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Sample records for legionella steelei sp

  1. Filamoeba sp. Isolated from Hot-water Piping System, a Host of Legionella Closely Related to Legionella micdadei

    Czech Academy of Sciences Publication Activity Database

    Dyková, Iva; Hrubá, L.; Kostka, Martin; Pecková, Hana

    2009-01-01

    Roč. 48, č. 4 (2009), s. 321-327 ISSN 0065-1583 R&D Projects: GA ČR GA524/09/0137; GA MŠk LC522 Institutional research plan: CEZ:AV0Z60220518 Keywords : Filamoeba sp. * intracellular * Legionella sp. Subject RIV: GJ - Animal Vermins ; Diseases, Veterinary Medicine Impact factor: 0.775, year: 2009

  2. 'Gatekeeper' unit defeats legionella.

    Science.gov (United States)

    Wolters, Jo

    2007-03-01

    Legionella presents an ongoing cause for concern in any sector where duty of care responsibilities prevail. Jo Wolters, microbiologist and sector head of TA-Aqua+ at Tour & Andersson, describes a recently developed method of legionella prevention and control.

  3. Occurrence and Control of Legionella in Recycled Water Systems.

    Science.gov (United States)

    Jjemba, Patrick K; Johnson, William; Bukhari, Zia; LeChevallier, Mark W

    2015-07-01

    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.

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

    Science.gov (United States)

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

    2012-11-01

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

  5. Occurrence and Control of Legionella in Recycled Water Systems

    OpenAIRE

    Jjemba, Patrick K.; Johnson, William; Bukhari, Zia; LeChevallier, Mark W.

    2015-01-01

    Legionella pneumophila is on the United States Environmental Protection Agency (USEPA) Candidate Contaminant list (CCL) as an important pathogen. It is commonly encountered in recycled water and is typically associated with amoeba, notably Naegleria fowleri (also on the CCL) and Acanthamoeba sp. No legionellosis outbreak has been linked to recycled water and it is important for the industry to proactively keep things that way. A review was conducted examine the occurrence of Legionella and it...

  6. [Studies on detection methods for Legionella species from environmental water].

    Science.gov (United States)

    Kasuga, O; Takagi, K; Tani, K; Kinumaki, A

    1999-01-01

    We investigated selective cultivation media and previous treatments of samples suitable for detection of Legionella species from environmental water and for elimination of co-existing microbes which gave rise to an interference with the evaluation of Legionella sp. growth. Twenty thousand U of polymyxin B (PL-B)/ml and 100 micrograms of oxytetracycline (OTC)/ml seem to be useful as additives to MWY selective agar medium. Both antibiotics markedly inhibited the growth of co-existing microbes with almost no influence on the growth of Legionella sp. In the studies on the resistance of 8 strains of Legionella sp., 24 strains of co-existing microbes and 2 standard strains of Staphylococcus aureus and Escherichia coli to acid treatment (0.2 M HCl-KCl, pH 2.2, 25 degrees C, 4 minutes) and heating (50 degrees C, 20 minutes), acid treatment or heating alone showed no inhibition on the growth of almost all strains examined. However, combination with acid treatment after heating resulted in an apparent extinction of almost all microbes except for Legionella sp., Seven strains from co-existing microbes showed an apparent growth inhibition against 8 strains of Legionella sp. with different serotypes and were all identified as Pseudomonas aeurginosa, which were all eliminated by means of the combination with acid treatment after heating. From these results, it was concluded that the combined pre-treatment of water samples with acid after heating and the addition of PL-B and OTC into the selective cultivation medium is an useful method for detection of Legionella sp. from environmental water.

  7. Legionella in habitations

    DEFF Research Database (Denmark)

    Krøjgaard, Louise Hjelmar

    Legionella is the causative agent of Legionnaires’ disease. The bacteria are widespread in nature and man-made water systems. In Denmark, approximately 120 cases are diagnosed each year, and the disease can be fatal. Legionella pneumophila is the species responsible for approximately 95% of cases....... The transmission pathway is through inhalation of contaminated water droplets mainly from technical systems such as hot water systems. Overall, the aim of this PhD thesis was to improve the background knowledge to accomplish risk assessment regarding Legionella in water systems. Based on a literature review...... and the results from the PhD work the following subjects were addressed: a) prevalence of Legionella in habitations, b) validation of the use of qPCR in risk assessment in hot water systems, c) clarifying risk factors mainly associated with Legionella in habitations, and d) discussion of interventions which could...

  8. Cooling towers and legionella; Koeltorens en legionella

    Energy Technology Data Exchange (ETDEWEB)

    Schalekamp, J.D. [Afdeling Medische Milieukunde, Gemeentelijke Gezondheidsdienst voor Rotterdam en omstreken, Rotterdam (Netherlands)

    2003-03-01

    A brief description is given of the principle of cooling towers, the prevention of legionella in cooling towers, and related regulations. Also the intermediate results of a survey on open cooling towers in the Rotterdam area are presented. Finally, attention is paid to the procedure to handle cases in which legionella has been detected. [Dutch] Jaarlijks vinden er op de wereld epidemieen van legionellose plaats, veroorzaakt door open koeltorens. in dit artikel wordt achtereenvolgens een korte beschrijving gegeven van het werkingsprincipe van koeltorens; de relatie tussen koeltorenbeheer en legionella in open koeltorens; en de huidige stand van zaken op het gebied van regelgeving. Daarnaast worden tussentijdse resultaten van een in Rotterdam gestarte inventarisatie van open koeltorens gegeven. Tenslotte wordt de procedure uiteengezet van de aanpak bij ziektegevallen waarbij legionellose wordt geconstateerd.

  9. Occurrence and Control of Legionella in Recycled Water Systems

    Directory of Open Access Journals (Sweden)

    Patrick K. Jjemba

    2015-07-01

    Full Text Available Legionella pneumophila is on the United States Environmental Protection Agency (USEPA Candidate Contaminant list (CCL as an important pathogen. It is commonly encountered in recycled water and is typically associated with amoeba, notably Naegleria fowleri (also on the CCL and Acanthamoeba sp. No legionellosis outbreak has been linked to recycled water and it is important for the industry to proactively keep things that way. A review was conducted examine the occurrence of Legionella and its protozoa symbionts in recycled water with the aim of developing a risk management strategy. The review considered the intricate ecological relationships between Legionella and protozoa, methods for detecting both symbionts, and the efficacy of various disinfectants.

  10. Alternative sources of Legionella bacteria

    NARCIS (Netherlands)

    van Heijnsbergen, H.H.L.

    2017-01-01

    Legionella bacteria can cause Legionnaires’ disease (LD) in humans. Symptoms of LD can range from mild disease to severe pneumonia with sometimes fatal outcome. In the Netherlands, the most important infective agent is Legionella pneumophila. L. pneumophila infection is associated with aquatic

  11. Compost facilities as a reservoir of Legionella pneumophila and other Legionella species.

    Science.gov (United States)

    Casati, S; Conza, L; Bruin, J; Gaia, V

    2010-07-01

    Data on the presence of Legionellae outside the aquatic environment are scarce. Alternative ecosystems that could act as a reservoir for Legionella spp. have been investigated to identify unconventional contaminated substrates that are able to produce bioaerosols. We considered eight green waste collection sites including three composting facilities. Legionella pneumophila sg 1-15, Legionella bozemanii, Legionella cincinnatiensis, Legionella jamestowniensis, Legionella micdadei and L. oakridgensis were isolated from samples taken at six of the eight sites. The degree of contamination ranged from 10(3) to 10(8) CFU/g. Compost facilities appear to comprise an important reservoir for Legionellae.

  12. Prevention of legionella. Special issue; Legionellapreventie. Themanummer

    Energy Technology Data Exchange (ETDEWEB)

    Van Wolferen, H. [TNO, Apeldoorn (Netherlands); Eland, A.; Meijer, J. [Procas, Rotterdam (Netherlands); Van den Brink, J. [Econosto Nederland, Capelle aan den IJssel (Netherlands); Tiedink, H. [Wilo Nederland, Beverwijk (Netherlands); Nuijten, O.W.W. [ISSO, Rotterdam (Netherlands); Brands, R. [Cumulus Nederland, Cuijk (Netherlands); Reijman, D.

    2008-09-15

    In seven articles attention is paid to several aspects with regard to the prevention of legionella in water heating systems: (1) an interview with an expert in this field (Will Scheffer); (2) regulations for alternative techniques; (3) experiences with collective water heaters; (4) the effect of pressure on legionella risk; (5) the prevention of legionella in cooling towers; (6) legionella safe adiabatic humidification of ambient air; (7) reducing the risk of legionella by maintenance and control. [Dutch] In 7 artikelen wordt aandacht besteed aan verschillende aspecten m.b.t. de voorkoming van legionella in verwarmingssystemen: (1) een interview met een expert op dit gebied (Will Scheffer); (2) beoordelingsrichtlijnen voor alternatieve technieken; (3) praktijkervaringen met collectieve leidingwaterinstallaties; (4) de invloed van drukverhoging op het risico van legionella; (5) legionellapreventie in koeltorens; (6) legionellaveilige adiabatische luchtbevochtiging; en (7) het beperken van het risico op legionella door onderhoud en beheer.

  13. Be prepared should Legionella strike.

    Science.gov (United States)

    Steel, Andrew

    2013-03-01

    When measures to prevent Legionella fail, and a positive result is returned from the laboratory, there is an imperative to act fast to stamp infection out, but taking the right action requires a rigorous approach. So says Andrew Steel, managing director of Airmec, a provider of essential air and water services solutions. Here he explains the legal and other obligations of health estates managers in taking all possible steps to prevent a Legionella outbreak, and, when they do face one, sets out the key remedial steps, and outlines what regulators or HSE personnel will expect to see as 'evidence' that sufficient 'duty of care' has been taken.

  14. sp

    African Journals Online (AJOL)

    Vihar

    adopted as the first line drug. SP has few untoward effects if used carefully in therapeutic doses. Nausea, vomiting, generalized body weakness; diarrhea, skin rashes and hematological reactions are some of the associated side effects. The drug can cause severe skin reactions such as Steven Johnson's syndrome. This.

  15. Occurrence of Legionella pneumophila in lakes serving as a cooling system of a power plant.

    Science.gov (United States)

    Walczak, Maciej; Kletkiewicz, Hanna; Burkowska, Aleksandra

    2013-12-01

    This study was aimed at determining whether Legionella pneumophila can be found in lakes serving as a natural cooling system of a power plant. Water samples were collected from five lakes forming the cooling system of the power plants Pątnów and Konin (Poland). The numbers of bacteria belonging to different phylogenetic groups (bacteria, Legionella sp. and L. pneumophila) were determined with the use of a molecular FISH method. The results of the present study show that thermally altered aquatic environments provide perfect conditions for the growth of L. pneumophila. These microorganisms were identified in the biofilm throughout the entire research period, and in the subsurface water layer in July and August. The percentage of L. pneumophila species in the Legionella genus was 11.55-12.42%.

  16. Legionella (Legionnaires' Disease and Pontiac Fever): Diagnosis

    Science.gov (United States)

    ... Program Application CDC Legionella Healthy Swimming CDC Vessel Sanitation Program Unexplained Respiratory Disease Outbreaks (URDO) European Legionnaires’ Disease Surveillance Network (ELDSNet) Diagnosis, Treatment, and Complications Language: English (US) ...

  17. Confirmed and Potential Sources of Legionella Reviewed

    NARCIS (Netherlands)

    van Heijnsbergen, Eri; Schalk, Johanna A C; Euser, Sjoerd M; Brandsema, Petra S; den Boer, Jeroen W; de Roda Husman, Ana Maria|info:eu-repo/dai/nl/139498281

    2015-01-01

    Legionella bacteria are ubiquitous in natural matrices and man-made systems. However, it is not always clear if these reservoirs can act as source of infection resulting in cases of Legionnaires' disease. This review provides an overview of reservoirs of Legionella reported in the literature, other

  18. Combined pericarditis and pneumonia caused by Legionella infection

    DEFF Research Database (Denmark)

    Svendsen, Jesper Hastrup; Jønsson, V; Niebuhr, U

    1987-01-01

    During a one year period acute pericarditis was diagnosed in 16 consecutive patients without acute infarction or malignancy. In two of these patients with both pericarditis and pneumonia Legionella infection was present. One case was caused by Legionella longbeachae and the other by both Legionella...... longbeachae and Legionella jordanis. When pericarditis is associated with pneumonia Legionella infection should be sought so that effective treatment with erythromycin may be started early....

  19. UV-C inactivation of Legionella rubrilucens.

    Science.gov (United States)

    Schmid, Julian; Hoenes, Katharina; Rath, Monika; Vatter, Petra; Hessling, Martin

    2017-01-01

    Background: Despite the great health significance of Legionella , there is only little information on their UV sensitivity. Besides Legionella pneumophila only L. longbeachae has been investigated so far. Methods: In this study L. rubrilucens has been spread on buffered charcoal yeast extract agar and irradiated with the 254 nm UV-C emission of a mercury vapor lamp. The disinfection success is measured by colony counting after incubation and comparison of the number of colonies on irradiated and unirradiated reference agar plates. Results: The average log-reduction dose is 1.08 mJ/cm 2 for free L. rubrilucens , which is at the lower end of the so far published Legionella log-reduction values, but all three Legionella species show similar UV-C sensitivities. Conclusion: The log-reduction dose of legionellae in amoebae has not been investigated, but with the observed high UV-C sensitivity for free Legionella , the idea of a future point-of-use disinfection by small UV-C LEDs in water-taps or shower heads appears to be realistic, even if legionellae are more resistant in amoebae.

  20. Activities of Tigecycline (GAR-936) against Legionella pneumophila In Vitro and in Guinea Pigs with L. pneumophila Pneumonia

    OpenAIRE

    Edelstein, Paul H.; Weiss, William J.; Edelstein, Martha A. C.

    2003-01-01

    The activities of tigecycline (Wyeth Research) against extracellular and intracellular Legionella pneumophila and for the treatment of guinea pigs with L. pneumophila pneumonia were studied. The tigecycline MIC at which 50% of strains are inhibited for 101 different Legionella sp. strains was 4 μg/ml versus 0.125 and 0.25 μg/ml for azithromycin and erythromycin, respectively. Tigecycline was about as active as erythromycin (tested at 1 μg/ml) against the F889 strain of L. pneumophila grown in...

  1. Legionella (Legionnaires' Disease and Pontiac Fever): Prevention

    Science.gov (United States)

    ... Program Application CDC Legionella Healthy Swimming CDC Vessel Sanitation Program Unexplained Respiratory Disease Outbreaks (URDO) European Legionnaires’ Disease Surveillance Network (ELDSNet) Prevention Language: English (US) Español (Spanish) Recommend ...

  2. Characterization of a Legionella micdadei mip mutant

    DEFF Research Database (Denmark)

    O'Connell, W A; Bangsborg, Jette Marie; Cianciotto, N P

    1995-01-01

    The pathogenesis of Legionella micdadei is dependent upon its ability to infect alveolar phagocytes. To better understand the basis of intracellular infection by this organism, we examined the importance of its Mip surface protein. In Legionella pneumophila, Mip promotes infection of both human m...... into the phagocyte. Similarly, the mutant was less able to parasitize Hartmannella amoebae. Taken together, these data argue that Mip specifically potentiates intracellular growth by L. micdadei....

  3. UV-C-Inaktivierung von Legionella rubrilucens

    OpenAIRE

    Schmid, J; Hoenes, K; Rath, M; Vatter, P; Hessling, M

    2017-01-01

    Background: Despite the great health significance of Legionella , there is only little information on their UV sensitivity. Besides Legionella pneumophila only L. longbeachae has been investigated so far. Methods: In this study L. rubrilucens has been spread on buffered charcoal yeast extract agar and irradiated with the 254 nm UV-C emission of a mercury vapor lamp. The disinfection success is measured by colony counting after incubation and comparison of the number of colonies on irradi...

  4. Viable Legionella pneumophila bacteria in natural soil and rainwater puddles

    NARCIS (Netherlands)

    van Heijnsbergen, E.; de Roda Husman, A. M.; Lodder, W. J.; Bouwknegt, M.; Docters van Leeuwen, A. E.; Bruin, J. P.; Euser, S. M.; den Boer, J. W.; Schalk, J. A C

    Aims: For the majority of sporadic Legionnaires' disease cases the source of infection remains unknown. Infection may possible result from exposure to Legionella bacteria in sources that are not yet considered in outbreak investigations. Therefore, potential sources of pathogenic Legionella

  5. Faster Legionella testing on horizon.

    Science.gov (United States)

    Pearson, Susan

    2011-08-01

    While the "traditional" way to measure Legionella quantitatively in water is based on a complex culture method where results can take up to 14 days, the last few years have seen the availability of very rapid real-time monitoring of the bacterium in water systems, with the development of quantitative polymerase chain reaction (qPCR), a process which gives results "within hours". To date, however, a lack of consensus on how to interpret such results in relation to those from culture has been a stumbling block, although, as Susan Pearson, a freelance journalist and public relations consultant specialising in medicine and the environment, reports, the positive results of a recent multi-centre European study mean this could soon all change.

  6. Legionella safety in cooling towers; Legionellaveiligheid in koeltorens

    Energy Technology Data Exchange (ETDEWEB)

    Kordes, B. [Kordes Advies, (Netherlands); De Bok, F. [KBBL Wijhe, (Netherlands); De Zeeuw, L. [Holland Environment Group, (Netherlands); Settels, P. [Safety, Health Services and Ergonomics, ING, (Netherlands); Oesterholt, F.; Wullings, B. [KWR Watercycle Research Institute, (Netherlands); Guiot, P. [Tevan, Gorinchem (Netherlands); Brands, R. [Cumulus Nederland, Cuijk (Netherlands); Nuijten, O. [Kennisinstituut ISSO, Rotterdam (Netherlands); Wijne, R. [Beer advocaten, Amsterdam (Netherlands)

    2010-04-15

    In 9 articles attention is paid to several aspects with regard to Legionella in cooling towers: representative sampling, the use of copper and silver ionization or hydrogen peroxide to prevent Legionella growth and biofilms, the use of a zero-tolerance model to control a cooling tower installation, detection of DNA of Legionella Pneumophila, legionella safety in air conditioners, the model Legionella risk analysis and control of cooling tower installations, legislation and regulations for the control of cooling tower installations with regard to the Dutch Occupational Health and Safety Act ('Arbo-wet'), and an article about a lawsuit for victims of a Legionella outbreak, caused by careless owners of a cooling tower in Amsterdam, Netherlands. [Dutch] In 9 artikelen wordt in deze aflevering aandacht besteed aan verschillende aspecten m.b.t. Legionella in koeltorens: representatieve monstername, de toepassing van koper en zilver-ionisatie of waterstofperoxide om de groei van Legionella en biofilms te voorkomen, het gebruik van een zero-tolerance model om een koeltoren installatie te controleren, detectie van DNA van Legionella Pneumophila, Legionella veiligheid in luchtbehandelingsinstallaties, het model Legionella risicoanalyse en beheersplan voor koeltoreninstallaties, de rol van de Arbo-wet, en een artikel over een rechtszaak voor slachtoffers van Legionella door onzorgvuldig beheer van een koeltoren in Amsterdam.

  7. Nucleotide sequence analysis of the Legionella micdadei mip gene, encoding a 30-kilodalton analog of the Legionella pneumophila Mip protein

    DEFF Research Database (Denmark)

    Bangsborg, Jette Marie; Cianciotto, N P; Hindersson, P

    1991-01-01

    After the demonstration of analogs of the Legionella pneumophila macrophage infectivity potentiator (Mip) protein in other Legionella species, the Legionella micdadei mip gene was cloned and expressed in Escherichia coli. DNA sequence analysis of the L. micdadei mip gene contained in the plasmid p...

  8. [Legionella contamination of hospital hot water supply systems in the light of research conducted in 2008-2010 as part of supervision by the Country Sanitary Inspector in Bydgoszcz].

    Science.gov (United States)

    Zborowska-Dobosz, Renata; Kuziemski, Arkadiusz; Maron, Monika; Bahn, Dorota; Owczarek, Aleksandra

    2011-01-01

    The aim of the study was to assess the level of Legionella sp. contamination in the hot water supply systems at the premises of inpatient healthcare facilities. In the years 2008-2010 the State Poviat Sanitary Inspector in Bydgoszcz tested the hot water supply systems in 8 hospitals. A total of 88 samples of hot water were collected in the years 2008-2010. The analysis involved temperature measurements and microbiological testing of the hot water. Contamination levels exceeding the applicable standards were discovered in 6 hospitals. The corrective measures introduced allowed for a significant improvement in each case. The hospital hot water systems revealed Legionella sp. contamination levels considerably exceeding the approved standards. Of the 88 water samples tested, 37 contained excessive numbers of Legionella sp. bacteria (i.e. above 100 CFU in 100 ml of water), which constituted 63.6% of the samples tested. In 6 of the 8 investigated hospitals the Legionella sp. contamination of the hot water supply system was found to be on the medium or high level. The analysis of temperature measurements revealed that the lowest temperature readings were associated with high bacterial colonization of the plumbing system. After the implementation of corrective measures, 50 control samples were collected, and in 37 of them the bacterial levels were below 100 CFU per 100 ml of water. The Legionella sp. contamination was found to be associated with low temperature of the hot water.

  9. Aerobiology of the built environment: Synergy between Legionella and fungi.

    Science.gov (United States)

    Alum, Absar; Isaacs, Galahad Zachariah

    2016-09-02

    The modern built environment (BE) design creates unique ecological niches ideal for the survival and mutual interaction of microbial communities. This investigation focused on the synergistic relations between Legionella and the fungal species commonly found in BEs and the impact of these synergistic relationships on the survival and transmission of Legionella. A field study was conducted to identify the types and concentrations of fungi in BEs. The fungal isolates purified from BEs were cocultured with Legionella to study their synergistic association. Cocultured Legionella cells were aerosolized in an air-tight chamber to evaluate the efficacy of ultraviolet (UV) to inactivate these cells. Aspergillus, Alternaria, and Cladosporium were the most common fungi detected in samples that tested positive for Legionella. After coculturing, Legionella cells were detected inside fungal hyphae. The microscopic observations of Legionella internalization in fungal hyphae were confirmed by molecular analyses. UV disinfection of the aerosolized Legionella cells that were cocultured with fungi indicated that fungal spores and propagules act as a shield against UV radiation. The shield effect of fungal spores on Legionella cells was quantified at >2.5 log10. This study provides the first evidence, to our knowledge, of Legionella cell presence inside fungi detected in an indoor environment. This symbiotic relationship with fungi results in longer survival of Legionella under ambient conditions and provides protection against UV rays. Copyright © 2016. Published by Elsevier Inc.

  10. Combined pericarditis and pneumonia caused by Legionella infection

    DEFF Research Database (Denmark)

    Svendsen, Jesper Hastrup; Jønsson, V; Niebuhr, U

    1987-01-01

    During a one year period acute pericarditis was diagnosed in 16 consecutive patients without acute infarction or malignancy. In two of these patients with both pericarditis and pneumonia Legionella infection was present. One case was caused by Legionella longbeachae and the other by both Legionel...... longbeachae and Legionella jordanis. When pericarditis is associated with pneumonia Legionella infection should be sought so that effective treatment with erythromycin may be started early.......During a one year period acute pericarditis was diagnosed in 16 consecutive patients without acute infarction or malignancy. In two of these patients with both pericarditis and pneumonia Legionella infection was present. One case was caused by Legionella longbeachae and the other by both Legionella...

  11. Survey of Legionella spp. in Mud Spring Recreation Area

    Science.gov (United States)

    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.

  12. Rapid Method for Enumeration of Viable Legionella pneumophila and Other Legionella spp. in Water

    Science.gov (United States)

    Delgado-Viscogliosi, Pilar; Simonart, Tristan; Parent, Virginie; Marchand, Grégory; Dobbelaere, Marie; Pierlot, Eric; Pierzo, Véronique; Menard-Szczebara, Florence; Gaudard-Ferveur, Elisabeth; Delabre, Karine; Delattre, Jean Marie

    2005-01-01

    A sensitive and specific method has been developed to enumerate viable L. pneumophila and other Legionella spp. in water by epifluorescence microscopy in a short period of time (a few hours). This method allows the quantification of L. pneumophila or other Legionella spp. as well as the discrimination between viable and nonviable Legionella. It simultaneously combines the specific detection of Legionella cells using antibodies and a bacterial viability marker (ChemChrome V6), the enumeration being achieved by epifluorescence microscopy. The performance of this immunological double-staining (IDS) method was investigated in 38 natural filterable water samples from different aquatic sources, and the viable Legionella counts were compared with those obtained by the standard culture method. The recovery rate of the IDS method is similar to, or higher than, that of the conventional culture method. Under our experimental conditions, the limit of detection of the IDS method was waters, the double-staining method may be an interesting alternative (not equivalent) to the conventional standard culture methods for enumerating viable Legionella when rapid detection is required. PMID:16000824

  13. Criterios microbiológicos y de muestreo establecidos en la legislación vigente para el control de Legionella Microbiological and sampling criteria established in the present Spanish legislation for the control of Legionella

    Directory of Open Access Journals (Sweden)

    Isabel Inza Rojas

    2006-12-01

    Full Text Available Se ha investigado retrospectivamente la relación existente entre las bacterias aerobias totales (BAT y la presencia de Legionella en un total de 576 muestras de agua procedentes de torres de refrigeración industriales. Los resultados revelan un menor porcentaje de muestras positivas para Legionella (19,8% por encima del nivel de 104 UFC/ml BAT establecido como límite de acción, que por debajo de este (80,2%, lo que indica que este limite no cumple su función en las instalaciones investigadas. Niveles de BAT superiores 100 y 1000 UFC/ml parecen ser más útiles para predecir la presencia de Legionella. Hemos observado una concentración de BAT significativamente más elevada en las muestras positivas para Legionella que en las negativas. Esto indica que las BAT son una buena herramienta para la validación de la eficacia del Plan de Autocontol (PA establecido en estas instalaciones. Estos resultados apoyan la importancia de seguir manteniendo el control de las BAT (a 36 ± 1 oC como un indicador de riego de presencia de Legionella y evidencian la necesidad de introducir un límite de acción más bajo para las BAT. Por otra parte se discuten algunos aspectos vinculados a la toma de muestras y a su transporte, y algunos tópicos vinculados al periodo de incubación de la enfermedad y a los métodos de tipado epidemiológico.We have retrospectively investigated the relationship between the presence of total aerobic bacteria (TAB and the presence of Legionella sp. in 576 water samples from industrial cooling towers. A lower percentage of positive samples for Legionella sp. (19.8% was encountered when the concentration of TAB was above 104 CFU/ml than below this concentration (80.2%. The concentration of 104 CFU/ml TAB is established in Spanish legislation as the action limit, above which a higher risk of Legionella positive samples is expected. Despite that, our results indicate that levels of TAB above 100 or 1000 CFU/ml seem more useful for

  14. Risk reduction for legionella; Risicobeperking legionellose

    Energy Technology Data Exchange (ETDEWEB)

    Van Olffen, E. [Tebodin Consultants and Engineers, The Hague (Netherlands)

    1999-06-01

    Simple technical measures suffice to fight infection diseases caused by legionella bacteria. Next to the fact that there is not any form of prevention present in cold water supply systems, there also appears to be a lack of information to users of such systems. In this article some of the necessary information is given. 5 refs.

  15. Comparison of Sofia Legionella FIA and BinaxNOW® Legionella urinary antigen card in two national reference centers.

    Science.gov (United States)

    Beraud, L; Gervasoni, K; Freydiere, A M; Descours, G; Ranc, A G; Vandenesch, F; Lina, G; Gaia, V; Jarraud, S

    2015-09-01

    The Sofia Legionella Fluorescence Immunoassay (FIA; Quidel) is a recently introduced rapid immunochromatographic diagnostic test for Legionnaires' disease using immunofluorescence technology designed to enhance its sensitivity. The aim of this study was to evaluate its performance for the detection of urinary antigens for Legionella pneumophila serogroup 1 in two National Reference Centers for Legionella. The sensitivity and specificity of the Sofia Legionella FIA test were determined in concentrated and nonconcentrated urine samples, before and after boiling, in comparison with the BinaxNOW® Legionella Urinary Antigen Card (UAC; Alere). Compared with BinaxNOW® Legionella UAC, the sensitivity of the Sofia Legionella test was slightly higher in nonconcentrated urine samples and was identical in concentrated urine samples. The specificity of the Sofia Legionella FIA test was highly reduced by the concentration of urine samples. In nonconcentrated samples, a lack of specificity was observed in 2.3 % of samples, all of them resolved by heat treatment. The Sofia Legionella FIA is a sensitive test for detecting Legionella urinary antigens with no previous urine concentration. However, all positive samples have to be re-tested after boiling to reach a high specificity. The reading is automatized on the Sofia analyzer, which can be connected to laboratory information systems, facilitating accurate and rapid reporting of results.

  16. In Vitro Activity of ABT-773 against Legionella pneumophila, Its Pharmacokinetics in Guinea Pigs, and Its Use to Treat Guinea Pigs with L. pneumophila Pneumonia

    OpenAIRE

    Edelstein, Paul H.; Higa, F.; Edelstein, Martha A. C.

    2001-01-01

    The activity of ABT-773 was studied against extracellular and intracellular Legionella pneumophila and for the treatment of guinea pigs with L. pneumophila pneumonia. The ABT-773 MIC at which 50% of isolates are inhibited (MIC50) for 20 different Legionella sp. strains was 0.016 μg/ml, whereas the MIC50s of clarithromycin and erythromycin were 0.032 and 0.125 μg/ml, respectively. ABT-773 (1 μg/ml) was bactericidal for two L. pneumophila strains grown in guinea pig alveolar macrophages. In con...

  17. Legionella bacteria in combustion air humidifiers; Legionella i luftuppfuktare foer foerbraenningsluft

    Energy Technology Data Exchange (ETDEWEB)

    Jeppesen, Jessica; Hansson, Helen; Cederfeldt, Ola; Axby, Fredrik

    2007-10-15

    Over the last couple of years several outbreaks of Legionnaires' disease has occurred around the world. The source of infection varies, but in many cases the bacteria has been traced back to cooling towers. The common denominator for most of the sources of infection has been water systems with high oxygen levels and a system temperature ranging from 20 to 40 deg C. If a water system furthermore has high levels of other bacteria/microorganisms, then it is also probable that favorable conditions for legionella bacteria exist within the system. These conditions consist well with conditions in combustion air humidifiers in district heating plants and therefore it can be suspected that legionella may thrive also in these types of air humidifiers. In order to examine if legionella bacteria exists in combustion air humidifiers, and in what quantity, ten different district heating plants where selected for a survey. The goal of the survey is to get a general picture of the extent of the occurrence of legionella bacteria in different types of air humidifiers. This was done by taking legionella tests in three different types of air humidifiers in district heating plants. The first step of the testing was conducted by measuring the level of legionella bacteria in water samples from the air humidifiers in the different plants. The water samples were taken from water that circulates in the air humidifiers. A few months later another water sample was taken from the same location. Two district heating plants, where legionella bacteria had been detected, were selected for further testing after compiling the results from the first two tests. The further testing involved fuel gas analysis to see if any of the plants fuel gas contained legionella bacteria. The test results show that a major part of the district heating plants in the survey have heterotrophic bacteria in their water systems, which implies the possibility of growth of legionella bacteria. About half of the plants

  18. Legionella (Legionnaires' Disease and Pontiac Fever): Causes and Transmission

    Science.gov (United States)

    ... Program Application CDC Legionella Healthy Swimming CDC Vessel Sanitation Program Unexplained Respiratory Disease Outbreaks (URDO) European Legionnaires’ Disease Surveillance Network (ELDSNet) Causes, How it Spreads, and People at ...

  19. Identification of mip-like genes in the genus Legionella

    DEFF Research Database (Denmark)

    Cianciotto, N P; Bangsborg, Jette Marie; Eisenstein, B I

    1990-01-01

    in the genus Legionella, a large panel of Legionella strains was examined by Southern hybridization and immunoblot analyses for the presence and expression of mip-related sequences. Strains representing all 14 serogroups of L. pneumophila contained a mip gene and expressed a 24-kilodalton Mip protein. Although...... with specific Mip antisera. Moreover, the cloned mip analog from Legionella micdadei encoded the cross-reactive protein. Thus, mip is conserved and specific to L. pneumophila, but mip-like genes are present throughout the genus, perhaps potentiating the intracellular infectivity of all Legionella species....

