluscs in the ecosystem. This is the first record of a diplostomid digenean in valenciid fishes and of representatives of the Posthodiplostomum-Ornithodiplostomum clade in a native Greek freshwater fish. Our findings are discussed in conjunction to fish conservation interventions, since parasites may contribute to the decline of endangered species.

Efficacy of albendazole:β-cyclodextrin citrate in the parenteral stage of Trichinella spiralis infection.

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Codina, Ana V; García, Agustina; Leonardi, Darío; Vasconi, María D; Di Masso, Ricardo J; Lamas, María C; Hinrichsen, Lucila I

2015-01-01

Albendazole-β-cyclodextrin citrate (ABZ:C-β-CD) inclusion complex in vivo antiparasitic activity was evaluated in the parenteral phase of Trichinella spiralis infection in mice. An equimolar complex of ABZ:C-β-CD was prepared by spray-drying and tested in CBi-IGE male mice orally infected with L1 infective larvae. Infected animals were treated with 50 or 30mg/kg albendazole, (ABZ) equivalent amounts of the ABZ:C-β-CD complex and non treated (controls). Mice received a daily dose on days 28, 29 and 30 post-infection. A week later, larval burden and percentage of encysted dead larvae were assessed in the host by counting viable and non-viable larvae in the tongue. Complexation of ABZ with C-β-CD increased the drug dissolution efficiency nearly eightfold. At 37 days p-i, the reduction percentage in muscle larval load was 35% in mice treated with 50mg/kg/day ABZ and 68% in those given the complex. Treatment with the lower dose showed a similar decrease in parasite burden. Treated animals showed a high percentage of nonviable larvae, the proportion being significantly higher in mice receiving the complex than in control animals (72-88% vs. 11%, P=0.0032). These data indicate that ABZ:C-β-CD increases bioavailability and effectiveness of ABZ against encapsulated Trichinella larvae, thus allowing the use of small doses. Copyright © 2015 Elsevier B.V. All rights reserved.

Pentastomiasis and other parasitic zoonoses from reptiles and amphibians.

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Pantchev, Nikola; Tappe, Dennis

2011-01-01

Reptiles are growing in popularity as pets.The colonization of reptiles and amphibians by parasites and the resulting disease conditions are the most common problems seen in captive animals.This review focuses on pentastomiasis and sparganosis, important parasitic zoonoses of reptiles and amphibians, respectively, and free living-amoebae. Humans are suitable accidental hosts for some pentastomid species (particularly Armillifer and Porocephalus). In geographical areas with special ethnics, such as in West and Central Africa, and East Asia, 8-45% of the human population can be affected. Usually the larvae are coincidentally found during abdominal surgeries. However, fatalities have been described. Extreme caution is necessary when handling infected reptiles. Ocular or cerebral sparganosis is not uncommonly found in humans in East Asia. This disease is caused by spargana, tapeworm larvae (plerocercoids) of Spirometra sp. The infection occurs when uncooked meat from reptiles or amphibians is applied to wounds or eyes and the parasites migrate directly to human tissue, or by consumption of contaminated food or water. As a consequence of the reptile's predatory behaviour, the full spectrum of endo- and ectoparasites from potential prey animals can be found as transiting parasites in the intestinal tract, e. g. Hymenolepis nana, Cryptosporidium (C.) muris, C parvum or Capillaria hepatica. Occasionally, free-living amoebae are also found in reptile faeces (Acanthamoeba, Naegleria, Hartmanella, Vahlkampfia or Echinamoeba sp.).

Provirophages and transpovirons as the diverse mobilome of giant viruses.

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Desnues, Christelle; La Scola, Bernard; Yutin, Natalya; Fournous, Ghislain; Robert, Catherine; Azza, Saïd; Jardot, Priscilla; Monteil, Sonia; Campocasso, Angélique; Koonin, Eugene V; Raoult, Didier

2012-10-30

A distinct class of infectious agents, the virophages that infect giant viruses of the Mimiviridae family, has been recently described. Here we report the simultaneous discovery of a giant virus of Acanthamoeba polyphaga (Lentille virus) that contains an integrated genome of a virophage (Sputnik 2), and a member of a previously unknown class of mobile genetic elements, the transpovirons. The transpovirons are linear DNA elements of ~7 kb that encompass six to eight protein-coding genes, two of which are homologous to virophage genes. Fluorescence in situ hybridization showed that the free form of the transpoviron replicates within the giant virus factory and accumulates in high copy numbers inside giant virus particles, Sputnik 2 particles, and amoeba cytoplasm. Analysis of deep-sequencing data showed that the virophage and the transpoviron can integrate in nearly any place in the chromosome of the giant virus host and that, although less frequently, the transpoviron can also be linked to the virophage chromosome. In addition, integrated fragments of transpoviron DNA were detected in several giant virus and Sputnik genomes. Analysis of 19 Mimivirus strains revealed three distinct transpovirons associated with three subgroups of Mimiviruses. The virophage, the transpoviron, and the previously identified self-splicing introns and inteins constitute the complex, interconnected mobilome of the giant viruses and are likely to substantially contribute to interviral gene transfer.

Proteomic analysis of the cyst stage of Entamoeba histolytica.

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Ibne Karim M Ali

Full Text Available The category B agent of bioterrorism, Entamoeba histolytica has a two-stage life cycle: an infective cyst stage, and an invasive trophozoite stage. Due to our inability to effectively induce encystation in vitro, our knowledge about the cyst form remains limited. This also hampers our ability to develop cyst-specific diagnostic tools.Three main aims were (i to identify E. histolytica proteins in cyst samples, (ii to enrich our knowledge about the cyst stage, and (iii to identify candidate proteins to develop cyst-specific diagnostic tools.Cysts were purified from the stool of infected individuals using Percoll (gradient purification. A highly sensitive LC-MS/MS mass spectrometer (Orbitrap was used to identify cyst proteins.A total of 417 non-redundant E. histolytica proteins were identified including 195 proteins that were never detected in trophozoite-derived proteomes or expressed sequence tag (EST datasets, consistent with cyst specificity. Cyst-wall specific glycoproteins Jacob, Jessie and chitinase were positively identified. Antibodies produced against Jacob identified cysts in fecal specimens and have potential utility as a diagnostic reagent. Several protein kinases, small GTPase signaling molecules, DNA repair proteins, epigenetic regulators, and surface associated proteins were also identified. Proteins we identified are likely to be among the most abundant in excreted cysts, and therefore show promise as diagnostic targets.The proteome data generated here are a first for naturally-occurring E. histolytica cysts, and they provide important insights into the infectious cyst form. Additionally, numerous unique candidate proteins were identified which will aid the development of new diagnostic tools for identification of E. histolytica cysts.

Effect of arabinogalactan proteins from the root caps of pea and Brassica napus on Aphanomyces euteiches zoospore chemotaxis and germination.

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Cannesan, Marc Antoine; Durand, Caroline; Burel, Carole; Gangneux, Christophe; Lerouge, Patrice; Ishii, Tadashi; Laval, Karine; Follet-Gueye, Marie-Laure; Driouich, Azeddine; Vicré-Gibouin, Maïté

2012-08-01

Root tips of many plant species release a number of border, or border-like, cells that are thought to play a major role in the protection of root meristem. However, little is currently known on the structure and function of the cell wall components of such root cells. Here, we investigate the sugar composition of the cell wall of the root cap in two species: pea (Pisum sativum), which makes border cells, and Brassica napus, which makes border-like cells. We find that the cell walls are highly enriched in arabinose and galactose, two major residues of arabinogalactan proteins. We confirm the presence of arabinogalactan protein epitopes on root cap cell walls using immunofluorescence microscopy. We then focused on these proteoglycans by analyzing their carbohydrate moieties, linkages, and electrophoretic characteristics. The data reveal (1) significant structural differences between B. napus and pea root cap arabinogalactan proteins and (2) a cross-link between these proteoglycans and pectic polysaccharides. Finally, we assessed the impact of root cap arabinogalactan proteins on the behavior of zoospores of Aphanomyces euteiches, an oomycetous pathogen of pea roots. We find that although the arabinogalactan proteins of both species induce encystment and prevent germination, the effects of both species are similar. However, the arabinogalactan protein fraction from pea attracts zoospores far more effectively than that from B. napus. This suggests that root arabinogalactan proteins are involved in the control of early infection of roots and highlights a novel role for these proteoglycans in root-microbe interactions.

Effect of Arabinogalactan Proteins from the Root Caps of Pea and Brassica napus on Aphanomyces euteiches Zoospore Chemotaxis and Germination12[C][W

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Cannesan, Marc Antoine; Durand, Caroline; Burel, Carole; Gangneux, Christophe; Lerouge, Patrice; Ishii, Tadashi; Laval, Karine; Follet-Gueye, Marie-Laure; Driouich, Azeddine; Vicré-Gibouin, Maïté

2012-01-01

Root tips of many plant species release a number of border, or border-like, cells that are thought to play a major role in the protection of root meristem. However, little is currently known on the structure and function of the cell wall components of such root cells. Here, we investigate the sugar composition of the cell wall of the root cap in two species: pea (Pisum sativum), which makes border cells, and Brassica napus, which makes border-like cells. We find that the cell walls are highly enriched in arabinose and galactose, two major residues of arabinogalactan proteins. We confirm the presence of arabinogalactan protein epitopes on root cap cell walls using immunofluorescence microscopy. We then focused on these proteoglycans by analyzing their carbohydrate moieties, linkages, and electrophoretic characteristics. The data reveal (1) significant structural differences between B. napus and pea root cap arabinogalactan proteins and (2) a cross-link between these proteoglycans and pectic polysaccharides. Finally, we assessed the impact of root cap arabinogalactan proteins on the behavior of zoospores of Aphanomyces euteiches, an oomycetous pathogen of pea roots. We find that although the arabinogalactan proteins of both species induce encystment and prevent germination, the effects of both species are similar. However, the arabinogalactan protein fraction from pea attracts zoospores far more effectively than that from B. napus. This suggests that root arabinogalactan proteins are involved in the control of early infection of roots and highlights a novel role for these proteoglycans in root-microbe interactions. PMID:22645070

Involvement of cyclin K posttranscriptional regulation in the formation of Artemia diapause cysts.

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Yang Zhao

Full Text Available BACKGROUND: Artemia eggs tend to develop ovoviviparously to yield nauplius larvae in good rearing conditions; while under adverse situations, they tend to develop oviparously and encysted diapause embryos are formed instead. However, the intrinsic mechanisms regulating this process are not well understood. PRINCIPAL FINDING: This study has characterized the function of cyclin K, a regulatory subunit of the positive transcription elongation factor b (P-TEFb in the two different developmental pathways of Artemia. In the diapause-destined embryo, Western blots showed that the cyclin K protein was down-regulated as the embryo entered dormancy and reverted to relatively high levels of expression once development resumed, consistent with the fluctuations in phosphorylation of position 2 serines (Ser2 in the C-terminal domain (CTD of the largest subunit (Rpb1 of RNA polymerase II (RNAP II. Interestingly, the cyclin K transcript levels remained constant during this process. In vitro translation data indicated that the template activity of cyclin K mRNA stored in the postdiapause cyst was repressed. In addition, in vivo knockdown of cyclin K in developing embryos by RNA interference eliminated phosphorylation of the CTD Ser2 of RNAP II and induced apoptosis by inhibiting the extracellular signal-regulated kinase (ERK survival signaling pathway. CONCLUSIONS/SIGNIFICANCE: Taken together, these findings reveal a role for cyclin K in regulating RNAP II activity during diapause embryo development, which involves the post-transcriptional regulation of cyclin K. In addition, a further role was identified for cyclin K in regulating the control of cell survival during embryogenesis through ERK signaling pathways.

A new species of Carcinonemertes, Carcinonemertes conanobrieni sp. nov. (Nemertea: Carcinonemertidae, an egg predator of the Caribbean spiny lobster, Panulirus argus.

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Lunden Alice Simpson

Full Text Available A new species of nemertean worm belonging to the genus Carcinonemertes is described from egg masses of the Caribbean spiny lobster Panulirus argus from the Florida Keys, Florida, USA. This is the first species of Carcinonemertes reported to infect P. argus or any other lobster species in the greater Caribbean and western Atlantic Ocean. Carcinonemertes conanobrieni sp. nov. varies in body color from a translucent white to a pale orange, with males ranging in total body length from 2.35 to 12.71 mm and females ranging from 0.292 to 16.73 mm. Among the traits that separate this new species from previously described species in the genus Carcinonemertes are a relatively wide stylet basis, minimal sexual size dimorphism, and a unique mucus sheath decorated with external hooks. Also, juvenile worms were found to encyst themselves next to lobster embryos and female worms lay both long strings of eggs wound throughout the lobster's setae as well as spherical cases that are attached to lobster embryos. The stylet length and stylet basis remain unchanged throughout ontogeny for both male and female worms. Maximum likelihood and Bayesian inference phylogenetic analyses separated this newly described species from all other species of Carcinonemertes with available COI sequences. Carcinonemertes spp. are voracious egg predators and have been tied to the collapse of various crustacean fisheries. The formal description of this new species represents the first step to understand putative impacts of this worm on the population health of one of the most lucrative yet already depressed crustacean fisheries.

The nature and origin of nucleus-like intracellular inclusions in Paleoproterozoic eukaryote microfossils.

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Pang, K; Tang, Q; Schiffbauer, J D; Yao, J; Yuan, X; Wan, B; Chen, L; Ou, Z; Xiao, S

2013-11-01

The well-known debate on the nature and origin of intracellular inclusions (ICIs) in silicified microfossils from the early Neoproterozoic Bitter Springs Formation has recently been revived by reports of possible fossilized nuclei in phosphatized animal embryo-like fossils from the Ediacaran Doushantuo Formation of South China. The revisitation of this discussion prompted a critical and comprehensive investigation of ICIs in some of the oldest indisputable eukaryote microfossils-the ornamented acritarchs Dictyosphaera delicata and Shuiyousphaeridium macroreticulatum from the Paleoproterozoic Ruyang Group of North China-using a suite of characterization approaches: scanning electron microscopy (SEM), transmission electron microscopy (TEM), and focused ion beam scanning electron microscopy (FIB-SEM). Although the Ruyang acritarchs must have had nuclei when alive, our data suggest that their ICIs represent neither fossilized nuclei nor taphonomically condensed cytoplasm. We instead propose that these ICIs likely represent biologically contracted and consolidated eukaryotic protoplasts (the combination of the nucleus, surrounding cytoplasm, and plasma membrane). As opposed to degradational contraction of prokaryotic cells within a mucoidal sheath-a model proposed to explain the Bitter Springs ICIs-our model implies that protoplast condensation in the Ruyang acritarchs was an in vivo biologically programmed response to adverse conditions in preparation for encystment. While the discovery of bona fide nuclei in Paleoproterozoic acritarchs would be a substantial landmark in our understanding of eukaryote evolution, the various processes (such as degradational and biological condensation of protoplasts) capable of producing nuclei-mimicking structures require that interpretation of ICIs as fossilized nuclei be based on comprehensive investigations. © 2013 John Wiley & Sons Ltd.

Entamoeba Clone-Recognition Experiments: Morphometrics, Aggregative Behavior, and Cell-Signaling Characterization.

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Espinosa, Avelina; Paz-Y-Miño-C, Guillermo; Hackey, Meagan; Rutherford, Scott

2016-05-01

Studies on clone- and kin-discrimination in protists have proliferated during the past decade. We report clone-recognition experiments in seven Entamoeba lineages (E. invadens IP-1, E. invadens VK-1:NS, E. terrapinae, E. moshkovskii Laredo, E. moshkovskii Snake, E. histolytica HM-1:IMSS and E. dispar). First, we characterized morphometrically each clone (length, width, and cell-surface area) and documented how they differed statistically from one another (as per single-variable or canonical-discriminant analyses). Second, we demonstrated that amebas themselves could discriminate self (clone) from different (themselves vs. other clones). In mix-cell-line cultures between closely-related (E. invadens IP-1 vs. E. invadens VK-1:NS) or distant-phylogenetic clones (E. terrapinae vs. E. moshkovskii Laredo), amebas consistently aggregated with same-clone members. Third, we identified six putative cell-signals secreted by the amebas (RasGap/Ankyrin, coronin-WD40, actin, protein kinases, heat shock 70, and ubiquitin) and which known functions in Entamoeba spp. included: cell proliferation, cell adhesion, cell movement, and stress-induced encystation. To our knowledge, this is the first multi-clone characterization of Entamoeba spp. morphometrics, aggregative behavior, and cell-signaling secretion in the context of clone-recognition. Protists allow us to study cell-cell recognition from ecological and evolutionary perspectives. Modern protistan lineages can be central to studies about the origins and evolution of multicellularity. © 2016 The Author(s) Journal of Eukaryotic Microbiology © 2016 International Society of Protistologists.

Pseudomonas aeruginosa adaptation to lungs of cystic fibrosis patients leads to lowered resistance to phage and protist enemies.

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Ville-Petri Friman

Full Text Available Pathogenic life styles can lead to highly specialized interactions with host species, potentially resulting in fitness trade-offs in other ecological contexts. Here we studied how adaptation of the environmentally transmitted bacterial pathogen, Pseudomonas aeruginosa, to cystic fibrosis (CF patients affects its survival in the presence of natural phage (14/1, ΦKZ, PNM and PT7 and protist (Tetrahymena thermophila and Acanthamoebae polyphaga enemies. We found that most of the bacteria isolated from relatively recently intermittently colonised patients (1-25 months, were innately phage-resistant and highly toxic for protists. In contrast, bacteria isolated from long time chronically infected patients (2-23 years, were less efficient in both resisting phages and killing protists. Moreover, chronic isolates showed reduced killing of wax moth larvae (Galleria mellonella probably due to weaker in vitro growth and protease expression. These results suggest that P. aeruginosa long-term adaptation to CF-lungs could trade off with its survival in aquatic environmental reservoirs in the presence of microbial enemies, while lowered virulence could reduce pathogen opportunities to infect insect vectors; factors that are both likely to result in poorer environmental transmission. From an applied perspective, phage therapy could be useful against chronic P. aeruginosa lung infections that are often characterized by multidrug resistance: chronic isolates were least resistant to phages and their poor growth will likely slow down the emergence of beneficial resistance mutations.

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

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Sophie Gryseels

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

High throughput sequencing and proteomics to identify immunogenic proteins of a new pathogen: the dirty genome approach.

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Greub, Gilbert; Kebbi-Beghdadi, Carole; Bertelli, Claire; Collyn, François; Riederer, Beat M; Yersin, Camille; Croxatto, Antony; Raoult, Didier

2009-12-23

With the availability of new generation sequencing technologies, bacterial genome projects have undergone a major boost. Still, chromosome completion needs a costly and time-consuming gap closure, especially when containing highly repetitive elements. However, incomplete genome data may be sufficiently informative to derive the pursued information. For emerging pathogens, i.e. newly identified pathogens, lack of release of genome data during gap closure stage is clearly medically counterproductive. We thus investigated the feasibility of a dirty genome approach, i.e. the release of unfinished genome sequences to develop serological diagnostic tools. We showed that almost the whole genome sequence of the emerging pathogen Parachlamydia acanthamoebae was retrieved even with relatively short reads from Genome Sequencer 20 and Solexa. The bacterial proteome was analyzed to select immunogenic proteins, which were then expressed and used to elaborate the first steps of an ELISA. This work constitutes the proof of principle for a dirty genome approach, i.e. the use of unfinished genome sequences of pathogenic bacteria, coupled with proteomics to rapidly identify new immunogenic proteins useful to develop in the future specific diagnostic tests such as ELISA, immunohistochemistry and direct antigen detection. Although applied here to an emerging pathogen, this combined dirty genome sequencing/proteomic approach may be used for any pathogen for which better diagnostics are needed. These genome sequences may also be very useful to develop DNA based diagnostic tests. All these diagnostic tools will allow further evaluations of the pathogenic potential of this obligate intracellular bacterium.

Detection of Balamuthia mandrillaris DNA by real-time PCR targeting the RNase P gene

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Lewin Astrid

2008-12-01

Full Text Available Abstract Background The free-living amoeba Balamuthia mandrillaris may cause fatal encephalitis both in immunocompromised and in – apparently – immunocompetent humans and other mammalian species. Rapid, specific, sensitive, and reliable detection requiring little pathogen-specific expertise is an absolute prerequisite for a successful therapy and a welcome tool for both experimental and epidemiological research. Results A real-time polymerase chain reaction assay using TaqMan® probes (real-time PCR was established specifically targeting the RNase P gene of B. mandrillaris amoebae. The assay detected at least 2 (down to 0.5 genomes of B. mandrillaris grown in axenic culture. It did not react with DNA from closely related Acanthamoeba (3 species, nor with DNA from Toxoplasma gondii, Leishmania major, Pneumocystis murina, Mycobacterium bovis (BCG, human brain, various mouse organs, or from human and murine cell lines. The assay efficiently detected B. mandrillaris DNA in spiked cell cultures, spiked murine organ homogenates, B. mandrillaris-infected mice, and CNS tissue-DNA preparations from 2 patients with proven cerebral balamuthiasis. This novel primer set was successfully combined with a published set that targets the B. mandrillaris 18S rRNA gene in a duplex real-time PCR assay to ensure maximum specificity and as a precaution against false negative results. Conclusion A real-time PCR assay for B. mandrillaris amoebae is presented, that is highly specific, sensitive, and reliable and thus suited both for diagnosis and for research.

Strongyle egg reappearance period after moxidectin treatment and its relationship with management factors in UK equine populations.

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Tzelos, Thomas; Barbeito, Jessica S G; Nielsen, Martin K; Morgan, Eric R; Hodgkinson, Jane E; Matthews, Jacqueline B

2017-04-15

Parasitic nematodes, particularly cyathostomins, are ubiquitous in grazing horses world-wide. Considerable burdens of cyathostomin larvae can encyst in the large intestinal wall. The most recommended treatment against these pathogenic stages is moxidectin. Information is required on how effective moxidectin is against cyathostomin populations in different regions. The objectives here were to determine the efficacy of moxidectin treatment and estimate the strongyle egg reappearance period (ERP) after treatment in several equine populations, to confirm the type of strongyle nematodes present and to identify other (i.e. management) factors associated with shortened ERP. Eight yards were recruited and moxidectin in combination with praziquantel administered to all horses (n=261). Faecal egg count (FEC) analysis was performed at weeks 0, 2, 6, 10 and 12 after treatment to determine efficacy and ERP. The ERP was estimated using two previously published methods. Morphological identification of cultured third stage larvae from the sample population was compared to a Strongylus vulgaris-specific end-point PCR to examine the presence of S. vulgaris in samples before and after treatment. Strongyle egg shedding patterns were also compared to worm management practices at each site. At 2 weeks post-treatment, moxidectin was highly effective (faecal egg count reduction range, 99.9-100%). The strongyle ERP ranged from 6 weeks to >12 weeks depending on the calculation method applied. Only cyathostomin larvae were detected by morphological identification. The results from the coprocultures and PCR showed that S. vulgaris was absent before and after treatment. Analysis revealed that regular faecal removal from pasture was associated with lower average FEC and lower prevalence of egg shedding. Copyright © 2017 Elsevier B.V. All rights reserved.

The common parasite Toxoplasma gondii induces prostatic inflammation and microglandular hyperplasia in a mouse model.

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Colinot, Darrelle L; Garbuz, Tamila; Bosland, Maarten C; Wang, Liang; Rice, Susan E; Sullivan, William J; Arrizabalaga, Gustavo; Jerde, Travis J

2017-07-01

Inflammation is the most prevalent and widespread histological finding in the human prostate, and associates with the development and progression of benign prostatic hyperplasia and prostate cancer. Several factors have been hypothesized to cause inflammation, yet the role each may play in the etiology of prostatic inflammation remains unclear. This study examined the possibility that the common protozoan parasite Toxoplasma gondii induces prostatic inflammation and reactive hyperplasia in a mouse model. Male mice were infected systemically with T. gondii parasites and prostatic inflammation was scored based on severity and focality of infiltrating leukocytes and epithelial hyperplasia. We characterized inflammatory cells with flow cytometry and the resulting epithelial proliferation with bromodeoxyuridine (BrdU) incorporation. We found that T. gondii infects the mouse prostate within the first 14 days of infection and can establish parasite cysts that persist for at least 60 days. T. gondii infection induces a substantial and chronic inflammatory reaction in the mouse prostate characterized by monocytic and lymphocytic inflammatory infiltrate. T. gondii-induced inflammation results in reactive hyperplasia, involving basal and luminal epithelial proliferation, and the exhibition of proliferative inflammatory microglandular hyperplasia in inflamed mouse prostates. This study identifies the common parasite T. gondii as a new trigger of prostatic inflammation, which we used to develop a novel mouse model of prostatic inflammation. This is the first report that T. gondii chronically encysts and induces chronic inflammation within the prostate of any species. Furthermore, T. gondii-induced prostatic inflammation persists and progresses without genetic manipulation in mice, offering a powerful new mouse model for the study of chronic prostatic inflammation and microglandular hyperplasia. © 2017 Wiley Periodicals, Inc.

Nematode cysts and larvae found in Achatina fulica Bowdich, 1822.

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Franco-Acuña, D O; Pinheiro, J; Torres, E J L; Lanfredi, R M; Brandolini, S V P B

2009-02-01

This study describes the morphology of the nematode cysts and larvae found in Achatina fulica (giant African snail) in Brazil. Sixty snails were collected in Mesquita, Rio de Janeiro State. Fourteen of the snails were naturally infected. The cysts were spherical, pink colored and measured 0.97 to 1.57 mm in diameter. In the majority of cases they had a single larvae involved in amorphous material. A total of 222 encysted larvae were recovered. Of these, 30 were utilized in the morphological study. The length of the larvae varied from 2.57 to 5.8 mm and they were classified as small--up to 3.5 mm; medium--from 3.53 to 4.5 mm; and large--greater than 4.52 mm. The average length of the larvae in the three groups was 2.85, 3.87 and 5.23 mm, respectively. The larval cuticle was white, shiny and transversally striated until the posterior end of the body. At the anterior end there is a mouth with three lips, with amphids and papillae, followed by a muscular esophagus with average length of 0.61 mm, terminating in an esophageal bulb and having a nerve ring in the middle third of the esophagus, and an intestine with an opening near the posterior end. The tail begins from this opening and has two types of ends: short and abrupt or long and gradually tapering. The difference in the tail end can suggest sexual dimorphism, although no primordial reproductive structures were observed. These characteristics were not sufficient to identify the larvae, so there is a need for further study.

Infectious bacterial pathogens, parasites and pathological correlations of sewage pollution as an important threat to farmed fishes in Egypt.

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Mahmoud, Mahmoud A; Abdelsalam, Mohamed; Mahdy, Olfat A; El Miniawy, Hala M F; Ahmed, Zakia A M; Osman, Ahmed H; Mohamed, Hussein M H; Khattab, A M; Zaki Ewiss, M A

2016-12-01

This paper is a part of a multi-disciplinary research "Application of Decentralized On-Site Water Treatment System in Egypt for Use in Agriculture and Producing Safe Fish and Animal Proteins". The project aimed to investigate the environmental impact of implementing sewage water before and after treatment using the effluent of the on-site decentralized Japanese' Johkasou system, in agriculture and producing fish protein. The aim is to establish such system in Egypt to strengthen the sanitary conditions of water resources. In the present study, the impact of the sewage pollution in some fish farms at El-Fayyum, Port Said and El-Dakahlia governorates in Egypt was carried out. Water and fish (Oreochromis niloticus and Mugil cephalus) samples were collected from private fish farms of such localities. Bacteriological and chemical examination of water samples revealed the existence of coliforms and many other bacterial species of significant human health hazards. The chemical parameters of water showed a marked deviation from normal levels while examination of fish flesh specimens indicated contamination with Streptococcus Sp., Staphylococcus Sp., and Salmonella in all examined localities. Other bacterial isolates of human health importance (Morganella morganii, Pseudomonas cepacia and Enterococcos durans) were identified. The parasitological examination revealed the presence of encysted metacercariae (EMC); Diplostomatidae, Prohemistomatidae and Heterphyidae. Moreover, two protozoan parasites (Mxyoboulus tilapiae and Ichthyophthirius multifilis) were also recorded. The histopathological examination revealed mild tissue reaction in case of bacterial infection and severe pathological lesions in different organs in case of EMC infection. Lamellar hyperplasia and mononuclear cell infiltration in branchial tissue was common findings. In skeletal muscles, atrophy of muscle fibres, myolysis and myophagia were detected. Copyright © 2016 Elsevier Ltd. All rights reserved.