  20. A new Legionella species, Legionella feeleii species nova, causes Pontiac fever in an automobile plant.

    Science.gov (United States)

    Herwaldt, L A; Gorman, G W; McGrath, T; Toma, S; Brake, B; Hightower, A W; Jones, J; Reingold, A L; Boxer, P A; Tang, P W

    1984-03-01

    From 15 to 21 August 1981, Pontiac fever affected 317 automobile assembly plant workers. Results of serologic tests were negative for Mycoplasma, Chlamydia, respiratory tract viruses, and previously described legionellae. A gram-negative, rod-shaped organism (WO-44C) that did not grow on blood agar, required L-cysteine for growth, and contained large amounts of branched-chain fatty acids was isolated from a water-based coolant. The organism did not react with antisera against other legionellae, and on DNA hybridization the organism was less than 10% related to other Legionella species. Geometric mean titers found by indirect fluorescent antibody testing to WO-44C were significantly higher in ill employees than in controls (p = 0.0001). Attack rates by department decreased linearly with the department's distance from the implicated coolant system. The etiologic agent apparently was a new Legionella species; we propose the name Legionella feeleii species nova (AATC 35072). This is the first outbreak of nonpneumonic legionellosis in which the etiologic agent is not L. pneumophila, serogroup 1.

  1. Research on Legionella pneumophila in hospital supply networks - DOI: 10.3395/reciis.v1i2.85en

    Directory of Open Access Journals (Sweden)

    Aldo Pacheco Ferreira

    2007-12-01

    Full Text Available The species of Legionella (gram-negative aerobic bacillus are widely distributed in water environments. The Legionellaceae family comprises a group of fastidious bacteria that requires special isolation techniques, and of which the Legionella pneumophila is the member which is most commonly identified as the cause of human diseases or of the worsening of health conditions. The purpose of this work is to verify conditions in randomly chosen hospitals from the public system for the development of Legionella pneumophila, and, thus, contribute to the debate on legislation and technical standardization in the network, with the purpose of controlling and preventing the presence of this bacillus in the Brazilian hospital system. At the same time, some physicochemical and colimetric parameters (total and thermo-tolerant coliforms were evaluated aiming at defining the total quality of water used. The results show the presence of coliforms and Legionella sp in the water used in the monitored hospitals. In this way, it is shown that the maintenance of the quality of water used in hospitals is fundamental, as well as the carrying out of biological tests for the detection of Legionella pneumophila so that the necessary safety of the hospital environment can be achieved.

  2. Laboratory studies of disinfectants against Legionella pneumophila.

    OpenAIRE

    Skaliy, P; Thompson, T A; Gorman, G W; Morris, G K; McEachern, H V; Mackel, D C

    1980-01-01

    Legionella pneumophila suspended in tap water was exposed to biocides recommended for inhibiting biological growth in cooling towers and evaporative condensers of air-conditioning systems. Chlorine, 2,2-dibromo-3-nitrilopropionamide, and a compound containing didecyldimethylammonium chloride and isopropanol were effective in destroying concentratiois of 10(5) to 10(6) viable cells per ml. Formulations consisting of 5-chloro-2-methyl-4-isothiazolin-3-one and 2-methyl-4-isothiazolin-3-one, diso...

  3. UV-C inactivation of Legionella rubrilucens

    Directory of Open Access Journals (Sweden)

    Schmid, Julian

    2017-04-01

    Full Text Available Background: Despite the great health significance of , there is only little information on their UV sensitivity. Besides only has been investigated so far. Methods: In this study has been spread on buffered charcoal yeast extract agar and irradiated with the 254 nm UV-C emission of a mercury vapor lamp. The disinfection success is measured by colony counting after incubation and comparison of the number of colonies on irradiated and unirradiated reference agar plates.Results: The average log-reduction dose is 1.08 mJ/cm for free which is at the lower end of the so far published Legionella log-reduction values, but all three species show similar sensitivities. Conclusion: The log-reduction dose of legionellae in amoebae has not been investigated, but with the observed high UV-C sensitivity for free , the idea of a future point-of-use disinfection by small UV-C LEDs in water-taps or shower heads appears to be realistic, even if legionellae are more resistant in amoebae.

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

    Science.gov (United States)

    1993-08-01

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

  5. Legionella pneumophila Airway Colonisation in Patients Admitted to Hospital

    Czech Academy of Sciences Publication Activity Database

    Jarešová, M.; Petříčková, Kateřina; Korčáková, L.; Bohmová, R.; Puchmajerová, J.; Zazula, R.; Stříž, I.; Totušek, P.; Hložánek, I.

    2003-01-01

    Roč. 12, - (2003), s. 25-29 ISSN 1016-4901 R&D Projects: GA ČR GA310/01/0095 Institutional research plan: CEZ:AV0Z5020903 Keywords : legionella * Legionella pneumophila * airway Subject RIV: EE - Microbiology, Virology

  6. Identification of mip-like genes in the genus Legionella

    DEFF Research Database (Denmark)

    Cianciotto, N P; Bangsborg, Jette Marie; Eisenstein, B I

    1990-01-01

    with specific Mip antisera. Moreover, the cloned mip analog from Legionella micdadei encoded the cross-reactive protein. Thus, mip is conserved and specific to L. pneumophila, but mip-like genes are present throughout the genus, perhaps potentiating the intracellular infectivity of all Legionella species....

  7. Combined pericarditis and pneumonia caused by Legionella infection

    DEFF Research Database (Denmark)

    Svendsen, J H; Jønsson, V; Niebuhr, U

    1987-01-01

    During a one year period acute pericarditis was diagnosed in 16 consecutive patients without acute infarction or malignancy. In two of these patients with both pericarditis and pneumonia Legionella infection was present. One case was caused by Legionella longbeachae and the other by both Legionel...

  8. Viable Legionella pneumophila bacteria in natural soil and rainwater puddles.

    Science.gov (United States)

    van Heijnsbergen, E; de Roda Husman, A M; Lodder, W J; Bouwknegt, M; Docters van Leeuwen, A E; Bruin, J P; Euser, S M; den Boer, J W; Schalk, J A C

    2014-09-01

    For the majority of sporadic Legionnaires' disease cases the source of infection remains unknown. Infection may possible result from exposure to Legionella bacteria in sources that are not yet considered in outbreak investigations. Therefore, potential sources of pathogenic Legionella bacteria--natural soil and rainwater puddles on roads--were studied in 2012. Legionella bacteria were detected in 30% (6/20) of soils and 3·9% (3/77) of rainwater puddles by amoebal coculture. Legionella pneumophila was isolated from two out of six Legionella positive soil samples and two out of three Legionella positive rainwater samples. Several other species were found including the pathogenic Leg. gormanii and Leg. longbeachae. Sequence types (ST) could be assigned to two Leg. pneumophila strains isolated from soil, ST710 and ST477, and one strain isolated from rainwater, ST1064. These sequence types were previously associated with Legionnaires' disease patients. Rainwater and soil may be alternative sources for Legionella. The detection of clinically relevant strains indicates that rainwater and soil are potential sources of Legionella bacteria and future research should assess the public health implication of the presence of Leg. pneumophila in rainwater puddles and natural soil. © 2014 The Authors. Journal of Applied Microbiology published by John Wiley & Sons Ltd on behalf of the Society for Applied Microbiology.

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

    OpenAIRE

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

    2013-01-01

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

  10. Legionella and non-Legionella bacteria in a biological treatment plant.

    Science.gov (United States)

    Fykse, Else Marie; Aarskaug, Tone; Thrane, Ingjerd; Blatny, Janet Martha

    2013-02-01

    Legionella pneumophila were previously identified in the aeration ponds (up to 10(10) CFU/L) of a biological wastewater treatment plant at Borregaard Ind. Ltd., Sarpsborg, Norway, and in air samples (up to 3300 CFU/m(3)) collected above the aeration ponds. After 3 outbreaks of Legionnaires' disease reported in this area in 2005 and 2008, the aeration ponds of the plant were shut down by the Norwegian authorities in September 2008. The aim of the present work was to analyze the Legionella and non-Legionella bacterial communities in the aeration ponds before and during the shutdown process and to identify potential human pathogens. The non-Legionella bacterial community was investigated in selected samples during the shutdown process by 16S rDNA sequencing of clone libraries (400 clones) and growth analysis. The concentration of L. pneumophila and Pseudomonas spp. DNA were monitored by quantitative PCR. Results showed a decrease in the concentration of L. pneumophila and Pseudomonas spp. during the shutdown. This was accompanied by a significant change in the composition of the bacterial community in the aeration ponds. This study demonstrated that several advanced analytical methods are necessary to characterize the bacterial population in complex environments, such as the industrial aeration ponds.

  11. Legionella - A threat to groundwater: Pathogen transport in recharge basin.

    Science.gov (United States)

    McBurnett, Lauren R; Holt, Nathan T; Alum, Absar; Abbaszadegan, Morteza

    2018-04-15

    This study elucidates the potential risk posed by Legionella during aquifer recharge practices. Experiments were conducted using pilot-scale column simulating infiltration of bacterial surrogate and pathogen, E. coli and Legionella pneumophila, under central Arizona recharge basin conditions. A column was packed with a loamy sand media collected from a recharge basin and was fitted with six sampling ports at soil depths of 15, 30, 60, 92, 122cm and acclimated for a month with tertiary treated wastewater. Transport of Legionella appeared to be delayed compared to E. coli. The breakthrough of E. coli and Legionella at 122cm depth occurred at 3 and 24h, respectively. Slow transport of Legionella is consistent with its pleomorphic nature and variation in size and shape under low nutrient conditions. Legionella persisted for a longer time in the column, but at lower concentrations. Given the novel results of this study, the transport of Legionella into groundwater aquifers can occur through engineering recharge basin conditions creating a potential public health risk. Copyright © 2017. Published by Elsevier B.V.

  12. Laboratory studies of disinfectants against Legionella pneumophila.

    Science.gov (United States)

    Skaliy, P; Thompson, T A; Gorman, G W; Morris, G K; McEachern, H V; Mackel, D C

    1980-01-01

    Legionella pneumophila suspended in tap water was exposed to biocides recommended for inhibiting biological growth in cooling towers and evaporative condensers of air-conditioning systems. Chlorine, 2,2-dibromo-3-nitrilopropionamide, and a compound containing didecyldimethylammonium chloride and isopropanol were effective in destroying concentratiois of 10(5) to 10(6) viable cells per ml. Formulations consisting of 5-chloro-2-methyl-4-isothiazolin-3-one and 2-methyl-4-isothiazolin-3-one, disodium ethylene bis(thiocarbamate) and sodium dimethyl dithiocarbamate, and a phenolic with pentachlorophenate and sodium salts of other chlorophenols were less effective. PMID:6252840

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

    Science.gov (United States)

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

    2015-01-01

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

  14. Legionella risk assessment in cruise ships and ferries

    Directory of Open Access Journals (Sweden)

    Pasqualina Laganà

    2017-06-01

    Legionella pneumophila sg 1 was isolated from the samples of shower and tap water in 7 (70% of the 10 ferries examined, and in 3 (33% of the 6 cruise ships examined, and L. pneumophila sg 2–14 in 8 (80% and 1 (16.7% of these ships, respectively. No Legionella contamination was found in whirlpool baths, air and ice samples. In conclusion, the data obtained confirm higher levels of Legionella contamination in local ferries and cruise ships, underlining the need to adopt corrective actions more specific for these smaller vessels.

  15. Factors Stimulating Propagation of Legionellae in Cooling Tower Water

    OpenAIRE

    Yamamoto, Hiroyuki; Sugiura, Minoru; Kusunoki, Shinji; Ezaki, Takayuki; Ikedo, Masanari; Yabuuchi, Eiko

    1992-01-01

    Our survey of cooling tower water demonstrated that the highest density of legionellae, ≥104 CFU/100 ml, appeared in water containing protozoa, ≥102 MPN/100 ml, and heterotrophic bacteria, ≥106 CFU/100 ml, at water temperatures between 25 and 35°C. Viable counts of legionellae were detected even in the winter samples, and propagation, up to 105 CFU/100 ml, occurs in summer. The counts of legionellae correlated positively with increases in water temperature, pH, and protozoan counts, but not w...

  16. Legionella-protozoa-nematode interactions in aquatic biofilms and influence of Mip on Caenorhabditis elegans colonization.

    Science.gov (United States)

    Rasch, Janine; Krüger, Stefanie; Fontvieille, Dominique; Ünal, Can M; Michel, Rolf; Labrosse, Aurélie; Steinert, Michael

    2016-09-01

    Legionella pneumophila, the causative agent of Legionnaireś disease, is naturally found in aquatic habitats. The intracellular life cycle within protozoa pre-adapted the "accidental" human pathogen to also infect human professional phagocytes like alveolar macrophages. Previous studies employing the model organism Caenorhabditis elegans suggest that also nematodes might serve as a natural host for L. pneumophila. Here, we report for the first time from a natural co-habitation of L. pneumophila and environmental nematode species within biofilms of a warm water spring. In addition, we identified the protozoan species Oxytricha bifaria, Stylonychia mytilus, Ciliophrya sp. which have never been described as potential interaction partners of L. pneumophila before. Modeling and dissection of the Legionella-protozoa-nematode interaction revealed that C. elegans ruptures Legionella-infected amoebal cells and by this means incorporate the pathogen. Further infection studies revealed that the macrophage infectivity potentiator (Mip) protein of L. pneumophila, which is known to bind collagen IV during human lung infection, promotes the colonization of the intestinal tract of L4 larvae of C. elegans and negatively influences the life span of the worms. The Mip-negative L. pneumophila mutant exhibited a 32-fold reduced colonization rate of the nematodes after 48h when compared to the wild-type strain. Taken together, these studies suggest that nematodes may serve as natural hosts for L. pneumophila, promote their persistence and dissemination in the environment, and co-evolutionarily pre-adapt the pathogen for interactions with extracellular constituents of human lung tissue. Copyright © 2016 Elsevier GmbH. All rights reserved.

  17. Legionella risk assessment in water installations

    Directory of Open Access Journals (Sweden)

    Paulino Pastor Pérez

    2005-12-01

    Full Text Available Legionelose is an illness produced as a consequence of the inhalation of bacteria of the specie Legionella, which occasionally may colonize some of the water installations of the buildings.This article describes a Legionella risk assessment algorithm particularized for every type of installation covering all the specific characteristics of every piece of equipment, but keeping a common similar structure that allows easiness of use and homogeneous results. This algorithm has been developed by the Experts Commission of the Ministry of Health.The main use of these kind of tools is to minimize as much as possible the subjectivity of the inspectors in the identification and assessment of risks associated to the installations. These types of algorithms are applicable in risks depending on a relative low number of parameters that are somehow easy to control, as it happens in the water installations in the buildings.It has been chosen to develop a global index rated from 0 to 100 (meaning 0 minimum risk and 100 maximum risk distributed in term in three sub indexes, which define the main risk aspects to be taken into account.Structural Index: design.Maintenance Index: maintenance quality.Operational Index: performance of the equipment.Covering all the different aspects of the life of the installations.

  18. Legionella (Legionnaires' Disease and Pontiac Fever): Fast Facts

    Science.gov (United States)

    ... Program Application CDC Legionella Healthy Swimming CDC Vessel Sanitation Program Unexplained Respiratory Disease Outbreaks (URDO) European Legionnaires’ Disease Surveillance Network (ELDSNet) Fast Facts Language: English (US) Español (Spanish) ...

  19. Legionella (Legionnaires' Disease and Pontiac Fever): Signs and Symptoms

    Science.gov (United States)

    ... Program Application CDC Legionella Healthy Swimming CDC Vessel Sanitation Program Unexplained Respiratory Disease Outbreaks (URDO) European Legionnaires’ Disease Surveillance Network (ELDSNet) Signs and Symptoms Language: English Español (Spanish) ...

  20. Legionella (Legionnaires' Disease and Pontiac Fever): Treatment and Complications

    Science.gov (United States)

    ... Program for Laboratories For Media Water System Maintenance Water Management Toolkit Identify Buildings at Increased Risk Considerations When Working with Legionella Consultants Surveillance & Reporting Resources Materials Guidelines, Standards, and Laws Publications File Formats ...

  1. Survey of Legionella Species Found in Thai Soil

    Directory of Open Access Journals (Sweden)

    Tatiana C. Travis

    2012-01-01

    Full Text Available Members of the Gram-negative genus Legionella are typically found in freshwater environments, with the exception of L. longbeachae, which is present in composts and potting mixes. When contaminated aerosols are inhaled, legionellosis may result, typically as either the more serious pneumonia Legionnaires’ disease or the less severe flu-like illness Pontiac fever. It is presumed that all species of the genus Legionella are capable of causing disease in humans. As a followup to a prior clinical study of legionellosis in rural Thailand, indigenous soil samples were collected proximal to cases’ homes and workplaces and tested for the presence of legionellae by culture. We obtained 115 isolates from 22/39 soil samples and used sequence-based methods to identify 12 known species of Legionella represented by 87 isolates.

  2. Multiplication of Legionella spp. in tap water containing Hartmannella vermiformis.

    Science.gov (United States)

    Wadowsky, R M; Wilson, T M; Kapp, N J; West, A J; Kuchta, J M; States, S J; Dowling, J N; Yee, R B

    1991-07-01

    A model was developed to study the multiplication of various Legionella spp. in tap water containing Hartmannella vermiformis. Tap water cultures prepared with the following components were suitable for the multiplication studies: Legionella spp., 10(3) CFU/ml; H. vermiformis, 10(4.4) cysts per ml; and killed Pseudomonas paucimobilis, 10(9) cells per ml. Cocultures were incubated at 37 degrees C for at least 1 week. The following legionellae multiplied in tap water cocultures in each replicate experiment: L. bozemanii (WIGA strain), L. dumoffii (NY-23 and TX-KL strains), L. micdadei (two environmental strains), and L. pneumophila (six environmental strains and one clinical isolate). Growth yield values for these strains were 0.6 to 3.5 log CFU/ml. Legionellae which did not multiply in replicate cocultures included L. anisa (one strain), L. bozemanii (MI-15 strain), L. micdadei (a clinical isolate), L. longbeachae, (one strain), and L. pneumophila (Philadelphia 1 strain). L. gormanii and an environmental isolate of L. pneumophila multiplied in only one of three experiments. None of the legionellae multiplied in tap water containing only killed P. paucimobilis. The mean growth yield (+/- standard deviation) of H. vermiformis in the cocultures was 1.2 +/- 0.1 log units/ml. H. vermiformis supports multiplication of only particular strains of legionellae, some of which are from diverse origins.

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

    Energy Technology Data Exchange (ETDEWEB)

    Hilbi, H.

    2006-07-01

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

  4. Métodos analíticos para el estudio de Legionella Methods for Legionella detection

    Directory of Open Access Journals (Sweden)

    Carmen Pelaz Antolín

    2006-12-01

    Full Text Available Los ensayos para la determinación de Legionella en muestras de agua son uno de los aspectos contemplados en la legislación española sobre prevención de legionelosis. La periodicidad de estos ensayos en función del tipo de instalación, los laboratorios que los realizan, y las acciones correctoras que derivan de ellos en función de los recuentos bacterianos son acciones incluidas en los planes de mantenimiento preventivo de las instalaciones consideradas de riesgo (Real Decreto 865/2003. La comparación de nuestra legislación con otras legislaciones o recomendaciones adoptadas en otros paises (Reino Unido, Francia, Australia, América permite conocer nuestro grado de exigencia en relación a algunos de los parámetros contemplados. El cultivo de la bacteria es el método de referencia para la detección de Legionella en muestras de agua y existen varios ensayos normalizados, como los estándares ISO 11731/98 y 2004 y NF T 90- 431/2003 (AFNOR. Para ayudar a la interpretación de los resultados, los ensayos deben reflejar el estándar en el que se basan y el límite de detección del método, que no debe ser superior a 100 ufc/L. Además, los laboratorios que realizan estos ensayos deben estar acreditados por nuestra entidad de acreditación ENAC. En los últimos años se han desarrollado métodos rápidos de detección de la bacteria basados en la amplificación de ADN cromosómico en muestras de agua mediante reacciones de PCR. El desarrollo científico de estos métodos va por delante del desarrollo reglamentario, y los ensayos de PCR no deben desplazar a los ensayos de cultivo en cumplimiento de las normativas vigentes, sino que deben complementarlo.Assays for Legionella detection in water samples are one of the aspects included in the Spanish legislation on prevention of Legionnaires ́ disease. The frequency of these assays, laboratories that carry out them, and the required actions that derive from them, regarding colony counts, are

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

    Science.gov (United States)

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

    2015-01-01

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

  6. Legionella risk assessment in cruise ships and ferries.

    Science.gov (United States)

    Laganà, Pasqualina; Gambuzza, Maria Elsa; Delia, Santi

    2017-06-12

    Introduction. The increasing development of marine traffic has led to a rise in the incidence of legionellosis among travellers. It occurs in similar environments, especially closed and crowded, and aboard ships Legionella survives and multiplies easily in water pipes, spreading into the environment through air conditioning systems and water distribution points. Although in recent years in the construction of cruise ships preventive measures aimed at curbing the proliferation of Legionella (design, materials, focus on the operation and maintenance of the water system), have been taken account, little or no attention has been paid to small ships which, in many cases, are old and not well maintained. Objective. The aim of the study was to evaluate the frequency and severity of Legionella contamination in ferries and cruise ships in order to adopt more specific control measures. Materials and method. A prevalence study was carried out on 10 ferries and 6 cruise ships docking or in transit across the port of Messina (Sicily, Italy). Water and air samples collected from many critical points were tested for qualitative and quantitative identification of Legionella. Results and conclusions. Legionella pneumophila sg 1 was isolated from the samples of shower and tap water in 7 (70%) of the 10 ferries examined, and in 3 (33%) of the 6 cruise ships examined, and L. pneumophila sg 2-14 in 8 (80%) and 1 (16.7%) of these ships, respectively. No Legionella contamination was found in whirlpool baths, air and ice samples. In conclusion, the data obtained confirm higher levels of Legionella contamination in local ferries and cruise ships, underlining the need to adopt corrective actions more specific for these smaller vessels.

  7. Utilization of polymerase chain reaction and selective media cultivation to identify Legionella in Taiwan spring water samples.

    Science.gov (United States)

    Huang, Shih-Wei; Hsu, Bing-Mu; Huang, Chin-Chun; Chen, Jung-Sheng

    2011-03-01

    In this study, we investigated the distribution of Legionella in sources of spring water and wastewater throughout Taiwan. Legionella were detected with culture method and PCR method from water samples, combined with molecular methods that allow the taxonomic identification of Legionella species. Factors associated with the distribution of Legionella including the physical-chemical and microbiological water quality characteristics were also measured. Legionella were detected in 18 of the 68 samples. Additionally, 16 of the 68 samples analyzed by polymerase chain reaction (PCR) and eight of the 68 samples evaluated using the culture method were found to be positive for Legionella. The Legionella species identified included Legionella pneumophila, unnamed Legionella genotypes, uncultured Legionella spp., and Legionella lytica. Legionella species were found in water with temperatures ranging from 10.0 to 50.6°C and with pH values ranging from 5.59 to 9.32. Significant differences (Mann-Whitney U test, P<0.05) were observed in the presence/absence of Legionella in samples with different pH values when Legionella were detected by the culture method. The results of this survey confirmed that Legionella is ubiquitous in the water of spring recreational areas in Taiwan. Long-term investigations of the occurrence of Legionella in hot spring recreational areas throughout Taiwan are needed.

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

    Science.gov (United States)

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

    2013-01-01

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

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

    Directory of Open Access Journals (Sweden)

    Lisa Conza

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

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

    Science.gov (United States)

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

    2013-01-01

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

  11. Biofilms: The Stronghold of Legionella pneumophila

    Directory of Open Access Journals (Sweden)

    Mena Abdel-Nour

    2013-10-01

    Full Text Available Legionellosis is mostly caused by Legionella pneumophila and is defined as a severe respiratory illness with a case fatality rate ranging from 5% to 80%. L. pneumophila is ubiquitous in natural and anthropogenic water systems. L. pneumophila is transmitted by inhalation of contaminated aerosols produced by a variety of devices. While L. pneumophila replicates within environmental protozoa, colonization and persistence in its natural environment are also mediated by biofilm formation and colonization within multispecies microbial communities. There is now evidence that some legionellosis outbreaks are correlated with the presence of biofilms. Thus, preventing biofilm formation appears as one of the strategies to reduce water system contamination. However, we lack information about the chemical and biophysical conditions, as well as the molecular mechanisms that allow the production of biofilms by L. pneumophila. Here, we discuss the molecular basis of biofilm formation by L. pneumophila and the roles of other microbial species in L. pneumophila biofilm colonization. In addition, we discuss the protective roles of biofilms against current L. pneumophila sanitation strategies along with the initial data available on the regulation of L. pneumophila biofilm formation.

  12. Severe Neonatal Legionella Pneumonia: Full Recovery After Extracorporeal Life Support.

    Science.gov (United States)

    Moscatelli, Andrea; Buratti, Silvia; Castagnola, Elio; Mesini, Alessio; Tuo, Pietro

    2015-10-01

    Legionella pneumophila is responsible for hospital or community-acquired pneumonia. Neonatal legionellosis is associated with rapidly severe clinical course and high mortality rates. We describe a case of hospital-acquired Legionella pneumonia in a newborn with undiagnosed tracheoesophageal fistula and acute respiratory failure requiring venovenous extracorporeal membrane oxygenation support before fistula repair. Standardized multiplex polymerase chain reaction assay allowed early diagnosis. Extracorporeal life support associated with appropriate antibiotic therapy, surfactant, and steroid therapy was effective in achieving complete recovery. This is the first report of successful neonatal extracorporeal life support for respiratory failure secondary to L pneumophila. Copyright © 2015 by the American Academy of Pediatrics.

  13. 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.

  14. Presence and Persistence of Viable, Clinically Relevant Legionella pneumophila Bacteria in Garden Soil in the Netherlands.

    Science.gov (United States)

    van Heijnsbergen, E; van Deursen, A; Bouwknegt, M; Bruin, J P; de Roda Husman, A M; Schalk, J A C

    2016-09-01

    Garden soils were investigated as reservoirs and potential sources of pathogenic Legionella bacteria. Legionella bacteria were detected in 22 of 177 garden soil samples (12%) by amoebal coculture. Of these 22 Legionella-positive soil samples, seven contained Legionella pneumophila Several other species were found, including the pathogenic Legionella longbeachae (4 gardens) and Legionella sainthelensi (9 gardens). The L. pneumophila isolates comprised 15 different sequence types (STs), and eight of these STs were previously isolated from patients according to the European Working Group for Legionella Infections (EWGLI) database. Six gardens that were found to be positive for L. pneumophila were resampled after several months, and in three gardens, L. pneumophila was again isolated. One of these gardens was resampled four times throughout the year and was found to be positive for L. pneumophila on all occasions. Tracking the source of infection for sporadic cases of Legionnaires' disease (LD) has proven to be hard. L. pneumophila ST47, the sequence type that is most frequently isolated from LD patients in the Netherlands, is rarely found in potential environmental sources. As L. pneumophila ST47 was previously isolated from a garden soil sample during an outbreak investigation, garden soils were investigated as reservoirs and potential sources of pathogenic Legionella bacteria. The detection of viable, clinically relevant Legionella strains indicates that garden soil is a potential source of Legionella bacteria, and future research should assess the public health implication of the presence of L. pneumophila in garden soil. Copyright © 2016 van Heijnsbergen et al.

  15. Acanthamoeba and Dictyostelium as Cellular Models for Legionella Infection

    Directory of Open Access Journals (Sweden)

    A. Leoni Swart

    2018-03-01

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

  16. Susceptibility of Legionella pneumophila to chlorine in tap water.

    Science.gov (United States)

    Kuchta, J M; States, S J; McNamara, A M; Wadowsky, R M; Yee, R B

    1983-11-01

    A study was conducted to compare the susceptibility of legionellae and coliforms to disinfection by chlorine. The chlorine residuals used were similar to concentrations that might be found in the distribution systems of large public potable water supplies. The effects of various chlorine concentrations, temperatures, and pH levels were considered. A number of different Legionella strains, both environmental and clinical, were tested. The results indicate that legionellae are much more resistant to chlorine than are coliform bacteria. At 21 degrees C, pH 7.6, and 0.1 mg of free chlorine residual per liter, a 99% kill of L. pneumophila was achieved within 40 min, compared with less than 1 min for Escherichia coli. The observed resistance is enhanced as conditions for disinfection become less optimal. The required contact time for the removal of L. pneumophilia was twice as long at 4 degrees C than it was at 21 degrees C. These data suggest that legionellae can survive low levels of chlorine for relatively long periods of time.

  17. Characterization of a major 31-kilodalton peptidoglycan-bound protein of Legionella pneumophila

    International Nuclear Information System (INIS)

    Butler, C.A.; Hoffman, P.S.

    1990-01-01

    A 31-kilodalton (kDa) protein was solubilized from the peptidoglycan (PG) fraction of Legionella pneumophila after treatment with either N-acetylmuramidase from the fungus Chalaropsis sp. or with mutanolysin from Streptomyces globisporus. The protein exhibited a ladderlike banding pattern by autoradiography when radiolabeled [(35S]cysteine or [35S]methionine) PG material was extensively treated with hen lysozyme. The banding patterns ranging between 31 and 45 kDa and between 55 and 60 kDa resolved as a single 31-kDa protein when the material was subsequently treated with N-acetylmuramidase. Analysis of the purified 31-kDa protein for diaminopimelic acid by gas chromatography revealed 1 mol of diaminopimelic acid per mol of protein. When outer membrane PG material containing the major outer membrane porin protein was treated with N-acetylmuramidase or mutanolysin, both the 28.5-kDa major outer membrane protein and the 31-kDa protein were solubilized from the PG material under reducing conditions. In the absence of 2-mercaptoethanol, a high-molecular-mass complex (100 kDa) was resolved. The results of this study indicate that a 31-kDa PG-bound protein is a major component of the cell wall of L. pneumophila whose function may be to anchor the major outer membrane protein to PG. Finally, a survey of other Legionella species and other serogroups of L. pneumophila suggested that PG-bound proteins may be a common feature of this genus

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

    Science.gov (United States)

    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

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

    Science.gov (United States)

    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).

  20. Spatial distribution of Legionella pneumophila MLVA-genotypes in a drinking water system.

    Science.gov (United States)

    Rodríguez-Martínez, Sarah; Sharaby, Yehonatan; Pecellín, Marina; Brettar, Ingrid; Höfle, Manfred; Halpern, Malka

    2015-06-15

    Bacteria of the genus Legionella cause water-based infections, resulting in severe pneumonia. To improve our knowledge about Legionella spp. ecology, its prevalence and its relationships with environmental factors were studied. Seasonal samples were taken from both water and biofilm at seven sampling points of a small drinking water distribution system in Israel. Representative isolates were obtained from each sample and identified to the species level. Legionella pneumophila was further determined to the serotype and genotype level. High resolution genotyping of L. pneumophila isolates was achieved by Multiple-Locus Variable number of tandem repeat Analysis (MLVA). Within the studied water system, Legionella plate counts were higher in summer and highly variable even between adjacent sampling points. Legionella was present in six out of the seven selected sampling points, with counts ranging from 1.0 × 10(1) to 5.8 × 10(3) cfu/l. Water counts were significantly higher in points where Legionella was present in biofilms. The main fraction of the isolated Legionella was L. pneumophila serogroup 1. Serogroup 3 and Legionella sainthelensis were also isolated. Legionella counts were positively correlated with heterotrophic plate counts at 37 °C and negatively correlated with chlorine. Five MLVA-genotypes of L. pneumophila were identified at different buildings of the sampled area. The presence of a specific genotype, "MLVA-genotype 4", consistently co-occurred with high Legionella counts and seemed to "trigger" high Legionella counts in cold water. Our hypothesis is that both the presence of L. pneumophila in biofilm and the presence of specific genotypes, may indicate and/or even lead to high Legionella concentration in water. This observation deserves further studies in a broad range of drinking water systems to assess its potential for general use in drinking water monitoring and management. Copyright © 2015 Elsevier Ltd. All rights reserved.

  1. Molecular Evolution of the dotA Gene in Legionella pneumophila

    OpenAIRE

    Ko, Kwan Soo; Hong, Seong Karp; Lee, Hae Kyung; Park, Mi-Yeoun; Kook, Yoon-Hoh

    2003-01-01

    The molecular evolution of dotA, which is related to the virulence of Legionella pneumophila, was investigated by comparing the sequences of 15 reference strains (serogroups 1 to 15). It was found that dotA has a complex mosaic structure. The whole dotA gene of Legionella pneumophila subsp. pneumophila serogroups 2, 6, and 12 has been transferred from Legionella pneumophila subsp. fraseri. A discrepancy was found between the trees inferred from the nucleotide and deduced amino acid sequences ...

  2. Lymphoid cell blastogenesis as an in vitro indicator of cellular immunity to Legionella pneumophila antigens.

    OpenAIRE

    Friedman, F; Widen, R; Klein, T; Friedman, H

    1984-01-01

    The lymphocyte blastogenic transformation assay was adapted to study responsiveness of lymphoid cells from animals and humans to Legionella pneumophila antigens in vitro. Spleen cells from guinea pigs after active immunization with Legionella vaccine, but not from normal animals, responded by blast cell transformation when stimulated in vitro with killed Legionella whole-cell vaccine, sonic extracts thereof, or a purified somatic antigen. The response was dose dependent. Similar lymphocyte bl...

  3. Recirculation, stagnation and ventilation: The 2014 legionella episode

    Science.gov (United States)

    Russo, Ana; Soares, Pedro M. M.; Gouveia, Célia M.; Cardoso, Rita M.; Trigo, Ricardo M.

    2017-04-01

    Legionella transmission through the atmosphere is unusual, but not unprecedented. A scientific paper published in 2006 reports a surge in Pas-de-Calais, France, in which 86 people have been infected by bacteria released by a cooling tower more than 6 km away [3]. Similarly, in Norway, in 2005, there was another case where contamination spread beyond 10 km, although more concentrated within a radius of 1 km from an industrial unit [2]. An unprecedented large Legionella outbreak occurred in November 2014 nearby Lisbon, Portugal. As of 7 November 2014, 375 individuals become hill and 12 died infected by the Legionella pneumophila bacteria, contracted by inhalation of steam droplets of contaminated water (aerosols). These droplets are so small that can carry the bacteria directly to the lungs, depositing it in the alveoli. One way of studying the propagation of legionella episodes is through the use of aerosol dispersion models. However, such approaches often require detailed 3D high resolution wind data over the region, which isn't often available for long periods. The likely impact of wind on legionella transmission can also be understood based on the analysis of special types of flow conditions such as stagnation, recirculation and ventilation [1, 4]. The Allwine and Whiteman (AW) approach constitutes a straightforward method to assess the assimilative and dispersal capacities of different airsheds [1,4], as it only requires hourly wind components. Thus, it has the advantage of not needing surface and upper air meteorological observations and a previous knowledge of the atmospheric transport and dispersion conditions. The objective of this study is to analyze if the legionella outbreak event which took place in November 2014 had extreme potential recirculation and/or stagnation characteristics. In order to accomplish the proposed objective, the AW approach was applied for a hindcast time-series covering the affected area (1989-2007) and then for an independent

  4. Virulence properties of the Legionella pneumophila cell envelope

    Directory of Open Access Journals (Sweden)

    Olga eShevchuk

    2011-04-01

    Full Text Available The bacterial envelope plays a crucial role in the pathogenesis of infectious diseases. In this review, we summarize the current knowledge of the structure and molecular composition of the Legionella pneumophila cell envelope. We describe LPS biosynthesis and the biological activities of membrane and periplasmic proteins and discuss their decisive functions during the pathogen-host interaction. In addition to adherence, invasion and intracellular survival of L. pneumophila, special emphasis is laid on iron acquisition, detoxification, key elicitors of the immune response and the diverse functions of outer membrane vesicles. The critical analysis of the literature reveals that the dynamics and phenotypic plasticity of the Legionella cell surface during the different metabolic stages requires more attention in the future.

  5. [Legionella pneumonia after infliximab in a patient with rheumatoid arthritis].

    Science.gov (United States)

    Giassi, Karina de Souza; Furlanetto, Vilson; Fialho, Sonia; Gomes Ribeiro, Giovana; Pereira, Ivânio Alves

    2014-01-01

    The antagonists of tumour necrosis factor (anti-TNF) have been successfully used in several chronic inflammatory diseases such as Rheumatoid Arthritis (RA), but some studies have observed the development of infections by intracellular pathogens in patients using anti-TNF. We report a case of a female patient with previous diagnosis of RA for 16 years that used several disease-modifying anti-rheumatic drugs (DMARDs) that resulted in treatment failure, and then was treated with infliximab. After fifteen days of the second dose, the patient developed ventilatory-dependent chest pain, dry cough and dyspnea. She was hospitalized, and the diagnosis of pneumonia by Legionella pneumophila was confirmed by the presence of Legionella antigen in an urine test. TNF is an inflammatory cytokine that also acts inhibiting the bacterial growth of intracellular pathogens, and its inhibition seems to increase susceptibility to these infections in some patients. Copyright © 2014 Elsevier Editora Ltda. All rights reserved.

  6. Interaction of legionella pneumophila and helicobacter pylori with bacterial species isolated from drinking water biofilms

    Directory of Open Access Journals (Sweden)

    Azevedo Nuno F

    2011-03-01

    Full Text Available Abstract Background It is well established that Legionella pneumophila is a waterborne pathogen; by contrast, the mode of Helicobacter pylori transmission remains unknown but water seems to play an important role. This work aims to study the influence of five microorganisms isolated from drinking water biofilms on the survival and integration of both of these pathogens into biofilms. Results Firstly, both pathogens were studied for auto- and co-aggregation with the species isolated from drinking water; subsequently the formation of mono and dual-species biofilms by L. pneumophila or H. pylori with the same microorganisms was investigated. Neither auto- nor co-aggregation was observed between the microorganisms tested. For biofilm studies, sessile cells were quantified in terms of total cells by SYTO 9 staining, viable L. pneumophila or H. pylori cells were quantified using 16 S rRNA-specific peptide nucleic acid (PNA probes and cultivable cells by standard culture techniques. Acidovorax sp. and Sphingomonas sp. appeared to have an antagonistic effect on L. pneumophila cultivability but not on the viability (as assessed by rRNA content using the PNA probe, possibly leading to the formation of viable but noncultivable (VBNC cells, whereas Mycobacterium chelonae increased the cultivability of this pathogen. The results obtained for H. pylori showed that M. chelonae and Sphingomonas sp. help this pathogen to maintain cultivability for at least 24 hours. Conclusions It appears that M. chelonae may have an important role in the survival of both pathogens in drinking water. This work also suggests that the presence of some microorganisms can decrease the cultivability of L. pneumophila but not the viability which indicates that the presence of autochthonous microorganisms can lead to misleading results when the safety of water is assessed by cultivable methods alone.

  7. Atypical Pneumonia: Updates on Legionella, Chlamydophila, and Mycoplasma Pneumonia.

    Science.gov (United States)

    Sharma, Lokesh; Losier, Ashley; Tolbert, Thomas; Dela Cruz, Charles S; Marion, Chad R

    2017-03-01

    Community-acquired pneumonia (CAP) has multiple causes and is associated with illness that requires admission to the hospital and mortality. The causes of atypical CAP include Legionella species, Chlamydophila, and Mycoplasma. Atypical CAP remains a diagnostic challenge and, therefore, likely is undertreated. This article reviews the advancements in the evaluation and treatment of patients and discusses current conflicts and controversies of atypical CAP. Copyright © 2016 Elsevier Inc. All rights reserved.

  8. Contamination of water reservoirs to Legionella in khorramabad hospitals

    Directory of Open Access Journals (Sweden)

    seyed hamed Mirhossaini

    2009-04-01

    Full Text Available Background: Legionella is a negative aquatic bacterium and one of the most common nosocomial pathogen. Hospital environment in case of growth, aerosol transmission system and endangered individuals are the high potential location for growth and prevalence of this agent. The suitable temperature in water reservoirs and water distribution system empowered the growth of this bactria. The purpose of this investigation is the study of legionella presence in khorramabad water distribution system. Materials and Methods: Sampling performed with fifteen-day periods of each cold and hot hospital water reservoirs and also cold and hot water taps in those hospital wards which have more pathogens. Each of samples concentrate high vulnerable membrane and from each sample 2 plates were cultured with BCYE and GVPC optional culture media and the growth of bacteria in third and seventh and tenth days were controlled and registered. Results: From 240 samples of five Khorramabad hospitals 41.7 percent of the samples were positive. The percent of positive samples of Ashayer, Tamin ejtemaee, Tohid, and Asalian were respectively 68.8, 45.5, 33.3, 9.1 and 36.4 percent and the residual mean chlorine of samples were respectively 0.38, 0.52, 0.46, 0.82 and 0.62mg/l. The most positive samples related to hot shower and the lowest value related to cold water taps. Conclusion: In spite of the fact that all hospitals used treated water, but from 240 collected samples, 100 samples in different sections of hospital were positive these results show direct relation between residual chlorine value and presence of legionella, by the manner that in 0.6 mg/l and higher values of residual chlorine none of samples were positive. So usually the residual chlorine value in water distribution system is not enough to legionella against.

  9. Sequence analysis of the Legionella micdadei groELS operon

    DEFF Research Database (Denmark)

    Hindersson, P; Høiby, N; Bangsborg, Jette Marie

    1991-01-01

    A 2.7 kb DNA fragment encoding the 60 kDa common antigen (CA) and a 13 kDa protein of Legionella micdadei was sequenced. Two open reading frames of 57,677 and 10,456 Da were identified, corresponding to the heat shock proteins GroEL and GroES, respectively. Typical -35, -10, and Shine-Dalgarno heat...

  10. Isolation of Legionella pneumophila from hospital cooling towers.

    Science.gov (United States)

    Nadarajah, M; Goh, K T

    1986-01-01

    Water from twenty cooling towers from three major hospitals (A, B, C) were screened for the presence of Legionella pneumophila. Three of the water specimens were found to be positive for the organism. Serogroup 1 was isolated from the two cooling towers in Hospital A, while serogroup 4 was isolated from one of the cooling towers in Hospital B, this being the first time L. pneumophila has been isolated in Singapore.

  11. Ecology of Legionella pneumophila within water distribution systems.

    OpenAIRE

    Stout, J E; Yu, V L; Best, M G

    1985-01-01

    The reservoir for hospital-acquired Legionnaires disease has been shown to be the potable water distribution system. We investigated the influence of the natural microbial population and sediment (scale and organic particulates) found in water systems as growth-promoting factors for Legionella pneumophila. Our in vitro experiments showed that: (i) water from hot-water storage tank readily supported the survival of L. pneumophila, (ii) the concentration of sediment was directly related to the ...

  12. Multiplication of Legionella pneumophila in unsterilized tap water.

    OpenAIRE

    Yee, R B; Wadowsky, R M

    1982-01-01

    Naturally occurring Legionella pneumophila, an environmental isolate which had not been grown on artificial medium, was tested for the ability to multiply in tap water. A showerhead containing L. pneumophila and non-Legionellaceae bacteria was immersed in nonsterile tap water supplying this fixture. Also L. pneumophila and non-Legionellaceae bacteria were sedimented from tap water from a surgical intensive care unit. This bacterial suspension was inoculated into tap water from our laboratory....

  13. α-Hydroxyketone Synthesis and Sensing by Legionella and Vibrio

    Directory of Open Access Journals (Sweden)

    Hubert Hilbi

    2012-03-01

    Full Text Available Bacteria synthesize and sense low molecular weight signaling molecules, termed autoinducers, to measure their population density and community complexity. One class of autoinducers, the α-hydroxyketones (AHKs, is produced and detected by the water-borne opportunistic pathogens Legionella pneumophila and Vibrio cholerae, which cause Legionnaires’ disease and cholera, respectively. The “Legionella quorum sensing” (lqs or “cholera quorum sensing” (cqs genes encode enzymes that produce and sense the AHK molecules “Legionella autoinducer-1” (LAI-1; 3-hydroxypentadecane-4-one or cholera autoinducer-1 (CAI-1; 3-hydroxytridecane-4-one. AHK signaling regulates the virulence of L. pneumophila and V. cholerae, pathogen-host cell interactions, formation of biofilms or extracellular filaments, expression of a genomic “fitness island” and competence. Here, we outline the processes, wherein AHK signaling plays a role, and review recent insights into the function of proteins encoded by the lqs and cqs gene clusters. To this end, we will focus on the autoinducer synthases catalysing the biosynthesis of AHKs, on the cognate trans-membrane sensor kinases detecting the signals, and on components of the down-stream phosphorelay cascade that promote the transmission and integration of signaling events regulating gene expression.

  14. Effect of bacterial interference on biofilm development by Legionella pneumophila.

    Science.gov (United States)

    Guerrieri, Elisa; Bondi, Moreno; Sabia, Carla; de Niederhäusern, Simona; Borella, Paola; Messi, Patrizia

    2008-12-01

    In the ecology of Legionella pneumophila a crucial role may be played by its relationship with the natural flora; thus we investigated the interactions between Legionella and other aquatic bacteria, particularly within biofilms. Among 80 aquatic bacteria screened for the production of bacteriocin-like substances (BLSs), 66.2% of them were active against L. pneumophila. The possible effect of some of these aquatic bacteria on the development and stability of L. pneumophila biofilms was studied. Pseudomonas fluorescens, the best BLS producer, showed the greatest negative effect on biofilm formation and strongly enhanced the detachment of Legionella. Pseudomonas aeruginosa, Burkholderia cepacia, Pseudomonas putida, Aeromonas hydrophila, and Stenotrophomonas maltophilia, although producing BLSs at different levels, were less active in the biofilm experiments. Acinetobacter lwoffii did not produce any antagonistic compound and was the only one able to strongly enhance L. pneumophila biofilm. Our results highlight that BLS production may contribute to determining the fate of L. pneumophila within ecological niches. The interactions observed in this study are important features of L. pneumophila ecology, which knowledge may lead to more effective measures to control the persistence of the germ in the environment.

  15. Ciliate Paramecium is a natural reservoir of Legionella pneumophila

    Science.gov (United States)

    Watanabe, Kenta; Nakao, Ryo; Fujishima, Masahiro; Tachibana, Masato; Shimizu, Takashi; Watarai, Masahisa

    2016-04-01

    Legionella pneumophila, the causative agent of Legionnaires’ disease, replicates within alveolar macrophages and free-living amoebae. However, the lifestyle of L. pneumophila in the environment remains largely unknown. Here we established a novel natural host model of L. pneumophila endosymbiosis using the ciliate Paramecium caudatum. We also identified Legionella endosymbiosis-modulating factor A (LefA), which contributes to the change in life stage from endosymbiosis to host lysis, enabling escape to the environment. We isolated L. pneumophila strains from the environment, and they exhibited cytotoxicity toward P. caudatum and induced host lysis. Acidification of the Legionella-containing vacuole (LCV) was inhibited, and enlarged LCVs including numerous bacteria were observed in P. caudatum infected with L. pneumophila. An isogenic L. pneumophila lefA mutant exhibited decreased cytotoxicity toward P. caudatum and impaired the modification of LCVs, resulting in the establishment of endosymbiosis between them. Our results suggest that L. pneumophila may have a mechanism to switch their endosymbiosis in protistan hosts in the environment.

  16. Multiplication of Legionella pneumophila in unsterilized tap water.

    Science.gov (United States)

    Yee, R B; Wadowsky, R M

    1982-06-01

    Naturally occurring Legionella pneumophila, an environmental isolate which had not been grown on artificial medium, was tested for the ability to multiply in tap water. A showerhead containing L. pneumophila and non-Legionellaceae bacteria was immersed in nonsterile tap water supplying this fixture. Also L. pneumophila and non-Legionellaceae bacteria were sedimented from tap water from a surgical intensive care unit. This bacterial suspension was inoculated into tap water from our laboratory. The legionellae in both suspensions multiplied in the tap water at 32, 37, and 42 degrees C. The non-Legionellaceae bacteria multiplied at 25, 32, and 37 degrees C. A water sample which was collected from the bottom of a hot water tank was found to contain L. pneumophila and non-Legionellaceae bacteria. These legionellae also multiplied when the water sample was incubated at 37 degrees C. These results indicate that L. pneumophila may multiply in warm water environments such as hot water plumbing fixtures, hot water tanks, and cooling towers.

  17. Water ecology of Legionella and protozoan: environmental and public health perspectives.

    Science.gov (United States)

    Borella, Paola; Guerrieri, Elisa; Marchesi, Isabella; Bondi, Moreno; Messi, Patrizia

    2005-01-01

    Ecological studies on Legionella spp. are essential to better understand their sources in the natural environments, the mechanism of their entry into man-made water systems and the factors enabling their survival and growth in aquatic habitats. Legionella spp. exhibits peculiar and multiple strategies to adapt to stressful environment conditions which normally impair other germ survival. These strategies include the ability to enter in a viable but non-cultivable (VBNC) state, to multiply intracellularly within a variety of protozoa, such as amoebae, to survive as free organisms within biofilms and to be enhanced/inhibited by the presence of other aquatic bacteria. The host-parasite interaction has been shown to be central in the pathogenesis and ecology of L. pneumophila. The bacterial-protozoan interaction contributes to the amplification of Legionella population in water systems, represents a shelter against unfavourable environmental conditions, acts as a reservoir of infection and contributes to virulence by priming the pathogen to infect human cells. Legionella is able to survive as free organism for long periods within biofilms which are widespread in man-made water systems. Biofilm provides shelter and nutrients, exhibits a remarkable resistance to biocide compounds and chlorination, thus representing ecological niches for legionella persistence in such environments. Further knowledge on biofilm-associated legionellae may lead to effective control measures to prevent legionellosis. Lastly, new perspectives in controlling legionella contamination can arise from investigations on aquatic bacteria able to inhibit legionella growth in natural and artificial water systems.

  18. Isolation, identification, characterization and antibiotic sensitivity profile of pathogenic Legionella pneumophila isolates from different water sources

    Directory of Open Access Journals (Sweden)

    Kannan Subbaram

    2017-05-01

    Conclusions: Serious and fatal L. pneumophila infections may be transmitted through water. Legionella can survive under various conditions in various water sources. L. pneumophila is the important pathogen causing human disease. Great challenge prevails to health care professionals because these Legionellae acquired antibiotic resistance to many routinely prescribed antibiotics.

  19. Detection of Legionella spp. from Domestic Water in the Prefecture of Arta, Greece

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    Dimitra Dimitriadi

    2014-01-01

    Full Text Available The aim of this research was the isolation of Legionella spp. from domestic water supply networks in the Prefecture of Arta. A total of 100 water samples, from 25 houses, were collected. Half of the samples concerned the cold water and half the hot water supply. Purpose was to detect colonization of the water networks with Legionella spp. >500 cfu/L by using the method of filtration (ISO 11731. Out of 100 samples, 6 samples from 3 houses were positive for Legionella spp. Legionella pneumophila serogroup 2–14 was isolated in 5 of 6 samples, whereas in the sixth sample Legionella anisa was identified. Only three of the samples had residual chloride over 0.2 mg/L, rate which is necessary for potable water, according to the Greek hygienic practice. Concerning the temperature of hot water, the mean temperature of the negative for Legionella samples was higher compared to the mean temperature of the positive for Legionella samples (49.9°C versus 45.5°C. It is estimated that there is risk of infection through the use of showers. The low concentration of chloride and the temperature, which was found within the limits favorable to developing Legionella spp. (20–45°C, provide fertile ground for proliferation of the bacteria.

  20. Atypical presentation of Legionella pneumonia among patients with underlying cancer: A fifteen-year review.

    Science.gov (United States)

    del Castillo, Maria; Lucca, Anabella; Plodkowski, Andrew; Huang, Yao-Ting; Kaplan, Janice; Gilhuley, Kathleen; Babady, N Esther; Seo, Susan K; Kamboj, Mini

    2016-01-01

    Immunocompromised patients, especially those receiving treatment with corticosteroids and cytotoxic chemotherapy are at increased risk for developing Legionella pneumonia. The aim of this study was to determine clinical and radiographic characteristics of pulmonary infection due to Legionella in persons undergoing treatment for cancer and stem cell transplant (SCT) recipients. Retrospective review of Legionella cases at MSKCC over a fifteen-year study period from January 1999 and December 2013. Cases were identified by review of microbiology records. During the study period, 40 cases of Legionella infection were identified; nine among these were due to non-pneumophila species. Most cases occurred during the summer. The majority [8/9, (89%)] of patients with non-pneumophila infection had underlying hematologic malignancy, compared to 18/31 (58%) with Legionella pneumophila infections. Radiographic findings were varied-nodular infiltrates mimicking invasive fungal infection were seen only among patients with hematologic malignancy and hematopoietic stem cell transplant (SCT) recipients and were frequently associated with non-pneumophila infections (50% vs 16%; P = 0.0594). All cases of nodular Legionella pneumonia were found incidentally or had an indolent clinical course. Legionella should be considered in the differential diagnosis of nodular lung lesions in immunocompromised patients, especially those with hematologic malignancy and SCT recipients. Most cases of nodular disease due to Legionella are associated with non-pneumophila infections. Copyright © 2015 The British Infection Association. Published by Elsevier Ltd. All rights reserved.

  1. Cross-reactive Legionella antigens and the antibody response during infection

    DEFF Research Database (Denmark)

    Bangsborg, Jette Marie; Shand, G; Pearlman, E

    1991-01-01

    In order to define cross-reactive Legionella antigens suitable for diagnostic purposes, we investigated sonicate antigens from two Legionella species, including two serogroups of L. pneumophila. The antigens were reacted with heterologous and homologous rabbit antisera in Western blot. Sera from...

  2. The presence and growth of Legionella species in thermostatic shower mixer taps: an exploratory field study

    NARCIS (Netherlands)

    Joost van Hoof; P.W.J.J. van der Wielen; E. van der Blom; O.W.W. Nuijten; L. Hornstra

    2014-01-01

    Legislation in the Netherlands requires routine analysis of drinking water samples for cultivable Legionella species from high-priority installations. A field study was conducted to investigate the presence of Legionella species in thermostatic shower mixer taps. Water samples and the interior of

  3. Detection and Quantification of Legionella pneumophila from Water Systems in Kuwait Residential Facilities

    Directory of Open Access Journals (Sweden)

    Qadreyah A. Al-Matawah

    2012-01-01

    Full Text Available The prevalence of Legionella pneumophilia in water systems of residential facilities in Kuwait was performed during the period from November 2007 to November 2011. A total of 204 water samples collected from faucets and showerheads in bathrooms (n = 82, taps in kitchens (n = 51, and water tanks (n = 71, from different locations of residential facilities in Kuwait were screened for Legionella pneumophila by the standard culture method and by real time polymerase chain reaction (RT-PCR. Out of the 204 samples, 89 (43.6% samples were positive for Legionella spp., 48 (23.5% samples were detected by the standard culture method, and 85 (41.7% were detected by RT-PCR. Of the culture positive Legionella samples, counts ranged between 10 to 2250 CFU/L. Serological typing of 48 Legionella isolates revealed that 6 (12.5% of these isolates belonged to Legionella pneumophila serogroup 1, 37 (77.1% isolates to Legionella pneumophila serogroup 3, and 1 isolate each (2.1% belonged to serogroups 4, 7, and 10. The minimum inhibitory concentration (MICs of the 46 environmental L. pneumophila isolates against the 10 antimicrobials commonly used for Legionella infection treatments were determined. Rifampicin was found to be the most active against L. pneumophila serogroups isolates in vitro.

  4. The role of biofilms and protozoa in Legionella pathogenesis: implications for drinking water

    Science.gov (United States)

    Current models to study Legionella pathogenesis include the use of primary macrophages and monocyte cell lines, various free-living protozoan species and murine models of pneumonia. However, there are very few studies of Legionella spp. pathogenesis aimed at associating the role ...

  5. Satellite growth of Legionella pneumophila with an environmental isolate of Flavobacterium breve.

    Science.gov (United States)

    Wadowsky, R M; Yee, R B

    1983-12-01

    Legionella pneumophila serogroup 1 was observed to satellite around colonies of Flavobacterium breve on an L-cysteine-deficient medium which did not support growth of legionellae. Both isolates were recovered from the hot water tanks of hospitals. Ferric PPi stimulated satellite growth between 0.01 and 0.1%.

  6. Satellite growth of Legionella pneumophila with an environmental isolate of Flavobacterium breve.

    OpenAIRE

    Wadowsky, R M; Yee, R B

    1983-01-01

    Legionella pneumophila serogroup 1 was observed to satellite around colonies of Flavobacterium breve on an L-cysteine-deficient medium which did not support growth of legionellae. Both isolates were recovered from the hot water tanks of hospitals. Ferric PPi stimulated satellite growth between 0.01 and 0.1%.

  7. The E. coli immunosorbent as used in serodiagnosis of Legionella infections studied by crossed immunoelectrophoresis

    DEFF Research Database (Denmark)

    Bangsborg, Jette Marie; Friis-Møller, A; Rechnitzer, C

    1988-01-01

    In this study we investigated an immunosorbent, E. coli blocking fluid (BF), proposed for use in the Legionella Indirect Fluorescent Antibody Test (IFA). With crossed immunoelectrophoresis (CIE) of clinically relevant Legionella species, only one heat-stable antigen (no. 1) cross...

  8. Risk of infection from Legionella associated with spray irrigation of reclaimed water.

    Science.gov (United States)

    Pepper, Ian L; Gerba, Charles P

    2018-04-02

    Legionella pneumophila has been detected in reclaimed water used for spray irrigation of turfgrass in public parks and golf courses. This study determined the risks of infection from exposure to various levels of Legionella in reclaimed waters considering: the method of spray application; and the duration and frequency of exposure. Evaluation of these factors resulted in a risk of infection greater than 1:10,000 for several scenarios when the number of Legionella in the reclaimed water exceeded 1000 colony-forming units (CFU) per ml. Most current guidelines for control of Legionella in distribution systems recommend that increased monitoring or remedial action be taken when Legionella levels exceed 1000 to 10,000 CFU/ml. Based upon our risk assessment, these guidelines seem appropriate for reclaimed water systems where spray irrigation is practiced. Copyright © 2018 Elsevier Ltd. All rights reserved.

  9. [Presence of Legionella spp. in household drinking water reservoirs in Resistencia, Chaco, Argentina. Preliminary report].

    Science.gov (United States)

    Lösch, Liliana S; Merino, Luis A

    Legionella spp. is an environmental bacterium that can survive in a wide range of physicochemical conditions and may colonize distribution systems of drinking water and storage tanks. Legionella pneumophila is the major waterborne pathogen that can cause 90% of Legionnaires' disease cases. The aim of this study was to detect the presence of Legionella spp. in household drinking water tanks in the city of Resistencia, Chaco. The detection of Legionella in water samples was performed by culture methods as set out in ISO 11731:1998. Thirty two water samples were analyzed and Legionella spp. was recovered in 12 (37.5%) of them. The monitoring of this microorganism in drinking water is the first step towards addressing the control of its spread to susceptible hosts. Copyright © 2016 Asociación Argentina de Microbiología. Publicado por Elsevier España, S.L.U. All rights reserved.

  10. Reduction of Legionella spp. in Water and in Soil by a Citrus Plant Extract Vapor

    Science.gov (United States)

    Kurzbach, Elena; Score, Jodie; Tejpal, Jyoti; Chi Tangyie, George; Phillips, Carol

    2014-01-01

    Legionnaires' disease is a severe form of pneumonia caused by Legionella spp., organisms often isolated from environmental sources, including soil and water. Legionella spp. are capable of replicating intracellularly within free-living protozoa, and once this has occurred, Legionella is particularly resistant to disinfectants. Citrus essential oil (EO) vapors are effective antimicrobials against a range of microorganisms, with reductions of 5 log cells ml−1 on a variety of surfaces. The aim of this investigation was to assess the efficacy of a citrus EO vapor against Legionella spp. in water and in soil systems. Reductions of viable cells of Legionella pneumophila, Legionella longbeachae, Legionella bozemanii, and an intra-amoebal culture of Legionella pneumophila (water system only) were assessed in soil and in water after exposure to a citrus EO vapor at concentrations ranging from 3.75 mg/liter air to 15g/liter air. Antimicrobial efficacy via different delivery systems (passive and active sintering of the vapor) was determined in water, and gas chromatography-mass spectrometry (GC-MS) analysis of the antimicrobial components (linalool, citral, and β-pinene) was conducted. There was up to a 5-log cells ml−1 reduction in Legionella spp. in soil after exposure to the citrus EO vapors (15 mg/liter air). The most susceptible strain in water was L. pneumophila, with a 4-log cells ml−1 reduction after 24 h via sintering (15 g/liter air). Sintering the vapor through water increased the presence of the antimicrobial components, with a 61% increase of linalool. Therefore, the appropriate method of delivery of an antimicrobial citrus EO vapor may go some way in controlling Legionella spp. from environmental sources. PMID:25063652

  11. El hospital ante un brote prolongado de legionelosis The hospital faced with a prolonged Legionella outbreak

    Directory of Open Access Journals (Sweden)

    Juan Antonio Fernández

    2004-08-01

    Full Text Available Objetivo: Describir los recursos asistenciales utilizados en pacientes hospitalizados por neumonía por Legionella durante un brote ocurrido en Alcoy y compararlos con los empleados en otras neumonías. Métodos y resultados: Mediante un protocolo, se analiza y compara a 177 pacientes con neumonía por Legionella con 180 pacientes ingresados por otras neumonías, y se describe su tratamiento y los recursos empleados. La claritromicina fue el antibiótico más utilizado en ambos grupos. La estancia hospitalaria fue similar, pero los requerimientos de ventilación mecánica y cuidados intensivos fueron superiores para Legionella. La hospitalización domiciliaria se utilizó con éxito en un 15,6% de los pacientes con Legionella y en un 11,3% de los demás. El uso de oxigenoterapia domiciliaria al alta fue menor para Legionella (7,8% que para el resto de neumonías (16,7%. Conclusiones: Se destaca la mayor necesidad de ventilación mecánica en la neumonía por Legionella y los buenos resultados de la hospitalización domiciliaria dentro de los nuevos sistemas de gestión.Objective: To describe the health resources used in patients hospitalized with Legionella pneumonia during an outbreak of Legionnaire's disease in Alcoy (Spain, and to compare them with those used in other forms of pneumonia. Methods and results: Using a clinical protocol, 177 Legionella pneumonia patients were compared with 180 patients hospitalized for other types of pneumonia. Data on therapy and the resources used were collected. The most common antibiotic treatment in both groups was clarithromycin, but intensive care and mechanical ventilation requirements were greater in Legionella pneumonia. Home-based hospital care was successfully used in 15.6% of patients with Legionella pneumonia and in 11.3% of those with other types of pneumonia. Home oxygen therapy after discharge was less frequent in the Legionella pneumonia group (7.8% than in the group with non-Legionella

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

    Science.gov (United States)

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

    2014-11-24

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

  13. Legionella shows a diverse secondary metabolism dependent on a broad spectrum Sfp-type phosphopantetheinyl transferase

    Directory of Open Access Journals (Sweden)

    Nicholas J. Tobias

    2016-11-01

    Full Text Available Several members of the genus Legionella cause Legionnaires’ disease, a potentially debilitating form of pneumonia. Studies frequently focus on the abundant number of virulence factors present in this genus. However, what is often overlooked is the role of secondary metabolites from Legionella. Following whole genome sequencing, we assembled and annotated the Legionella parisiensis DSM 19216 genome. Together with 14 other members of the Legionella, we performed comparative genomics and analysed the secondary metabolite potential of each strain. We found that Legionella contains a huge variety of biosynthetic gene clusters (BGCs that are potentially making a significant number of novel natural products with undefined function. Surprisingly, only a single Sfp-like phosphopantetheinyl transferase is found in all Legionella strains analyzed that might be responsible for the activation of all carrier proteins in primary (fatty acid biosynthesis and secondary metabolism (polyketide and non-ribosomal peptide synthesis. Using conserved active site motifs, we predict some novel compounds that are probably involved in cell-cell communication, differing to known communication systems. We identify several gene clusters, which may represent novel signaling mechanisms and demonstrate the natural product potential of Legionella.

  14. Legionella shows a diverse secondary metabolism dependent on a broad spectrum Sfp-type phosphopantetheinyl transferase.