Contact lens-associated microbial keratitis Ceratites microbianas relacionadas a lente de contato

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Aline Silveira Moriyama

2008-12-01

Full Text Available PURPOSE: Contact lens-associated microbial keratitis is a severe condition with sight-threatening potential and increasing incidence. Information regarding the etiological agents is essential in guiding management and may vary geographically. The aim of this study was to analyze the microbiological results of corneal scrapings collected from patients presenting with contact lens-associated microbial keratitis. METHODS: Retrospective analysis of the records of all patients who were clinically diagnosed with contact lens-associated microbial keratitis and had corneal scrapings sent to the Laboratory of Ocular Microbiology, UNIFESP/EPM during a 5-year period from January 2002 to December 2007. RESULTS: The etiological agent was identified in 239 patients. Bacterial isolates accounted for 166 (69.46% cases, Acanthamoeba for 95 (39.75% cases and fungi for 4 (1.67% cases. Among the bacterial infections, coagulase-negative Staphylococcus was demonstrated in 74 cases, while Pseudomonas spp was found in 32 patients. All coagulase negative Staphylococcus and Pseudomonas were susceptible to ciprofloxacin and ofloxacin. Resistance to gentamicin was documented in a single case of Pseudomonas. Fourth-generation flouoroquinolone resistance was not observed among Pseudomonas cases. CONCLUSION: Coagulase-negative Staphylococcus was the most frequent isolate, and such data must be considered when determining empiric treatment. Second-generation fluoroquinolones ciprofloxacin and ofloxacin and fourth-generation fluoroquinolones moxifloxacin and gatifloxacin showed a good antibacterial profile and therefore could be good options for initial management.OBJETIVO: A ceratite microbiana associada ao uso de lente de contato é uma condição clínica grave, com risco de perda visual e cuja incidência vem aumentando progressivamente. Os principais agentes etiológicos podem variar geograficamente e informações referentes aos agentes causais são essenciais para

Evidence that the intra-amoebal Legionella drancourtii acquired a sterol reductase gene from eukaryotes

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Fournier Pierre-Edouard

2009-03-01

Full Text Available Abstract Background Free-living amoebae serve as a natural reservoir for some bacteria that have evolved into «amoeba-resistant» bacteria. Among these, some are strictly intra-amoebal, such as Candidatus "Protochlamydia amoebophila" (Candidatus "P. amoebophila", whose genomic sequence is available. We sequenced the genome of Legionella drancourtii (L. drancourtii, another recently described intra-amoebal bacterium. By comparing these two genomes with those of their closely related species, we were able to study the genetic characteristics specific to their amoebal lifestyle. Findings We identified a sterol delta-7 reductase-encoding gene common to these two bacteria and absent in their relatives. This gene encodes an enzyme which catalyses the last step of cholesterol biosynthesis in eukaryotes, and is probably functional within L. drancourtii since it is transcribed. The phylogenetic analysis of this protein suggests that it was acquired horizontally by a few bacteria from viridiplantae. This gene was also found in the Acanthamoeba polyphaga Mimivirus genome, a virus that grows in amoebae and possesses the largest viral genome known to date. Conclusion L. drancourtii acquired a sterol delta-7 reductase-encoding gene of viridiplantae origin. The most parsimonious hypothesis is that this gene was initially acquired by a Chlamydiales ancestor parasite of plants. Subsequently, its descendents transmitted this gene in amoebae to other intra-amoebal microorganisms, including L. drancourtii and Coxiella burnetii. The role of the sterol delta-7 reductase in prokaryotes is as yet unknown but we speculate that it is involved in host cholesterol parasitism.

Identifying Corneal Infections in Formalin-Fixed Specimens Using Next Generation Sequencing.

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Li, Zhigang; Breitwieser, Florian P; Lu, Jennifer; Jun, Albert S; Asnaghi, Laura; Salzberg, Steven L; Eberhart, Charles G

2018-01-01

We test the ability of next-generation sequencing, combined with computational analysis, to identify a range of organisms causing infectious keratitis. This retrospective study evaluated 16 cases of infectious keratitis and four control corneas in formalin-fixed tissues from the pathology laboratory. Infectious cases also were analyzed in the microbiology laboratory using culture, polymerase chain reaction, and direct staining. Classified sequence reads were analyzed with two different metagenomics classification engines, Kraken and Centrifuge, and visualized using the Pavian software tool. Sequencing generated 20 to 46 million reads per sample. On average, 96% of the reads were classified as human, 0.3% corresponded to known vectors or contaminant sequences, 1.7% represented microbial sequences, and 2.4% could not be classified. The two computational strategies successfully identified the fungal, bacterial, and amoebal pathogens in most patients, including all four bacterial and mycobacterial cases, five of six fungal cases, three of three Acanthamoeba cases, and one of three herpetic keratitis cases. In several cases, additional potential pathogens also were identified. In one case with cytomegalovirus identified by Kraken and Centrifuge, the virus was confirmed by direct testing, while two where Staphylococcus aureus or cytomegalovirus were identified by Centrifuge but not Kraken could not be confirmed. Confirmation was not attempted for an additional three potential pathogens identified by Kraken and 11 identified by Centrifuge. Next generation sequencing combined with computational analysis can identify a wide range of pathogens in formalin-fixed corneal specimens, with potential applications in clinical diagnostics and research.

Folic acid is a potent chemoattractant of free-living amoebae in a new and amazing species of protist, Vahlkampfia sp.

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Maeda, Yasuo; Mayanagi, Taira; Amagai, Aiko

2009-03-01

Folic acid (folate; vitamin Bc) is well recognized as essential for the proper metabolism of the essential amino acid methionine as well as for the synthesis of adenine and thymine. A folate deficiency has been Implicated in a wide variety of disorders from Alzheimer's disease to depression and neural tube defects. In the cellular slime molds, including Dictyostelium, vegetative growth-phase cells are known to chemotactically move toward folate that is secreted by bacterial food sources such as Escherichia coli. Intracellular folate signal transductlon, including G proteins, Ca(2+)channels, and the PIP3 pathway, has been reported in D. discoideum. To our surprise, the genuine chemoattractant(s) of free-living protozoan amoebae have remained to be determined, possibly because of lack of a pertinent method for assaying chemotaxis. We recently isolated a primitive free-living amoeba from the soil of Costa Rica and identified it as a new species of the genus Vahlkampfia belonging to Subclass Gymnamoebia, which includes Entamoeba and Acanthamoeba. The amoebae can grow and multiply quite rapidly, engulfing nearby bacteria such as E. coli. Importantly, we have demonstrated here using a quite simple but finely designed chemotaxis assay that the Vahlkampfia amoebae exhibit chemotaxis toward higher folate concentrations. Riboflavin and cyanocobalamin were also found to serve as positive chemoattractants. Among these chemoattractants, folate is of particular importance because its function seems to be evolutionarily conserved as a potent chemoattractant of amoeboid cells in a wide range of organisms as well as in the Protista and cellular slime molds.

Relationship between Organic Carbon and Opportunistic Pathogens in Simulated Glass Water Heaters

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Krista Williams

2015-06-01

Full Text Available Controlling organic carbon levels in municipal water has been hypothesized to limit downstream growth of bacteria and opportunistic pathogens in premise plumbing (OPPPs. Here, the relationships between influent organic carbon (0–15,000 µg ozonated fulvic acid /L and the number of total bacteria [16S rRNA genes and heterotrophic plate counts (HPCs] and a wide range of OPPPs (gene copy numbers of Acanthamoeba polyphaga, Vermamoeba vermiformis, Legionella pneumophila, and Mycobacterium avium were examined in the bulk water of 120-mL simulated glass water heaters (SGWHs. The SGWHs were operated at 32–37 °C, which is representative of conditions encountered at the bottom of electric water heaters, with water changes of 80% three times per week to simulate low use. This design presented advantages of controlled and replicated (triplicate conditions and avoided other potential limitations to OPPP growth in order to isolate the variable of organic carbon. Over seventeen months, strong correlations were observed between total organic carbon (TOC and both 16S rRNA gene copy numbers and HPC counts (avg. R2 > 0.89. Although M. avium gene copies were occasionally correlated with TOC (avg. R2 = 0.82 to 0.97, for 2 out of 4 time points and over a limited TOC range (0–1000 µg/L, no other correlations were identified between other OPPPs and added TOC. These results suggest that reducing organic carbon in distributed water is not adequate as a sole strategy for controlling OPPPs, although it may have promise in conjunction with other approaches.

Relationship between Organic Carbon and Opportunistic Pathogens in Simulated Glass Water Heaters.

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Williams, Krista; Pruden, Amy; Falkinham, Joseph O; Edwards, Marc; Williams, Krista; Pruden, Amy; Falkinham, Joseph O; Edwards, Marc

2015-06-09

Controlling organic carbon levels in municipal water has been hypothesized to limit downstream growth of bacteria and opportunistic pathogens in premise plumbing (OPPPs). Here, the relationships between influent organic carbon (0-15,000 µg ozonated fulvic acid /L) and the number of total bacteria [16S rRNA genes and heterotrophic plate counts (HPCs)] and a wide range of OPPPs (gene copy numbers of Acanthamoeba polyphaga, Vermamoeba vermiformis, Legionella pneumophila, and Mycobacterium avium) were examined in the bulk water of 120-mL simulated glass water heaters (SGWHs). The SGWHs were operated at 32-37 °C, which is representative of conditions encountered at the bottom of electric water heaters, with water changes of 80% three times per week to simulate low use. This design presented advantages of controlled and replicated (triplicate) conditions and avoided other potential limitations to OPPP growth in order to isolate the variable of organic carbon. Over seventeen months, strong correlations were observed between total organic carbon (TOC) and both 16S rRNA gene copy numbers and HPC counts (avg. R2 > 0.89). Although M. avium gene copies were occasionally correlated with TOC (avg. R2 = 0.82 to 0.97, for 2 out of 4 time points) and over a limited TOC range (0-1000 µg/L), no other correlations were identified between other OPPPs and added TOC. These results suggest that reducing organic carbon in distributed water is not adequate as a sole strategy for controlling OPPPs, although it may have promise in conjunction with other approaches.

Antiprotozoal activities of benzimidazoles and correlations with beta-tubulin sequence.

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Katiyar, S K; Gordon, V R; McLaughlin, G L; Edlind, T D

1994-01-01

Benzimidazoles have been widely used since the 1960s as anthelmintic agents in veterinary and human medicine and as antifungal agents in agriculture. More recently, selected benzimidazole derivatives were shown to be active in vitro against two protozoan parasites, Trichomonas vaginalis and Giardia lamblia, and clinical studies with AIDS patients have suggested that microsporidia are susceptible as well. Here, we first present in vitro susceptibility data for T. vaginalis and G. lamblia using an expanded set of benzimidazole derivatives. Both parasites were highly susceptible to four derivatives, including mebendazole, flubendazole, and fenbendazole (50% inhibitory concentrations of 0.005 to 0.16 microgram/ml). These derivatives also had lethal activity that was time dependent: 90% of T. vaginalis cells failed to recover following a 20-h exposure to mebendazole at 0.17 microgram/ml. G. lamblia, but not T. vaginalis, was highly susceptible to five additional derivatives. Next, we examined in vitro activity of benzimidazoles against additional protozoan parasites: little or no activity was observed against Entamoeba histolytica, Leishmania major, and Acanthamoeba polyphaga. Since the microtubule protein beta-tubulin has been identified as the benzimidazole target in helminths and fungi, potential correlations between benzimidazole activity and beta-tubulin sequence were examined. This analysis included partial sequences (residues 108 to 259) from the organisms mentioned above, as well as the microsporidia Encephalitozoon hellem and Encephalitozoon cuniculi and the sporozoan Cryptosporidium parvum. beta-tubulin residues Glu-198 and, in particular, Phe-200 are strong predictors of benzimidazole susceptibility; both are present in Encephalitozoon spp. but absent in C. parvum. PMID:7811023

High throughput sequencing and proteomics to identify immunogenic proteins of a new pathogen: the dirty genome approach.

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Gilbert Greub

Full Text Available BACKGROUND: With the availability of new generation sequencing technologies, bacterial genome projects have undergone a major boost. Still, chromosome completion needs a costly and time-consuming gap closure, especially when containing highly repetitive elements. However, incomplete genome data may be sufficiently informative to derive the pursued information. For emerging pathogens, i.e. newly identified pathogens, lack of release of genome data during gap closure stage is clearly medically counterproductive. METHODS/PRINCIPAL FINDINGS: We thus investigated the feasibility of a dirty genome approach, i.e. the release of unfinished genome sequences to develop serological diagnostic tools. We showed that almost the whole genome sequence of the emerging pathogen Parachlamydia acanthamoebae was retrieved even with relatively short reads from Genome Sequencer 20 and Solexa. The bacterial proteome was analyzed to select immunogenic proteins, which were then expressed and used to elaborate the first steps of an ELISA. CONCLUSIONS/SIGNIFICANCE: This work constitutes the proof of principle for a dirty genome approach, i.e. the use of unfinished genome sequences of pathogenic bacteria, coupled with proteomics to rapidly identify new immunogenic proteins useful to develop in the future specific diagnostic tests such as ELISA, immunohistochemistry and direct antigen detection. Although applied here to an emerging pathogen, this combined dirty genome sequencing/proteomic approach may be used for any pathogen for which better diagnostics are needed. These genome sequences may also be very useful to develop DNA based diagnostic tests. All these diagnostic tools will allow further evaluations of the pathogenic potential of this obligate intracellular bacterium.

Utilization of organic nitrogen by arbuscular mycorrhizal fungi-is there a specific role for protists and ammonia oxidizers?

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Bukovská, Petra; Bonkowski, Michael; Konvalinková, Tereza; Beskid, Olena; Hujslová, Martina; Püschel, David; Řezáčová, Veronika; Gutiérrez-Núñez, María Semiramis; Gryndler, Milan; Jansa, Jan

2018-04-01

Arbuscular mycorrhizal (AM) fungi can significantly contribute to plant nitrogen (N) uptake from complex organic sources, most likely in concert with activity of soil saprotrophs and other microbes releasing and transforming the N bound in organic forms. Here, we tested whether AM fungus (Rhizophagus irregularis) extraradical hyphal networks showed any preferences towards certain forms of organic N (chitin of fungal or crustacean origin, DNA, clover biomass, or albumin) administered in spatially discrete patches, and how the presence of AM fungal hyphae affected other microbes. By direct 15 N labeling, we also quantified the flux of N to the plants (Andropogon gerardii) through the AM fungal hyphae from fungal chitin and from clover biomass. The AM fungal hyphae colonized patches supplemented with organic N sources significantly more than those receiving only mineral nutrients, organic carbon in form of cellulose, or nothing. Mycorrhizal plants grew 6.4-fold larger and accumulated, on average, 20.3-fold more 15 N originating from the labeled organic sources than their nonmycorrhizal counterparts. Whereas the abundance of microbes (bacteria, fungi, or Acanthamoeba sp.) in the different patches was primarily driven by patch quality, we noted a consistent suppression of the microbial abundances by the presence of AM fungal hyphae. This suppression was particularly strong for ammonia oxidizing bacteria. Our results indicate that AM fungi successfully competed with the other microbes for free ammonium ions and suggest an important role for the notoriously understudied soil protists to play in recycling organic N from soil to plants via AM fungal hyphae.

Detailed transcriptome description of the neglected cestode Taenia multiceps.