    Science.gov (United States)

    Tobias, Nicholas J; Ahrendt, Tilman; Schell, Ursula; Miltenberger, Melissa; Hilbi, Hubert; Bode, Helge B

    2016-01-01

    Several members of the genus Legionella cause Legionnaires' disease, a potentially debilitating form of pneumonia. Studies frequently focus on the abundant number of virulence factors present in this genus. However, what is often overlooked is the role of secondary metabolites from Legionella . Following whole genome sequencing, we assembled and annotated the Legionella parisiensis DSM 19216 genome. Together with 14 other members of the Legionella , we performed comparative genomics and analysed the secondary metabolite potential of each strain. We found that Legionella contains a huge variety of biosynthetic gene clusters (BGCs) that are potentially making a significant number of novel natural products with undefined function. Surprisingly, only a single Sfp-like phosphopantetheinyl transferase is found in all Legionella strains analyzed that might be responsible for the activation of all carrier proteins in primary (fatty acid biosynthesis) and secondary metabolism (polyketide and non-ribosomal peptide synthesis). Using conserved active site motifs, we predict some novel compounds that are probably involved in cell-cell communication, differing to known communication systems. We identify several gene clusters, which may represent novel signaling mechanisms and demonstrate the natural product potential of Legionella .

  15. Fatal pneumonia by Legionella in a farmer with hypersensitivity pneumonitis Neumonía por Legionella con desenlace fatal en un granjero con neumonitis por hipersensibilidad

    Directory of Open Access Journals (Sweden)

    Vega García López

    2009-12-01

    Full Text Available Background: The retrospective investigation of a fatal sporadic Legionnaires' disease identified an unknown case of occupational hypersensitivity pneumonitis in a swine breeder. Methods: Chest high-resolution computed tomography, bronchoalveolar lavage, lung biopsy, arterial gasometry, pulmonary function tests and autopsy were performed. It was studied the presence of Legionella by serology and risk water samples were analyzed to identify the Legionella's source. Results: HP and Legionella pneumophila pneumonia diagnostics were confirmed. Lung fibrosis, a restrictive functional pattern, decreased diffusion, hypoxemia and bronchoalveolar lavage lymphocytosis were evidenced. Legionella´s source was detected in a shower and a positive serology in the patient. Autopsy verified pulmonary fibrosis and the septic shock leaded to Legionella causing the death. Conclusions: Chronic cough and pulmonary infiltrates in a farmer should suspect the presence of hypersensitivity pneumonitis. Later diagnosis carries a worse prognosis, the offending antigens exposure can't be avoided and fibrotic stage enhanced opportunity infection disease.Introducción: La investigación retrospectiva sobre un fallecimiento aislado por Legionelosis, hizo aflorar un caso de neumonitis por hipersensibilidad en un granjero cuidador de cerdos. Métodos: Se realizaron las siguientes pruebas: tomografía axial computerizada de alta resolución, lavado broncoalveolar, biopsia pulmonar, gasometría arterial, pruebas de función respiratoria y autopsia. Se estudió la presencia de Legionella por serología y se analizaron las muestras de fuentes de riesgo para identificar el foco de Legionella. Resultados: El estudio confirmó los diagnósticos de neumonitis por hipersensibilidad y neumonía por Legionella pneumophila. Las pruebas realizadas objetivaron la fibrosis pulmonar, un patrón respiratorio funcional restrictivo, un descenso de la difusión pulmonar, hipoxemia y la presencia de

  16. Legionella pneumophila transcriptome during intracellular multiplication in human macrophages

    Directory of Open Access Journals (Sweden)

    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.

  17. AUTOMATED DEAD-END ULTRAFILTRATION FOR ENHANCED SURVEILLANCE OF LEGIONELLA 2 PNEUMOPHILA AND LEGIONELLA SPP. IN COOLING TOWER WATERS

    Energy Technology Data Exchange (ETDEWEB)

    Brigmon, R.; Leskinen, S.; Kearns, E.; Jones, W.; Miller, R.; Betivas, C.; Kingsley, M.; Lim, D.

    2011-10-10

    Detection of Legionella pneumophila in cooling towers and domestic hot water systems involves concentration by centrifugation or membrane filtration prior to inoculation onto growth media or analysis using techniques such as PCR or immunoassays. The Portable Multi-use Automated Concentration System (PMACS) was designed for concentrating microorganisms from large volumes of water in the field and was assessed for enhancing surveillance of L. pneumophila at the Savannah River Site, SC. PMACS samples (100 L; n = 28) were collected from six towers between August 2010 and April 2011 with grab samples (500 ml; n = 56) being collected before and after each PMACS sample. All samples were analyzed for the presence of L. pneumophila by direct fluorescence immunoassay (DFA) using FITC-labeled monoclonal antibodies targeting serogroups 1, 2, 4 and 6. QPCR was utilized for detection of Legionella spp. in the same samples. Counts of L. pneumophila from DFA and of Legionella spp. from qPCR were normalized to cells/L tower water. Concentrations were similar between grab and PMACS samples collected throughout the study by DFA analysis (P = 0.4461; repeated measures ANOVA). The same trend was observed with qPCR. However, PMACS concentration proved advantageous over membrane filtration by providing larger volume, more representative samples of the cooling tower environment, which led to reduced variability among sampling events and increasing the probability of detection of low level targets. These data highlight the utility of the PMACS for enhanced surveillance of L. pneumophila by providing improved sampling of the cooling tower environment.

  18. Hijacking of the Host Ubiquitin Network by Legionella pneumophila

    Directory of Open Access Journals (Sweden)

    Jiazhang Qiu

    2017-12-01

    Full Text Available Protein ubiquitination is critical for regulation of numerous eukaryotic cellular processes such as protein homeostasis, cell cycle progression, immune response, DNA repair, and vesicular trafficking. Ubiquitination often leads to the alteration of protein stability, subcellular localization, or interaction with other proteins. Given the importance of ubiquitination in the regulation of host immunity, it is not surprising that many infectious agents have evolved strategies to interfere with the ubiquitination network with sophisticated mechanisms such as functional mimicry. The facultative intracellular pathogen Legionella pneumophila is the causative agent of Legionnaires' disease. L. pneumophila is phagocytosed by macrophages and is able to replicate within a niche called Legionella-containing vacuole (LCV. The biogenesis of LCV is dependent upon the Dot/Icm type IV secretion system which delivers more than 330 effector proteins into host cytosol. The optimal intracellular replication of L. pneumophila requires the host ubiquitin-proteasome system. Furthermore, membranes of the bacterial phagosome are enriched with ubiquitinated proteins in a way that requires its Dot/Icm type IV secretion system, suggesting the involvement of effectors in the manipulation of the host ubiquitination machinery. Here we summarize recent advances in our understanding of mechanisms exploited by L. pneumophila effector proteins to hijack the host ubiquitination pathway.

  19. Opportunistic Pathogens Mycobacterium Avium Complex (MAC) and Legionella spp. Colonise Model Shower.

    Science.gov (United States)

    Whiley, Harriet; Giglio, Steven; Bentham, Richard

    2015-07-24

    Legionella spp. and Mycobacterium avium complex (MAC) are opportunistic pathogens of public health concern. Hot water systems, including showers, have been identified as a potential source of infection. This paper describes the colonization of Legionella and MAC on the flexible tubing within a model potable shower system, utilizing thermostatic mixing and a flexible shower head. A MAC qPCR method of enumeration was also developed. MAC and Legionella spp. were detected within the biofilm at maximum concentrations of 7.0 × 104 and 2.0 × 103 copies/cm2 PVC tubing respectively. No significant changes were observed between sample of the flexible shower tubing that dried between uses and those that remained filled with water. This suggested the "unhooking" showerheads and allowing them to dry is not an effective method to reduce the risk of Legionella or MAC colonisation.

  20. Legionella pneumonia associated with severe acute respiratory distress syndrome and diffuse alveolar hemorrhage - A rare association

    Directory of Open Access Journals (Sweden)

    Muhammad Kashif

    2017-01-01

    Full Text Available Legionella pneumophila is a common, usually underreported and undiagnosed cause of community acquired pneumonia which can lead to significant morbidity and mortality. Diffuse alveolar hemorrhage rarely have been associated with legionella infection. We present a 61-year-old man with hypertension, diabetes mellitus and obesity admitted with severe acute respiratory distress syndrome. He was found to have Legionella pneumonia with associated diffuse alveolar hemorrhage diagnosed with bronchoscopic sequential bronchoalveolar lavage. He was successfully managed with antibiotics, lung protective strategies and intravenous pulse dose steroids. This patient highlights the unusual association of Legionella infection and diffuse alveolar hemorrhage. Additionally, the case re-enforces the need for early and aggressive evaluation and management of patients presenting with pneumonia and progressive hypoxia despite adequate treatment.

  1. Uncovering the Legionella genus effector repertoire - strength in diversity and numbers

    Science.gov (United States)

    Burstein, David; Amaro, Francisco; Zusman, Tal; Lifshitz, Ziv; Cohen, Ofir; Gilbert, Jack A; Pupko, Tal; Shuman, Howard A; Segal, Gil

    2016-01-01

    Infection by the human pathogen Legionella pneumophila relies on the translocation of ~300 virulence proteins, termed effectors, which manipulate host-cell processes. However, almost no information exists regarding effectors in other Legionella pathogens. Here we sequenced, assembled and characterized the genomes of 38 Legionella species, and predicted their effector repertoire using a previously validated machine-learning approach. This analysis revealed a treasure trove of 5,885 predicted effectors. The effector repertoire of different Legionella species was found to be largely non-overlapping, and only seven core-effectors were shared among all species studied. Species-specific effectors had atypically low GC content, suggesting exogenous acquisition, possibly from their natural protozoan hosts. Furthermore, we detected numerous novel conserved effector domains, and discovered new domain combinations, which allowed inferring yet undescribed effector functions. The effector collection and network of domain architectures described here can serve as a roadmap for future studies of effector function and evolution. PMID:26752266

  2. Recreational Vehicle Water Tanks as a Possible Source for Legionella Infections

    Directory of Open Access Journals (Sweden)

    Christine M. Litwin

    2013-01-01

    Full Text Available We investigated recreational vehicle (RV water reservoirs in response to a case of pneumonia in which Legionella pneumophila was cultured both from the patient and a RV reservoir in which he travelled. Water samples processed and cultured at the CDC according to standard protocol were positive for Legionella spp. in 4/17 (24% faucets, 1/11 (9% water tanks from 4/20 (20% RVs from three different campsites. Legionella spp. that were isolated included L. pneumophila (serogroups 1 and 6, L. anisa, L. feeleii, and L. quateriensis. Environmental controls from the potable water of the three campsites were culture-negative. A survey of maintenance practices by the RV users at the campsites revealed that chlorine disinfection of the water tanks was rarely performed. To prevent the possibility of Legionella infections, RV owners should implement regular chlorine disinfection of their water tanks and follow the recommended maintenance guidelines according to their owner's manuals.

  3. Recreational vehicle water tanks as a possible source for legionella infections.

    Science.gov (United States)

    Litwin, Christine M; Asebiomo, Bankole; Wilson, Katherine; Hafez, Michael; Stevens, Valerie; Fliermans, Carl B; Fields, Barry S; Fisher, John F

    2013-01-01

    We investigated recreational vehicle (RV) water reservoirs in response to a case of pneumonia in which Legionella pneumophila was cultured both from the patient and a RV reservoir in which he travelled. Water samples processed and cultured at the CDC according to standard protocol were positive for Legionella spp. in 4/17 (24%) faucets, 1/11 (9%) water tanks from 4/20 (20%) RVs from three different campsites. Legionella spp. that were isolated included L. pneumophila (serogroups 1 and 6), L. anisa, L. feeleii, and L. quateriensis. Environmental controls from the potable water of the three campsites were culture-negative. A survey of maintenance practices by the RV users at the campsites revealed that chlorine disinfection of the water tanks was rarely performed. To prevent the possibility of Legionella infections, RV owners should implement regular chlorine disinfection of their water tanks and follow the recommended maintenance guidelines according to their owner's manuals.

  4. Epidemiology and Ecology of Opportunistic Premise Plumbing Pathogens: Legionella pneumophila, Mycobacterium avium, and Pseudomonas aeruginosa

    Science.gov (United States)

    BACKGROUND: Legionella pneumophila, Mycobacterium avium, and Pseudomonas aeruginosa are opportunistic premise plumbing pathogens (OPPPs) that persist and grow in household plumbing, habitats they share with humans. Infections caused by these OPPPs involve individuals with preexis...

  5. Invasion of Eukaryotic Cells by Legionella Pneumophila: A Common Strategy for all Hosts?

    OpenAIRE

    Paul S Hoffman

    1997-01-01

    Legionella pneumophila is an environmental micro-organism capable of producing an acute lobar pneumonia, commonly referred to as Legionnaires’ disease, in susceptible humans. Legionellae are ubiquitous in aquatic environments, where they survive in biofilms or intracellularly in various protozoans. Susceptible humans become infected by breathing aerosols laden with the bacteria. The target cell for human infection is the alveolar macrophage, in which the bacteria abrogate phagolysosomal fusio...

  6. Severe Pneumonia Caused by Legionella pneumophila: Differential Diagnosis and Therapeutic Considerations.

    Science.gov (United States)

    Chahin, Abdullah; Opal, Steven M

    2017-03-01

    Severe legionella pneumonia poses a diagnostic challenge and requires early intervention. Legionnaire's disease can have several presenting signs, symptoms, and laboratory abnormalities that suggest that Legionella pneumophila is the pathogen, but none of these are sufficient to distinguish L pneumophila pneumonia from other respiratory pathogens. L pneumophila is primarily an intracellular pathogen and needs treatment with antibiotics that efficiently enter the intracellular space. Copyright © 2016 Elsevier Inc. All rights reserved.

  7. Comparison of slide agglutination test and direct immunofluorescence assay for identification of Legionella isolates.

    OpenAIRE

    Thacker, W L; Wilkinson, H W; Benson, R F

    1983-01-01

    It is technically impractical for many clinical laboratories to use the direct immunofluorescence assay for identifying and serogrouping clinical isolates of Legionella. We compared the results obtained with the direct immunofluorescence assay with the results of a simple and less-demanding slide agglutination test for identifying 15 serogroups representing seven Legionella species. The slide agglutination test was in complete agreement with the direct immunofluorescence assay, and the serogr...

  8. Exploring the Legionella pneumophila positivity rate in hotel water samples from Antalya, Turkey.

    Science.gov (United States)

    Sepin Özen, Nevgün; Tuğlu Ataman, Şenay; Emek, Mestan

    2017-05-01

    The genus Legionella is a fastidious Gram-negative bacteria widely distributed in natural waters and man made water supply systems. Legionella pneumophila is the aetiological agent of approximately 90% of reported Legionellosis cases, and serogroup 1 is the most frequent cause of infections. Legionnaires' disease is often associated with travel and continues to be a public health concern at present. The correct water management quality practices and rapid methods for analyzing Legionella species in environmental water is a key point for the prevention of Legionnaires' disease outbreaks. This study aimed to evaluate the positivity rates and serotyping of Legionella species from water samples in the region of Antalya, Turkey, which is an important tourism center. During January-December 2010, a total of 1403 samples of water that were collected from various hotels (n = 56) located in Antalya were investigated for Legionella pneumophila. All samples were screened for L. pneumophila by culture method according to "ISO 11731-2" criteria. The culture positive Legionella strains were serologically identified by latex agglutination test. A total of 142 Legionella pneumophila isolates were recovered from 21 (37.5%) of 56 hotels. The total frequency of L. pneumophila isolation from water samples was found as 10.1%. Serological typing of 142 Legionella isolates by latex agglutination test revealed that strains belonging to L. pneumophila serogroups 2-14 predominated in the examined samples (85%), while strains of L. pneumophila serogroup 1 were less numerous (15%). According to our knowledge, our study with the greatest number of water samples from Turkey demonstrates that L. pneumophila serogroups 2-14 is the most common isolate. Rapid isolation of L. pneumophila from environmental water samples is essential for the investigation of travel related outbreaks and the possible resources. Further studies are needed to have epidemiological data and to determine the types of L

  9. Cerebellar involvement that occurred during treatment of Legionella pneumonia: A case report

    Directory of Open Access Journals (Sweden)

    Ozlem Alici

    2013-06-01

    Full Text Available Legionnaires’ disease can appear with different levels of severity. A case of a previously healthy lady with communityacquiredpneumonia who progressed to severe acute respiratory distress syndrome and developed cerebellar dysfunctionis reported. In patients presenting with neurological symptoms after an episode of pneumonia, Legionella infectionshould be considered. J Microbiol Infect Dis 2013; 3(2: 83-85Key words: Legionella, cerebellar dysfunction, dysarthria, ataxia

  10. Microbial and physicochemical parameters associated with Legionella contamination in hot water recirculation systems.

    Science.gov (United States)

    Serrano-Suárez, Alejandra; Dellundé, Jordi; Salvadó, Humbert; Cervero-Aragó, Sílvia; Méndez, Javier; Canals, Oriol; Blanco, Silvia; Arcas, Antoni; Araujo, Rosa

    2013-08-01

    Hot water recirculation systems (HWRS) in hotels and nursing homes, which are common in countries such as Spain, have been related to outbreaks of legionellosis. To establish the relationships of microbial and physicochemical parameters, especially protozoa, with the occurrence of Legionella in HWRS, 231 samples from hotels and nursing homes were analysed for Legionella, protozoa, heterotrophic plate counts (HPC) at 22 and 37 °C, Pseudomonas, metals, temperature and others. Legionella pneumophila was the dominant species isolated, and 22 % were sg. 1. The sampling method became particularly important in order to define which factors were involved on the occurrence of Legionella. Results showed that the bacteria and the accompanying microbiota were more abundant in the first flush water whose temperature was lower. The bacteria occurred in those samples with high HPC and were inversely correlated with high temperatures. Multivariate regression showed that a concentration above 1 × 10(5) CFU/100 mL of HPC at 37 °C, Fe above 0.095 ppm and the presence of protozoa increased significantly the risk of Legionella colonization, while univariant regression showed that the presence of Cu above 0.76 ppm and temperature above 55 °C diminished it. Therefore, to reduce the risk associated with Legionella occurrence in HWRS these parameters should be taken into consideration.

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

    Science.gov (United States)

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

    2014-01-01

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

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

    Science.gov (United States)

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

    2014-08-01

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

  13. Interactive Effects of Corrosion, Copper, and Chloramines on Legionella and Mycobacteria in Hot Water Plumbing.

    Science.gov (United States)

    Rhoads, William J; Pruden, Amy; Edwards, Marc A

    2017-06-20

    Complexities associated with drinking water plumbing systems can result in undesirable interactions among plumbing components that undermine engineering controls for opportunistic pathogens (OPs). In this study, we examine the effects of plumbing system materials and two commonly applied disinfectants, copper and chloramines, on water chemistry and the growth of Legionella and mycobacteria across a transect of bench- and pilot-scale hot water experiments carried out with the same municipal water supply. We discovered that copper released from corrosion of plumbing materials can initiate evolution of >1100 times more hydrogen (H 2 ) from water heater sacrificial anode rods than does presence of copper dosed as soluble cupric ions. H 2 is a favorable electron donor for autotrophs and causes fixation of organic carbon that could serve as a nutrient for OPs. Dosed cupric ions acted as a disinfectant in stratified stagnant pipes, inhibiting culturable Legionella and biofilm formation, but promoted Legionella growth in pipes subject to convective mixing. This difference was presumably due to continuous delivery of nutrients to biofilm on the pipes under convective mixing conditions. Chloramines eliminated culturable Legionella and prevented L. pneumophila from recolonizing biofilms, but M. avium gene numbers increased by 0.14-0.76 logs in the bulk water and were unaffected in the biofilm. This study provides practical confirmation of past discrepancies in the literature regarding the variable effects of copper on Legionella growth, and confirms prior reports of trade-offs between Legionella and mycobacteria if chloramines are applied as secondary disinfectant residual.

  14. Legionella detection by culture and qPCR: Comparing apples and oranges.

    Science.gov (United States)

    Whiley, Harriet; Taylor, Michael

    2016-01-01

    Legionella spp. are the causative agent of Legionnaire's disease and an opportunistic pathogen of significant public health concern. Identification and quantification from environmental sources is crucial for identifying outbreak origins and providing sufficient information for risk assessment and disease prevention. Currently there are a range of methods for Legionella spp. quantification from environmental sources, but the two most widely used and accepted are culture and real-time polymerase chain reaction (qPCR). This paper provides a review of these two methods and outlines their advantages and limitations. Studies from the last 10 years which have concurrently used culture and qPCR to quantify Legionella spp. from environmental sources have been compiled. 26/28 studies detected Legionella at a higher rate using qPCR compared to culture, whilst only one study detected equivalent levels of Legionella spp. using both qPCR and culture. Aggregating the environmental samples from all 28 studies, 2856/3967 (72%) tested positive for the presence of Legionella spp. using qPCR and 1331/3967 (34%) using culture. The lack of correlation between methods highlights the need to develop an acceptable standardized method for quantification that is sufficient for risk assessment and management of this human pathogen.

  15. Legionella Persistence in Manufactured Water Systems: Pasteurization Potentially Selecting for Thermal Tolerance

    Directory of Open Access Journals (Sweden)

    Harriet Whiley

    2017-07-01

    Full Text Available Legionella is an opportunistic waterborne pathogen of increasing public health significance. Pasteurization, otherwise known as super-heat and flush (increasing water temperature to above 70°C and flushing all outlets, has been identified as an important mechanism for the disinfection of Legionella in manufactured water systems. However, several studies have reported that this procedure was ineffective at remediating water distribution systems as Legionella was able to maintain long term persistent contamination. Up to 25% of L. pneumophila cells survived heat treatment of 70°C, but all of these were in a viable but non-culturable state. This demonstrates the limitations of the culture method of Legionella detection currently used to evaluate disinfection protocols. In addition, it has been demonstrated that pasteurization and nutrient starvation can select for thermal tolerant strains, where L. pneumophila was consistently identified as having greater thermal tolerance compared to other Legionella species. This review demonstrates that further research is needed to investigate the effectiveness of pasteurization as a disinfection method. In particular, it focuses on the potential for pasteurization to select for thermal tolerant L. pneumophila strains which, as the primary causative agent of Legionnaires disease, have greater public health significance compared to other Legionella species.

  16. Frequency of legionella contamination in conditional & water distribution systems of Tehran hospitals

    Directory of Open Access Journals (Sweden)

    Davod Esmaieli

    2008-09-01

    Full Text Available Background: Legionella species are ubiquitous in natural aquatic environments, capable of existing in waters with varied temperatures, PH levels, and nutrient and oxygen contents. Of 49 known legionella species, 20 species have been linked to pneumonia in humans. Contamination by legionella has occurred in the distribution systems of many hospitals. Aerosol-generating systems such as faucets, showerheads, cooling towers, and nebulizers are responsible for their transmission from water to air. Methods: A total of 113 water samples were gathered from different wards of 32 hospitals in different geographical regions of Tehran city. These samples were concentrated by filtration, treated with the acid and temperature buffers, and isolated on a BCYE agar culture medium. Results: A total of 22 hospitals out of 33 (26.5% were contaminated by legionella species, and 30 samples (26.5% out of 113 were positive. Chlorine concentration and pH level of the water samples were 0.18-2.2 mg/l and 6.6-7.6, respectively. Conclusion: The high rate of waste water contamination in Tehran hospitals with Legionella indicates the resistance of this microorganism to chlorine and other disinfectants, or inadequate disinfection process, representing the insufficiency of the current decontamination of hospital water distribution system. Thus identifying legionella species and their controlling in water distribution system of hospitals is of great importance.

  17. Detection of airborne Legionella while showering using liquid impingement and fluorescent in situ hybridization (FISH).

    Science.gov (United States)

    Deloge-Abarkan, Magali; Ha, Thi-Lan; Robine, Enric; Zmirou-Navier, Denis; Mathieu, Laurence

    2007-01-01

    Aerosols of water contaminated with Legionella bacteria constitute the only mode of exposure for humans. However, the prevention strategy against this pathogenic bacteria risk is managed through the survey of water contamination. No relationship linked the Legionella bacteria water concentration and their airborne abundance. Therefore, new approaches in the field of the metrological aspects of Legionella bioaerosols are required. This study was aimed at testing the main principles for bioaerosol collection (solid impaction, liquid impingement and filtration) and the in situ hybridization (FISH) method, both in laboratory and field assays, with the intention of applying such methodologies for airborne Legionella bacteria detection while showering. An aerosolization chamber was developed to generate controlled and reproducible L. pneumophila aerosols. This tool allowed the identification of the liquid impingement method as the most appropriate one for collecting airborne Legionella bacteria. The culturable fraction of airborne L. pneumophila recovered with the liquid impingement principle was 4 and 700 times higher compared to the impaction and filtration techniques, respectively. Moreover, the concentrations of airborne L. pneumophila in the impinger fluid were on average 7.0 x 10(5) FISH-cells m(-3) air with the fluorescent in situ hybridization (FISH) method versus 9.0 x 10(4) CFU m(-3) air with the culture method. These results, recorded under well-controlled conditions, were confirmed during the field experiments performed on aerosols generated by hot water showers in health institutions. This new approach may provide a more accurate characterization of aerobiocontamination by Legionella bacteria.

  18. Ecology of Legionella within water cooling circuits of nuclear power plants along the French Loire River

    International Nuclear Information System (INIS)

    Jakubek, Delphine

    2012-01-01

    The cooling circuits of nuclear power plants, by their mode of operating, can select thermophilic microorganisms including the pathogenic organism Legionella pneumophila. To control the development of this genus, a disinfection treatment of water cooling systems with monochloramine can be used. To participate in the management of health and environmental risks associated with the physico-chemical and microbiological modification of water collected from the river, EDF is committed to a process of increasing knowledge about the ecology of Legionella in cooling circuits and its links with its environment (physical, chemical and microbiological) supporting or not their proliferation. Thus, diversity and dynamics of culturable Legionella pneumophila were determined in the four nuclear power plants along the Loire for a year and their links with physico-chemical and microbiological parameters were studied. This study revealed a high diversity of Legionella pneumophila subpopulations and their dynamic seems to be related to the evolution of a small number of subpopulations. Legionella subpopulations seem to maintain strain-specific relationships with biotic parameters and present different sensitivities to physico-chemical variations. The design of cooling circuits could impact the Legionella community. The use of monochloramine severely disrupts the ecosystem but does not select biocide tolerant subpopulations. (author)

  19. Incidence of Legionella and heterotrophic bacteria in household rainwater tanks in Azumino, Nagano prefecture, Japan.

    Science.gov (United States)

    Kobayashi, Michiko; Oana, Kozue; Kawakami, Yoshiyuki

    2014-01-01

    Many administrative agencies in Japan are encouraging installation of household rainwater-storage tanks for more effective use of natural rainwater. Water samples were collected periodically from 43 rainwater tanks from 40 households and tested for the presence of Legionella species and the extent of heterotrophic bacteria in Azumino city, Nagano prefecture, Japan. PCR assays indicated the presence of Legionella spp. in 12 (30%) of the 43 tank water samples. Attempts were made to identify correlations between PCR positive samples, topography, pH, chemical oxygen demand (COD), atmospheric temperature and the numbers of heterotrophic bacteria. Between June and October, 2012, the numbers of heterotrophic bacteria in rainwater tanks and the values of COD positively correlated with the presence of Legionella species. In most of the Legionella-positive cases, heterotrophic bacterial cell counts were >10(4) CFU/mL. Moreover, Legionella species were less frequently detected when the COD value was >5 mg KMnO(4)/L. Therefore, at least in Azumino, Japan between June and October 2012, both heterotrophic bacterial counts and COD values may be considered index parameters for the presence of Legionella cells in rainwater tanks. Much more accumulation of such data is needed to verify the accuracy of these findings. © 2013 The Societies and Wiley Publishing Asia Pty Ltd.

  20. A Multiplex PCR for Detection of Mycoplasma pneumoniae, Chlamydophila pneumoniae, Legionella pneumophila, and Bordetella pertussis in Clinical Specimens

    National Research Council Canada - National Science Library

    McDonough, E. A; Barrozo, C. P; Russell, K. L; Metzgar, D

    2005-01-01

    A multiplex PCR was developed that is capable of detecting four of the most important bacterial agents of atypical pneumophia, Mycaplasma pneumoniae, Chlamydophia pneumoniae, Legionella pneumophila...

  1. A multicenter evaluation of the Biotest legionella urinary antigen EIA

    DEFF Research Database (Denmark)

    Harrison, Timothy; Uldum, Søren; Alexiou-Daniel, Stella

    1998-01-01

    OBJECTIVES: To undertake a multicenter study to evaluate the Biotest legionella urinary antigen enzyme immunoassay (EIA) performance against those EIAs already in use in 14 European laboratories. METHODS: Each laboratory examined urine specimens from appropriate patients using both their current...... assay and the Biotest EIA. Each examined: a standard panel of 12 coded urine samples (distributed by Biotest); a panel of 10 coded urine samples provided as part of a European external quality assurance (EQA) scheme; urine samples from patients with proven legionnaires' disease (LD); urine samples from...... patients with pneumonia of microbiologically proven cause other than LD; and urine samples submitted for routine examination. Thus, the performance of the Biotest assay (in comparison with current EIAs), its specificity and utility, and the inter-laboratory agreement were assessed. RESULTS: Inter...

  2. Influence of aquatic microorganisms on Legionella pneumophila survival.

    Science.gov (United States)

    Guerrieri, Elisa; Bondi, Moreno; Borella, Paola; Messi, Patrizia

    2007-07-01

    The ability of aquatic bacteria Pseudomonas fluorescens SSD (Ps-D) and Pseudomonas putida SSC (Ps-C) to support the persistence of Legionella pneumophila (Lp-1) in an artificial water microcosm was investigated for 42 day, at two different incubation temperatures. At 4 degrees C, individually suspended Lp-1 was no longer detectable just after 24 hours, while in co-cultures with Pseudomonas, Lp1 showed a better survival capability. At 30 degrees C, Lp-1 alone displayed high survival rates over the entire period of observation. When Lp-1 was inoculated with Ps-C and Ps-D, its count showed a marked decrease, followed by a gradual and costant decline.

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

    Science.gov (United States)

    Tyndall, R L; Domingue, E L

    1982-10-01

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

  4. Cocultivation of Legionella pneumophila and free-living amoebae

    Energy Technology Data Exchange (ETDEWEB)

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

    1982-10-01

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

  5. Legionella pneumophila pangenome reveals strain-specific virulence factors

    Directory of Open Access Journals (Sweden)

    Peris-Bondia Francesc

    2010-03-01

    Full Text Available Abstract Background Legionella pneumophila subsp. pneumophila is a gram-negative γ-Proteobacterium and the causative agent of Legionnaires' disease, a form of epidemic pneumonia. It has a water-related life cycle. In industrialized cities L. pneumophila is commonly encountered in refrigeration towers and water pipes. Infection is always via infected aerosols to humans. Although many efforts have been made to eradicate Legionella from buildings, it still contaminates the water systems. The town of Alcoy (Valencian Region, Spain has had recurrent outbreaks since 1999. The strain "Alcoy 2300/99" is a particularly persistent and recurrent strain that was isolated during one of the most significant outbreaks between the years 1999-2000. Results We have sequenced the genome of the particularly persistent L. pneumophila strain Alcoy 2300/99 and have compared it with four previously sequenced strains known as Philadelphia (USA, Lens (France, Paris (France and Corby (England. Pangenome analysis facilitated the identification of strain-specific features, as well as some that are shared by two or more strains. We identified: (1 three islands related to anti-drug resistance systems; (2 a system for transport and secretion of heavy metals; (3 three systems related to DNA transfer; (4 two CRISPR (Clustered Regularly Interspaced Short Palindromic Repeats systems, known to provide resistance against phage infections, one similar in the Lens and Alcoy strains, and another specific to the Paris strain; and (5 seven islands of phage-related proteins, five of which seem to be strain-specific and two shared. Conclusions The dispensable genome disclosed by the pangenomic analysis seems to be a reservoir of new traits that have mainly been acquired by horizontal gene transfer and could confer evolutionary advantages over strains lacking them.

  6. Annual variations and effects of temperature on Legionella spp. and other potential opportunistic pathogens in a bathroom.

    Science.gov (United States)

    Lu, Jingrang; Buse, Helen; Struewing, Ian; Zhao, Amy; Lytle, Darren; Ashbolt, Nicholas

    2017-01-01

    Opportunistic pathogens (OPs) in drinking water, like Legionella spp., mycobacteria, Pseudomonas aeruginosa, and free-living amobae (FLA) are a risk to human health, due to their post-treatment growth in water systems. To assess and manage these risks, it is necessary to understand their variations and environmental conditions for the water routinely used. We sampled premise tap (N cold  = 26, N hot  = 26) and shower (N shower  = 26) waters in a bathroom and compared water temperatures to levels of OPs via qPCR and identified Legionella spp. by 16S ribosomal RNA (rRNA) gene sequencing. The overall occurrence and cell equivalent quantities (CE L -1 ) of Mycobacterium spp. were highest (100 %, 1.4 × 10 5 ), followed by Vermamoeba vermiformis (91 %, 493), Legionella spp. (59 %, 146), P. aeruginosa (14 %, 10), and Acanthamoeba spp. (5 %, 6). There were significant variations of OP's occurrence and quantities, and water temperatures were associated with their variations, especially for Mycobacterium spp., Legionella spp., and V. vermiformis. The peaks observed for Legionella, mainly consisted of Legionella pneumophila sg1 or Legionella anisa, occurred in the temperature ranged from 19 to 49 °C, while Mycobacterium spp. and V. vermiformis not only co-occurred with Legionella spp. but also trended to increase with increasing temperatures. There were higher densities of Mycobacterium in first than second draw water samples, indicating their release from faucet/showerhead biofilm. Legionella spp. were mostly at detectable levels and mainly consisted of L. pneumophila, L. anisa, Legionella donaldsonii, Legionella tunisiensis, and an unknown drinking water isolate based on sequence analysis. Results from this study suggested potential health risks caused by opportunistic pathogens when exposed to warm shower water with low chlorine residue and the use of Mycobacterium spp. as an indicator of premise pipe biofilm and the control management of those potential

  7. A new oligonucleotide microarray for detection of pathogenic and non-pathogenic Legionella spp.

    Directory of Open Access Journals (Sweden)

    Boyang Cao

    Full Text Available Legionella pneumophila has been recognized as the major cause of legionellosis since the discovery of the deadly disease. Legionella spp. other than L. pneumophila were later found to be responsible to many non-pneumophila infections. The non-L. pneumophila infections are likely under-detected because of a lack of effective diagnosis. In this report, we have sequenced the 16S-23S rRNA gene internal transcribed spacer (ITS of 10 Legionella species and subspecies, including L. anisa, L. bozemanii, L. dumoffii, L. fairfieldensis, L. gormanii, L. jordanis, L. maceachernii, L. micdadei, L. pneumophila subspp. fraseri and L. pneumophila subspp. pasculleii, and developed a rapid oligonucleotide microarray detection technique accordingly to identify 12 most common Legionella spp., which consist of 11 pathogenic species of L. anisa, L. bozemanii, L. dumoffii, L. gormanii, L. jordanis, L. longbeachae, L. maceachernii, L. micdadei, and L. pneumophila (including subspp. pneumophila, subspp. fraseri, and subspp. pasculleii and one non-pathogenic species, L. fairfieldensis. Twenty-nine probes that reproducibly detected multiple Legionella species with high specificity were included in the array. A total of 52 strains, including 30 target pathogens and 22 non-target bacteria, were used to verify the oligonucleotide microarray assay. The sensitivity of the detection was at 1.0 ng with genomic DNA or 13 CFU/100 mL with Legionella cultures. The microarray detected seven samples of air conditioner-condensed water with 100% accuracy, validating the technique as a promising method for applications in basic microbiology, clinical diagnosis, food safety, and epidemiological surveillance. The phylogenetic study based on the ITS has also revealed that the non-pathogenic L. fairfieldensis is the closest to L. pneumophila than the nine other pathogenic Legionella spp.

  8. Survival and multiplication of Legionella pneumophila in municipal drinking water systems.

    Science.gov (United States)

    States, S J; Conley, L F; Kuchta, J M; Oleck, B M; Lipovich, M J; Wolford, R S; Wadowsky, R M; McNamara, A M; Sykora, J L; Keleti, G

    1987-05-01

    Studies were conducted to investigate the survival and multiplication of Legionella spp. in public drinking water supplies. An attempt was made, over a period of several years, to isolate legionellae from a municipal system. Sampling sites included the river water supply, treatment plant, finished water reservoir system, mains, and distribution taps. Despite the use of several isolation techniques, Legionella spp. could not be detected in any of the samples other than those collected from the river. It was hypothesized that this was due to the maintenance of a chlorine residual throughout the system. To investigate the potential for Legionella growth, additional water samples, collected from throughout the system, were dechlorinated, pasteurized, and inoculated with Legionella pneumophila. Subsequent growth indicated that many of these samples, especially those collected from areas affected by an accumulation of algal materials, exhibited a much greater ability to support Legionella multiplication than did river water prior to treatment. Chemical analyses were also performed on these samples. Correlation of chemical data and experimental growth results indicated that the chemical environment significantly affects the ability of the water to support multiplication, with turbidity, organic carbon, and certain metals being of particular importance. These studies indicate that the potential exists for Legionella growth within municipal systems and support the hypothesis that public water supplies may contaminate the plumbing systems of hospitals and other large buildings. The results also suggest that useful methods to control this contamination include adequate treatment plant filtration, maintenance of a chlorine residual throughout the treatment and distribution network, and effective covering of open reservoirs.

  9. The Sensitization of Legionella pneumophila to Some Antibiotics by Reserpine and Anti-Legionella Effects of Different Benzofuranone Derivatives

    Directory of Open Access Journals (Sweden)

    Mohsen Khaleghi

    2015-10-01

    Full Text Available Background: Legionella pneumophila is  a  dangerous pathogenic bacterium can cause serious infectious diseases especially in hospitalized immuno-compromised patients. This bacterium is shown to be resistant against different antibiotics. Resistance against a wide range of antibiotics is usually mediated by efflux pump in bacteria. Efflux pumps are proteinaceous transporters localized in the cytoplasmic membrane of all kinds of cells which excreted antibiotics outside the cells. However, synthesis of new anti-Legionella compounds or selection of resistant modulating agents are useful strategy to combat with L. pneumophilain the future.Methods: In this study the antibacterial activity of some benzofuranone derivatives have been investigated by disk diffusion method against L. pneumophila. Also the sensitivity of this test strain was evaluated against 19 antibiotics and the combination effect of reserpine at a sub-inhibitory concentration was further studied with these antibiotics using disk diffusion method with some modifications.Conclusion: Among the different synthetic compounds which were tested against L. pneumophila, the most antibacterial activity was observed for compounds 1j and 1m which contain hydroxyl and methoxy groups on the C-6 and C-7 positionsagainst L. pneumophila. To evaluate whether efflux pumps are active in L. pneumophila or not an efflux inhibitor (reserpine was tested in combination of different antibiotics against this test strain. Reserpine significantly enhanced the antibacterial activities of kanamycin, nitrofurantoin, co-trimoxazole, erythromycin, ofloxacillin, gentamycin, rifampin, ciprofloxacin, nalidixic acid, minocycline, tobramycin, and amikacin against L. pneumophila which shows the resistances to these antibiotics are mediated by efflux system in this bacterium.

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

    Science.gov (United States)

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

    2014-01-01

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

  11. Rate of Legionella pneumophila colonization in hospital hot water network after time flow taps installation.

    Science.gov (United States)

    Totaro, M; Valentini, P; Costa, A L; Giorgi, S; Casini, B; Baggiani, A

    2018-01-01

    In hospital water systems legionellae may be resistant to disinfectants in pipework, which is a problem particularly in areas where there is low flow or stagnation of water. We evaluated legionella colonization of a water network of an Italian hospital after time flow taps (TFTs) installation in proximity to dead legs. The water volume flushed was 64 L/day from May 2016, and 192 L/day from December 2016. Before TFTs installation, Legionella pneumophila sg2-14 was detected in all points (4 × 10 4  ± 3.1 × 10 4  cfu/L). All sites remained positive (2.9 × 10 4  ± 1.9 × 10 4  cfu/L) through November 2016. From December 2016 legionella persisted in one point only (2 × 10 2 to 6.8 × 10 3  cfu/L). TFTs with chemical disinfection may reduce legionella colonization associated with dead legs. Copyright © 2017 The Healthcare Infection Society. Published by Elsevier Ltd. All rights reserved.

  12. Legionella Antibodies in a Danish Hospital Staff with Known Occupational Exposure

    International Nuclear Information System (INIS)

    Rudbeck, M.; Uldum, S.A.; Rudbeck, M.; Viskum, S.; Molbak, K.

    2010-01-01

    Although legionnaires' disease frequently is acquired in health care institutions, little is known about the occupational risk of Legionella infection among health care workers. The aim of the present cross-sectional study was to analyse antibody levels among exposed hospital workers and to determine the correlation between antibodies to Legionella and self-reported symptoms. The study included 258 hospital employees and a reference group of 708 healthy blood donors. Hospital workers had a higher prevalence of Legionella antibody titres (=1 : 128) than blood donors (odds ratio 3.4; 95% CI 2.4-4.8). Antibody levels were not higher among staff members at risk of frequent aerosol exposure than among less exposed employees. There was no consistent association between a history of influenza-like symptom complex and the presence of antibodies. The results indicate that hospital workers have a higher risk of Legionella infections than the general population. However, since no excess morbidity was associated with sero positivity, most Legionella infections may be asymptomatic.

  13. Hot water systems as sources of Legionella pneumophila in hospital and nonhospital plumbing fixtures.

    Science.gov (United States)

    Wadowsky, R M; Yee, R B; Mezmar, L; Wing, E J; Dowling, J N

    1982-05-01

    Samples obtained from plumbing systems of hospitals, nonhospital institutions and homes were cultured for Legionella spp. by plating the samples directly on a selective medium. Swab samples were taken from the inner surfaces of faucet assemblies (aerators, spouts, and valve seats), showerheads, and shower pipes. Water and sediment were collected from the bottom of hot-water tanks. Legionella pneumophila serogroups 1, 5, and 6 were recovered from plumbing fixtures of the hospitals and nonhospital institutions and one of five homes. The legionellae (7 to 13,850 colony-forming units per ml) were also present in water and sediment from hot-water tanks maintained at 30 to 54 degrees C, but not in those maintained at 71 and 77 degrees C. Legionella micdadei was isolated from one tank. Thus legionellae are present in hot-water tanks which are maintained at warm temperatures or whose design results in warm temperatures at the bottom of the tanks. We hypothesize that hot-water tanks are a breeding site and a major source of L. pneumophila for the contamination of plumbing systems. The existence of these bacteria in the plumbing systems and tanks was not necessarily associated with disease. The extent of the hazard of this contamination needs to be delineated.

  14. An alkaline approach to treating cooling towers for control of Legionella pneumophila.

    Science.gov (United States)

    States, S J; Conley, L F; Towner, S G; Wolford, R S; Stephenson, T E; McNamara, A M; Wadowsky, R M; Yee, R B

    1987-08-01

    Earlier field and laboratory studies have shown that Legionella species survive and multiply in the pH range 5.5 to 9.2. Additionally, the technical feasibility of operating cooling towers at elevated alkalinities and pH has previously been documented by published guidelines. The guidelines indicate that these conditions facilitate corrosion control and favor chlorine persistence which enhances the effectiveness of continuous chlorination in biofouling control. This information suggests that control of Legionella species in cooling towers can be accomplished by operating the towers under alkaline conditions. To test this possibility, we collected water samples over a period of months from a hospital cooling tower. The samples were analyzed for a variety of chemical parameters. Subsamples were pasteurized and inoculated with non-agar-passaged Legionella pneumophila which had been maintained in tap water. Correlation of subsequent Legionella growth with corresponding pH and alkalinity values revealed statistically significant inverse associations. These data support the hypothesis that operating cooling towers outside of the optimal conditions for Legionella growth (e.g., at elevated alkalinities and a pH greater than 9) may be a useful approach to controlling growth in this habitat.

  15. Hot water systems as sources of Legionella pneumophila in hospital and nonhospital plumbing fixtures

    Energy Technology Data Exchange (ETDEWEB)

    Wadowsky, R.M.; Yee, R.B.; Mezmar, L.; Wing, E.J.; Dowling, J.N.

    1982-05-01

    Samples obtained from plumbing systems of hospitals, nonhospital institutions, and homes were cultured for Legionella spp. by plating the samples directly on a selective medium. Swab samples were taken from the inner surfaces of faucet assemblies (aerators, spouts, and valve seats), showerheads, and shower pipes. Water and sediment were collected from the bottom of hot-water tanks. Legionella pnenumophila serogroups 1.5, and 6 were recovered from plubming fixtures of the hospitals and nonhospital institutions and one of five homes. The legionellae (7 to 13,850 colony-forming units per ml) were also present in water and sediment from hot-water tanks maintained at 30 to 54/sup 0/C, but not in those maintained at 71 and 77/sup 0/C. Legionella micdadei was isolated from one tank. Thus legionellae are present in hot-water tanks which are maintained at warm temperatures or whose design results in warm temperatures at the bottom of the tanks. We hypothesize that hot-water tanks are a breeding site and a major source of L. pneumophila for the contamination of plumbing systems. The existence of these bacteria in the plumbing systems and tanks was not necessarily associated with disease. The extent of the hazard of this contamination needs to be delineated.

  16. Alkaline approach to treating cooling towers for control of Legionella pneumophila

    Energy Technology Data Exchange (ETDEWEB)

    States, S.J.; Conley, L.F.; Towner, S.G.; Wolford, R.S.; Stephenson, T.E.; McNamara, M.; Wadowsky, R.M.; Yee, R.B.

    1987-08-01

    Earlier field and laboratory studies have shown that Legionella species survive and multiply in the pH range 5.5 to 9.2. Additionally, the technical feasibility of operating cooling towers at elevated alkalinities and pH has previously been documented by published guidelines. The guidelines indicate that these conditions facilitate corrosion control and favor chlorine persistence which enhances the effectiveness of continuous chlorination in biofouling control. This information suggest that control of Legionella species in cooling towers can be accomplished by operating the towers under alkaline conditions. To test this possibility, we collected water samples over a period of months from a hospital cooling tower. The samples were analyzed for a variety of chemical parameters. Subsamples were pasteurized and inoculated with non-agar-passaged Legionella pneumophila which had been maintained in tap water. Correlation of subsequent Legionella growth with corresponding pH and alkalinity values revealed statistically significant inverse associations. These data support the hypothesis that operating cooling towers outside of the optimal conditions for Legionella growth (e.g., at elevated alkalinities and a pH greater than 9) may be a useful approach to controlling growth in this habitat.

  17. Hyperoxia accelerates Fas-mediated signaling and apoptosis in the lungs of Legionella pneumophila pneumonia

    Directory of Open Access Journals (Sweden)

    Tanabe Yoshinari

    2011-04-01

    Full Text Available Abstract Background Oxygen supplementation is commonly given to the patients with severe pneumonia including Legionella disease. Recent data suggested that apoptosis may play an important role, not only in the pathogenesis of Legionella pneumonia, but also in oxygen-induced tissue damage. In the present study, the lethal sensitivity to Legionella pneumonia were compared in the setting of hyperoxia between wild-type and Fas-deficient mice. Findings C57BL/6 mice and B6.MRL-Faslpr mice characterized with Fas-deficiency were used in this study. After intratracheal administration of L. pneumophila, mice were kept in hyperoxic conditions (85-90% O2 conc. in an airtight chamber for 3 days. Bone-marrow derived macrophages infected with L. pneumophila were also kept in hyperoxic conditions. Caspase activity and cytokine production were determined by using commercially available kits. Smaller increases of several apoptosis markers, such as caspase-3 and -8, were demonstrated in Fas-deficient mice, even though the bacterial burdens in Fas-deficient and wild type mice were similar. Bone-marrow derived macrophages from Fas-deficient mice were shown to be more resistant to Legionella-induced cytotoxicity than those from wild-type mice under hyperoxia. Conclusions These results demonstrated that Fas-mediated signaling and apoptosis may be a crucial factor in the pathogenesis of Legionella pneumonia in the setting of hyperoxia.

  18. Automobile windshield washer fluid: A potential source of transmission for Legionella.

    Science.gov (United States)

    Schwake, David Otto; Alum, Absar; Abbaszadegan, Morteza

    2015-09-01

    Epidemiological evidence suggesting driving cars to be a risk factor for legionellosis has prompted public health studies to investigate vehicle windshield washer fluid as a novel transmission source of this disease. The goal of the current study was to investigate whether or not windshield washer fluid could serve as a potential source of transmission for Legionella. A wide variation in the survival of L. pneumophila was observed when incubated in different washer fluids at 25 and 37 °C, however, one brand tested supported Legionella survival similar to or greater than sterilized deionized water. In addition, 1 L of tap water contained in a washer fluid reservoir was able to support population growth and survival of Legionella for several months. In a field study examining the windshield washer fluid of 12 elementary school buses, Legionella were detected from 84% of samples at a high concentration of 8.1×10(4) CFU/mL. Culturable cells were also detected in aerosolized washer fluid during washer fluid spray. By demonstrating survival in certain windshield washer fluids, growth within washer fluid reservoirs, and the presence of viable cells in bus washer fluid spray, we have provided evidence suggesting the potential for a novel route of Legionella exposure. Copyright © 2015 Elsevier B.V. All rights reserved.

  19. Legionella species colonization of water distribution systems, pools and air conditioning systems in cruise ships and ferries

    Directory of Open Access Journals (Sweden)

    Gourgoulianis Kostantinos

    2008-11-01

    Full Text Available Abstract Background Legionnaires' disease continues to be a public health concern in passenger ships. This study was scheduled in order to investigate Legionella spp. colonization of water distribution systems (WDS, recreational pools, and air-conditioning systems on board ferries and cruise ships in an attempt to identify risk factors for Legionella spp. colonization associated with ship water systems and water characteristics. Methods Water systems of 21 ferries and 10 cruise ships including WDS, air conditioning systems and pools were investigated for the presence of Legionella spp. Results The 133 samples collected from the 10 cruise ships WDS, air conditioning systems and pools were negative for Legionella spp. Of the 21 ferries WDS examined, 14 (66.7% were legionellae-positive. A total of 276 samples were collected from WDS and air conditioning systems. Legionella spp. was isolated from 37.8% of the hot water samples and 17.5% of the cold water samples. Of the total 96 positive isolates, 87 (90.6% were L. pneumophila. Legionella spp. colonization was positively associated with ship age. The temperature of the hot water samples was negatively associated with colonization of L. pneumophila serogroup (sg 1 and that of L. pneumophila sg 2 to 14. Increases in pH ≥7.8 and total plate count ≥400 CFU/L, correlated positively with the counts of L. pneumophila sg 2 to 14 and Legionella spp. respectively. Free chlorine of ≥0.2 mg/L inhibited colonization of Legionella spp. Conclusion WDS of ferries can be heavily colonized by Legionella spp. and may present a risk of Legionnaires' disease for passengers and crew members. Guidelines and advising of Legionnaires' disease prevention regarding ferries are needed, in particular for operators and crew members.

  20. Influenceof antimicrobial therapy on the sensitivity of legionella PCR - three case report

    Directory of Open Access Journals (Sweden)

    Renato Eržen

    2007-02-01

    Full Text Available Background: L. pneumophila is a possible causative agent of severe community and hospital acquired pneumonia. Available diagnostic tests have a lot of shortcomings. Detection of Legionella DNA with the PCR technique is a promissing new method allowing diagnosis within clinically useful time.Methods: Three cases of severe Legionella pneumonia were followed with the PCR technique, urinary antigen and serologic methods.Results: Diagnostic tests have different sensitivity and specificity and also different shortcomings. Serologic tests are unable to provide diagnosis within clinically useful time. Urinary antigen is specific just for L. pneumophila serotype 1. Antimicrobial treatment has impact on the sensitivity of Legionella PCR.Conclusions: Detection of L. pneumophila infection with the PCR technique in addition to urinary antigen testing is likely to be the best diagnostic tool for the detection of all species within a time frame that will affect clinical management.

  1. Review of Various Solutions for avoiding critical levels of Legionella Bacteria in Domestic Hot Water System

    DEFF Research Database (Denmark)

    Yang, Xiaochen; Li, Hongwei; Svendsen, Svend

    2013-01-01

    Low temperature district heating (DH) is designed as 55/25oC for supply/return temperature to fulfill the low energy demand of future buildings. However, to secure the safety of domestic hot water, the supply temperature has to be kept around 60oC to avoid the existence of legionella, which...... reproduces rapidly at the temperature around 25oC- 45 oC. After several outbreaks of pheumonia and fever caused by legionella bacteria, most countries require 60 oC in the network and 50-55 oC at the faucets with periodic flush by hot water above 60 oC as disinfection solution. That makes obstacles of low...... temperature DH implementation. Therefore, effective solution of legionella bacteria is in urgent demand. To select optimal disinfection treatments for certain cases which are quite different in dimension or purpose of use, various methods were reviewed, including shock hyperchlorination, super heating...

  2. Application of ozonation process for the removal of Legionella pneumophila from water

    Directory of Open Access Journals (Sweden)

    Mohammad Safaee

    2015-09-01

    Full Text Available Background: Legionella pneumophila mortality and morbidity is a health concern worldwide. Due to the role of water in transmission of Legionenlla, several techniques have been used for water disinfection. This research was aimed to analyze the efficacy of ozonation process and the effects of bacterial density, contact time and pH on the removal of Legionella pneumophila from water. Methods: Legionella pneumophila was isolated from hospital water line and spiked into sterile drinking water with 300, 700 and 1000 CFU/ml densities. Ozonation was conducted within 1 L batch glass reactor with injection of 5 mg/h and contact time of 5 to 30 minutes at pH = 5, 7 and 9. Legionella culture was performed in supplemented BCYE containing GVPC and thermal treatment. After ozonation, the developed colonies were identified via biochemical and morphological tests. Results: In pH =5, the contact time 25 min and pH= 7 as well as the contact time 30 min, increase of legionella density from 300 to 1000 CFU/ml led to the reduction of removal efficiency from 100 to 87% and 100 to 82%, respectively. In pH=9 and contact time 20 min with the same bacterial density, 300 to 1000 CFU/ml, the disinfection efficacy was decreased from 100 to 91.5 %. Conclusion: Ozonation is an appropriate technique for elimination of legionella from water. The increased bacterial density led to the reduction of removal efficiency. The lowest and highest performance rates were obtained in pH=7 and 9, respectively.

  3. Is Venice an ideal habitat for Legionella pneumophila?

    Science.gov (United States)

    Ziprani, C; Selle, V; Palazzi, B; Capretta, F; Ghio, L; Flora, M E; Baldo, V

    2012-09-01

    Legionella bacterium manifests itself in Legionnaire's disease and Pontiac fever, it is mainly found and transmitted by aerosol produced in cooling towers, water distribution plants and medical equipment, and it affects mainly elder persons in poor health. The population of Venice Local Health Unit was divided in two areas of study and the incidence of legionellosis in residents of Venice historical centre (Distretto Sanitario 1) and in residents of the mainland and coastal areas (Distretti Sanitari 2, 3, 4) was calculated. The cases were those notified to the Public Health Unit by law, and the population of residents was that of the eligible for health care in the archives of the Local Health Unit. Only cases of legionellosis in residents who had not travelled in the 10 days previous of the onset of disease, and not related to nosocomial clusters were considered. The standardized incidence ratio was then calculated and confidence interval were defined by Poisson distribution. Given the population of the two areas, 59801 in Distretto Sanitario 1 and 237555 in Distretti 2, 3, 4, the raw incidence of disease is respectively 87 per 100000 and 20 per 100000 in time 2002-2010. The standardized incidence ratio for the population of Distretto Sanitario 1 vs the remaining population is 4.3. The difference in risk of getting the disease in this two residential areas geographically very close, is probably related to the different buildings' characteristics, old and difficult to maintain in Venice historical centre.

  4. Growth of Legionella pneumophila in Acanthamoeba castellanii enhances invasion.

    Science.gov (United States)

    Cirillo, J D; Falkow, S; Tompkins, L S

    1994-01-01

    Legionella pneumophila is considered to be a facultative intracellular parasite. Therefore, the ability of these bacteria to enter, i.e., invade, eukaryotic cells is expected to be a key pathogenic determinant. We compared the invasive ability of bacteria grown under standard laboratory conditions with that of bacteria grown in Acanthamoeba castellanii, one of the protozoan species that serves as a natural host for L. pneumophila in the environment. Amoeba-grown L. pneumophila cells were found to be at least 100-fold more invasive for epithelial cells and 10-fold more invasive for macrophages and A. castellanii than were L. pneumophila cells grown on agar. Comparison of agar- and amoeba-grown L. pneumophila cells by light and electron microscopy demonstrated dramatic differences in the morphology and structure of the bacteria. Analyses of protein expression in the two strains of bacteria suggest that these phenotypic differences may be due to the expression of new proteins in amoeba-grown L. pneumophila cells. In addition, the amoeba-grown bacteria were found to enter macrophages via coiling phagocytosis at a higher frequency than agar-grown bacteria did. Replication of L. pneumophila in protozoans present in domestic water supplies may be necessary to produce bacteria that are competent to enter mammalian cells and produce human disease. Images PMID:8039895

  5. Legionella pneumophila prevents proliferation of its natural host Acanthamoeba castellanii

    Science.gov (United States)

    Mengue, Luce; Régnacq, Matthieu; Aucher, Willy; Portier, Emilie; Héchard, Yann; Samba-Louaka, Ascel

    2016-01-01

    Legionella pneumophila is a ubiquitous, pathogenic, Gram-negative bacterium responsible for legionellosis. Like many other amoeba-resistant microorganisms, L. pneumophila resists host clearance and multiplies inside the cell. Through its Dot/Icm type IV secretion system, the bacterium injects more than three hundred effectors that modulate host cell physiology in order to promote its own intracellular replication. Here we report that L. pneumophila prevents proliferation of its natural host Acanthamoeba castellanii. Infected amoebae could not undergo DNA replication and no cell division was observed. The Dot/Icm secretion system was necessary for L. pneumophila to prevent the eukaryotic proliferation. The absence of proliferation was associated with altered amoebal morphology and with a decrease of mRNA transcript levels of CDC2b, a putative regulator of the A. castellanii cell cycle. Complementation of CDC28-deficient Saccharomyces cerevisiae by the CDC2b cDNA was sufficient to restore proliferation of CDC28-deficient S. cerevisiae and suggests for the first time that CDC2b from A. castellanii could be functional and a bona fide cyclin-dependent kinase. Hence, our results reveal that L. pneumophila impairs proliferation of A. castellanii and this effect could involve the cell cycle protein CDC2b. PMID:27805070

  6. An investigation of virulence factors of Legionella pneumophila environmental isolates

    Directory of Open Access Journals (Sweden)

    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.

  7. Rash, disseminated intravascular coagulation and legionella: Episode 10 and a rewind into the past

    Directory of Open Access Journals (Sweden)

    Prashanth M. Thalanayar

    2015-01-01

    Full Text Available Legionella pneumophila is the most common cause of legionellosis and is one of the organisms causing atypical pneumonia. We report the presentation of disseminated intravascular coagulation (DIC and skin rash in a single case of severe Legionella pneumonia. The unique clinical presentation of a diffuse rash diagnosed as purpura fulminans and the unpredictable variations encountered during the diagnostic work-up of the case make this write-up crucial. This article synthesizes all reported cases of L. pneumonia associated with cutaneous manifestations as well as cases presenting with DIC. Furthermore, this manuscript illustrates the correlation between cutaneous and coagulopathic manifestations, and morbidity and mortality from L. pneumonia.

  8. Prediction of the origin of French Legionella pneumophila strains using a mixed-genome microarray.

    Science.gov (United States)

    Den Boer, Jeroen W; Euser, Sjoerd M; Nagelkerke, Nico J; Schuren, Frank; Jarraud, Sophie; Etienne, Jerome

    2013-07-01

    Legionella is a water and soil bacterium that can infect humans, causing a pneumonia known as Legionnaires' disease. The pneumonia is almost exclusively caused by the species L. pneumophila, of which serogroup 1 is responsible for 90% of patients. Within serogroup 1, large differences in prevalence in clinical isolates have been described. A recent study, using a Dutch Legionella strain collection, identified five virulence associated markers. In our study, we verify whether these five Dutch markers can predict the patient or environmental origin of a French Legionella strain collection. In addition, we identify new potential virulence markers and verify whether these can predict better. A total of 219 French patient isolates and environmental strains were compared using a mixed-genome micro-array. The micro-array data were analysed to identify predictive markers, using a Random Forest algorithm combined with a logistic regression model. The sequences of the identified markers were compared with eleven known Legionella genomes, using BlastN and BlastX; the functionality for each of the predictive markers was checked in the literature. The five Dutch markers insufficiently predicted the patient or environmental origin of the French Legionella strains. Subsequent analyses identified four predictive markers for the French collection that were used for the logistic regression model. This model showed a negative predictive value of 91%. Three of the French markers differed from the Dutch markers, one showed considerable overlap and was found in one of the Legionella genomes (Lorraine strain). This marker encodes for a structural toxin protein RtxA, described for L. pneumophila as a factor involved in virulence and entry in both human cells and amoebae. The combination of a mixed-genome micro-array and statistical analysis using a Random Forest algorithm has identified virulence markers in a consistent way. The Lorraine strain and related Dutch and French Legionella

  9. Detection of Legionella bozemanae, a New Cause of Septic Arthritis, by PCR Followed by Specific Culture

    DEFF Research Database (Denmark)

    Just, Søren Andreas; Knudsen, John Bonde; Uldum, Søren Anker

    2012-01-01

    Legionella bozemanae is a rare isolate in clinical specimens. We describe a case of joint infection due to L. bozemanae in an immunocompromised patient with dermatomyositis. Without the use of PCR screening or culture on specialized medium, the organism would not have been detected.......Legionella bozemanae is a rare isolate in clinical specimens. We describe a case of joint infection due to L. bozemanae in an immunocompromised patient with dermatomyositis. Without the use of PCR screening or culture on specialized medium, the organism would not have been detected....

  10. Survey of Naegleria and its resisting bacteria-Legionella in hot spring water of Taiwan using molecular method.

    Science.gov (United States)

    Huang, Shih-Wei; Hsu, Bing-Mu

    2010-05-01

    Naegleria is a free-living amoebae existing in soil and aquatic environments. Within the genus Naegleria, N. fowleri is most recognized as potential human pathogen causing primary amoebic meningoencephalitis (PAM). Furthermore, the Naegleria spp. can serve as vehicles for facultative pathogens, such as Legionella. In this study, we identified Naegleria and Legionella based on the PCR amplification with a genus-specific primer pair and investigated the distribution of Naegleria and Legionella at five spring recreation areas in Taiwan. In this study of hot spring and other water sources in Taiwan, five Naegleria spp. were detected in 15 (14.2%) of the water samples. The most frequently detected was N. lovaniensis (n = 6), followed by N. australiensis (n = 5), and then N. clarki (n = 2). N. americana and N. pagei were detected once, respectively. The pathogenic species N. fowleri was not detected; however, N. australiensis considered to be a potential pathogen species in humans was found. Legionella spp., an endosymbiont of Naegleria, was detected in 19 (17.9%) of the water samples in this study. Overall, 5.7% of the water samples contained both Naegleria and Legionella. The Legionella spp. identified were L. pneumophila and L. erythra. Results of this survey confirm the existence of Naegleria and Legionella in Taiwan spring recreation areas. It should be considered a potential threat for health associated with human activities in spring recreation areas of Taiwan.

  11. Impact of non-Legionella bacteria on the uptake and intracellular replication of Legionella pneumophila in Acanthamoeba castellanii and Naegleria lovaniensis.

    Science.gov (United States)

    Declerck, P; Behets, J; Delaedt, Y; Margineanu, A; Lammertyn, E; Ollevier, F

    2005-11-01

    In aquatic environments, Legionella pneumophila survives, in association with other bacteria, within biofilms by multiplying in free-living amoebae. The precise mechanisms underlying several aspects of the uptake and intracellular replication of L. pneumophila in amoebae, especially in the presence of other bacteria, remain unknown. In the present study, we examined the competitive effect of selected non-Legionella bacteria (Escherichia coli, Aeromonas hydrophila, Flavobacterium breve, and Pseudomonas aeruginosa) on the uptake of L. pneumophila serogroup 1 by the amoebae Acanthamoeba castellanii and Naegleria lovaniensis. We also investigated their possible influence on the intracellular replication of L. pneumophila in both amoeba species. Our results showed that the non-Legionella bacteria did not compete with L. pneumophila for uptake, suggesting that the amoeba hosts took in L. pneumophila through a specific and presumably highly efficient uptake mechanism. Living and heat-inactivated P. aeruginosa best supported the replication of L. pneumophila in N. lovaniensis and A. castellanii, respectively, whereas for both amoeba species, E. coli yielded the lowest number of replicated L. pneumophila. Furthermore, microscopic examination showed that 100% of the A. castellanii and only 2% of the N. lovaniensis population were infected with L. pneumophila at the end of the experiment. This study clearly shows the influence of some non-Legionella bacteria on the intracellular replication of L. pneumophila in A. castellanii and N. lovaniensis. It also demonstrates the different abilities of the two tested amoeba species to serve as a proper host for the replication and distribution of the human pathogen in man-made aquatic environments such as cooling towers, shower heads, and air conditioning systems with potential serious consequences for human health.