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Wu, Xuhang; Fu, Yan; Yang, Deying; Zhang, Runhui; Zheng, Wanpeng; Nie, Huaming; Xie, Yue; Yan, Ning; Hao, Guiying; Gu, Xiaobin; Wang, Shuxian; Peng, Xuerong; Yang, Guangyou

2012-01-01

The larval stage of Taenia multiceps, a global cestode, encysts in the central nervous system (CNS) of sheep and other livestock. This frequently leads to their death and huge socioeconomic losses, especially in developing countries. This parasite can also cause zoonotic infections in humans, but has been largely neglected due to a lack of diagnostic techniques and studies. Recent developments in next-generation sequencing provide an opportunity to explore the transcriptome of T. multiceps. We obtained a total of 31,282 unigenes (mean length 920 bp) using Illumina paired-end sequencing technology and a new Trinity de novo assembler without a referenced genome. Individual transcription molecules were determined by sequence-based annotations and/or domain-based annotations against public databases (Nr, UniprotKB/Swiss-Prot, COG, KEGG, UniProtKB/TrEMBL, InterPro and Pfam). We identified 26,110 (83.47%) unigenes and inferred 20,896 (66.8%) coding sequences (CDS). Further comparative transcripts analysis with other cestodes (Taenia pisiformis, Taenia solium, Echincoccus granulosus and Echincoccus multilocularis) and intestinal parasites (Trichinella spiralis, Ancylostoma caninum and Ascaris suum) showed that 5,100 common genes were shared among three Taenia tapeworms, 261 conserved genes were detected among five Taeniidae cestodes, and 109 common genes were found in four zoonotic intestinal parasites. Some of the common genes were genes required for parasite survival, involved in parasite-host interactions. In addition, we amplified two full-length CDS of unigenes from the common genes using RT-PCR. This study provides an extensive transcriptome of the adult stage of T. multiceps, and demonstrates that comparative transcriptomic investigations deserve to be further studied. This transcriptome dataset forms a substantial public information platform to achieve a fundamental understanding of the biology of T. multiceps, and helps in the identification of drug targets and

A new oomycete species parasitic in nematodes, Chlamydomyzium dictyuchoides sp. nov.: developmental biology and phylogenetic studies.

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Beakes, Gordon W; Glockling, Sally L; James, Timothy Y

2014-07-01

The genus Chlamydomyzium is a little studied holocarpic oomycete parasite of nematodes of uncertain phylogenetic and taxonomic position. A new holocarpic species, Chlamydomyzium dictyuchoides, is described which has usually refractile cytoplasm and a dictyuchoid pattern of spore release. This new species infects bacteriotrophic rhabditid nematodes and was isolated from diverse geographical locations. Infection was initiated by zoospore encystment on the host surface and direct penetration of the cuticle. A sparsely branched, constricted, refractile thallus was formed which eventually occupied almost the entire host body cavity, often accompanied by complete dissolution of the host cuticle. Walled primary cysts formed throughout the thallus and each cyst released a single zoospore via an individual exit papillum, leaving a characteristic dictyuchoid wall net behind. At later stages of infection some thalli formed thick-walled stellate resting spores in uniseriate rows. Resting spore formation appeared to be parthenogenetic and was not accompanied by the formation of antheridial compartments. These spores had ooplast-like vacuoles and thick multi-layered walls, both of which suggest they were oospores. The maximum likelihood tree of sequences of the small ribosomal subunit (SSU) gene placed this new isolate in a clade before the main saprolegnialean and peronosporalean lines diverge. A second undescribed Chlamydomyzium sp., which has direct spore release forms a paraphyletic clade, close to C. dictyuchoides and Sapromyces. The fine structure of other documented Chlamydomyzium species was compared, including an undescribed (but sequenced) isolate, SL02, from Japan, Chlamydomyzium anomalum and Chlamydomyzium oviparasiticum. Chlamydomyzium as currently constituted is a paraphyletic genus that is part of a group of phylogenetically problematic early diverging clades that lie close to both the Leptomitales and Rhipidiales. Crown Copyright © 2014. Published by Elsevier

Ultrastructural and immunohistochemical studies on Trichomonas vaginalis adhering to and phagocytizing genitourinary epithelial cells

Institute of Scientific and Technical Information of China (English)

陈文列; 陈金富; 钟秀容; 梁平; 林炜

2004-01-01

Background Trichomonas vaginalis (T. vaginalis) belongs to a common sexually transmitted disease pathogen causing genitourinary trichomoniasis in both sexes. We investigated the pathogenetic mechanism of genitourinary trichomoniasis.Methods Cultured T. vaginalis bodies were injected into the vaginas of rats, or incubated with genitourinary epithelial cells of female subjects, male subjects, and sperm. The ultrastructural and microscopic changes were observed via transmission and scanning electron microscopy and through microscopic histochemistry.Results Groups of T.vaginalis adhered to PAS positive columnar cells at the surface of stratified epithelium in the middle and upper portions of the vaginas. They also traversed under these cells. The parasites were shown to be PAS, cathepsin D, and actin positive, and they could release hydrolase into the cytoplasm of adhered epithelial cells. In the amebiform T.vaginalis, microfilaments were arranged into reticular formation. Similar phenomena were found during the interaction of T.vaginalis with host cells, both in vitro and in vivo. Usually several protozoa adhered to an epithelial cell and formed polymorphic pseudopodia or surface invaginations to surround and phagocytize the microvilli or other parts of the epithelial cytoplasm. Adhesion and phagocytosis of sperm by the protozoa occurred at 15-30 minutes of incubation. Digestion of sperm was found at 45-75 minutes and was complete at 90-105 minutes.Conclusions T.vaginalis tends to parasitize at the fornix of the vagina, because this is the site where columnar cells are rich in mucinogen granules and their microvilli are helpful for adhesion and nibbling. T.vaginalis possesses some invading and attacking abilities. Shape change, canalization, encystation, phagocytosis, digestion, the cell coat, cytoskeleton, and lysosome all play important roles in the process of adhesion. They have two methods of phagocytosis: nibbling and ingestion. Genitourinary epithelium may be

Detailed transcriptome description of the neglected cestode Taenia multiceps.

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Xuhang Wu

Full Text Available BACKGROUND: The larval stage of Taenia multiceps, a global cestode, encysts in the central nervous system (CNS of sheep and other livestock. This frequently leads to their death and huge socioeconomic losses, especially in developing countries. This parasite can also cause zoonotic infections in humans, but has been largely neglected due to a lack of diagnostic techniques and studies. Recent developments in next-generation sequencing provide an opportunity to explore the transcriptome of T. multiceps. METHODOLOGY/PRINCIPAL FINDINGS: We obtained a total of 31,282 unigenes (mean length 920 bp using Illumina paired-end sequencing technology and a new Trinity de novo assembler without a referenced genome. Individual transcription molecules were determined by sequence-based annotations and/or domain-based annotations against public databases (Nr, UniprotKB/Swiss-Prot, COG, KEGG, UniProtKB/TrEMBL, InterPro and Pfam. We identified 26,110 (83.47% unigenes and inferred 20,896 (66.8% coding sequences (CDS. Further comparative transcripts analysis with other cestodes (Taenia pisiformis, Taenia solium, Echincoccus granulosus and Echincoccus multilocularis and intestinal parasites (Trichinella spiralis, Ancylostoma caninum and Ascaris suum showed that 5,100 common genes were shared among three Taenia tapeworms, 261 conserved genes were detected among five Taeniidae cestodes, and 109 common genes were found in four zoonotic intestinal parasites. Some of the common genes were genes required for parasite survival, involved in parasite-host interactions. In addition, we amplified two full-length CDS of unigenes from the common genes using RT-PCR. CONCLUSIONS/SIGNIFICANCE: This study provides an extensive transcriptome of the adult stage of T. multiceps, and demonstrates that comparative transcriptomic investigations deserve to be further studied. This transcriptome dataset forms a substantial public information platform to achieve a fundamental understanding of

Characterization of Amoeboaphelidium protococcarum, an Algal Parasite New to the Cryptomycota Isolated from an Outdoor Algal Pond Used for the Production of Biofuel

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Letcher, Peter M.; Lopez, Salvador; Schmieder, Robert; Lee, Philip A.; Behnke, Craig; Powell, Martha J.; McBride, Robert C.

2013-01-01

Mass culture of algae for the production of biofuels is a developing technology designed to offset the depletion of fossil fuel reserves. However, large scale culture of algae in open ponds can be challenging because of incidences of infestation with algal parasites. Without knowledge of the identity of the specific parasite and how to control these pests, algal-based biofuel production will be limited. We have characterized a eukaryotic parasite of Scenedesmus dimorphus growing in outdoor ponds used for biofuel production. We demonstrated that as the genomic DNA of parasite FD01 increases, the concentration of S. dimorphus cells decreases; consequently, this is a highly destructive pathogen. Techniques for culture of the parasite and host were developed, and the endoparasite was identified as the Aphelidea, Amoeboaphelidium protococcarum. Phylogenetic analysis of ribosomal sequences revealed that parasite FD01 placed within the recently described Cryptomycota, a poorly known phylum based on two species of Rozella and environmental samples. Transmission electron microscopy demonstrated that aplanospores of the parasite produced filose pseudopodia, which contained fine fibers the diameter of actin microfilaments. Multiple lipid globules clustered and were associated with microbodies, mitochondria and a membrane cisternae, an arrangement characteristic of the microbody-lipid globule complex of chytrid zoospores. After encystment and attachment to the host cells, the parasite injected its protoplast into the host between the host cell wall and plasma membrane. At maturity the unwalled parasite occupied the entire host cell. After cleavage of the protoplast into aplanospores, a vacuole and lipids remained in the host cell. Amoeboaphelidium protococcarum isolate FD01 is characteristic of the original description of this species and is different from strain X-5 recently characterized. Our results help put a face on the Cryptomycota, revealing that the phylum is more

The brine shrimp Artemia: adapted to critical life conditions

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Gonzalo M Gajardo

2012-06-01

Full Text Available The brine shrimp Artemia is a micro-crustacean, well adapted to the harsh conditions that severely hypersaline environments impose on survival and reproduction. Adaptation to these conditions has taken place at different functional levels or domains, from the individual (molecular-cellular-physiological to the population level. Such conditions are experienced by very few equivalent macro-planktonic organisms; thus, Artemia can be considered a model animal extremophile offering a unique suite of adaptations that are the focus of this review. The most obvious is a highly efficient osmoregulation system to withstand up to 10 times the salt concentration of ordinary seawater. Under extremely critical environmental conditions, for example when seasonal lakes dry out, Artemia takes refuge by producing a highly resistant encysted gastrula embryo (cyst capable of severe dehydration enabling an escape from population extinction. Cysts can be viewed as gene banks that store a genetic memory of historical population conditions. Their occurrence is due to the evolved ability of females to perceive forthcoming unstable environmental conditions expressed by their ability to switch reproductive mode, producing either cysts (oviparity when environmental conditions become deleterious or free-swimming nauplii (ovoviviparity that are able to maintain the population under suitable conditions.At the population level the trend is for conspecific populations to be fragmented into locally adapted populations, whereas species are restricted to salty lakes in particular regions (regional endemism. The Artemia model depicts adaptation as a complex response to critical life conditions, integrating and refining past and present experiences at all levels of organization. Although we consider an invertebrate restricted to a unique environment, the processes to be discussed are of general biological interest. Finally, we highlight the benefits of understanding the stress

Prevalence of Clonorchis sinensis infection in freshwater fishes in northeastern China.

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Zhang, Y; Chang, Q C; Zhang, Y; Na, L; Wang, W T; Xu, W W; Gao, D Z; Liu, Z X; Wang, C R; Zhu, X Q

2014-08-29

The prevalence of Clonorchis sinensis infection in freshwater fishes was surveyed in Heilongjiang Province, northeastern China, between August 2011 and September 2013. Thirteen species of freshwater fish (n=3221) and one species of shrimp (n=93) were collected from Songhua river, Nenjiang river and other lakes or ponds in 37 sites of 15 representative cities in Heilongjiang Province. They were individually examined by digestion technique, and the C. sinensis metacercariae were identified morphologically followed by confirmation using sequences of the second internal transcribed spacer of ribosomal DNA. Ten of the 13 examined species of freshwater fishes were infected with C. sinensis metacercariae, while all shrimps were negative. The overall prevalence of C. sinensis infection in 3221 examined freshwater fishes was 19.96%, with 42.57% (272/639) in Pseudorasbora parva, 22.55% (83/368) in Hemicculter leuciclus, 20.44% (121/592) in Carassius auratus, 17.71% (68/384) in Saurogobio dabryi, 10.85% (23/212) in Rhodeus ocellatus, 10.54% (48/455) in Phoxinus lagowskii, 8.20% (21/256) in Perccottus glehnii, 6.25% (5/80) in Misgurnus anguillicaudatus, 4.55% (1/22) in Xenocypris davidi, and 1.49% (1/67) in Cyprinus carpio. The average infection intensity in P. parva was 103.3 encysted metacercariae per gram of fish meat in Zhaoyuan city. The average prevalence of C. sinensis infection in Songhua river, Nenjiang river and lakes or ponds were 31.96% (503/1574), 11.30% (102/903) and 7.93% (59/744), respectively. The prevalence of C. sinensis infection in Zhaoyuan city (43.68%) was the highest among all sampling locations. These results revealed a high-prevalence of C. sinensis infection in freshwater fishes in Heilongjiang Province, northeastern China, posing significant public health concern. Copyright © 2014 Elsevier B.V. All rights reserved.

Microbial adhesion to silicone hydrogel lenses: a review.

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Willcox, Mark D P

2013-01-01

Microbial adhesion to contact lenses is believed to be one of the initiating events in the formation of many corneal infiltrative events, including microbial keratitis, that occur during contact lens wear. The advent of silicone hydrogel lenses has not reduced the incidence of these events. This may partly be related to the ability of microbes to adhere to these lenses. The aim of this study was to review the published literature on microbial adhesion to contact lenses, focusing on adhesion to silicone hydrogel lenses. The literature on microbial adhesion to contact lenses was searched, along with associated literature on adverse events that occur during contact lens wear. Particular reference was paid to the years 1995 through 2012 because this encompasses the time when the first clinical trials of silicone hydrogel lenses were reported, and their commercial availability and the publication of epidemiology studies on adverse events were studied. In vitro studies of bacterial adhesion to unworn silicone hydrogel lens have shown that generally, bacteria adhere to these lenses in greater numbers than to the hydroxyethyl methacrylate-based soft lenses. Lens wear has different effects on microbial adhesion, and this is dependent on the type of lens and microbial species/genera that is studied. Biofilms that can be formed on any lens type tend to protect the bacteria and fungi from the effects on disinfectants. Fungal hyphae can penetrate the surface of most types of lenses. Acanthamoeba adhere in greater numbers to first-generation silicone hydrogel lenses compared with the second-generation or hydroxyethyl methacrylate-based soft lenses. Microbial adhesion to silicone hydrogel lenses occurs and is associated with the production of corneal infiltrative events during lens wear.

Evaluation of a TaqMan Array Card for Detection of Central Nervous System Infections.