  12. Environmental surveillance of Legionella pneumophila in two Italian hospitals

    Directory of Open Access Journals (Sweden)

    Marina Tesauro

    2010-01-01

    Full Text Available The aim of this study was to identify the most effective disinfection protocol to reduce the presence of Legionella pneumophila in the water system of two Italian hospitals. From 2004 to 2009, 271 samplings of hot water were carried out in 11 hospital units to detect the presence of L. pneumophila. Additionally, water samples collected from one boiler outlet and the hot water recirculation were tested. From 2004 to 2009, L. pneumophila was present in 37% of the samples. Of these, 68.3% and 18.8% were positive for serogroups 2-14 and 1, respectively. Furthermore, 12.9% of the samples were positive for both serogroups. Finally, a maximal count of 10(4 CFU/L was measured in the most distal sites. To reduce L. pneumophila colonization, a two-year long hyperchlorination (2004-2006 was carried out. Moreover, from June 2005 until now, continuous maintenance of boilers and tanks, substitution of the shower heads and increase of the boiler outlet temperature to 60 ºC were performed. All these treatments led to a marked reduction of L. pneumophila colonization in the short but not in the medium-long term. Only the use of chlorine dioxide led, after four years, to a reduction of the loads of L. pneumophila to values below 100 CFU/L. However, in the distal sites a persistent degree of colonization (maximum value 700 CFU/L, average 600 CFU/L was observed probably due to the presence of L. pneumophila in the stagnant water in dead legs. In conclusion, data show that long-term chlorination of hot water sources together with carefully aimed maintenance of water pipes can lead to an effective reduction of L. pneumophila concentration in hospital water systems.

  13. Environmental surveillance of Legionella pneumophila in two Italian hospitals.

    Science.gov (United States)

    Tesauro, Marina; Bianchi, Annalisa; Consonni, Michela; Pregliasco, Fabrizio; Galli, Maria Gabriella

    2010-01-01

    The aim of this study was to identify the most effective disinfection protocol to reduce the presence of Legionella pneumophila in the water system of two Italian hospitals. From 2004 to 2009, 271 samplings of hot water were carried out in 11 hospital units to detect the presence of L. pneumophila. Additionally, water samples collected from one boiler outlet and the hot water recirculation were tested. From 2004 to 2009, L. pneumophila was present in 37% of the samples. Of these, 68.3% and 18.8% were positive for serogroups 2-14 and 1, respectively. Furthermore, 12.9% of the samples were positive for both serogroups. Finally, a maximal count of 10(4) CFU/L was measured in the most distal sites. To reduce L. pneumophila colonization, a two-year long hyperchlorination (2004-2006) was carried out. Moreover, from June 2005 until now, continuous maintenance of boilers and tanks, substitution of the shower heads and increase of the boiler outlet temperature to 60 °C were performed. All these treatments led to a marked reduction of L. pneumophila colonization in the short but not in the medium-long term. Only the use of chlorine dioxide led, after four years, to a reduction of the loads of L. pneumophila to values below 100 CFU/L. However, in the distal sites a persistent degree of colonization (maximum value 700 CFU/L, average 600 CFU/L) was observed probably due to the presence of L. pneumophila in the stagnant water in dead legs. In conclusion, data show that long-term chlorination of hot water sources together with carefully aimed maintenance of water pipes can lead to an effective reduction of L. pneumophila concentration in hospital water systems.

  14. Mutational paths towards increased fluoroquinolone resistance in Legionella pneumophila.

    Science.gov (United States)

    Almahmoud, Iyad; Kay, Elisabeth; Schneider, Dominique; Maurin, Max

    2009-08-01

    Fluoroquinolone resistance has been poorly studied in Legionella pneumophila, an intracellular pathogen responsible for legionellosis. Our goal was to further characterize molecular mechanisms involved in fluoroquinolone resistance in this species. Eight independent lineages were founded from a common fluoroquinolone-susceptible L. pneumophila ancestor and propagated by serial passages in moxifloxacin-containing culture medium. We identified the substituted mutations that affected the DNA topoisomerase II-encoding genes, determined the order of substitution of the mutations leading to the stepwise MIC increases of moxifloxacin over evolutionary time and demonstrated their direct involvement in the resistance process. Adaptation occurred through parallel stepwise increases in the moxifloxacin MICs up to 512-fold the MIC for the parental strain. Mutations affected the topoisomerase II-encoding genes gyrA, parC and gyrB, reflecting a high degree of genetic parallelism across the independent lineages. During evolution, the T83I change in GyrA occurred first, followed by G78D or S80R in ParC and D87N in GyrA, or S464Y or D426N in GyrB. By constructing isogenic strains, we showed that the progressive increase in resistance was linked to a precise order of mutation substitution, but also to the co-existence of several subpopulations of bacteria bearing different mutations. Specific mutational trajectories were identified, strongly suggesting that intermolecular epistatic interactions between DNA topoisomerases underlie the mechanism of fluoroquinolone resistance in L. pneumophila. Our results suggest that L. pneumophila has strong potential to become resistant to fluoroquinolone compounds and warrant further investigation of resistance in clinical and environmental strains of this pathogen.

  15. Tunge- og stemmebåndsparese efter endotrakeal intubation for Legionella-pneumoni

    DEFF Research Database (Denmark)

    Sønnichsen, Rikke; Lauritsen, Anne Oberg

    2013-01-01

    Extracranial involvement of the hypoglossal nerve and recurrent laryngeal branch of the vagal nerve can be a complication of anaesthetic airway management (Tapia's syndrome) or focal involvement due to Legionella infection. We present a patient with bilateral hypoglossal and unilateral recurrent ...

  16. Legionella - Combating their spreading in domestic water systems; Legionellen - Bekaempfung ihrer Verbreitung in Hausinstallationen

    Energy Technology Data Exchange (ETDEWEB)

    Haas, R.

    2003-07-01

    This comprehensive article discusses the ways in which water systems that are infected by legionella bacteria can be cleaned up and the very high demands placed on companies that conduct the disinfection. The various species of legionella bacteria are listed and their rates of multiplication at various temperatures presented. Also, the time taken to kill them off at various temperatures is examined. The history of the legionnaires illness, the discovery of its cause and various other topics concerned such as risk factors, endangered locations - such as hospitals, nursing homes, prisons and residential estates with centralised hot water systems - are examined, as is the presence of legionella in aerosols and biofilms. Ways of avoiding and combating legionella using thermal, chemical as well as physical methods such UV-light and ozone disinfection and membrane filters are discussed. Recommendations are made on which materials should or should not be used in the hot water systems. Specific recommendations are made concerning the design of hot water boilers, temperature controllers and the installations and fittings to be used. Methods of ensuring that hot water systems are sufficiently protected, such as by avoiding long pipes and standing water, are discussed.

  17. Receptor-mediated uptake of Legionella pneumophila by Acanthamoeba castellanii and Naegleria lovaniensis.

    Science.gov (United States)

    Declerck, P; Behets, J; De Keersmaecker, B; Ollevier, F

    2007-12-01

    Investigation of the attachment and uptake of Legionella pneumophila by Acanthamoeba castellanii and Naegleria lovaniensis, as these are two critical steps in the subsequent bacterial survival in both amoeba hosts. Initially, the mode of Legionella uptake was examined using inhibitors of microfilament-dependent and receptor-mediated uptake phagocytosis. Secondly, the minimum saccharide structure to interfere with L. pneumophila uptake was determined by means of selected saccharides. Bacterial attachment and uptake by each of the amoeba species occurred through a receptor-mediated endocytosis, which required de novo synthesis of host proteins. Legionella pneumophila showed a high affinity to the alpha1-3D-mannobiose domain of the mannose-binding receptor located on A. castellanii. In contrast, L. pneumophila bacteria had a high affinity for the GalNAcbeta1-4Gal domain of the N-acetyl-D-galactosamine receptor of N. lovaniensis. Our data pointed to a remarkable adaptation of L. pneumophila to invade different amoeba hosts, as the uptake by both amoeba species is mediated by two different receptor families. The fact that L. pneumophila is taken up by two different amoeba species using different receptor families adds further complexity to the host-parasite interaction process, as 14 amoeba species are known to be appropriate Legionella hosts.

  18. Characterization of an acetyltransferase that detoxifies aromatic chemicals in Legionella pneumophila

    DEFF Research Database (Denmark)

    Kubiak, Xavier Jean Philippe; Dervins-Ravault, Delphine; Pluvinage, Benjamin

    2012-01-01

    Legionella pneumophila is an opportunistic pathogen and the causative agent of Legionnaires' disease. Despite being exposed to many chemical compounds in its natural and man-made habitats (natural aquatic biotopes and man-made water systems), L. pneumophila is able to adapt and survive in these e...

  19. Analysis and research on promising solutions of low temperature district heating without risk of legionella

    DEFF Research Database (Denmark)

    Yang, Xiaochen; Li, Hongwei; Fog, Jette M.

    2014-01-01

    resources. The most crucial restriction for applying low temperature district heating is the worry about the breakout of legionella, which exists preferably in low temperature hot water systems. Several novel techniques such as electric tracing and flat station were investigated for such dilemma. The pros...

  20. Multiple regression as a preventive tool for determining the risk of Legionella spp.

    Directory of Open Access Journals (Sweden)

    Enrique Gea-Izquierdo

    2012-04-01

    Full Text Available To determine the interrelationship between health & hygiene conditions for prevention of legionellosis, the compositionof materials used in water distribution systems, the water origin and Legionella pneumophila risk. Material and methods. Include adescriptive study and multiple regression analysis on a sample of golf course sprinkler irrigation systems (n=31 pertaining to hotelslocated on the Costa del Sol (Malaga, Spain. The study was carried out in 2009. Results. Presented a significant lineal relation, withall the independent variables contributing significantly (p<0.05 to the model’s fit. The relationship between water type and the risk ofLegionella, as well as the material composition and the latter, is lineal and positive. In contrast, the relationship between health-hygieneconditions and Legionella risk is lineal and negative. Conclusion. The characterization of Legionella pneumophila concentration, asdefined by the risk in water and through use of the predictive method, can contribute to the consideration of new influence variables inthe development of the agent, resulting in improved control and prevention of the disease.

  1. Legionella detection in oropharyngeal aspirates of transplant patients prior to surgery

    Czech Academy of Sciences Publication Activity Database

    Jarešová, M.; Hložánek, I.; Stříž, I.; Petříčková, Kateřina; Kocmoud, Z.

    2006-01-01

    Roč. 25, - (2006), s. 63-64 ISSN 0934-9723 R&D Projects: GA ČR GA310/01/0095; GA MŠk LN00B030 Institutional research plan: CEZ:AV0Z50200510 Keywords : legionella * organ transplantation * pcr Subject RIV: EE - Microbiology, Virology Impact factor: 2.330, year: 2006

  2. Occurrence of Legionella pneumophila and Hartmannella vermiformis in fresh water environments and their interactions in biofilms

    NARCIS (Netherlands)

    Kuiper, M.W.

    2006-01-01

    Legionella pneumophila, the causative agent of Legionnaires’ disease, is widespread in natural fresh water environments and is also frequently found in man-made water systems. Microbial biofilms and protozoa are known to play a major role in the proliferation of L. pneumophila. Biofilms provide

  3. Isolation of Legionella pneumophila from hospital cooling towers in Johor, Malaysia.

    Science.gov (United States)

    Abdul Samad, B H; Suhaili, M R; Baba, N; Rajasekaran, G

    2004-08-01

    Water-based cooling towers and their water supply at two hospitals in Johor were surveyed for the presence Legionella pneumophila. L. pneumophila were grown from 19 (76%) out of 25 collected water samples. One hospital cooling tower was contaminated with L. pneumophila serogroup 1.

  4. Predictors of positive or negative legionella urinary antigen test in community-acquired pneumonia.

    Science.gov (United States)

    Roed, Torsten; Schønheyder, Henrik C; Nielsen, Henrik

    2015-07-01

    Legionella pneumonia remains a diagnostic challenge. The legionella urinary antigen test (LUT) primarily detects Legionella pneumophila serogroup 1, accounting for 64% of Danish cases, and is often the only legionella test performed. We aimed to identify variables predictive of a positive or negative test result and to explore how the LUT was used in clinical practice. The study was an audit-based cohort study. LUT-positive patients were compared with three randomly selected age- and gender-matched LUT-negative referent patients admitted at a Danish university hospital during 2003-2013. Data were extracted from charts and databases. Positive and negative likelihood ratios (LR+ and LR-) were calculated. For CURB-65 and sepsis, sensitivity analyses were made due to incomplete data. In all, 25 cases were compared with 75 referents. Factors associated with LUT positivity included recent travel outside Scandinavia (LR + 5.3), Na(+) 200 mg/L (LR + 3.5), temperature > 39 °C (LR + 3.5), and CURB-65 score ≥ 3 (LR + 3.0-15.0, depending on the model). Decreasing the likelihood of LUT positivity were CRP LUT. The LUT is often used inappropriately and should be accompanied by PCR analysis.

  5. Genetic relatedness of Legionella longbeachae isolates from human and environmental sources in Australia

    International Nuclear Information System (INIS)

    Lanser, J.A.; Doyle, R.; Sangster, N.; Steele, T.W.; Adams, M.

    1990-01-01

    The genetic relatedness of Legionella longbeachae isolated in Australia since 1987 was investigated by restriction fragment length polymorphism (RFLP) analysis and allozyme electrophoresis. Three radiolabeled probes were used in Southern hybridizations for the RFLP studies. They were Escherichia coli 16S and 23S rRNA and cloned fragments of L. longbeachae selected empirically from genomal banks in lambda and a cosmid. The legionellae included in the study comprised 11 Legionella longbeachae serogroup 1 organisms isolated form humans, 28 L. longbeachae serogroup 1 isolates from environmental sources, 3 L. longbeachae serogroup 2 environmental isolates. These were compared with the American Type Culture Collection reference strains of both serogroups and some other related Legionella species. Results of allozyme and RFLP analysis showed that all the isolates from humans and all but three of the environmental L. longbeachae serogroup 1 isolates were closely related. They were also closely related to L. longbeachae serogroup 1 ATCC 33462. There was wider variation among the three L. longbeachae serogroup 2 environmental isolates. One of these was closely related to L. longbeachae serogroup 2 ATCC 33484. RFLP studies with the rRNA probe provided the most discrimination among isolates but did not distinguish between the two serogroups

  6. Field inversion gel electrophoretic analysis of Legionella pneumophila strains associated with nosocomial legionellosis in children.

    Science.gov (United States)

    Green, M; Wald, E R; Dashefsky, B; Barbadora, K; Wadowsky, R M

    1996-01-01

    Two nosocomial cases of Legionnaires' disease occurred in children. Legionella pneumophila serogroup 1 was isolated from both patients and 30 of 39 plumbing system sites in the hospital. The patient and hospital environmental isolates yielded identical field inversion gel electrophoretic patterns which differed from patterns observed with epidemiologically unrelated strains.

  7. Septic Arthritis Caused by Legionella dumoffii in a Patient with Systemic Lupus Erythematosus-Like Disease

    NARCIS (Netherlands)

    Flendrie, M.; Jeurissen, S.M.F.; Franssen, M.; Kwa, D.; Klaassen, C.; Vos, F.

    2011-01-01

    We describe a patient with systemic lupus erythematosus (SLE)-like disease on immunosuppressive treatment who developed septic arthritis of the knee involving Legionella dumoffii. Cultures initially remained negative. A broad-range 16S PCR using synovial fluid revealed L. dumoffii rRNA genes, a

  8. Prevalence study of Legionella spp. contamination in ferries and cruise ships

    Directory of Open Access Journals (Sweden)

    Masia Maria

    2006-04-01

    Full Text Available Abstract Background In the last years, international traffic volume has significantly increased, raising the risk for acquisition of infectious diseases. Among travel-associated infections, increased incidence of legionellosis has been reported among travellers. Aim of our study was: to describe the frequency and severity of Legionella spp. contamination in ferries and cruise ships; to compare the levels of contamination with those indicated by the Italian ministerial guidelines for control and prevention of legionellosis, in order to assess health risks and to adopt control measures. Method A prevalence study was carried out on 9 ships docked at the seaports of northern Sardinia in 2004. Water samples were collected from critical sites: passenger cabins, crew cabins, kitchens, coffee bars, rooms of the central air conditioning system. It was performed a qualitative and quantitative identification of Legionella spp. and a chemical, physical and bacteriological analysis of water samples. Results Forty-two percent (38/90 water samples were contaminated by Legionella spp.. Positive samples were mainly drawn from showers (24/44, washbasins (10/22. L. pneumophila was isolated in 42/44 samples (95.5%, followed by L. micdadei (4.5%. Strains were identified as L. pneumophila serogroup 6 (45.2%; 19 samples, 2–14 (42.9%, 5 (7.1% and 3 (4.8%. Legionella spp. load was high; 77.8% of the water samples contained > 104 CFU/L. Low residual free chlorine concentration (0–0,2 mg/L was associated to a contamination of the 50% of the water samples. Conclusion Legionella is an ubiquitous bacterium that could create problems for public health. We identified Legionella spp. in 6/7 ferries. Microbial load was predominantly high (> 104 CFU/L or ranging from 103 to 104 CFU/L. It is matter of concern when passengers are subjects at risk because of Legionella spp. is an opportunist that can survive in freshwater systems; high bacterial load might be an important

  9. A quantitative model of intracellular growth of Legionella pneumophila in Acanthamoeba castellanii.

    Science.gov (United States)

    Moffat, J F; Tompkins, L S

    1992-01-01

    A model of intracellular growth for Legionella pneumophila in Acanthamoeba castellanii has been developed and provides a quantitative measure of survival and replication after entry. In this model, Acanthamoeba monolayers were incubated with bacteria in tissue culture plates under nutrient-limiting conditions. Gentamicin was used to kill extracellular bacteria following the period of incubation, and the number of intracellular bacteria was determined following lysis of amebae. Intracellular growth of virulent L. pneumophila and other wild-type Legionella species was observed when the assay was performed at 37 degrees C. At room temperature, none of the Legionella strains tested grew intracellularly, while an avirulent L. pneumophila strain was unable to replicate in this assay at either temperature. The effect of nutrient limitation on A. castellanii during the assay prevented multiplication of the amebae and increased the level of infection by Legionella spp. The level of infection of the amebae was directly proportional to the multiplicity of infection with bacteria; at an inoculum of 1.03 x 10(7) bacteria added to wells containing 1.10 x 10(5) amebae (multiplicity of infection of 100), approximately 4.4% of A. castellanii cells became infected. Cytochalasin D reduced the uptake of bacteria by the amebae primarily by causing amebae to lift off the culture dish, reducing the number of target hosts; methylamine also reduced the level of initial infection, yet neither inhibitor was able to prevent intracellular replication of Legionella spp. Consequently, once the bacteria entered the cell, only lowered temperature could restrict replication. This model of intracellular growth provides a one-step growth curve and should be useful to study the molecular basis of the host-parasite interaction. PMID:1729191

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

    Science.gov (United States)

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

    2014-11-15

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

  11. Contamination of the cold water distribution system of health care facilities by Legionella pneumophila: do we know the true dimension?

    Science.gov (United States)

    Arvand, M; Jungkind, K; Hack, A

    2011-04-21

    German water guidelines do not recommend routine assessment of cold water for Legionella in healthcare facilities, except if the water temperature at distal sites exceeds 25°C. This study evaluates Legionella contamination in cold and warm water supplies of healthcare facilities in Hesse, Germany, and analyses the relationship between cold water temperature and Legionella contamination. Samples were collected from four facilities, with cases of healthcare-associated Legionnaires' disease or notable contamination of their water supply. Fifty-nine samples were from central lines and 625 from distal sites, comprising 316 cold and 309 warm water samples. Legionella was isolated from central lines in two facilities and from distal sites in four facilities. 17% of all central and 32% of all distal samples were contaminated. At distal sites, cold water samples were more frequently contaminated with Legionella (40% vs 23%, p contamination rate. 35% of cold water samples under 20 °C at collection were contaminated. Our data highlight the importance of assessing the cold water supply of healthcare facilities for Legionella in the context of an intensified analysis.

  12. [Evaluation of a Legionella outbreak emerged in a recently opening hotel].

    Science.gov (United States)

    Erdoğan, Haluk; Arslan, Hande

    2013-04-01

    Legionnaires' disease (LD) is a systemic infection caused by Legionella species especially colonized in the water systems. Hotels are common locations in which waterwork-associated sporadic or epidemic legionellosis can be detected. The aim of this study was to evaluate a small Legionella outbreak emerged in a recently opened 600-bed hotel in Alanya, a touristic county in Mediterranean part of Turkey. A 66 years old male patient who stayed in this hotel opened on May 15th, 2009, was admitted to our hospital on May 21st, 2009 with the complaints of high fever, headache and diarrhea lasting for three days. Since chest X-ray revealed non-homogenous density increase in left middle and inferior zone, the patient was diagnosed as atypical pneumoniae and LD was confirmed by positive urinary Legionella antigen test (Card test, BinaxNOW®Legionella Urinary Antigen Test; Alere Co, USA) result. Following the identification of the index case, the records of our hospital were reviewed and revealed another case being treated with the diagnosis of community acquired pneumonia who was also the guest of the same hotel. This patient was then diagnosed as LD by positive urinary antigen test. Since new cases were identified during the following days (May 22, 25 and 26) the Antalya County Health Department and hotel management were informed about a cluster of LD. In addition subsequent investigation for environmental surveillance and water sampling were conducted. The LD diagnosis and environmental inspections were performed according to the procedure described in the guideline from "Turkish Ministry of Health Travel-Associated Legionnaires' Disease Control Programme". Five definitive cases and one presumptive case of LD were identified during the outbreak period (May 20-26, 2009). All of the cases were successfully treated (intravenous ciprofloxacin or levofloxacin or clarithromycin), however one patient died due to sudden death during sleep after being discharged. Since sputum

  13. Effect of heat shock on hot water plumbing microbiota and Legionella pneumophila control.

    Science.gov (United States)

    Ji, Pan; Rhoads, William J; Edwards, Marc A; Pruden, Amy

    2018-02-09

    Heat shock is a potential control strategy for Legionella pneumophila in hot water plumbing systems. However, it is not consistently effective, with little understanding of its influence on the broader plumbing microbiome. Here, we employed a lab-scale recirculating hot water plumbing rig to compare the pre- and post-"heat shock" (i.e., 40 → 60 → 40 °C) microbiota at distal taps. In addition, we used a second plumbing rig to represent a well-managed system at 60 °C and conducted a "control" sampling at 60 °C, subsequently reducing the temperature to 40 °C to observe the effects on Legionella and the microbiota under a simulated "thermal disruption" scenario. According to 16S rRNA gene amplicon sequencing, in the heat shock scenario, there was no significant difference or statistically significant, but small, difference in the microbial community composition at the distal taps pre- versus post-heat shock (both biofilm and water; weighted and unweighted UniFrac distance matrices). While heat shock did lead to decreased total bacteria numbers at distal taps, it did not measurably alter the richness or evenness of the microbiota. Quantitative PCR measurements demonstrated that L. pneumophila relative abundance at distal taps also was not significantly different at 2-month post-heat shock relative to the pre-heat shock condition, while relative abundance of Vermamoeba vermiformis, a known Legionella host, did increase. In the thermal disruption scenario, relative abundance of planktonic L. pneumophila (quantitative PCR data) increased to levels comparable to those observed in the heat shock scenario within 2 months of switching long-term operation at 60 to 40 °C. Overall, water use frequency and water heater temperature set point exhibited a stronger effect than one-time heat shock on the microbial composition and Legionella levels at distal taps. While heat shock may be effective for instantaneous Legionella control and reduction in total

  14. Molecular characterization of Legionella pneumophila-induced interleukin-8 expression in T cells

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    Mukaida Naofumi

    2010-01-01

    Full Text Available Abstract Background Legionella pneumophila is the causative agent of human Legionnaire's disease. During infection, the bacterium invades macrophages and lung epithelial cells, and replicates intracellularly. However, little is known about its interaction with T cells. We investigated the ability of L. pneumophila to infect and stimulate the production of interleukin-8 (IL-8 in T cells. The objective of this study was to assess whether L. pneumophila interferes with the immune system by interacting and infecting T cells. Results Wild-type L. pneumophila and flagellin-deficient Legionella, but not L. pneumophila lacking a functional type IV secretion system Dot/Icm, replicated in T cells. On the other hand, wild-type L. pneumophila and Dot/Icm-deficient Legionella, but not flagellin-deficient Legionella or heat-killed Legionella induced IL-8 expression. L. pneumophila activated an IL-8 promoter through the NF-κB and AP-1 binding regions. Wild-type L. pneumophila but not flagellin-deficient Legionella activated NF-κB, p38 mitogen-activated protein kinase (MAPK, Jun N-terminal kinase (JNK, and transforming growth factor β-associated kinase 1 (TAK1. Transfection of dominant negative mutants of IκBα, IκB kinase, NF-κB-inducing kinase, TAK1, MyD88, and p38 MAPK inhibited L. pneumophila-induced IL-8 activation. Inhibitors of NF-κB, p38 MAPK, and JNK blocked L. pneumophila-induced IL-8 expression. In addition, c-Jun, JunD, cyclic AMP response element binding protein, and activating transcription factor 1, which are substrates of p38 MAPK and JNK, bound to the AP-1 site of the IL-8 promoter. Conclusions Taken together, L. pneumophila induced a flagellin-dependent activation of TAK1, p38 MAPK, and JNK, as well as NF-κB and AP-1, which resulted in IL-8 production in human T cells, presumably contributing to the immune response in Legionnaire's disease.

  15. Ecological behaviour of three serogroups of Legionella pneumophila within a model plumbing system.

    Science.gov (United States)

    Messi, P; Anacarso, I; Bargellini, A; Bondi, M; Marchesi, I; de Niederhäusern, S; Borella, P

    2011-02-01

    Three Legionella pneumophila strains isolated from water samples and belonging to serogroups (sgs) 1, 6 and 9 were analysed for their capacity to colonise an experimental model simulating a domestic hot water distribution system. Ecological factors that could influence the persistence of the sgs such as intracellular life within protozoan hosts and bacterial interference by the production of antagonistic compounds were also studied. Viable counts of L. pneumophila increased both in the planktonic and in the sessile phases. Sg 6 showed a marked prevalence during the whole experiment and exhibited the highest host infection efficiency. Sg 1 was significantly less represented, but showed the highest capacity to reproduce in the protozoan hosts. Sg 9 was poorly represented and less adapted to intracellular life. Among the 14 bacteria constantly isolated in the system, five (35.7%) produced antagonistic substances against Legionella, with differences according to the bacterial strain and L. pneumophila sgs.

  16. Micro- and macromethod assays for the ecological study of Legionella pneumophila.

    Science.gov (United States)

    Guerrieri, Elisa; Bondi, Moreno; Ciancio, Claudia; Borella, Paola; Messi, Patrizia

    2005-11-01

    The survival of a strain of Legionella pneumophila (Lp-1) inoculated in artificial water microcosms was investigated with and without an amoebal host and varying environmental conditions, such as biofilm formation, amount of nutrients and incubation temperature. The results obtained using short (micromethod) and long (macromethod) term methods showed that L. pneumophila Lp-1 dies rapidly at 4 degrees C in the "macromethod" assay. When the same temperature (4 degrees C) was applied to the "micromethod" assay, L. pneumophila Lp-1 survived for three weeks, although it progressively decreased. At an incubation temperature of 30 degrees C, the aquatic environment was more favourable and better survival emerged in the "macromethod"; in contrast, this favourable temperature condition did not improve the survival of L. pneumophila Lp-1 cultured with the "micromethod". The role of the protozoa Acanthamoeba polyphaga proved to be indispensable for legionella survival only when environmental conditions become unfavourable.

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

    Directory of Open Access Journals (Sweden)

    Renaud Bigot

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

  18. Legionella bacteria in shower aerosols increase the risk of Pontiac fever among older people in retirement homes.

    Science.gov (United States)

    Bauer, M; Mathieu, L; Deloge-Abarkan, M; Remen, T; Tossa, P; Hartemann, P; Zmirou-Navier, D

    2008-10-01

    828 elderly subjects residing in nursing homes were followed up during 4 months to ascertain incidence of symptoms associated with Pontiac fever (PF) in a non-epidemic setting. The exposure situation was inhalation of Legionella bacteria while showering. An audit of the hot water system in all institutions allowed ascribing each subject to a water quality area wherefrom one shower was sampled for Legionella assays at the end of the follow-up period. Legionella were detected in water and aerosols using the culture (CFU, colony forming units) and in situ hybridization (FISH) techniques. Among 32 Pontiac-like episodes, 29 cases complied with the operational definition of PF elaborated for this study. Incidence density was 0.11 case/person-year (95% CI 0.07 to 0.15). Water concentrations greater than 105 Legionella FISH/l and 104 Legionella CFU/l were associated with an increased risk of PF (respectively RR 2.23, p = 0.05 and RR 2.39, p = 0.11, with significant dose-response patterns: p for trend <0.04). The condition also seems associated with aerosol concentrations above 103 Legionella FISH/l of air. A significantly higher risk of Pontiac-like episodes (RR 6.24, 95% CI 2.12 to 18.38) was seen for elderly subjects receiving corticosteroid therapy. The water and threshold values identified in this research could be used to inform guidance measures aimed at protecting institutionalised older people from Legionnaires' disease. Immunosuppressive therapy in the same population group can significantly enhance susceptibility to Legionella bacteria.

  19. Presence of Legionella spp. in Hot Water Networks of Different Italian Residential Buildings: A Three-Year Survey.

    Science.gov (United States)

    Totaro, Michele; Valentini, Paola; Costa, Anna Laura; Frendo, Lorenzo; Cappello, Alessia; Casini, Beatrice; Miccoli, Mario; Privitera, Gaetano; Baggiani, Angelo

    2017-10-26

    Although the European reports highlight an increase in community-acquired Legionnaires' disease cases, the risk of Legionella spp. in private houses is underestimated. In Pisa (Italy) we performed a three-year survey on Legionella presence in 121 buildings with an independent hot water production (IB); 64 buildings with a central hot water production (CB); and 35 buildings with a solar thermal system for hot water production (TB). From all the 220 buildings Legionella spp. was researched in two hot water samples collected either at the recirculation point or on the first floor and on the last floor, while the potable water quality was analysed in three cold water samples collected at the inlet from the aqueduct network, at the exit from the autoclave, and at the most remote tap. Legionella pneumophila sg1, Legionella pneumophila sg2-16, and non- pneumophila Legionella species were detected in 26% of the hot water networks, mostly in CB and TB. In these buildings we detected correlations between the presence of Legionella and the total chlorine concentration decrease and/or the increase of the temperature. Cold water resulted free from microbiological hazards, with the exception of Serratia liquefaciens and Enterobacter cloacae isolated at the exit from two different autoclaves. We observed an increase in total microbial counts at 22 °C and 37 °C between the samples collected at the most remote taps compared to the ones collected at the inlet from the aqueduct. The study highlights a condition of potential risk for susceptible categories of population and supports the need for measures of risk assessment and control.