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Onyango, Clayton O; Loparev, Vladimir; Lidechi, Shirley; Bhullar, Vinod; Schmid, D Scott; Radford, Kay; Lo, Michael K; Rota, Paul; Johnson, Barbara W; Munoz, Jorge; Oneko, Martina; Burton, Deron; Black, Carolyn M; Neatherlin, John; Montgomery, Joel M; Fields, Barry

2017-07-01

Infections of the central nervous system (CNS) are often acute, with significant morbidity and mortality. Routine diagnosis of such infections is limited in developing countries and requires modern equipment in advanced laboratories that may be unavailable to a number of patients in sub-Saharan Africa. We developed a TaqMan array card (TAC) that detects multiple pathogens simultaneously from cerebrospinal fluid. The 21-pathogen CNS multiple-pathogen TAC (CNS-TAC) assay includes two parasites ( Balamuthia mandrillaris and Acanthamoeba ), six bacterial pathogens ( Streptococcus pneumonia e, Haemophilus influenzae , Neisseria meningitidis , Mycoplasma pneumoniae , Mycobacterium tuberculosis , and Bartonella ), and 13 viruses (parechovirus, dengue virus, Nipah virus, varicella-zoster virus, mumps virus, measles virus, lyssavirus, herpes simplex viruses 1 and 2, Epstein-Barr virus, enterovirus, cytomegalovirus, and chikungunya virus). The card also includes human RNase P as a nucleic acid extraction control and an internal manufacturer control, GAPDH (glyceraldehyde-3-phosphate dehydrogenase). This CNS-TAC assay can test up to eight samples for all 21 agents within 2.5 h following nucleic acid extraction. The assay was validated for linearity, limit of detection, sensitivity, and specificity by using either live viruses (dengue, mumps, and measles viruses) or nucleic acid material (Nipah and chikungunya viruses). Of 120 samples tested by individual real-time PCR, 35 were positive for eight different targets, whereas the CNS-TAC assay detected 37 positive samples across nine different targets. The CNS-TAC assays showed 85.6% sensitivity and 96.7% specificity. Therefore, the CNS-TAC assay may be useful for outbreak investigation and surveillance of suspected neurological disease. Copyright © 2017 American Society for Microbiology.

Identification of eukaryotic open reading frames in metagenomic cDNA libraries made from environmental samples.

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Grant, Susan; Grant, William D; Cowan, Don A; Jones, Brian E; Ma, Yanhe; Ventosa, Antonio; Heaphy, Shaun

2006-01-01

Here we describe the application of metagenomic technologies to construct cDNA libraries from RNA isolated from environmental samples. RNAlater (Ambion) was shown to stabilize RNA in environmental samples for periods of at least 3 months at -20 degrees C. Protocols for library construction were established on total RNA extracted from Acanthamoeba polyphaga trophozoites. The methodology was then used on algal mats from geothermal hot springs in Tengchong county, Yunnan Province, People's Republic of China, and activated sludge from a sewage treatment plant in Leicestershire, United Kingdom. The Tenchong libraries were dominated by RNA from prokaryotes, reflecting the mainly prokaryote microbial composition. The majority of these clones resulted from rRNA; only a few appeared to be derived from mRNA. In contrast, many clones from the activated sludge library had significant similarity to eukaryote mRNA-encoded protein sequences. A library was also made using polyadenylated RNA isolated from total RNA from activated sludge; many more clones in this library were related to eukaryotic mRNA sequences and proteins. Open reading frames (ORFs) up to 378 amino acids in size could be identified. Some resembled known proteins over their full length, e.g., 36% match to cystatin, 49% match to ribosomal protein L32, 63% match to ribosomal protein S16, 70% to CPC2 protein. The methodology described here permits the polyadenylated transcriptome to be isolated from environmental samples with no knowledge of the identity of the microorganisms in the sample or the necessity to culture them. It has many uses, including the identification of novel eukaryotic ORFs encoding proteins and enzymes.

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

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

2013-01-01

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

Extensive in silico analysis of Mimivirus coded Rab GTPase homolog suggests a possible role in virion membrane biogenesis

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Amrutraj eZade

2015-09-01

Full Text Available Rab GTPases are the key regulators of intracellular membrane trafficking in eukaryotes. Many viruses and intracellular bacterial pathogens have evolved to hijack the host Rab GTPase functions, mainly through activators and effector proteins, for their benefit. Acanthamoeba polyphaga mimivirus (APMV is one of the largest viruses and belongs to the monophyletic clade of nucleo-cytoplasmic large DNA viruses (NCLDV. The inner membrane lining is integral to the APMV virion structure. APMV assembly involves extensive host membrane modifications, like vesicle budding and fusion, leading to the formation of a membrane sheet that is incorporated into the virion. Intriguingly, APMV and all group I members of the Mimiviridae family code for a putative Rab GTPase protein. APMV is the first reported virus to code for a Rab GTPase (encoded by R214 gene. Our thorough in silico analysis of the subfamily specific (SF region of Mimiviridae Rab GTPase sequences suggests that they are related to Rab5, a member of the group II Rab GTPases, of lower eukaryotes. Because of their high divergence from the existing three isoforms, A, B and C of the Rab5-family, we suggest that Mimiviridae Rabs constitute a new isoform, Rab5D. Phylogenetic analysis indicated probable horizontal acquisition from a lower eukaryotic ancestor followed by selection and divergence. Furthermore, interaction network analysis suggests that vps34 (a Class III P13K homolog, coded by APMV L615, Atg-8 and dynamin (host proteins are recruited by APMV Rab GTPase during capsid assembly. Based on these observations, we hypothesize that APMV Rab plays a role in the acquisition of inner membrane during virion assembly.

Queratitis infecciosa no viral: factores predisponentes, agentes etiológicos y diagnóstico de laboratorio Non viral infectious keratitis: predisposing factors, etiologic agents and laboratory diagnosis

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Federico Nicola

2005-12-01

Full Text Available Las queratitis infecciosas poseen una elevada morbilidad, poniendo en riesgo la visión en casos graves. Dada la eficaz protección que brinda el epitelio corneal, para que ocurra una infección se requiere la presencia de factores condicionantes. El principal predisponente para las queratitis infecciosas es el uso de lentes de contacto, seguido por traumatismos y cirugías oculares y luego diversas afecciones locales o generales. Los agentes etiológicos abarcan una enorme diversidad de microorganismos, incluyendo bacterias, micobacterias, virus, hongos y parásitos. Para poder instaurar un tratamiento acotado se necesita un diagnóstico etiológico, lo que requiere una correcta toma de muestra y un exhaustivo análisis microbiológico.Infectious keratitis cause significant morbidity and, if it is not promptly and appropriately treated, can lead to severe ocular disability. Almost all cases of keratitis are associated to predisposing conditions. In occident, the main risk factor is contact lens wear, but previous ocular surgery or trauma are also important, as well as various ocular surface diseases. An enormous diversity of etiologic agents for infectious keratitis exist, including virus, bacteria, mycobacteria, fungi and parasites. This review provides literature and personal based information about main predisposing factors, etiologic agents and pathophysiology of infectious keratitis, excluding those of viral origin. Focus is made on microbiologic procedures, describing stains and media that should be used, and highlighting their utility. A special mention on particular situations is made, including laboratory diagnosis of Acanthamoeba keratitis, utility of lens cases analysis, keratitis in patients with previous treatment, as well as molecular biology techniques described in ophthalmology.

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

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Lisa Conza

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

Epidemiological profile of infectious keratitis

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Roberta Farias

Full Text Available Abstract Objective: To evaluate the epidemiologic aspects of cornea ulcers in a referred ophthalmology private practice center. Methods: Retrospective study over electronic files of patients treated for corneal ulcers during a period of 7 years by the same physician (RJMF at Centro de Olhos São Francisco, between june 2007 and june 2014. These patients were evaluated for risk factors such as: trauma causes, co-morbidities and contact lenses use. They were also evaluated by the microorganism found at the smears, treatment and patient's outcome. Patients that didn't return with the smears results or that didn´t have a complete follow-up were excluded from the study. Results: A total of 242 patients were fully treated during this period. 55 patients were excluded and 187 patients were included. 28.88% (54/187 were positive for gram negative bacteria, 27.81% (52/187 for filamentous fungi, 16.04% (29/187 for gram positive bacteria and only 1.07% for acanthamoeba. Twenty-four patients had negative cultures and 21 patients had combined infections (bacteria + fungi or other combination. Corneal collagen Cross-linking was effective on melting arrest in 16 of the 16 patients that we have submitted to this procedure during 2008 and 2009. Among all the patients that had corneal infection and were previously using contact lenses, 81% were infected by pseudomonas or pseudomonas associated with other gram negative bacteria. Filamentous fungi had a strong association with physical trauma. While use of saline solution for contact lenses cleaning had a strong association with gram negative bacteria and biological trauma, a strong association with combined infection. Conclusion: Gram negative bacteria (Pseudomonas followed by filamentous fungi (Fusarium sp were the most frequent etiologic agent found in our study. Cross-link was effective on corneal melting arrest. Pseudomonas aeruginosa was the most frequent isolated agent on hydrophilic contact lenses wearers.

Risk factors, microbiological findings, and clinical outcomes in cases of microbial keratitis admitted to a tertiary referral center in ireland.

LENUS (Irish Health Repository)

Saeed, Ayman

2012-02-01

AIM: To identify the risk factors for, and to report the microbiological findings and clinical outcomes of, severe microbial keratitis (MK). METHODS: This was a retrospective study of all cases of presumed MK admitted to a tertiary referral center over a 2-year period (September 2001 to August 2003). Data recorded included demographic data, details relating to possible risk factors, results of microbiological studies, clinical findings at presentation, and clinical and visual outcomes. RESULTS: Ninety patients were admitted with a diagnosis of presumed MK during the study period. The mean age of patients was 45 +\\/- 32 years, and the male to female ratio was 47:43 (52.2%:47.7%). Predisposing risk factors for MK included contact lens wear (37; 41.1%), anterior segment disease (19; 21.1%), ocular trauma (13; 14.4%), systemic disease (5; 5.6%), and previous ocular surgery (1; 1.1%). Cultured organisms included gram-negative bacteria (17; 51.5%), gram-positive bacteria (11, 33.3%), acanthamoeba (2; 6.1%), and fungi (1; 3%). Visual acuity improved significantly after treatment [mean best-corrected visual acuity (+\\/-standard deviation) at presentation: 0.76 (+\\/-0.11); mean best-corrected visual acuity at last follow-up: 0.24 (+\\/-0.07); P < 0.001]. Secondary surgical procedures were required in 18 (20%) cases, and these included punctal cautery (1; 1.1%), tissue glue repair of corneal perforation (2; 2.2%), tarsorrhaphy (9; 9.9%), Botulinum toxin-induced ptosis (1; 1.1%), penetrating keratoplasty (3; 3.3%), and evisceration (2; 2.2%). CONCLUSIONS: Contact lens wear remains a significant risk factor for severe MK. MK remains a threat to vision and to the eye, but the majority of cases respond to prompt and appropriate antimicrobial therapy.

Active and adaptive Legionella CRISPR-Cas reveals a recurrent challenge to the pathogen.

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Rao, Chitong; Guyard, Cyril; Pelaz, Carmen; Wasserscheid, Jessica; Bondy-Denomy, Joseph; Dewar, Ken; Ensminger, Alexander W

2016-10-01

Clustered regularly interspaced short palindromic repeats with CRISPR-associated gene (CRISPR-Cas) systems are widely recognized as critical genome defense systems that protect microbes from external threats such as bacteriophage infection. Several isolates of the intracellular pathogen Legionella pneumophila possess multiple CRISPR-Cas systems (type I-C, type I-F and type II-B), yet the targets of these systems remain unknown. With the recent observation that at least one of these systems (II-B) plays a non-canonical role in supporting intracellular replication, the possibility remained that these systems are vestigial genome defense systems co-opted for other purposes. Our data indicate that this is not the case. Using an established plasmid transformation assay, we demonstrate that type I-C, I-F and II-B CRISPR-Cas provide protection against spacer targets. We observe efficient laboratory acquisition of new spacers under 'priming' conditions, in which initially incomplete target elimination leads to the generation of new spacers and ultimate loss of the invasive DNA. Critically, we identify the first known target of L. pneumophila CRISPR-Cas: a 30 kb episome of unknown function whose interbacterial transfer is guarded against by CRISPR-Cas. We provide evidence that the element can subvert CRISPR-Cas by mutating its targeted sequences - but that primed spacer acquisition may limit this mechanism of escape. Rather than generally impinging on bacterial fitness, this element drives a host specialization event - with improved fitness in Acanthamoeba but a reduced ability to replicate in other hosts and conditions. These observations add to a growing body of evidence that host range restriction can serve as an existential threat to L. pneumophila in the wild. © 2016 The Authors Cellular Microbiology Published by John Wiley & Sons Ltd.

Survey of Legionella spp. in Mud Spring Recreation Area

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Hsu, B.-M.; Ma, P.-H.; Su, I.-Z.; Chen, N.-S.

2009-04-01

Legionella genera are parasites of FLA, and intracellular bacterial replication within the FLA plays a major role in the transmission of disease. At least 13 FLA species—including Acanthamoeba spp., Naegleria spp., and Hartmannella spp.—support intracellular bacterial replication. In the study, Legionellae were detected with microbial culture or by direct DNA extraction and analysis from concentrated water samples or cultured free-living amoebae, combined with molecular methods that allow the taxonomic identification of these pathogens. The water samples were taken from a mud spring recreation area located in a mud-rock-formation area in southern Taiwan. Legionella were detected in 15 of the 34 samples (44.1%). Four of the 34 samples analyzed by Legionella culture were positive for Legionella, five of 34 were positive for Legionella when analyzed by direct DNA extraction and analysis, and 11 of 34 were positive for amoebae-resistant Legionella when analyzed by FLA culture. Ten samples were shown to be positive for Legionella by one analysis method and five samples were shown to be positive by two analysis methods. However, Legionella was detected in no sample by all three analysis methods. This suggests that the three analysis methods should be used together to detect Legionella in aquatic environments. In this study, L. pneumophila serotype 6 coexisted with A. polyphaga, and two uncultured Legionella spp. coexisted with either H. vermiformis or N. australiensis. Of the unnamed Legionella genotypes detected in six FLA culture samples, three were closely related to L. waltersii and the other three were closely related to L. pneumophila serotype 6. Legionella pneumophila serotype 6, L. drancourtii, and L. waltersii are noted endosymbionts of FLA and are categorized as pathogenic bacteria. This is significant for human health because these Legionella exist within FLA and thus come into contact with typically immunocompromised people.

Genome of Phaeocystis globosa virus PgV-16T highlights the common ancestry of the largest known DNA viruses infecting eukaryotes

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Santini, Sebastien; Jeudy, Sandra; Bartoli, Julia; Poirot, Olivier; Lescot, Magali; Abergel, Chantal; Barbe, Valérie; Wommack, K. Eric; Noordeloos, Anna A. M.; Brussaard, Corina P. D.; Claverie, Jean-Michel

2013-01-01

Large dsDNA viruses are involved in the population control of many globally distributed species of eukaryotic phytoplankton and have a prominent role in bloom termination. The genus Phaeocystis (Haptophyta, Prymnesiophyceae) includes several high-biomass-forming phytoplankton species, such as Phaeocystis globosa, the blooms of which occur mostly in the coastal zone of the North Atlantic and the North Sea. Here, we report the 459,984-bp-long genome sequence of P. globosa virus strain PgV-16T, encoding 434 proteins and eight tRNAs and, thus, the largest fully sequenced genome to date among viruses infecting algae. Surprisingly, PgV-16T exhibits no phylogenetic affinity with other viruses infecting microalgae (e.g., phycodnaviruses), including those infecting Emiliania huxleyi, another ubiquitous bloom-forming haptophyte. Rather, PgV-16T belongs to an emerging clade (the Megaviridae) clustering the viruses endowed with the largest known genomes, including Megavirus, Mimivirus (both infecting acanthamoeba), and a virus infecting the marine microflagellate grazer Cafeteria roenbergensis. Seventy-five percent of the best matches of PgV-16T–predicted proteins correspond to two viruses [Organic Lake phycodnavirus (OLPV)1 and OLPV2] from a hypersaline lake in Antarctica (Organic Lake), the hosts of which are unknown. As for OLPVs and other Megaviridae, the PgV-16T sequence data revealed the presence of a virophage-like genome. However, no virophage particle was detected in infected P. globosa cultures. The presence of many genes found only in Megaviridae in its genome and the presence of an associated virophage strongly suggest that PgV-16T shares a common ancestry with the largest known dsDNA viruses, the host range of which already encompasses the earliest diverging branches of domain Eukarya. PMID:23754393

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.

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Jitender P Dubey

Full Text Available 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 of the same age, strain, and sex, after oral inoculation with oocysts (natural route and natural stage of infection of 11 T. gondii strains of seven genotypes. Rats were euthanized at two months post inoculation (p.i. to investigate whether the parasite genotype affects the distribution, location, tissue cyst size, or lesions. Tissue cysts were enumerated in different regions of the brains, both in histological sections as well in saline homogenates. Tissue cysts were found in all regions of the brain. The tissue cyst density in different brain regions varied extensively between rats with many regions highly infected in some animals. Overall, the colliculus was most highly infected although there was a large amount of variability. The cerebral cortex, thalamus, and cerebellum had higher tissue cyst densities and two strains exhibited tropism for the colliculus and olfactory bulb. Histologically, lesions were confined to the brain and eyes. Tissue cyst rupture was frequent with no clear evidence for reactivation of tachyzoites. Ocular lesions were found in 23 (25% of 92 rat eyes at two months p.i. The predominant lesion was focal inflammation in the retina. Tissue cysts were seen in the sclera of one and in the optic nerve of two rats. The choroid was not affected. Only tissue cysts, not active tachyzoite infections, were detected. Tissue cysts were seen in histological sections of tongue of 20 rats but not in myocardium and leg muscle.This study reevaluated

Characterization of Amoeboaphelidium protococcarum, an algal parasite new to the cryptomycota isolated from an outdoor algal pond used for the production of biofuel.

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Peter M Letcher

Full Text Available Mass culture of algae for the production of biofuels is a developing technology designed to offset the depletion of fossil fuel reserves. However, large scale culture of algae in open ponds can be challenging because of incidences of infestation with algal parasites. Without knowledge of the identity of the specific parasite and how to control these pests, algal-based biofuel production will be limited. We have characterized a eukaryotic parasite of Scenedesmus dimorphus growing in outdoor ponds used for biofuel production. We demonstrated that as the genomic DNA of parasite FD01 increases, the concentration of S. dimorphus cells decreases; consequently, this is a highly destructive pathogen. Techniques for culture of the parasite and host were developed, and the endoparasite was identified as the Aphelidea, Amoeboaphelidium protococcarum. Phylogenetic analysis of ribosomal sequences revealed that parasite FD01 placed within the recently described Cryptomycota, a poorly known phylum based on two species of Rozella and environmental samples. Transmission electron microscopy demonstrated that aplanospores of the parasite produced filose pseudopodia, which contained fine fibers the diameter of actin microfilaments. Multiple lipid globules clustered and were associated with microbodies, mitochondria and a membrane cisternae, an arrangement characteristic of the microbody-lipid globule complex of chytrid zoospores. After encystment and attachment to the host cells, the parasite injected its protoplast into the host between the host cell wall and plasma membrane. At maturity the unwalled parasite occupied the entire host cell. After cleavage of the protoplast into aplanospores, a vacuole and lipids remained in the host cell. Amoeboaphelidium protococcarum isolate FD01 is characteristic of the original description of this species and is different from strain X-5 recently characterized. Our results help put a face on the Cryptomycota, revealing that the

Trichostatin A effects on gene expression in the protozoan parasite Entamoeba histolytica

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Singh Upinder

2007-07-01

transcription in cultured parasites. In contrast, the effect of TSA on E. histolytica gene expression is more substantial and includes genes involved in the encystation pathway. These observations will allow further dissection of the effects of histone acetylation and the genetic pathways regulating stage conversion in this pathogenic parasite.

A assimilação dos imigrantes como questão nacional

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Giralda Seyferth

1997-04-01

Full Text Available A campanha de nacionalização instituída durante o Estado Novo (1937-1945 interferiu na vida cotidiana dos imigrantes e descendentes estabelecidos no Brasil - denominados alienígenas em razão das etnicidades prevalecentes e das culturas diferenciadas - exigindo sua assimilação (enquanto sinônimo de caldeamento em nome da unidade nacional. O presente artigo focaliza aspectos do discurso militar sobre a campanha e seus objetivos de assimilação forçada, a partir de textos produzidos por oficiais do Exército que trabalharam como agentes da nacionalização no Vale do Itajaí (SC - região considerada paradigma de "enquistamento", afastada dos princípios da "brasilidade". Procura mostrar que, na condução da campanha pelos militares, prevaleceu uma concepção de Estado-nação que negou legitimidade a quaisquer formas de pertencimento étnico, conforme parâmetros próprios da ideologia nacionalista brasileira gestada desde o século XIX.The nationalization campaign promoted during the Estado Novo (1937-1945 interfered in the daily lives of immigrants and their descendents in Brazil - labeled as aliens because of the prevailing ethnicities and the differentiated cultures - requiring their assimilation (as a synonym for miscegenation in the name of national unity. This study focuses on aspects of the military discourse related to this campaign and its objectives of forced assimilation, based on documents produced by Army officers working as agents for nationalization in the Itajaí Valley, in the State of Santa Catarina, a region viewed as a paradigm of "encystment", straying from the principles of "Brazilianness". It seeks to demonstrate how - in the campaign by the military - a concept of nation-state prevailed which denied legitimacy to any form of ethnic belonging, in accordance with the very parameters of Brazilian nationalist ideology as fomented since the 19th century.

Shortened strongyle-type egg reappearance periods in naturally infected horses treated with moxidectin and failure of a larvicidal dose of fenbendazole to reduce fecal egg counts.

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Rossano, M G; Smith, A R; Lyons, E T

2010-10-29

Deworming horses with anthelmintics that have activity against encysted small strongyle larvae (L(3) and L(4)) is a common practice in parasite control programs. The two drugs currently available for this use are moxidectin (MOX) administered in a single dose of 0.4 mg/kg and fenbendazole (FBZ) given at the larvicidal dose (10mg/kg for 5 days). Here, we report the efficacy of MOX and the larvicidal dose of FBZ for reducing counts of strongyle-type eggs per gram of feces in naturally infected horses. Fecal egg counts (FECs) of 15 yearlings were observed following deworming. On day 0, 6 of the 15 yearlings were administered a larvicidal dose of FBZ; 14 days later, all 15 yearlings received MOX at a single dose of 0.4 mg/kg. Feces were collected on day 0 for pre-treatment egg counts. Feces were collected at weekly intervals thereafter during FEC observation periods. FECs of FBZ-treated horses were compared at day 0 and 14 days post-treatment. The difference in means pre- and post-treatment with FBZ was not statistically significant (p=0.65). On days 0 and 42 of the MOX treatment observation period the mean FEC of the yearlings that had not received the FBZ treatment did not differ significantly from that of the FBZ-treated yearlings. MOX was effective in reducing fecal egg counts to 0 EPG for 21 days. At day 35 all but 2 of the yearlings had some eggs present (range=4-361 EPG) and at day 42 all but 1 yearling had eggs present (range=3-432 EPG). At day 42 the group mean FEC reduction had fallen from 100% to 67%. Results of this study do not support the use of the larvicidal dose of FBZ for small strongyle control. Larger field studies will be needed to investigate whether egg reappearance periods are shortening for MOX-treated horses. Copyright © 2010 Elsevier B.V. All rights reserved.

Amoebal endosymbiont Parachlamydia acanthamoebae Bn9 can grow in immortal human epithelial HEp-2 cells at low temperature; an in vitro model system to study chlamydial evolution.

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Yamane, Chikayo; Yamazaki, Tomohiro; Nakamura, Shinji; Matsuo, Junji; Ishida, Kasumi; Yamazaki, Sumire; Oguri, Satoshi; Shouji, Natsumi; Hayashi, Yasuhiro; Yoshida, Mitsutaka; Yimin; Yamaguchi, Hiroyuki

2015-01-01

Ancient chlamydiae diverged into pathogenic and environmental chlamydiae 0.7-1.4 billion years ago. However, how pathogenic chlamydiae adapted to mammalian cells that provide a stable niche at approximately 37 °C, remains unknown, although environmental chlamydiae have evolved as endosymbionts of lower eukaryotes in harsh niches of relatively low temperatures. Hence, we assessed whether an environmental chlamydia, Parachlamydia Bn9, could grow in human HEp-2 cells at a low culture temperature of 30 °C. The assessment of inclusion formation by quantitative RT-PCR revealed that the numbers of bacterial inclusion bodies and the transcription level of 16SrRNA significantly increased after culture at 30 °C compared to at 37 °C. Confocal microscopy showed that the bacteria were located close to HEp-2 nuclei and were actively replicative. Transmission electron microscopy also revealed replicating bacteria consisting of reticular bodies, but with a few elementary bodies. Cytochalasin D and rifampicin inhibited inclusion formation. Lactacystin slightly inhibited bacterial inclusion formation. KEGG analysis using a draft genome sequence of the bacteria revealed that it possesses metabolic pathways almost identical to those of pathogenic chlamydia. Interestingly, comparative genomic analysis with pathogenic chlamydia revealed that the Parachlamydia similarly possess the genes encoding Type III secretion system, but lacking genes encoding inclusion membrane proteins (IncA to G) required for inclusion maturation. Taken together, we conclude that ancient chlamydiae had the potential to grow in human cells, but overcoming the thermal gap was a critical event for chlamydial adaptation to human cells.

New strategy for rapid diagnosis and characterization of fungal infections: the example of corneal scrapings.

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Pablo Goldschmidt

Full Text Available PURPOSE: The prognosis of people infected with Fungi especially immunocompromised depends on rapid and accurate diagnosis to capitalize on time administration of specific treatments. However, cultures produce false negative results and nucleic-acid amplification techniques require complex post-amplification procedures to differentiate relevant fungal types. The objective of this work was to develop a new diagnostic strategy based on real-time polymerase-chain reaction high-resolution melting analysis (PCR-HRM that a detects yeasts and filamentous Fungi, b differentiates yeasts from filamentous Fungi, and c discriminates among relevant species of yeasts. METHODS: PCR-HRM detection limits and specificity were assessed with a isolated strains; b human blood samples experimentally infected with Fungi; c blood experimentally infected with other infectious agents; d corneal scrapings from patients with suspected fungal keratitis (culture positive and negative and e scrapings from patients with suspected bacterial, viral or Acanthamoeba infections. The DNAs were extracted and mixed with primers diluted in the MeltDoctor® HRM Master Mix in 2 tubes, the first for yeasts, containing the forward primer CandUn (5'CATGCCTGTTTGAGCGTC and the reverse primer FungUn (5'TCCTCCGCTT ATTGATATGCT and the second for filamentous Fungi, containing the forward primer FilamUn (5'TGCCTGTCCGAGCGTCAT and FungUn. Molecular probes were not necessary. The yields of DNA extraction and the PCR inhibitors were systematically monitored. RESULTS: PCR-HRM detected 0.1 Colony Forming Units (CFU/µl of yeasts and filamentous Fungi, differentiated filamentous Fungi from yeasts and discriminated among relevant species of yeasts. PCR-HRM performances were higher than haemoculture and sensitivity and specificity was 100% for culture positive samples, detecting and characterizing Fungi in 7 out 10 culture negative suspected fungal keratitis. CONCLUSIONS: PCR-HRM appears as a new, sensitive

Factors influencing the sporulation and cyst formation of Aphanomyces invadans, etiological agent of ulcerative mycosis in Atlantic menhaden, Brevoortia tyrannus

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Kiryu, Y.; Blazer, V.S.; Vogelbein, W.K.; Kator, H.; Shields, J.D.

2005-01-01

Oomycete infections caused by Aphanomyces invadans occur in freshwater and estuarine fishes around the world. Along the east coast of the USA, skin ulcers caused by A. invadans are prevalent in Atlantic menhaden, Brevoortia tyrannus. From laboratory observations low salinities appear crucial to transmission of the pathogen. To better understand aspects of transmission, we characterized sporulation and cyst formation of secondary zoospores of two isolates of A. invadans at different salinities and temperatures. Sporulation occurred only at low salinities. At room temperature (ca. 20-22 C), using "pond water" augmented with artificial sea salts, the endemic strain WIC and the Thailand strain PA7 of A. invadans produced free-swimming secondary zoospores at salinities of 0, 1 and 2 psu (practical salinity unit = ???), but not at 4 psu or higher. Secondary zoospores of another species, ATCC-62427 (Aphanomyces sp.), were observed at 1, 2, 4 and 8 psu but not at 0 and 12 psu. Secondary zoospores of all three isolates, especially WIC, were abundant and motile 1-2 d post-sporulation. Sporulation was temperature dependent and occurred over a relatively narrow range. No sporulation occurred at 4, 30 or 35 C for either WIC or PA7. For both strains zoospore production within 1-3 d after the initiation of sporulation was more prolific at 25 C than at 20 and 15 C. At 15 C production of zoospores was sustained over 11 d for WIC and 5 d for PA7. At room temperature single WIC secondary zoospores remained motile 12-18 h. Salinities exceeding 4 psu or vigorous shaking caused immediate cyst formation of WIC secondary zoospores. Exposure to menhaden tissue, but not tissues of other fishes to secondary zoospores (WIC), caused rapid (2 h) cyst formation. Cysts were capable of excysting when transferred to 1 psu water within 2-3 h of cyst formation. Cysts that had remained encysted in 6.5 psu for 24 h did not excyst when transferred to 1 psu water. Salinity and temperature requirements

Origin of the cell nucleus, mitosis and sex: roles of intracellular coevolution.

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Cavalier-Smith, Thomas

2010-02-04

in naked growing cells, probably as indirect consequences of the origin of phagotrophy. The first eukaryote had 1-2 cilia and also walled resting cysts; I outline how encystation may have promoted the origin of meiotic sex. I also explain why many alternative ideas are inadequate. Nuclear pore complexes are evolutionary chimaeras of endomembrane- and mitosis-related chromatin-associated proteins. The keys to understanding eukaryogenesis are a proper phylogenetic context and understanding organelle coevolution: how innovations in one cell component caused repercussions on others.