  20. Legionella pneumophila infection of Drosophila S2 cells induces only minor changes in mitochondrial dynamics.

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    Elizabeth Wen Sun

    Full Text Available During infection of cells by Legionella pneumophila, the bacterium secretes a large number of effector proteins into the host cell cytoplasm, allowing it to alter many cellular processes and make the vacuole and the host cell into more hospitable environments for bacterial replication. One major change induced by infection is the recruitment of ER-derived vesicles to the surface of the vacuole, where they fuse with the vacuole membrane and prevent it from becoming an acidified, degradative compartment. However, the recruitment of mitochondria to the region of the vacuole has also been suggested by ultrastructural studies. In order to test this idea in a controlled and quantitative experimental system, and to lay the groundwork for a genome-wide screen for factors involved in mitochondrial recruitment, we examined the behavior of mitochondria during the early stages of Legionella pneumophila infection of Drosophila S2 cells. We found that the density of mitochondria near vacuoles formed by infection with wild type Legionella was not different from that found in dotA(- mutant-infected cells during the first 4 hours after infection. We then examined 4 parameters of mitochondrial motility in infected cells: velocity of movement, duty cycle of movement, directional persistence and net direction. In the 4 hours following infection, most of these measures were indistinguishable between wild type and dotA(-.infection. However, wild type Legionella did induce a modest shift in the velocity distribution toward faster movement compared dotA(- infection, and a small downward shift in the duty cycle distribution. In addition, wild type infection produced mitochondrial movement that was biased in the direction of the bacterial vacuole relative to dotA-, although not enough to cause a significant accumulation within 10 um of the vacuole. We conclude that in this host cell, mitochondria are not strongly recruited to the vacuole, nor is their motility

  1. Legionella pneumophila-Derived Outer Membrane Vesicles Promote Bacterial Replication in Macrophages.

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    Anna Lena Jung

    2016-04-01

    Full Text Available The formation and release of outer membrane vesicles (OMVs is a phenomenon of Gram-negative bacteria. This includes Legionella pneumophila (L. pneumophila, a causative agent of severe pneumonia. Upon its transmission into the lung, L. pneumophila primarily infects and replicates within macrophages. Here, we analyzed the influence of L. pneumophila OMVs on macrophages. To this end, differentiated THP-1 cells were incubated with increasing doses of Legionella OMVs, leading to a TLR2-dependent classical activation of macrophages with the release of pro-inflammatory cytokines. Inhibition of TLR2 and NF-κB signaling reduced the induction of pro-inflammatory cytokines. Furthermore, treatment of THP-1 cells with OMVs prior to infection reduced replication of L. pneumophila in THP-1 cells. Blocking of TLR2 activation or heat denaturation of OMVs restored bacterial replication in the first 24 h of infection. With prolonged infection-time, OMV pre-treated macrophages became more permissive for bacterial replication than untreated cells and showed increased numbers of Legionella-containing vacuoles and reduced pro-inflammatory cytokine induction. Additionally, miRNA-146a was found to be transcriptionally induced by OMVs and to facilitate bacterial replication. Accordingly, IRAK-1, one of miRNA-146a's targets, showed prolonged activation-dependent degradation, which rendered THP-1 cells more permissive for Legionella replication. In conclusion, L. pneumophila OMVs are initially potent pro-inflammatory stimulators of macrophages, acting via TLR2, IRAK-1, and NF-κB, while at later time points, OMVs facilitate L. pneumophila replication by miR-146a-dependent IRAK-1 suppression. OMVs might thereby promote spreading of L. pneumophila in the host.

  2. Geostatistics – a tool applied to the distribution of Legionella pneumophila in a hospital water system

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    Pasqualina Laganà

    2015-12-01

    Full Text Available [b]Introduction.[/b] Legionnaires’ disease is normally acquired by inhalation of legionellae from a contaminated environmental source. Water systems of large buildings, such as hospitals, are often contaminated with legionellae and therefore represent a potential risk for the hospital population. The aim of this study was to evaluate the potential contamination of [i]Legionella pneumophila[/i] (LP in a large hospital in Italy through georeferential statistical analysis to assess the possible sources of dispersion and, consequently, the risk of exposure for both health care staff and patients. [b]Materials and Method. [/b]LP serogroups 1 and 2–14 distribution was considered in the wards housed on two consecutive floors of the hospital building. On the basis of information provided by 53 bacteriological analysis, a ‘random’ grid of points was chosen and spatial geostatistics or [i]FAIk Kriging[/i] was applied and compared with the results of classical statistical analysis. [b]Results[/b]. Over 50% of the examined samples were positive for [i]Legionella pneumophila[/i]. LP 1 was isolated in 69% of samples from the ground floor and in 60% of sample from the first floor; LP 2–14 in 36% of sample from the ground floor and 24% from the first. The iso-estimation maps show clearly the most contaminated pipe and the difference in the diffusion of the different [i]L. pneumophila[/i] serogroups. [b]Conclusion.[/b] Experimental work has demonstrated that geostatistical methods applied to the microbiological analysis of water matrices allows a better modeling of the phenomenon under study, a greater potential for risk management and a greater choice of methods of prevention and environmental recovery to be put in place with respect to the classical statistical analysis.

  3. Prevalence of Legionella species isolated from shower water in public bath facilities in Toyama Prefecture, Japan.

    Science.gov (United States)

    Kanatani, Jun-Ichi; Isobe, Junko; Norimoto, Shiho; Kimata, Keiko; Mitsui, Chieko; Amemura-Maekawa, Junko; Kura, Fumiaki; Sata, Tetsutaro; Watahiki, Masanori

    2017-05-01

    We investigated the prevalence of Legionella spp. isolated from shower water in public bath facilities in Toyama Prefecture, Japan. In addition, we analyzed the genetic diversity among Legionella pneumophila isolates from shower water as well as the genetic relationship between isolates from shower water and from stock strains previously analyzed from sputum specimens. The isolates were characterized using serogrouping, 16S rRNA gene sequencing, and sequence-based typing. Legionella spp. were isolated from 31/91 (34.1%) samples derived from 17/37 (45.9%) bath facilities. Isolates from shower water and bath water in each public bath facility were serologically or genetically different, indicating that we need to isolate several L. pneumophila colonies from both bath and shower water to identify public bath facilities as sources of legionellosis. The 61 L. pneumophila isolates from shower water were classified into 39 sequence types (STs) (index of discrimination = 0.974), including 19 new STs. Among the 39 STs, 12 STs match clinical isolates in the European Working Group for Legionella Infections database. Notably, ST505 L. pneumophila SG 1, a strain frequently isolated from patients with legionellosis and from bath water in this area, was isolated from shower water. Pathogenic L. pneumophila strains including ST505 strain were widely distributed in shower water in public bath facilities, with genetic diversity showing several different origins. This study highlights the need to isolate several L. pneumophila colonies from both bath water and shower water to identify public bath facilities as infection sources in legionellosis cases. Copyright © 2017 Japanese Society of Chemotherapy and The Japanese Association for Infectious Diseases. Published by Elsevier Ltd. All rights reserved.

  4. Contamination of Hospital Water Supplies in Gilan, Iran, with Legionella pneumophila, Escherichia coli, and Pseudomonas aeruginosa

    OpenAIRE

    Ahmadi Jalali Moghadam, Masoumeh; Honarmand, Hamidreza; Asfaram Meshginshahr, Sajad

    2015-01-01

    This study is designed to determine the contamination degree of hospital water supplies with Pseudomonas aeruginosa, Legionella pneumophila, and E. coli in Gilan, Iran. Samples were collected directly into sterile containers and concentrated by centrifuge. Half part of any sample transferred to yeast extract broth and the second part transferred to Trypticase Soy Broth and incubated for 3 days. DNA was extracted by using commercial kit. Four rounds of PCR were performed as follows: multiplex ...

  5. Identification and DNA fingerprinting of Legionella strains by randomly amplified polymorphic DNA analysis.

    OpenAIRE

    Bansal, N S; McDonell, F

    1997-01-01

    The randomly amplified polymorphic DNA (RAPD) technique was used in the development of a fingerprinting (typing) and identification protocol for Legionella strains. Twenty decamer random oligonucleotide primers were screened for their discriminatory abilities. Two candidate primers were selected. By using a combination of these primers, RAPD analysis allowed for the differentiation between all different species, between the serogroups, and further differentiation between subtypes of the same ...

  6. Sensitivity of Legionella pneumophila to sunlight in fresh and marine waters.

    OpenAIRE

    Dutka, B J

    1984-01-01

    Studies were carried out to assess the sunlight sensitivity of Legionella pneumophila suspended in fresh and marine waters. Comparison studies on sunlight sensitivity of lake water bacteria, Pseudomonas aeruginosa, Escherichia coli and Streptococcus faecalis, were also undertaken. The effects of full sunlight and polyacrylic-screened sunlight were monitored in the study. Results indicate that L. pneumophila cells are slightly more sensitive to sunlight in seawater than in fresh water. Enumera...

  7. Activity of Six Essential Oils Extracted from Tunisian Plants against Legionella pneumophila.

    Science.gov (United States)

    Chaftar, Naouel; Girardot, Marion; Quellard, Nathalie; Labanowski, Jérôme; Ghrairi, Tawfik; Hani, Khaled; Frère, Jacques; Imbert, Christine

    2015-10-01

    The aim of this study was to investigate the composition of six essential oils extracted from Tunisian plants, i.e., Artemisia herba-alba Asso, Citrus sinensis (L.) Osbeck, Juniperus phoenicea L., Rosmarinus officinalis L., Ruta graveolens L., and Thymus vulgaris L., and to evaluate their activity against Legionella pneumophila (microdilution assays). Eight Legionella pneumophila strains were studied, including the two well-known serogroup 1 Lens and Paris strains as controls and six environmental strains isolated from Tunisian spas belonging to serogroups 1, 4, 5, 6, and 8. The essential oils were generally active against L. pneumophila. The activities of the A. herba-alba, C. sinensis, and R. officinalis essential oils were strain-dependent, whereas those of the J. phoenicea and T. vulgaris oils, showing the highest anti-Legionella activities, with minimum inhibitory concentrations (MICs) lower than 0.03 and lower than or equal to 0.07 mg/ml, respectively, were independent of the strains' serogroup. Moreover, the microorganisms treated with T. vulgaris essential oil were shorter, swollen, and less electron-dense compared to the untreated controls. Isoborneol (20.91%), (1S)-α-pinene (18.30%) β-phellandrene (8.08%), α-campholenal (7.91%), and α-phellandrene (7.58%) were the major components isolated from the J. phoenicea oil, while carvacrol (88.50%) was the main compound of the T. vulgaris oil, followed by p-cymene (7.86%). This study highlighted the potential interest of some essential oils extracted from Tunisian plants as biocides to prevent the Legionella risk. Copyright © 2015 Verlag Helvetica Chimica Acta AG, Zürich.

  8. A Preliminary study on the prevalence of Legionella pneumophila infection in Lebanon

    International Nuclear Information System (INIS)

    Hamze, M.; Mallat, H.; Mokhbat, J.

    2009-01-01

    After the first outbreak of Legionnaire disease was described for the first time in1976, legionellosis has been increasingly recognized in association with many outbreaks, both community and hospital acquired. This disease remains a model of water borne transmission. In Lebanon, no report has yet been published on the prevalence of Legionella pneumophila infections. This study aims to evaluate the prevalence of legionellosis in hospitalized patients with community acquired pneumonia (CAP) and in patients with nosocomial pneumonia (HAP). The study was conducted between April 2004 and August 2007. Eleven community hospitals participated to this endeavour. Laboratory evaluation was conducted in two centers (Middle East Health Institute, Bsalim, Lebanon and Nini Hospital, Tripoli, Lebanon). Two hundred and forty two urinary samples and ninety one pulmonary samples from 242 patients were analyzed. The age range was 16 to 71 years. Two hundred and fifteen were from community acquired pneumonias and twenty seven were nosocomially acquired in intensive care units. The urinary antigen was investigated in 242 patients; the Binax kit ® was used in this research. In lung secretions of 91 patients, the presence of Legionella pneumophila was searched for using direct immunofluorescence method and culture. Among the 215 urinary samples from patients with CAP, three were positive for Legionella antigen with a prevalence of 1.4 %. Among the 27 patients with HAP, one was positive for Legionella antigen with a prevalence of 3.7%. This preliminary study reveals the fact that legionellosis is indeed present in Lebanon, hence the importance of considering it in the work-up and the management of patients with pneumonia. Hospital laboratories should therefore routin ely search for this pathogen through at least urinary antigen detection. Health authorities should also enforce microbiological regulations concerning water sanitation in hospitals and in the community to prevent the

  9. Effect of non-Legionellaceae bacteria on the multiplication of Legionella pneumophila in potable water.

    OpenAIRE

    Wadowsky, R M; Yee, R B

    1985-01-01

    A naturally occurring suspension of Legionella pneumophila and associated microbiota contained three unidentified non-Legionellaceae bacteria which supported satellite growth of a subculture of L. pneumophila on an L-cysteine-deficient medium and another bacterium which did not support growth of the subculture. Washed suspensions containing 10(3), 10(5), 10(7), or 10(8) CFU of a mixture of isolates of these non-Legionellaceae bacteria failed to support the multiplication of an isolate of agar...

  10. Subversion of Cell-Autonomous Immunity and Cell Migration by Legionella pneumophila Effectors

    OpenAIRE

    Simon, Sylvia; Hilbi, Hubert

    2015-01-01

    Bacteria trigger host defense and inflammatory processes, such as cytokine production, pyroptosis, and the chemotactic migration of immune cells toward the source of infection. However, a number of pathogens interfere with these immune functions by producing specific so-called “effector” proteins, which are delivered to host cells via dedicated secretion systems. Air-borne Legionella pneumophila bacteria trigger an acute and potential fatal inflammation in the lung termed Legionnaires’ diseas...

  11. Anti-Legionella dumoffii Activity of Galleria mellonella Defensin and Apolipophorin III

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    Małgorzata Cytryńska

    2012-12-01

    Full Text Available The gram-negative bacterium Legionella dumoffii is, beside Legionella pneumophila, an etiological agent of Legionnaires’ disease, an atypical form of pneumonia. The aim of this study was to determine the antimicrobial activity of Galleria mellonella defense polypeptides against L. dumoffii. The extract of immune hemolymph, containing a mixture of defense peptides and proteins, exhibited a dose-dependent bactericidal effect on L. dumoffii. The bacterium appeared sensitive to a main component of the hemolymph extract, apolipophorin III, as well as to a defense peptide, Galleria defensin, used at the concentrations 0.4 mg/mL and 40 μg/mL, respectively. L. dumoffii cells cultured in the presence of choline were more susceptible to both defense factors analyzed. A transmission electron microscopy study of bacterial cells demonstrated that Galleria defensin and apolipophorin III induced irreversible cell wall damage and strong intracellular alterations, i.e., increased vacuolization, cytoplasm condensation and the appearance of electron-white spaces in electron micrographs. Our findings suggest that insects, such as G. mellonella, with their great diversity of antimicrobial factors, can serve as a rich source of compounds for the testing of Legionella susceptibility to defense-related peptides and proteins.

  12. Direct targeting of membrane fusion by SNARE mimicry: Convergent evolution of Legionella effectors.

    Science.gov (United States)

    Shi, Xingqi; Halder, Partho; Yavuz, Halenur; Jahn, Reinhard; Shuman, Howard A

    2016-08-02

    Legionella pneumophila, the Gram-negative pathogen causing Legionnaires' disease, infects host cells by hijacking endocytic pathways and forming a Legionella-containing vacuole (LCV) in which the bacteria replicate. To promote LCV expansion and prevent lysosomal targeting, effector proteins are translocated into the host cell where they alter membrane traffic. Here we show that three of these effectors [LegC2 (Legionella eukaryotic-like gene C2)/YlfB (yeast lethal factor B), LegC3, and LegC7/YlfA] functionally mimic glutamine (Q)-SNARE proteins. In infected cells, the three proteins selectively form complexes with the endosomal arginine (R)-SNARE vesicle-associated membrane protein 4 (VAMP4). When reconstituted in proteoliposomes, these proteins avidly fuse with liposomes containing VAMP4, resulting in a stable complex with properties resembling canonical SNARE complexes. Intriguingly, however, the LegC/SNARE hybrid complex cannot be disassembled by N-ethylmaleimide-sensitive factor. We conclude that LegCs use SNARE mimicry to divert VAMP4-containing vesicles for fusion with the LCV, thus promoting its expansion. In addition, the LegC/VAMP4 complex avoids the host's disassembly machinery, thus effectively trapping VAMP4 in an inactive state.

  13. Detection of Legionella, L. pneumophila and Mycobacterium Avium Complex (MAC along Potable Water Distribution Pipelines

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    Harriet Whiley

    2014-07-01

    Full Text Available Inhalation of potable water presents a potential route of exposure to opportunistic pathogens and hence warrants significant public health concern. This study used qPCR to detect opportunistic pathogens Legionella spp., L. pneumophila and MAC at multiple points along two potable water distribution pipelines. One used chlorine disinfection and the other chloramine disinfection. Samples were collected four times over the year to provide seasonal variation and the chlorine or chloramine residual was measured during collection. Legionella spp., L. pneumophila and MAC were detected in both distribution systems throughout the year and were all detected at a maximum concentration of 103 copies/mL in the chlorine disinfected system and 106, 103 and 104 copies/mL respectively in the chloramine disinfected system. The concentrations of these opportunistic pathogens were primarily controlled throughout the distribution network through the maintenance of disinfection residuals. At a dead-end and when the disinfection residual was not maintained significant (p < 0.05 increases in concentration were observed when compared to the concentration measured closest to the processing plant in the same pipeline and sampling period. Total coliforms were not present in any water sample collected. This study demonstrates the ability of Legionella spp., L. pneumophila and MAC to survive the potable water disinfection process and highlights the need for greater measures to control these organisms along the distribution pipeline and at point of use.

  14. Legionella RavZ Plays a Role in Preventing Ubiquitin Recruitment to Bacteria-Containing Vacuoles

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    Tomoko Kubori

    2017-08-01

    Full Text Available Bacterial pathogens like Salmonella and Legionella establish intracellular niches in host cells known as bacteria-containing vacuoles. In these vacuoles, bacteria can survive and replicate. Ubiquitin-dependent selective autophagy is a host defense mechanism to counteract infection by invading pathogens. The Legionella effector protein RavZ interferes with autophagy by irreversibly deconjugating LC3, an autophagy-related ubiquitin-like protein, from a phosphoglycolipid phosphatidylethanolamine. Using a co-infection system with Salmonella, we show here that Legionella RavZ interferes with ubiquitin recruitment to the Salmonella-containing vacuoles. The inhibitory activity is dependent on the same catalytic residue of RavZ that is involved in LC3 deconjugation. In semi-permeabilized cells infected with Salmonella, external addition of purified RavZ protein, but not of its catalytic mutant, induced removal of ubiquitin associated with Salmonella-containing vacuoles. The RavZ-mediated restriction of ubiquitin recruitment to Salmonella-containing vacuoles took place in the absence of the host system required for LC3 conjugation. These observations suggest the possibility that the targets of RavZ deconjugation activity include not only LC3, but also ubiquitin.

  15. Amino Acid Uptake and Metabolism of Legionella pneumophila Hosted by Acanthamoeba castellanii.

    Science.gov (United States)

    Schunder, Eva; Gillmaier, Nadine; Kutzner, Erika; Eisenreich, Wolfgang; Herrmann, Vroni; Lautner, Monika; Heuner, Klaus

    2014-07-25

    Legionella pneumophila survives and replicates within a Legionella-containing vacuole (LCV) of amoebae and macrophages. Less is known about the carbon metabolism of the bacteria within the LCV. We have now analyzed the transfer and usage of amino acids from the natural host organism Acanthamoeba castellanii to Legionella pneumophila under in vivo (LCV) conditions. For this purpose, A. castellanii was 13C-labeled by incubation in buffer containing [U-(13)C(6)]glucose. Subsequently, these 13C-prelabeled amoebae were infected with L. pneumophila wild type or some mutants defective in putative key enzymes or regulators of carbon metabolism. 13C-Isotopologue compositions of amino acids from bacterial and amoebal proteins were then determined by mass spectrometry. In a comparative approach, the profiles documented the efficient uptake of Acanthamoeba amino acids into the LCV and further into L. pneumophila where they served as precursors for bacterial protein biosynthesis. More specifically, A. castellanii synthesized from exogenous [U-13C6]glucose unique isotopologue mixtures of several amino acids including Phe and Tyr, which were also observed in the same amino acids from LCV-grown L. pneumophila. Minor but significant differences were only detected in the isotopologue profiles of Ala, Asp, and Glu from the amoebal or bacterial protein fractions, respectively, indicating partial de novo synthesis of these amino acids by L. pneumophila. The similar isotopologue patterns in amino acids from L. pneumophila wild type and the mutants under study reflected the robustness of amino acid usage in the LCV of A. castellannii.

  16. Identification of Legionella rubrilucens isolated from a hot spring for foot-soaking in Niigata, Japan.

    Science.gov (United States)

    Furuhata, Katsunori; Edagawa, Akiko; Miyamoto, Hiroshi; Morimoto, Yo; Fukuyama, Masafumi

    2012-06-01

    In May 2011, strain HYNE-20 (=JCM 17837) was isolated from a sample of hot spring water from a foot spa in Niigata, Japan, by a plating method using glycine vancomycin polymyxin B cycloheximide α-ketoglutarate (GVPCα) medium at 36°C for 7 d. The 16S rDNA sequences (1,469bp) of this strain (accession number: AB638719) had high (99.7%) similarity to Legionella rubrilucens, and we identified that this strain was indeed Legionella rubrilucens. When this strain was cultured on buffered charcoal yeast extract α-ketoglutarate (BCYEα) agar at 36°C for 7 d, it exhibited red autofluorescence under UV light (365 nm) . The dominant cellular fatty acids of the strain HYNE-20 were 16:1ω7c (29.9%) , and the guanine-plus-cytosine (G+C) content of DNA was 49.0 mol%. This is the first report that Legionella rubrilucens was isolated from a hot spring for foot soaking.

  17. SP. Pescado

    Directory of Open Access Journals (Sweden)

    Renato Gendre

    2003-12-01

    Full Text Available Nell'occhiello di un articolo dal titolo Il Peru dei de[Jini rosa e de/la grande pioggia si legge: "da una partenza  in aereo al «pescado»  che ti  sfamera."1 Questa parola spagnola, giustamente chiusa tra caporali, a noi pare molto interes­ sante, perche, nonostante l'apparenza, non ha nulla da spartire sotto i1 profilo se­ mantico con l'it. pescato. lnfatti, tutti i piu importanti dizionari della lingua italiana, di ieri e di oggi, etimologici e non 2, registrano  accanto a pescata,  ii lemma pescato, 3 ma lo spiegano come "quantita di pesce catturato nel corso di una battuta o di una stagione di pesca",4 mentre lo sp. pescado  indica i1 "pesce (solo nel senso di: pesGe pescato da mangiare [...]".s

  18. Detection of Legionella-contaminated aerosols in the vicinity of a bio-trickling filter of a breeding sow facility - A pilot study.

    Science.gov (United States)

    Walser, Sandra M; Brenner, Bernhard; Wunderlich, Anika; Tuschak, Christian; Huber, Stefanie; Kolb, Stefanie; Niessner, Reinhard; Seidel, Michael; Höller, Christiane; Herr, Caroline E W

    2017-01-01

    The urbanization of agricultural areas results in a reduction of distances between residential buildings and livestock farms. In the public debate, livestock farming is increasingly criticized due to environmental disturbance and odor nuisance originating from such facilities. One method to reduce odor and ammonia is by exhaust air treatment, for example, by biological exhaust air purification processes with bio-trickling filters filled with tap water. Higher temperatures in the summer time and the generation of biofilms are ideal growth conditions for Legionella. However, there are no studies on the presence of Legionella in the water of bio-trickling filters and the release of Legionella-containing aerosols. Therefore, the aim of this study was to investigate Legionella in wash water and emitted bioaerosols of a bio-trickling filter system of a breeding sow facility. For this purpose, measurements were carried out using a cyclone sampler. In addition, samples of wash water were taken. Legionella were not found by culture methods. However, using molecular biological methods, Legionella spp. could be detected in wash water as well as in bioaerosol samples. With antibody-based methods, Legionella pneumophila were identified. Further studies are needed to investigate the environmental health relevance of Legionella-containing aerosols emitted by such exhaust air purification systems. Copyright © 2016 Elsevier B.V. All rights reserved.

  19. A STUDY ON LEGIONELLA PNEUMOPHILA, WATER CHEMISTRY, AND ATMOSPHERIC CONDITIONS IN COOLING TOWERS AT THE SAVANNAH RIVER SITE

    Energy Technology Data Exchange (ETDEWEB)

    Smith, C.; Brigmon, R.

    2009-10-20

    Legionnaires disease is a pneumonia caused by the inhalation of the bacterium Legionella pneumophila. The majority of illnesses have been associated with cooling towers since these devices can harbor and disseminate the bacterium in the aerosolized mist generated by these systems. Historically, Savannah River Site (SRS) cooling towers have had occurrences of elevated levels of Legionella in all seasons of the year and in patterns that are difficult to predict. Since elevated Legionella in cooling tower water are a potential health concern a question has been raised as to the best control methodology. In this work we analyze available chemical, biological, and atmospheric data to determine the best method or key parameter for control. The SRS 4Q Industrial Hygiene Manual, 4Q-1203, 1 - G Cooling Tower Operation and the SRNL Legionella Sampling Program, states that 'Participation in the SRNL Legionella Sampling Program is MANDATORY for all operating cooling towers'. The resulting reports include L. pneumophila concentration information in cells/L. L. pneumophila concentrations >10{sup 7} cells/L are considered elevated and unsafe so action must be taken to reduce these densities. These remedial actions typically include increase biocide addition or 'shocking'. Sometimes additional actions are required if the problem persists including increase tower maintenance (e.g. cleaning). Evaluation of 14 SRS cooling towers, seven water quality parameters, and five Legionella serogroups over a three-plus year time frame demonstrated that cooling tower water Legionella densities varied widely though out this time period. In fact there was no one common consistent significant variable across all towers. The significant factors that did show up most frequently were related to suspended particulates, conductivity, pH, and dissolved oxygen, not chlorine or bromine as might be expected. Analyses of atmospheric data showed that there were more frequent significant

  20. Experimental human-like model to assess the part of viable Legionella reaching the thoracic region after nebulization.

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    Jérémie Pourchez

    Full Text Available The incidence of Legionnaires' disease (LD in European countries and the USA has been constantly increasing since 1998. Infection of humans occurs through aerosol inhalation. To bridge the existing gap between the concentration of Legionella in a water network and the deposition of bacteria within the thoracic region (assessment of the number of viable Legionella, we validated a model mimicking realistic exposure through the use of (i recent technology for aerosol generation and (ii a 3D replicate of the human upper respiratory tract. The model's sensitivity was determined by monitoring the deposition of (i aerosolized water and Tc99m radio-aerosol as controls, and (ii bioaerosols generated from both Escherichia coli and Legionella pneumophila sg 1 suspensions. The numbers of viable Legionella prior to and after nebulization were provided by culture, flow cytometry and qPCR. This study was designed to obtain more realistic data on aerosol inhalation (vs. animal experimentation and deposition at the thoracic region in the context of LD. Upon nebulization, 40% and 48% of the initial Legionella inoculum was made of cultivable and non-cultivable cells, respectively; 0.7% of both populations reached the filter holder mimicking the thoracic region in this setup. These results are in agreement with experimental data based on quantitative microbial risk assessment methods and bring new methods that may be useful for preventing LD.

  1. [Identification of Systemic Contaminations with Legionella Spec. in Drinking Water Plumbing Systems: Sampling Strategies and Corresponding Parameters].

    Science.gov (United States)

    Völker, S; Schreiber, C; Müller, H; Zacharias, N; Kistemann, T

    2017-05-01

    After the amendment of the Drinking Water Ordinance in 2011, the requirements for the hygienic-microbiological monitoring of drinking water installations have increased significantly. In the BMBF-funded project "Biofilm Management" (2010-2014), we examined the extent to which established sampling strategies in practice can uncover drinking water plumbing systems systemically colonized with Legionella. Moreover, we investigated additional parameters that might be suitable for detecting systemic contaminations. We subjected the drinking water plumbing systems of 8 buildings with known microbial contamination (Legionella) to an intensive hygienic-microbiological sampling with high spatial and temporal resolution. A total of 626 drinking hot water samples were analyzed with classical culture-based methods. In addition, comprehensive hygienic observations were conducted in each building and qualitative interviews with operators and users were applied. Collected tap-specific parameters were quantitatively analyzed by means of sensitivity and accuracy calculations. The systemic presence of Legionella in drinking water plumbing systems has a high spatial and temporal variability. Established sampling strategies were only partially suitable to detect long-term Legionella contaminations in practice. In particular, the sampling of hot water at the calorifier and circulation re-entrance showed little significance in terms of contamination events. To detect the systemic presence of Legionella,the parameters stagnation (qualitatively assessed) and temperature (compliance with the 5K-rule) showed better results. © Georg Thieme Verlag KG Stuttgart · New York.

  2. A detection method for Legionella spp in (cooling) water: fluorescent in situ hybridisation (FISH) on whole bacteria.

    Science.gov (United States)

    Declerck, P; Verelst, L; Duvivier, L; Van Damme, A; Ollevier, F

    2003-01-01

    Although traditional culture methods are appropriate for detection of Legionella species, such culture takes several days. Rapid detection (FISH) on whole bacteria. Water samples were filtered and the concentrated bacteria were immediately detected (without culture) with a fluorescence microscope following appropriate labelling. The detection level was very high and quantification was possible. For the detection of all Legionella spp. the probe LEG705 was used, complementary to a 16S rRNA sequence conserved in all Legionella spp. For specific detection of L. pneumophila the probe LEGPNE1 was used. This probe is designed against a variable domain of the 16S rRNA sequence from L. pneumophila. CY3 and FLUOS labels were tested and CY3 showed clearly detectable bacteria with minimum background staining. This FISH technique is very sensitive, fast, reliable and individual bacteria are easily detected.

  3. Health risks from exposure to Legionella in reclaimed water aerosols: Toilet flushing, spray irrigation, and cooling towers.

    Science.gov (United States)

    Hamilton, Kerry A; Hamilton, Mark T; Johnson, William; Jjemba, Patrick; Bukhari, Zia; LeChevallier, Mark; Haas, Charles N

    2018-05-01

    The use of reclaimed water brings new challenges for the water industry in terms of maintaining water quality while increasing sustainability. Increased attention has been devoted to opportunistic pathogens, especially Legionella pneumophila, due to its growing importance as a portion of the waterborne disease burden in the United States. Infection occurs when a person inhales a mist containing Legionella bacteria. The top three uses for reclaimed water (cooling towers, spray irrigation, and toilet flushing) that generate aerosols were evaluated for Legionella health risks in reclaimed water using quantitative microbial risk assessment (QMRA). Risks are compared using data from nineteen United States reclaimed water utilities measured with culture-based methods, quantitative PCR (qPCR), and ethidium-monoazide-qPCR. Median toilet flushing annual infection risks exceeded 10 -4 considering multiple toilet types, while median clinical severity infection risks did not exceed this value. Sprinkler and cooling tower risks varied depending on meteorological conditions and operational characteristics such as drift eliminator performance. However, the greatest differences between risk scenarios were due to 1) the dose response model used (infection or clinical severity infection) 2) population at risk considered (residential or occupational) and 3) differences in laboratory analytical method. Theoretical setback distances necessary to achieve a median annual infection risk level of 10 -4 are proposed for spray irrigation and cooling towers. In both cooling tower and sprinkler cases, Legionella infection risks were non-trivial at potentially large setback distances, and indicate other simultaneous management practices could be needed to manage risks. The sensitivity analysis indicated that the most influential factors for variability in risks were the concentration of Legionella and aerosol partitioning and/or efficiency across all models, highlighting the importance of

  4. Detection methods for legionellae. Possibilities and limitations of detection methods; Niet alle detectiemethoden geschikt voor legionellagroei. Mogelijkheden en beperkingen detectiemethoden

    Energy Technology Data Exchange (ETDEWEB)

    Scheffer, W.

    2011-06-15

    This article provides an overview of existing and new methods for detecting legionella in water. It offers insight in the characteristics and applications of existing and new detection methods. However, it is impossible to assess the value of alternative methods at the moment. That will require additional research. [Dutch] Een overzicht is gemaakt van bestaande en nieuwe methoden om legionella in water aan te tonen. Daarmee wordt inzicht verkregen in de kenmerken en toepassingsmogelijkheden van bestaande en nieuwe detectiemethoden. De waarde van alternatieve methoden in een installatie kan echter nog niet worden beoordeeld. Daarvoor is nader onderzoek nodig.

  5. Draft genome sequences of six neonatal meningitis-causing escherichia coli isolates (SP-4, SP-5, SP-13, SP-16, SP-46, and SP-65)

    Science.gov (United States)

    Neonatal meningitis Escherichia coli isolates (SP-4, SP-5, SP-13, SP-16, SP-46, and SP-65) were recovered from infants in the Netherlands from 1989 to 1997. Here, we report the draft genome sequences for these six E. coli isolates, which are currently being used to validate food safety processing te...