The metabolism of 32P-CP-PLLA seed implanted in the liver and its damage to the normal liver tissue: a study in the experimental dogs

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Tan Zhongbao; Liu Lu; Guo Jinhe; Zhu Guangyu; Wang Fuan; Nie Qi; Gao Hailin; Teng Gaojun

2010-01-01

Objective: To investigate the effects of intratumoral implantation of 32 P -CP-PLLA seeds on the normal canine liver tissue and to explore the metabolism of 32 P-CP-PLLA seeds implanted in the liver of experimental dogs. Methods: Twelve beagles were enrolled in this study. The dogs were randomly and equally divided into four groups: group A (185 MBq), group B (370 MBq), group C (740 MBq) and group D (0 MBq). By using laparotomy procedure 32 P-CP-PLLA seeds were implanted into dog's liver. CT scan was performed before operation as well as before the dog was sacrificed. All dogs were sacrificed three months after the implantation. Before the procedure and 1, 2, 4, 8 and 12 weeks after the procedure the blood tests and serum biochemical tests were conducted. One dog from group B and group C was selected respectively and was fed in a metabolic cage. Within one month after the procedure the cpm in feces and in urine was determined every 24 hours. One dog was picked out from each of the three groups and was punctured to get its liver tissue for pathologic exam each time at 1, 2, 4, 8 and 12 weeks after the implantation, and SPECT imaging was also performed at the same time. Pathologic study, both macroscopic and microscopic (including optical and electronic microscopy) was made to observe the liver damage after the dog was sacrificed. The statistical analysis was processed by using SPSS 13.0 software and the measuring data were expressed with mean ± standard deviation (x ± s). Results: Two months after the procedure, serological examination found that the serum alkaline phosphatase (BKP) in both group B and group C was significantly higher than that in other groups, the difference was statistically significant (P 32 P-CP-PLLA seeds was manifested as a spherical lesion which was encysted by a layer of fibrous tissue with an edematous zone peripherally. Conclusion: The implantation of 32 P-CP-PLLA seeds in dog's liver causes only localized hepatic damage with no general

Effect of antitubercular treatment on ovarian function in female genital tuberculosis with infertility

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Jai Bhagwan Sharma

2016-01-01

Full Text Available AIM: To evaluate the effect of antitubercular therapy (ATT on an ovarian function such as ovarian reserve, ovarian dimensions, and ovarian stromal blood flow. SETTINGS AND DESIGN: Prospective study design. MATERIALS AND METHODS: Fifty infertile women with female genital tuberculosis (FGTB without tubo-ovarian masses diagnosed by positive acid-fast bacilli culture or epithelioid granuloma on endometrial aspirate or positive polymerase chain reaction with positive findings on laparoscopy or hysteroscopy were recruited. The ovarian function tests were performed on day 2/3 as follicle-stimulating hormone (FSH levels and anti-Mullerian hormone (AMH levels. Ovarian dimensions (length, width, and depth were measured using a transvaginal ultrasound. Mean antral follicle count (AFC and ovarian stromal blood flow (peak systolic velocity [PSV], pulsatility index (PI, and resistive index [RI] were measured using a transvaginal ultrasound. All women were started on ATT for 6 months by directly observed treatment strategy. After completion of ATT, all the parameters were repeated. RESULTS: There was a significant increase in AMH (2.68 ± 0.97 ng/ml to 2.8 ± 1.03 ng/ml pre- to post-ATT, nonsignificant increase in FSH (7.16 ± 2.34 mIU/ml to 7.26 ± 2.33 mIU/ml post-ATT, significant increase in mean AFC (7.40 ± 2.12-8.14 ± 2.17, PSV in the right ovary (6.015-6.11 cm/s and left ovary (6.05-6.08 cm/s, PI in the right ovary (0.935-0.951 cm/s and left ovary (0.936-0.957 cm/s, and RI in the right ovary (0.62 ± 0.01-0.79 ± 0.02 and left ovary (0.65 ± 0.02-0.84 ± 0.01 with ATT. There was no significant change in mean ovarian dimensions (ovarian length, breadth, and width and summed ovarian volume with ATT. On laparoscopy, tubercles were seen in 27 (54% women. Caseous nodules and encysted ascites were seen in 8% cases each. CONCLUSION: ATT improves the ovarian function (AMH and AFC and ovarian blood flow in women with FGTB.

Origin of the cell nucleus, mitosis and sex: roles of intracellular coevolution

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Cavalier-Smith Thomas

2010-02-01

advantages. These successive changes took place in naked growing cells, probably as indirect consequences of the origin of phagotrophy. The first eukaryote had 1-2 cilia and also walled resting cysts; I outline how encystation may have promoted the origin of meiotic sex. I also explain why many alternative ideas are inadequate. Conclusion Nuclear pore complexes are evolutionary chimaeras of endomembrane- and mitosis-related chromatin-associated proteins. The keys to understanding eukaryogenesis are a proper phylogenetic context and understanding organelle coevolution: how innovations in one cell component caused repercussions on others. Reviewers This article was reviewed by Anthony Poole, Gáspár Jékely and Eugene Koonin.

Giardia y giardiasis Giardia and giardiasis

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Hugo D. Luján

2006-02-01

responses and produce acute, chronic and/or recurrent infections. During the last years, important advances in the knowledge of the molecular basis underlying the mechanisms of encystation and antigenic variation in Giardia suggest that novel chemotherapeutic and/or immunoprophylactic reagents against this important human pathogen could soon be available.

Extinction Risk of Phytoplankton Species to Potential Killing Mechanisms at the Cretaceous-Paleogene Boundary

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Bralower, T. J.; Schueth, J.; Jiang, S.

2013-05-01

The impact at Chicxulub caused catastrophic changes in marine habitats including extended darkness, ocean acidification and eutrophication. These changes were devastating to some groups of phytoplankton at the base of the marine food chain while others escaped virtually unscathed. For example, diatoms had ~85% survival across the boundary and dinoflagellates actually increased in diversity. These non-calcareous plankton most likely survived due to their adaptation to high-stress environments and their ability to form spores and resting cysts. The calcareous nannoplankton, however, were decimated with approximately 85% of genera and 93% of species going extinct. Nannoplankton generally lack the ability to encyst and thus, as a group, would have been susceptible to darkness, ocean acidification and eutrophication. However, we still do not fully understand why certain nannofossil taxa survived while others went extinct. Extinction risk, the projected susceptibility of a taxon to extinction based on its ecology and ability to adapt, is a concept that is widely applied to extant species and higher order fossil groups, but not to phytoplankton. This concept is a useful for probing the selectivity of ancient species to mass extinction. Determining the extinction risk of latest Maastrichtian nannoplankton species would be a step towards understanding the selection of survivors. The deep-sea record contains a remarkable archive of nannoplankton extinction and recovery across the Cretaceous-Paleogene boundary. The recovery was geologically extended, enabling detailed comparisons between the ocean basins. A large, global database of assemblages had led to the discovery that the Northern Hemisphere oceans suffered higher nannoplankton extinction rates than the Southern Hemisphere with an ecological "crisis" that lasted for approximately 350 thousand years after the impact. In addition, incumbency played a major role in the origination of new species. Since extinction almost

INFLUENCIA DE LAS CONDICIONES DE INCUBACIÓN SOBRE LA ACTIVACIÓN DE ZOOSPORAS DE Spongospora subterranea, EN BUSCA DE UN INÓCULO PARA EL ESTUDIO DE LA SARNA POLVOSA INFLUENCE OF INCUBATION CONDITIONS ON THE ACTIVATION OF ZOOSPORES OF Spongospora subterranea, SEARCHING FOR AN INOCULUM TO THE STUDY OF THE POWDERY SCAB

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Edgar Beltrán Sánchez

2009-12-01

Full Text Available La sarna polvosa es una enfermedad que decrece la producción de papa. Es causada por el parásito obligado Spongospora subterranea f. sp. subterranea, el cual produce grupos de zoosporas enquistadas llamados quistosoros. Cuando las zoosporas están activas pueden infectar raíces y tubérculos iniciando un nuevo ciclo reproductivo. Para estudiar la influencia de las condiciones de incubación sobre la activación de las zoosporas, suspensiones acuosas de quistosoros fueron incubadas bajo diferentes condiciones. Los quistosoros y las zoosporas activas fueron diariamente contados utilizando microscopio óptico. El mayor número de zoosporas activas se encontró al cuarto día de incubación, mientras que el menor número de quistosoros se registró al quinto día de incubación. Esto sugiere que las zoosporas permanecen activas por un corto periodo. El número de zoosporas activas por quistosoro fue significativamente (PThe powdery scab is a disease that decreases potato production. It is caused by Spongospora subterranea f. sp. subterranea. This obligate parasite produces cystosori, which are groups of encysted zoospores. When these zoospores are active, they can infect roots and tubers initiating a new replication cycle. To investigate the influence of the incubation conditions on the zoospore activation, suspensions of cystosori in water were incubated under different conditions. The cystosori and the active zoospores were daily counted under microscope. The higher number of active zoospores was found after four days of incubation, meanwhile the lower number of cystosori was found after five days of incubation. This suggests that zoospores are active for a short period. The number of active zoospores per cystosorus was significantly (P<0.05 higher in tap water than in distillated water, and in presence of light than in dark. More zoospores per cystosorus were activated at 19 °C than at 5 °C, but potato roots did not have a significant (P<0

Organic Nitrogen-Driven Stimulation of Arbuscular Mycorrhizal Fungal Hyphae Correlates with Abundance of Ammonia Oxidizers

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Bukovská, Petra; Gryndler, Milan; Gryndlerová, Hana; Püschel, David; Jansa, Jan

2016-01-01

Large fraction of mineral nutrients in natural soil environments is recycled from complex and heterogeneously distributed organic sources. These sources are explored by both roots and associated mycorrhizal fungi. However, the mechanisms behind the responses of arbuscular mycorrhizal (AM) hyphal networks to soil organic patches of different qualities remain little understood. Therefore, we conducted a multiple-choice experiment examining hyphal responses to different soil patches within the root-free zone by two AM fungal species (Rhizophagus irregularis and Claroideoglomus claroideum) associated with Medicago truncatula, a legume forming nitrogen-fixing root nodules. Hyphal colonization of the patches was assessed microscopically and by quantitative real-time PCR (qPCR) using AM taxon-specific markers, and the prokaryotic and fungal communities in the patches (pooled per organic amendment treatment) were profiled by 454-amplicon sequencing. Specific qPCR markers were then designed and used to quantify the abundance of prokaryotic taxa showing the strongest correlation with the pattern of AM hyphal proliferation in the organic patches as per the 454-sequencing. The hyphal density of both AM fungi increased due to nitrogen (N)-containing organic amendments (i.e., chitin, DNA, albumin, and clover biomass), while no responses as compared to the non-amended soil patch were recorded for cellulose, phytate, or inorganic phosphate amendments. Abundances of several prokaryotes, including Nitrosospira sp. (an ammonium oxidizer) and an unknown prokaryote with affiliation to Acanthamoeba endosymbiont, which were frequently recorded in the 454-sequencing profiles, correlated positively with the hyphal responses of R. irregularis to the soil amendments. Strong correlation between abundance of these two prokaryotes and the hyphal responses to organic soil amendments by both AM fungi was then confirmed by qPCR analyses using all individual replicate patch samples. Further

Microbiological quality in the Zacatepec's aquifer, Morelos, Mexico; Calidad microbiologica del acuifero de Zacatepec, Morelos, Mexico

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Ramirez, Elizabeth; Robles, Esperanza; Sainz, Ma. Guadalupe; Ayala, Reynaldo; Campoy, Emelia [Facultad de Estudios Superiores, Iztacala, Universidad Nacional Autonoma de Mexico, Tlalnepantla, Estado de Mexico (Mexico)]. E-mail: erf@servidor.unam.mx

2009-11-15

Groundwater is one of the main supply sources of drinking water in Mexico, as they support a poor biological community, because of their low contain of nutrients. However in the recent years, human activities and urban growth are the main factors of aquifer contamination. Therefore, water may contain chemical contaminants and microorganisms that can cause several diseases; such is the case of free-living amoebae (FLA). The objective of this research is to determine the microbiological quality (total coliforms, fecal coliforms and free-living amoebae) of Zacatepec aquifer, in the state of Morelos. Monthly samplings of thirteen wells of the aquifer were carried out during one year. The following physicochemical parameters were measure in situ: pH, dissolved oxygen (DO) and temperature. Total and fecal coliforms were determined by the membrane filter technique. Free-living amoebae were cultured on non-nutritive agar medium (NNE); the identification was carried out taking in account their morphological features. All the wells were polluted with total coliforms, one with a geometric mean of 107 CFU/100 mL. Fecal coliforms were absent in four wells; nevertheless, one presented a geometric mean of 107 CFU/100 mL. The most polluted well was number 3, and the months with more bacteria were April and July. Free-living amoebae were isolated in all the wells, the highest isolation number was found in well number 2, while the lowest were found in wells 4, 5, and 10. December and January were the months with the highest number of amoebae, and April with the lowest. Twenty two species belonging to 16 genera were isolated; the most frequent was Hartmannella with 38 %; this amoeba has not been reported as pathogenic, but it has been associated to ocular and brain infections in humans. Of the pathogenic amoebae, Acanthamoeba was isolated, but with a low frequency (6.7 %). Physicochemical parameters had low seasonal variation, pH was close to neutrality in a range of 6.4 to 7

How to help the patient motivate himself?

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Grimaldi, A

2012-03-01

forgetting to play the devils' advocate. Lastly, the expression of the patient's "identity self" through discussion groups, drawing or writing workshops, or "living theatre" may enable expression of an encysted wound. While progressing in that direction, the patient will modify his representation of the disease and its treatment, and enhance his understanding of who he is and how he functions (metacognition). This is the objective of therapeutic education. However, it is necessary for caregivers to demonstrate real empathy that is not only cognitive but also emotional. Copyright © 2012 Elsevier Masson SAS. All rights reserved.

Lack of association between parenchymal neurocysticercosis and HLA Class I and Class II antigens

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Eni Picchioni Bompeixe

1999-03-01

Full Text Available Neurocysticercosis, caused by encysted larvae of the tapeworm Taenia solium, is the most common infection of the central nervous system and a major public health problem in many countries. Prevalence in the region of Curitiba, located in the southern Brazilian State of Paraná, is one of the highest in the world. The genetics of host susceptibility to neurocysticercosis (NCC is still obscure. To investigate if major histocompatibility complex (MHC genes influence individual susceptibility to NCC, we performed a case-control association analysis. Fifty-two Caucasoid patients and 149 matched controls were typed for antigens of the HLA-A, B, C, DR and DQ loci. All patients had computerized tomography and clinical features compatible with parenchymal NCC. Indirect immunofluorescence of cerebrospinal fluid showed that 19 (37% of the patients presented anti-cysticercus antibodies at titers ³ 1:10. Frequencies of HLA specificities in the whole group of patients and in the subgroup with antibodies in cerebrospinal fluid were compared to those of the control group. No significant difference was found. These results do not support the hypothesis of HLA gene participation in susceptibility to parenchymal neurocysticercosis.A neurocisticercose, causada pelo cisticerco, a larva do cestóide Taenia solium, é a infecção mais comum do sistema nervoso central e constitui importante problema de saúde pública em muitos países. A sua prevalência na região de Curitiba, localizada no Estado do Paraná, foi estimada em 9%, situando-se entre as mais elevadas do mundo. Os aspectos genéticos de suscetibilidade à neurocisticercose (NCC ainda são pouco conhecidos. Com o objetivo de investigar se genes do MHC influenciam a suscetibilidade individual à NCC, realizamos uma análise de associação caso-controle. Cinqüenta e dois pacientes caucasóides e 149 indivíduos-controle pareados foram tipados para antígenos dos locos HLA-A, B, C, DR e DQ. Todos os

Enhanced bioavailability and anthelmintic efficacy of mebendazole in redispersible microparticles with low-substituted hydroxypropylcellulose

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Torre-Iglesias PM

2014-09-01

–∞ values. Likewise, after oral administration of the RDM-1:2.5 and RDM-1:5 the AUC0–∞ were 2.67- and 2.97-fold higher, respectively, compared to those of pure MBZ. Therapeutic activity, assessed on the Trichinella spiralis life cycle, showed that RDM-1:5 was the most effective in reducing the number of parasites (4.56-fold as compared to pure MBZ, on the encysted stage.Conclusion: The MBZ: L-HPC RDM might be an effective way of improving oral bioavailability and therapeutic activity using low doses of MBZ (5 mg/kg, which implies a low degree of toxicity for humans. Keywords: benzimidazole carbamates, redispersible microparticles, in vitro dissolution, pharmacokinetics, anthelmintic activity, Trichinella spiralis

Diagnosis and treatment outcome of mycotic keratitis at a tertiary eye care center in eastern india

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Rautaraya Bibhudutta

2011-12-01

Full Text Available Abstract Background Mycotic keratitis is an important cause of corneal blindness world over including India. Geographical location and climate are known to influence the profile of fungal diseases. While there are several reports on mycotic keratitis from southern India, comprehensive clinico-microbiological reports from eastern India are few. The reported prevalence of mycotic keratitis are 36.7%,36.3%,25.6%,7.3% in southern, western, north- eastern and northern India respectively. This study reports the epidemiological characteristics, microbiological diagnosis and treatment outcome of mycotic keratitis at a tertiary eye care center in eastern India. Methods A retrospective review of medical and microbiology records was done for all patients with laboratory proven fungal keratitis. Results Between July 2006 and December 2009, 997 patients were clinically diagnosed as microbial keratitis. While no organisms were found in 25.4% (253/997 corneal samples, 23.4% (233/997 were bacterial, 26.4% (264/997 were fungal (45 cases mixed with bacteria, 1.4% (14/997 were Acanthamoeba with or without bacteria and 23.4% (233/997 were microsporidial with or without bacteria. Two hundred fifteen of 264 (81.4%, 215/264 samples grew fungus in culture while 49 corneal scrapings were positive for fungal elements only in direct microscopy. Clinical diagnosis of fungal keratitis was made in 186 of 264 (70.5% cases. The microscopic detection of fungal elements was achieved by 10% potassium hydroxide with 0.1% calcoflour white stain in 94.8%(238/251 cases. Aspergillus species (27.9%, 60/215 and Fusarium species (23.2%, 50/215 were the major fungal isolates. Concomitant bacterial infection was seen in 45 (17.1%, 45/264 cases of mycotic keratitis. Clinical outcome of healed scar was achieved in 94 (35.6%, 94/264 cases. Fifty two patients (19.7%, 52/264 required therapeutic PK, 9 (3.4%, 9/264 went for evisceration, 18.9% (50/264 received glue application with bandage

This Issue at A Glance

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Hormoz Chams

2014-05-01

Full Text Available In the retrospective investigation titled “Risk factors evaluation of threshold retinopathy of prematurity” at a tertiary ophthalmic center in Tehran, Iran, the authors have studied 859 premature neonates during one year (2008-2009. 791 eyes have had retinopathy of prematurity (ROP and 7.4% of them presented threshold retinopathy (TROP. The investigators aim has been to present and compare the risk factors of ROP and TROP and also to emphasize that the proposed criteria1 for investigation of ROP and TROP are not very precise and valuable for the developing countries. For example they remark that 33% of their TROP had birth weight of more than 1,500 grams. In this investigation the most important risk factors for TROP have been delay in initial examination, low weight, low gestation age, and duration of oxygen therapy. In another presentation, the authors have investigated “The prevalence of viral conjunctivitis in patients who referred to eye specialist hospitals in Tehran, Iran”. They received 150 swap samples and after DNA extraction, and multiplex real-time PCR they found 14.6% of samples were positive for adenovirus and 3.3% positive for herpes simplex 1 (HSV-1. The results for herpes simplex 2 (HSV-2 and varicella-Zoster (VZV were negative. In other investigations2,3 of viral conjunctivitis adenoviruses, HSV-1, HSV-2 and VZV have been reported to be the most important causes of viral conjunctivitis. The authors propose a more extensive investigation on number of patients and the emerging viruses particularly in this part of the world. Rahimi et al in their presentation “Clinical outcomes in acanthamoeba keratitis treated with polyhexamethylene biguanide as monotherapy” have studied 27 eyes of 25 patients presenting Acanthamoeba keratitis (AK. They emphasize the importance of confocal biomicroscopy to find the cysts or trophozoites of the amoeba, indicating 96.3% of positive results compared to 81.5% of positivity in the culture

Evidence from bioinformatics, expression and inhibition studies of phosphoinositide-3 kinase signalling in Giardia intestinalis

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Crompton Mark R

2006-05-01

Full Text Available Abstract Background Giardia intestinalis is a parasitic protozoan and major cause of diarrhoeal disease. Disease transmission is dependent on the ability of the parasite to differentiate back and forth between an intestine-colonising trophozoite and an environmentally-resistant infective cyst. Our current understanding of the intracellular signalling mechanisms that regulate parasite replication and differentiation is limited, yet such information could suggest new methods of disease control. Phosphoinositide-3 kinase (PI3K signalling pathways have a central involvement in many vital eukaryotic processes, such as regulation of cell growth, intracellular membrane trafficking and cell motility. Here we present evidence for the existence of functional PI3K intracellular signalling pathways in G. intestinalis. Results We have identified and characterised two genes, Gipi3k1 and Gipi3k2, which encode putative PI3Ks. Both genes are expressed in trophozoites and encysting cells, suggesting a possible role of GiPI3K1 and GiPI3K2 in regulating giardial growth and differentiation. Extensive nucleotide and amino acid sequence characterisation predicts that both encoded PI3Ks are functional as indicated by the presence of highly conserved structural domains and essential catalytic residues. The inhibitory effect of the PI3K inhibitor LY294002 on trophozoite proliferation also supports their functionality. Phylogenetic analysis supports the identity of GiPI3K1 as a Class I isoform and GiPI3K2 as a Class III isoform. In addition, giardial genes encoding putative homologues of phosphoinositide-metabolising enzymes such as PTEN, MTM, PIPkin and PI 5-phosphatase as well as downstream effectors with phosphoinositide-binding domains have been identified, placing GiPI3K1 and GiPI3K2 in a broader signalling context. Compared with twenty-six PI3Ks from other organisms, GiPI3K1 and GiPI3K2 are unique in that they contain large insertions within their highly conserved

Laparoscopic findings in female genital tuberculosis.

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Sharma, Jai Bhagwan; Roy, Kallol K; Pushparaj, M; Kumar, S; Malhotra, N; Mittal, S

2008-10-01

To evaluate the laparoscopic findings in genital tuberculosis (TB). A total of 85 women of genital TB, who underwent diagnostic laparoscopy for infertility or chronic pelvic pain were enrolled in this retrospective study conducted in our unit at All India Institute of Medical Sciences, New Delhi, India from September 2004 to 2007. The mean age was 28.2 years and the mean parity was 0.24. Most women were from poor socioeconomic status (68.1%). Past history of TB was seen in 29 (34.1%) women with pulmonary TB in 19 (22.35%) women and extrapulmonary in 10 (11.7%) women. Most women presented with infertility (90.6% primary 72.9%; secondary 17.6%) while the rest had chronic pelvic pain (9.4%). The mean duration of infertility was 6.2 years. A total of 49 (57.6%) women had normal menses, while hypomenorrhea, oligomenorrhea, secondary amenorrhea and menorrhagia were seen in 25 (30.1%), 3 (3.5%), 5 (5.9%), and 2 (2.4%) women respectively. Diagnosis of genital TB was made by histopathological evidence of TB granuloma in 16 (18.8%) (Endometrial biopsy in 12.9%, laparoscopy biopsy in 5.9%) women, demonstration of acid fast bacilli (AFB) on microscopy in 2(2.3%), positive AFB culture in 2 (2.3%), positive polymerase chain reaction (PCR) in 55 (64.7%) and laparoscopic findings of genital TB in 40 (47.1%). The various findings on laparoscopy were tubercles on peritoneum (12.9%) or ovary (1.2%), tubovarian masses (7.1%), caseous nodules (5.8%), encysted ascitis in 7.1% women. Various grades of pelvic adhesions were seen in 56(65.8%) women. The various findings on fallopian tubes were normal looking tubes in (7.1%), inability to visualize in 12(14.1%), presence of tubercles on tubes in 3 (3.52%), caseous granuloma in 3 (3.52%), hydrosalpinx in 15 (17.6%) (Right tube 11.7%, left tube 5.9%), pyosalphinx in 3 (3.5%) on right tube and 2 (2.35%) in left tube, beaded tube in 3 (3.5%) on right tube, 4 (4.7%) in left tube with tobacco pouch appearance in 2 (2.35%) women. The right tube

8th European Conference on Rare Diseases & Orphan Products (ECRD 2016

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Michael Schlander

2016-11-01

Full Text Available Table of contents O1 The European Social Preferences Measurement (ESPM study project: social cost value analysis, budget impact, commercial life cycle revenue management, and the economics of biopharmaceutical Research & Development (R&D Michael Schlander, Søren Holm, Erik Nord, Jeff Richardson, Silvio Garattini, Peter Kolominsky-Rabas, Deborah Marshall, Ulf Persson, Maarten Postma, Steven Simoens, Oriol de Solà Morales, Keith Tolley, Mondher Toumi, Harry Telser O2 Newborn Screening: the potential and the challenges James R Bonham O3 Untreatable disease outcomes - how would we measure them? Helmut Hintner, Anja Diem, Martin Laimer O4 Taking Integrated Care Forward: Experiences from Canada to inspire service provision for people living with rare disease in Europe Réjean Hébert O5 Listening to the patient’s voice: social media listening for safety and benefits in rare diseases Nabarun Dasgupta, Carrie E. Pierce, Melissa Jordan O6 Via Opta: Mobile apps making visually impaired patients’ lives easier Barbara Bori, Mohanad Fors, Emilie Prazakova O7 A report of the IRDiRC “Small Population Clinical Trial” Task Force Simon Day O8 HAE patient identification and diagnosis: An innovative, ‘game changing’ collaboration Thomas J. Croce Jr. O9 Co-creating with the community: primary packaging & administration for people with haemophilia Jonas Fransson, Philip Wood O10 Go with Gaucher, taking forward the next generation. How to involve young people to create a new generation of patient advocates Anne-Grethe Lauridsen, Joanne Higgs, Vesna Stojmirova Aleksovska P1 ODAK – Orphan Drug for Acanthamoeba Keratitis Christina Olsen, Ritchie Head, Antonio Asero, Vincenzo Papa, Christa van Kan, Loic Favennec, Silvana Venturella, Michela Salvador, Alan Krol P5 Rare Navigators help people living with rare diseases to manage the social – and healthcare systems Stephanie J. Nielsen, Birthe B. Holm P6 The eAcademy for Tay-Sachs & Sandhoff disease app

Health and condition of endangered young-of-the-year Lost River and Shortnose suckers relative to water quality in Upper Klamath Lake, Oregon, 2014–2015

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Burdick, Summer M.; Conway, Carla M.; Elliott, Diane G.; Hoy, Marshal S.; Dolan-Caret, Amari; Ostberg, Carl O.

2017-10-19

2015; however, hyperplastic and hypertrophic gill tissue and trichodinids on the gills were observed more frequently in 2014. There were more suckers with normal liver color and texture in 2014 than in 2015. The prevalence of suckers with liver inflammation was greater in 2014 and only observed in suckers collected after August 5, whereas liver inflammation occurred intermittently in 2015. Liver glycogen among suckers decreased in late-August 2014 and increased from early August to mid-September 2015. Lost River suckers had greater whole-body triglyceride content but a larger proportion with an absence of visceral fat observed in 2014 than in 2015. In contrast, shortnose suckers were similar between years in regard to both whole-body triglyceride and visceral fat. Black-spot-forming parasites (trematode metacercariae) were observed in a higher prevalence on shortnose suckers but not Lost River suckers in 2014 than in 2015. Opercular deformities were less prevalent in both species in 2014 than in 2015.Neither gross nor histological examination revealed a high prevalence of abnormalities in suckers that clearly indicate a primary mechanism for juvenile mortality in Upper Klamath Lake. Histological abnormalities were almost always focal and minimal or mild except where associated with parasites. Mild to severe focal abnormalities associated with Lernaea sp. attachment sites and encysted digenean (trematode) metacercariae are unlikely to be associated with mortality. Severe and diffuse inflammation and hyperplasia of the gills associated with Ichthyobodo sp. on one Lost River sucker, may indicate a potential cause of mortality. High mortality may have primarily occurred outside our study period (for example, in spring or over winter), or was caused by a factor that could not be detected with our methods (for example, predation). Alternatively, abnormalities in a small percentage of passively captured suckers in Upper Klamath Lake may indicate health-related issues that

Experimental life cycle of Lagochilascaris minor Leiper, 1909 Ciclo evolutivo experimental de Lagochilascaris minor, Leiper 1909

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Dulcinéa Maria Barbosa Campos

1992-08-01

Full Text Available The life cycle of Lagochilascaris minor was studied using material collected from human lesion and applying the experimental model: rodents (mice, hamsters, and carnivorae (cats, dogs. In mice given infective eggs, orally, hatch of the third stage larvae was noted in the gut wall, with migration to liver, lungs, skeletal musculature and subcutaneous tissue becoming, soon after, encysted. In cats infected with skinned carcasses of mice (60 to 235 days of infection it was observed: hatch of third stage larvae from the nodules (cysts in the stomach, migration through the oesophagus, pharynx, trachea, related tissues (rhino-oropharynx, and cervical lymphonodes developing to the mature stage in any of these sites on days 9-20 post inoculation (P.I.. There was no parasite development up to the mature stage in cats inoculated orally with infective eggs, which indicates that the life cycle of this parasite includes an obligatory intermediate host. In one of the cats (fed carcass of infected mice necropsied on day 43 P.I., it was observed the occurence of the self-infective cycle of L. minor in the lung tissues and in the cervical region which was characterized by the finding of eggs in different stages of development, third stage larvae and mature worms. It's believed that some component of the carnivorae gastrointestinal tracts may preclude the development of third stage larvae from L. minor eggs what explains the interruption of the life cycle in animals fed infective eggs. It's also pointed out the role of the intermediate host in the first stages of the life cycle of this helminth.A partir de material colhido de lesões humanas estudou-se o ciclo evolutivo de Lagochilascaris minor empregando-se o modelo experimental: roedores (camundongos, hamster e carnívoros (gatos, cão. Em camundongos inoculados com ovos infectantes, por via oral, observou-se eclosão de larvas de 3º estágio na parede do intestino, migração das mesmas para o fígado, pulm