  6. Necessity and effect of combating Legionella pneumophila in municipal shower systems.

    Science.gov (United States)

    Wiik, Ragnhild; Krøvel, Anne Vatland

    2014-01-01

    The objective was to obtain research-based, holistic knowledge about necessity and effect of practiced measures against L. pneumophila in municipal shower systems in Stavanger, Norway. The effects of hot water treatment and membrane-filtering were investigated and compared to no intervention at all. The studies were done under real-world conditions. Additionally, a surveillance pilot study of municipal showers in Stavanger was performed. The validity of high total plate count (TPC) as an indication of L. pneumophila was evaluated. A simplified method, named "dripping method", for detection and quantification of L. pneumophila was developed. The sensitivity of the dripping method is 5 colony-forming units of L. pneumophila/ml. The transference of L. pneumophila from shower water to aerosols was studied. Interviews and observational studies among the stakeholders were done in order to identify patterns of communication and behavior in a Legionella risk perspective. No substantial effects of the measures against L. pneumophila were demonstrated, except for a distally placed membrane filter. No significant positive correlation between TPC and L. pneumophila concentrations were found. L. pneumophila serogroup 2-14 was demonstrated in 21% of the 29 buildings tested in the surveillance pilot. Relatively few cells of L. pneumophila were transferred from shower water to aerosols. Anxiety appeared as the major driving force in the risk governance of Legionella. In conclusion, the risk of acquiring Legionnaires' disease from municipal shower systems is evaluated as low and uncertain. By eliminating ineffective approaches, targeted Legionella risk governance can be practiced. Risk management by surveillance is evaluated as appropriate.

  7. INVESTIGATION OF LEGIONELLA SPECIES IN TEHRAN’S HOSPITAL WATER SUPPLIES

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    MOHAMMAD RAFIEE

    2015-03-01

    Full Text Available The present work was performed to investigate the presence of Legionella spp. and its common species in hospital water supplies. Considering the drawback of culture method, polymerase chain reaction (PCR assays were developed to detect the gene 16S rRNA regardless of the bacterial serotype. Four well-established DNA extraction protocols (freeze & thaw and phenol-chloroform as two manual protocols and two commercial kits were examined and critiqued to release DNA from bacterial cells. A total of 45 samples were collected from seven distinct hospitals’ sites during a period of 10 months. The PCR assay was exploited to amplify a 654-bp fragment of the 16S rRNA gene. Legionella were detected in 13 samples (28.9% by all of the methods applied for DNA extraction. Considerable differences were noted in the yield of extracted nucleic acids. Legionella were not detected in any of the samples when DNA extraction by freeze & thaw was used. Omitting this method and comparing manual protocol with commercial kits, Kappa coefficient was calculated as 0.619 with p < 0.05. Although no meaningful differences were found between the kits, DNA extraction with Bioneer kit displayed a higher sensitivity than classical Qiagen. Showerheads and cold-water taps were the most and least contaminated sources with 55.5 and 9 percent positive samples, respectively. Moreover two positive samples were identified for species by DNA sequencing and submitted to the Gene Bank database with accession Nos. FJ480932 and FJ480933.

  8. Hospital-wide Eradication of a Nosocomial Legionella pneumophila Serogroup 1 Outbreak.

    Science.gov (United States)

    Bartley, Paul B; Ben Zakour, Nouri L; Stanton-Cook, Mitchell; Muguli, Raghuram; Prado, Luis; Garnys, Vyt; Taylor, Katherine; Barnett, Timothy C; Pinna, Glen; Robson, Jennifer; Paterson, David L; Walker, Mark J; Schembri, Mark A; Beatson, Scott A

    2016-02-01

    Two proven nosocomial cases of Legionella pneumonia occurred at the Wesley Hospital (Brisbane, Australia) in May 2013. To trace the epidemiology of these cases, whole genome sequence analysis was performed on Legionella pneumophila isolates from the infected patients, prospective isolates collected from the hospital water distribution system (WDS), and retrospective patient isolates available from the Wesley Hospital and other local hospitals. Legionella pneumophila serogroup 1 isolates were cultured from patient sputum (n = 3), endobronchial washings (n = 3), pleural fluid (n = 1), and the Wesley Hospital WDS (n = 39). Whole genome sequencing and de novo assembly allowed comparison with the L. pneumophila Paris reference strain to infer phylogenetic and epidemiological relationships. Rapid disinfection of the hospital WDS with a chlorinated, alkaline detergent and subsequent superchlorination followed by maintenance of residual free chlorine, combined with removal of redundant plumbing, was instituted. The 2011 and 2013 L. pneumophila patient isolates were serogroup 1 and closely related to all 2013 hospital water isolates based on single nucleotide polymorphisms and mobile genetic element profiles, suggesting a single L. pneumophila population as the source of nosocomial infection. The L. pneumophila population has evolved to comprise 3 clonal variants, each associated with different parts of the hospital WDS. This study provides an exemplar for the use of clinical and genomic epidemiological methods together with a program of rapid, effective remedial biofilm, plumbing and water treatment to characterize and eliminate a L. pneumophila population responsible for nosocomial infections. © The Author 2015. Published by Oxford University Press for the Infectious Diseases Society of America. All rights reserved. For permissions, e-mail journals.permissions@oup.com.

  9. Legionella Effector AnkX Disrupts Host Cell Endocytic Recycling in a Phosphocholination-Dependent Manner

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    Samual C. Allgood

    2017-09-01

    Full Text Available The facultative intracellular bacterium Legionella pneumophila proliferates within amoebae and human alveolar macrophages, and it is the causative agent of Legionnaires' disease, a life-threatening pneumonia. Within host cells, L. pneumophila establishes a replicative haven by delivering numerous effector proteins into the host cytosol, many of which target membrane trafficking by manipulating the function of Rab GTPases. The Legionella effector AnkX is a phosphocholine transferase that covalently modifies host Rab1 and Rab35. However, a detailed understanding of the biological consequence of Rab GTPase phosphocholination remains elusive. Here, we broaden the understanding of AnkX function by presenting three lines of evidence that it interferes with host endocytic recycling. First, using immunogold transmission electron microscopy, we determined that GFP-tagged AnkX ectopically produced in mammalian cells localizes at the plasma membrane and tubular membrane compartments, sites consistent with targeting the endocytic recycling pathway. Furthermore, the C-terminal region of AnkX was responsible for association with the plasma membrane, and we determined that this region was also able to bind the phosphoinositide lipids PI(3P and PI(4P in vitro. Second, we observed that mCherry-AnkX co-localized with Rab35, a regulator of recycling endocytosis and with major histocompatibility class I protein (MHC-I, a key immunoregulatory protein whose recycling from and back to the plasma membrane is Rab35-dependent. Third, we report that during infection of macrophages, AnkX is responsible for the disruption of endocytic recycling of transferrin, and AnkX's phosphocholination activity is critical for this function. These results support the hypothesis that AnkX targets endocytic recycling during host cell infection. Finally, we have demonstrated that the phosphocholination activity of AnkX is also critical for inhibiting fusion of the Legionella

  10. Amino Acid Uptake and Metabolism of Legionella pneumophila Hosted by Acanthamoeba castellanii*

    Science.gov (United States)

    Schunder, Eva; Gillmaier, Nadine; Kutzner, Erika; Herrmann, Vroni; Lautner, Monika; Heuner, Klaus; Eisenreich, Wolfgang

    2014-01-01

    Legionella pneumophila survives and replicates within a Legionella-containing vacuole (LCV) of amoebae and macrophages. Less is known about the carbon metabolism of the bacteria within the LCV. We have now analyzed the transfer and usage of amino acids from the natural host organism Acanthamoeba castellanii to Legionella pneumophila under in vivo (LCV) conditions. For this purpose, A. castellanii was 13C-labeled by incubation in buffer containing [U-13C6]glucose. Subsequently, these 13C-prelabeled amoebae were infected with L. pneumophila wild type or some mutants defective in putative key enzymes or regulators of carbon metabolism. 13C-Isotopologue compositions of amino acids from bacterial and amoebal proteins were then determined by mass spectrometry. In a comparative approach, the profiles documented the efficient uptake of Acanthamoeba amino acids into the LCV and further into L. pneumophila where they served as precursors for bacterial protein biosynthesis. More specifically, A. castellanii synthesized from exogenous [U-13C6]glucose unique isotopologue mixtures of several amino acids including Phe and Tyr, which were also observed in the same amino acids from LCV-grown L. pneumophila. Minor but significant differences were only detected in the isotopologue profiles of Ala, Asp, and Glu from the amoebal or bacterial protein fractions, respectively, indicating partial de novo synthesis of these amino acids by L. pneumophila. The similar isotopologue patterns in amino acids from L. pneumophila wild type and the mutants under study reflected the robustness of amino acid usage in the LCV of A. castellannii. PMID:24904060

  11. Molecular typing of Legionella pneumophila serogroup 1 clinical strains isolated in Italy.

    Science.gov (United States)

    Fontana, Stefano; Scaturro, Maria; Rota, Maria Cristina; Caporali, Maria Grazia; Ricci, Maria Luisa

    2014-07-01

    Molecular typing methods for discriminating different bacterial isolates are essential epidemiological tools in prevention and control of Legionella infections and outbreaks. A selection of 56 out of 184 Legionella pneumophila serogroup 1 (Lp1) clinical isolates, collected from different Italian regions between 1987 and 2012, and stored at the National Reference Laboratory for Legionella, were typed by monoclonal antibody (MAb) subgrouping, amplified fragment length polymorphism (AFLP) and sequence based typing (SBT). These strains were isolated from 39 community (69.6%), 14 nosocomial (25%) and 3 travel associated (5.4%) Legionnaires'disease cases. MAb typing results showed a prevalence of MAb 3/1 positive isolates (75%) with the Philadelphia subgroup representing 35.7%, followed by Knoxville (23.2%), Benidorm (12.5%), Allentown/France (1.8%), Allentown/France-Philadelphia (1.8%). The remaining 25% were MAb 3/1 negative, namely 11 Olda (19.6%), 2 Oxford (3.6%) and 1 Bellingham (1.8%) subgroups. AFLP analysis detected 20 different genomic profiles. SBT analysis revealed 32 different sequence types (STs) with high diversity of STs (IODSTs=0.952) 12 of which were never described before. ST1 and ST23 were most frequently isolated as observed worldwide. A helpful analysis of data from SBT, MAb subgrouping and AFLP is provided, as well as a comparison to the Lp1 types investigated from other countries. This study describes the first Italian Lp1 strains database, providing molecular epidemiology data useful for future epidemiological investigations, especially of travel associated Legionnaires' diseases (TALD) cases, Italy being the country associated with the highest number of clusters. Copyright © 2014 Elsevier GmbH. All rights reserved.

  12. The occurrence of antibodies against Legionella pneumophila in patients with autoimmune rheumatic diseases.

    Science.gov (United States)

    Sikora, Agnieszka; Koszarny, Arkadiusz; Kozioł-Montewka, Maria; Majdan, Maria; Paluch-Oleś, Jolanta; Kozioł, Małgorzata M

    2015-01-01

    Patients with autoimmune rheumatic diseases are more susceptible to infection, owing to the underlying disease itself or to its treatment. Most commonly, infections affect the respiratory and urinary tracts. One of the etiological factors of infections in these patients is the bacteria of the genus Legionella. The aim of the study was to assess the prevalence of anti-Legionella pneumophila (L. pneumophila) antibodies in patients with autoimmune rheumatic diseases and to analyze individual and environmental risk factors for the development of Legionella infection in patients with positive antibody results. The study group consisted of 165 patients with autoimmune rheumatic diseases and 100 healthy subjects. Serum samples were tested for the presence of specific antibodies in the immunoglobulin (Ig) M and IgG classes against L. pneumophila serogroups 1 to 7 (SG 1-7) and the IgG class for serogroup 1 (SG 1). Antibodies against L. pneumophila were found in 7 patients (4%): 5 cases with antibody positivity only in the IgG class and 2 cases with antibody positivity in both classes. In patients with positive IgG antibodies for SG 1-7, specific antibodies for L. pneumophila SG 1 were not detected. In the control group, positive results were obtained in 9 cases (9%): IgM positivity in 6 (6%) and IgG positivity in 3 (3%). The frequency of antibodies to L. pneumophila in our patients is comparable to that in healthy individuals. L. pneumophila should be recognized as a potential pathogen in patients with autoimmune rheumatic diseases. Primary disease condition, immunosuppressive therapy, and other risk factors should not be ignored in these patients.

  13. 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.

  14. Colonization of components of a model hot water system by Legionella pneumophila.

    Science.gov (United States)

    Schofield, G M; Locci, R

    1985-02-01

    A model hot water distribution network was seeded with a virulent strain of Legionella pneumophila serotype 1. Ten weeks after inoculation, components of the system, which include aluminium discs, copper, stainless steel, silicone tubing, rubber and glass beads, were examined for colonization by L. pneumophila. The samples were stained with fluorescein-labelled antibodies to the strain and were examined with scanning electron microscopy. Colonization, which was accompanied by copious quantities of a slime-like debris, was heaviest on the rubber and least on the copper. Adherence to silicone tubing and stainless steel was observed.

  15. Epidemiology of Legionella in environments for recreational, tourism or physical activities

    Directory of Open Access Journals (Sweden)

    Vincenzo Romano Spica

    2004-12-01

    Full Text Available

    Legionella is a waterborne microorganism responsible of severe pneumonia. Legionnaire’s disease and Pontiac fever are clinical syndromes related to contaminated environments.

    Outbreak sources have been identified in sport facilities, thermal springs, spas, hotels, campsites, cruise ships and other structures for recreational or tourism purposes. Infection spreads by inhalation of contaminated aerosols from showers or whirlpool baths. Exposure to properly maintained swimming pools does not represent a major risk factor. Several prevention strategies and decontamination procedures are available.

  16. Antimicrobial Activity of Solithromycin against Clinical Isolates of Legionella pneumophila Serogroup 1

    OpenAIRE

    Mallegol, Julia; Fernandes, Prabhavathi; Melano, Roberto G.; Guyard, Cyril

    2014-01-01

    The activity of solithromycin was evaluated against clinical Legionella pneumophila serogroup 1 (Lp1) isolates (n = 196) collected in Ontario, Canada, from 1980 to 2011. Its in vitro activity was compared to that of azithromycin (AZM) using the broth microdilution method. Solithromycin had a MIC50 of ≤0.015 μg/ml and a MIC90 of 0.031 μg/ml, making its activity at least 8-fold to 32-fold higher than that of AZM (MIC50 and MIC90, 0.125 μg/ml and 1 μg/ml, respectively). Ninety-nine percent of th...

  17. Crystallization and preliminary crystallographic analysis of an aminoglycoside kinase from Legionella pneumophila

    International Nuclear Information System (INIS)

    Lemke, Christopher T.; Hwang, Jiyoung; Xiong, Bing; Cianciotto, Nicholas P.; Berghuis, Albert M.

    2005-01-01

    Two crystal forms of the antibiotic resistance enzyme APH(9)-Ia from L. pneumophila are reported. 9-Aminoglycoside phosphotransferase type Ia [APH(9)-Ia] is a resistance factor in Legionella pneuemophila, the causative agent of legionnaires’ disease. It is responsible for providing intrinsic resistance to the antibiotic spectinomycin. APH(9)-Ia phosphorylates one of the hydroxyl moieties of spectinomycin in an ATP-dependent manner, abolishing the antibiotic properties of this drug. Here, the crystallization and preliminary X-ray studies of this enzyme in two crystal forms is reported. One of the these crystal forms provides diffraction data to a resolution of 1.7 Å

  18. Water Safety and Legionella in Health Care: Priorities, Policy, and Practice.

    Science.gov (United States)

    Gamage, Shantini D; Ambrose, Meredith; Kralovic, Stephen M; Roselle, Gary A

    2016-09-01

    Health care facility water distribution systems have been implicated in the transmission of pathogens such as Legionella and nontuberculous mycobacteria to building occupants. These pathogens are natural inhabitants of water at low numbers and can amplify in premise plumbing water, especially if conditions are conducive to their growth. Because patients and residents in health care facilities are often at heightened risk for opportunistic infections, a multidisciplinary proactive approach to water safety is important to balance the various water priorities in health care and prevent water-associated infections in building occupants. Published by Elsevier Inc.

  19. Optimization of Pulsed-Field Gel Electrophoresis for Legionella pneumophila Subtyping▿ †

    OpenAIRE

    Zhou, Haijian; Ren, Hongyu; Zhu, Bingqing; Kan, Biao; Xu, Jianguo; Shao, Zhujun

    2010-01-01

    A total of 32 strains of Legionella pneumophila were used to optimize pulsed-field gel electrophoresis (PFGE) for subtyping of L. pneumophila. Twenty-six isolates of L. pneumophila with various origins and 11 isolates from five different water systems were used as the panels. For optimization of electrophoretic parameters (EPs) of SfiI PFGE, 26 isolates were analyzed with SfiI digestion, using four EPs yielding the same D value. The EP of a switch time of 5 to 50 s for 21 h had the smallest s...

  20. Molecular detection of Acanthamoeba spp., Naegleria fowleri and Vermamoeba (Hartmannella) vermiformis as vectors for Legionella spp. in untreated and solar pasteurized harvested rainwater.

    Science.gov (United States)

    Dobrowsky, Penelope H; Khan, Sehaam; Cloete, Thomas E; Khan, Wesaal

    2016-10-10

    Legionella spp. employ multiple strategies to adapt to stressful environments including the proliferation in protective biofilms and the ability to form associations with free-living amoeba (FLA). The aim of the current study was to identify Legionella spp., Acanthamoeba spp., Vermamoeba (Hartmannella) vermiformis and Naegleria fowleri that persist in a harvested rainwater and solar pasteurization treatment system. Pasteurized (45 °C, 65 °C, 68 °C, 74 °C, 84 °C and 93 °C) and unpasteurized tank water samples were screened for Legionella spp. and the heterotrophic plate count was enumerated. Additionally, ethidium monoazide quantitative polymerase chain reaction (EMA-qPCR) was utilized for the quantification of viable Legionella spp., Acanthamoeba spp., V. vermiformis and N. fowleri in pasteurized (68 °C, 74 °C, 84 °C and 93 °C) and unpasteurized tank water samples, respectively. Of the 82 Legionella spp. isolated from unpasteurized tank water samples, Legionella longbeachae (35 %) was the most frequently isolated, followed by Legionella norrlandica (27 %) and Legionella rowbothamii (4 %). Additionally, a positive correlation was recorded between the heterotrophic plate count vs. the number of Legionella spp. detected (ρ = 0.710, P = 0.048) and the heterotrophic plate count vs. the number of Legionella spp. isolated (ρ = 0.779, P = 0.0028) from the tank water samples collected. Solar pasteurization was effective in reducing the gene copies of viable V. vermiformis (3-log) and N. fowleri (5-log) to below the lower limit of detection at temperatures of 68-93 °C and 74-93 °C, respectively. Conversely, while the gene copies of viable Legionella and Acanthamoeba were significantly reduced by 2-logs (P = 0.0024) and 1-log (P = 0.0015) overall, respectively, both organisms were still detected after pasteurization at 93 °C. Results from this study indicate that Acanthamoeba spp. primarily acts as the vector and aids in

  1. Comparative and functional genomics of Legionella identified eukaryotic like proteins as key players in host-pathogen interactions

    Directory of Open Access Journals (Sweden)

    Laura eGomez-Valero

    2011-10-01

    Full Text Available Although best known for its ability to cause severe pneumonia in people whose immune defenses are weakened, Legionella pneumophila and Legionella longbeachae are two species of a large genus of bacteria that are ubiquitous in nature, where they parasitize protozoa. Adaptation to the host environment and exploitation of host cell functions are critical for the success of these intracellular pathogens. The establishment and publication of the complete genome sequences of L. pneumophila and L. longbeachae isolates paved the way for major breakthroughs in understanding the biology of these organisms. In this review we present the knowledge gained from the analyses and comparison of the complete genome sequences of different L. pneumophila and L. longbeachae strains. Emphasis is given on putative virulence and Legionella life cycle related functions, such as the identification of an extended array of eukaryotic-like proteins, many of which have been shown to modulate host cell functions to the pathogen's advantage. Surprisingly, many of the eukaryotic domain proteins identified in L. pneumophila as well as many substrates of the Dot/Icm type IV secretion system essential for intracellular replication are different between these two species, although they cause the same disease. Finally, evolutionary aspects regarding the eukaryotic like proteins in Legionella are discussed.

  2. Colonization of hospital water systems by Legionella pneumophila, Pseudomonas aeroginosa, and Acinetobacter in ICU wards of Tehran hospitals

    Directory of Open Access Journals (Sweden)

    Somayeh Yaslianifard

    2012-01-01

    Full Text Available Background: Nosocomial infection caused by non-Enterobacteriaceae gram negative bacteria (GNB-NE is increasing in intensive care units (ICU. Aim: The objective of this study was to determine whether potable water in ICU wards at Tehran hospitals is contaminated with L. pneomophila, P. aeroginosa and Acinetobacter spp. Materials and Methods: A total of 52 water samples from shower bath and taps water in seven hospitals of Tehran were collected. The water sample concentrated by filtering through millipore cellulose filters and cultured on BCYE agar and tryptic soya agar media. The presence of Legionella pneumophila was confirmed by real time PCR assay using primers-probe designed for the mip gene. Results: Legionella pneumophila, Pseudomonas aeroginosa and Acinetobacter were isolated from 5 (9.6%, 6 (11.4% and 1 (1.8% of the hospital water systems, respectively. This study demonstrated the presence of Legionella, Pseudomonas and Acinetobacter in water system in ICU wards of different hospitals in Tehran. Conclusions: Hot water from shower heads could be a potential source of infection for Legionella pneumophila. Water was also proved to contain Pseudomonas aeruginonsa, the main GNB-NE causing nosocomila pneumonia at Tehran hospitals. Care should be taken concerning cleanliness and decontamination of water supplies at ICUs for pathogenic organisms.

  3. Legionella pneumophila: the paradox of a highly sensitive opportunistic waterborne pathogen able to persist in the environment

    Directory of Open Access Journals (Sweden)

    Jean-Marc eBerjeaud

    2016-04-01

    Full Text Available Legionella pneumophila, the major causative agent of Legionnaires’ disease, is found in freshwater environments in close association with free-living amoebae and multispecies biofilms, leading to persistence, spread, biocide resistance, and elevated virulence of the bacterium. Indeed, legionellosis outbreaks are mainly due to the ability of this bacterium to colonize and persist in water facilities, despite harsh physical and chemical treatments. However, these treatments are not totally efficient and, after a lag period, L. pneumophila may be able to quickly re-colonize these systems. Several natural compounds (biosurfactants, antimicrobial peptides… with anti-Legionella properties have recently been described in the literature, highlighting their specific activities against this pathogen. In this review, we first consider this hallmark of Legionella to resist killing, in regard to its biofilm or host-associated life style. Then, we focus more accurately on natural anti-Legionella molecules described so far, which could provide new eco-friendly and alternative ways to struggle against this important pathogen in plumbing.

  4. Community-acquired pneumonia due to Legionella pneumophila, the utility of PCR, and a review of the antibiotics used

    Directory of Open Access Journals (Sweden)

    Paul Zarogoulidis

    2011-01-01

    Full Text Available Paul Zarogoulidis1,2, Ioanna Alexandropoulou1,2, Gioulia Romanidou3, Theocharis G Konstasntinidis1,2, Eirini Terzi3, S Saridou1, Athanasios Stefanis1, Kostas Zarogoulidis1,2, TC Constantinidis1,21Regional Laboratory of Public Health, East Macedonia-Thrace, Komotini, Greece; 2Laboratory of Hygiene and Environmental Protection, Medical School, Democritus University of Thrace, Komotini, Greece; 3Department of Internal Medicine, General Municipal Hospital (“Sismanoglio” of Komotini, Komotini, GreeceIntroduction: There are at least 40 types of Legionella bacteria, half of which are capable of producing disease in humans. The Legionella pneumophila bacterium, the root cause of Legionnaires’ disease, causes 90% of legionellosis cases.Case presentation: We describe the case of a 60-year-old woman with a history of diabetes mellitus and arterial hypertension who was admitted to our hospital with fever and symptoms of respiratory infection, diarrhea, and acute renal failure. We used real-time polymerase chain reaction (PCR to detect L. pneumophila DNA in peripheral blood and serum samples and urine antigen from a patient with pneumonia. Legionella DNA was detected in all two sample species when first collected.Conclusion: Since Legionella is a cause of 2% to 15% of all community-acquired pneumonias that require hospitalization, legionellosis should be taken into account in an atypical pulmonary infection and not be forgotten. Moreover, real-time PCR should be considered a useful diagnostic method.Keywords: Legionnaires’ disease, Lionella pneumophila

  5. Detection of protozoan hosts for Legionella pneumophila in engineered water systems by using a biofilm batch test

    NARCIS (Netherlands)

    Valster, R.M.; Wullings, B.A.; Kooij, van der D.

    2010-01-01

    Legionella pneumophila proliferates in aquatic habitats within free-living protozoa, 17 species of which have been identified as hosts by using in vitro experiments. The present study aimed at identifying protozoan hosts for L. pneumophila by using a biofilm batch test (BBT). Samples (600 ml)

  6. Effect of temperature and colonization of Legionella pneumophila and Vermamoeba vermiformis on bacterial community composition of copper drinking water biofilms.

    Science.gov (United States)

    Buse, Helen Y; Ji, Pan; Gomez-Alvarez, Vicente; Pruden, Amy; Edwards, Marc A; Ashbolt, Nicholas J

    2017-07-01

    It is unclear how the water-based pathogen, Legionella pneumophila (Lp), and associated free-living amoeba (FLA) hosts change or are changed by the microbial composition of drinking water (DW) biofilm communities. Thus, this study characterized the bacterial community structure over a 7-month period within mature (> 600-day-old) copper DW biofilms in reactors simulating premise plumbing and assessed the impact of temperature and introduction of Lp and its FLA host, Vermamoeba vermiformis (Vv), co-cultures (LpVv). Sequence and quantitative PCR (qPCR) analyses indicated a correlation between LpVv introduction and increases in Legionella spp. levels at room temperature (RT), while at 37°C, Lp became the dominant Legionella spp. qPCR analysis suggested Vv presence may not be directly associated with Lp biofilm growth at RT and 37°C, but may contribute to or be associated with non-Lp legionellae persistence at RT. Two-way PERMANOVA and PCoA revealed that temperature was a major driver of microbiome diversity. Biofilm community composition also changed over the seven-month period and could be associated with significant shifts in dissolved oxygen, alkalinity and various metals in the influent DW. Hence, temperature, biofilm age, DW quality and transient intrusions/amplification of pathogens and FLA hosts may significantly impact biofilm microbiomes and modulate pathogen levels over extended periods. © 2016 The Authors. Microbial Biotechnology published by John Wiley & Sons Ltd and Society for Applied Microbiology.

  7. Free-living protozoa in drinking water supplies: community composition and role as hosts for Legionella pneumophila

    NARCIS (Netherlands)

    Valster, R.M.

    2011-01-01

    Free-living protozoa in drinking water supplies: community composition and role as hosts for Legionella pneumophila


    Free-living protozoa, which feed on bacteria, play an important role in the communities of microorganisms and invertebrates in drinking water supplies and in (warm)

  8. Septic arthritis due to Legionella cincinnatiensis: case report and review of the literature.

    Science.gov (United States)

    Banderet, Florian; Blaich, Annette; Soleman, Evelin; Gaia, Valeria; Osthoff, Michael

    2017-08-01

    Legionella spp. are an important cause of pulmonary and rarely extrapulmonary infections. L. cincinnatiensis has only been implicated in five cases to date. We herein report the first case of L. cincinnatiensis septic arthritis in a 90-year old lady with a past medical history of chronic kidney disease. She developed septic arthritis of her left wrist after having received intraarticular corticosteroid injections and oral corticosteroids administered for presumed chondrocalcinosis. Appropriate antimicrobial treatment of L. cincinnatiensis septic arthritis was delayed until identification of this organism in joint biopsies by broad-range bacterial PCR targeting the 16S rRNA gene with subsequent rDNA sequence analysis and by culture on special media. Reviewing all reported cases of septic arthritis caused by Legionella spp. other than L. cincinnatiensis it is notable that diagnosis was established by PCR in the majority of cases and only subsequently confirmed by special culture. Although most patients were immunosuppressed, outcome was favourable. Treatment consisted of a fluoroquinolone alone or in combination with rifampicin or a macrolide. Our case highlights the need for a high index of suspicion for infections with unusual/fastidious organisms when symptoms are suggestive of septic arthritis but conventional methods fail to identify a causative organism.

  9. Legionella spp. Risk Assessment in Recreational and Garden Areas of Hotels.

    Science.gov (United States)

    Papadakis, Antonios; Chochlakis, Dimosthenis; Sandalakis, Vassilios; Keramarou, Maria; Tselentis, Yannis; Psaroulaki, Anna

    2018-03-26

    Several Travel-associated Legionnaires' disease (TALD) cases occur annually in Europe. Except from the most obvious sites (cooling towers and hot water systems), infections can also be associated with recreational, water feature, and garden areas of hotels. This argument is of great interest to better comprehend the colonization and to calculate the risk to human health of these sites. From July 2000-November 2017, the public health authorities of the Island of Crete (Greece) inspected 119 hotels associated with TALD, as reported through the European Legionnaires' Disease Surveillance Network. Five hundred and eighteen samples were collected from decorative fountain ponds, showers near pools and spas, swimming pools, spa pools, garden sprinklers, drip irrigation systems (reclaimed water) and soil. Of those, 67 (12.93%), originating from 43 (35.83%) hotels, tested positive for Legionella ( Legionella pneumophila serogroups 1, 2, 3, 6, 7, 8, 13, 14, 15 and non-pneumophila species ( L. anisa , L. erythra , L. taurinensis , L. birminghamensis , L. rubrilucens ). A Relative Risk (R.R.) > 1 ( p Safety Plan implementation (R.R.: 3.96). High risk (≥10⁴ CFU/L) was estimated for pool showers (16.42%), garden sprinklers (7.46%) and pool water (5.97%).

  10. Legionella spp. Risk Assessment in Recreational and Garden Areas of Hotels

    Directory of Open Access Journals (Sweden)

    Antonios Papadakis

    2018-03-01

    Full Text Available Several Travel-associated Legionnaires’ disease (TALD cases occur annually in Europe. Except from the most obvious sites (cooling towers and hot water systems, infections can also be associated with recreational, water feature, and garden areas of hotels. This argument is of great interest to better comprehend the colonization and to calculate the risk to human health of these sites. From July 2000–November 2017, the public health authorities of the Island of Crete (Greece inspected 119 hotels associated with TALD, as reported through the European Legionnaires’ Disease Surveillance Network. Five hundred and eighteen samples were collected from decorative fountain ponds, showers near pools and spas, swimming pools, spa pools, garden sprinklers, drip irrigation systems (reclaimed water and soil. Of those, 67 (12.93%, originating from 43 (35.83% hotels, tested positive for Legionella (Legionella pneumophila serogroups 1, 2, 3, 6, 7, 8, 13, 14, 15 and non-pneumophila species (L. anisa, L. erythra, L. taurinensis, L. birminghamensis, L. rubrilucens. A Relative Risk (R.R. > 1 (p < 0.0001 was calculated for chlorine concentrations of less than 0.2 mg/L (R.R.: 54.78, star classification (<4 (R.R.: 4.75 and absence of Water Safety Plan implementation (R.R.: 3.96. High risk (≥104 CFU/L was estimated for pool showers (16.42%, garden sprinklers (7.46% and pool water (5.97%.

  11. Influence of Legionella pneumophila and other water bacteria on the survival and growth of Acanthamoeba polyphaga.

    Science.gov (United States)

    Anacarso, I; Guerrieri, E; Bondi, M; de Niederhäusern, S; Iseppi, R; Sabia, C; Contri, M; Borella, P; Messi, P

    2010-10-01

    We investigated in solid medium, in water microcosm co-cultures and by light and transmission electron microscopy the influence of Legionella pneumophila Lp-1, Pseudomonas aeruginosa ATCC 27853, Burkholderia cepacia ATCC 25416 and Pseudomonas fluorescens SSD35 on the growth and survival of Acanthamoeba polyphaga. The infection with L. pneumophila was microscopically characterized by the presence of few bacteria inside protozoa at 4th h, and by the beginning of disruptive effects in late phase of trial. In water microcosm studies, performed at different temperature, the more significant interactions were observed at 30°C. In these conditions, L. pneumophila caused a marked reduction in trophozoite and cyst counts from the 4th day until the end of incubation (11 days). B. cepacia showed, by microscopic observation, few and generally single rods within protozoan phagosomes and caused a light reduction of trophozoite viability and cyst formation in co-cultures. A more invasive type of endocytosis, characterized by an early invasion with the presence of a high bacteria number inside amoebae, was observed for Pseudomonas strains. P. fluorescens produced a violent lysis of the host, whereas P. aeruginosa did not cause lysis or suffering. These results underline that water bacteria other than legionella are capable of intracellular survival in Acanthamoeba, influencing the protozoa viable cycle.

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

    Science.gov (United States)

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

    1998-10-01

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

  13. Contamination of Hospital Water Supplies in Gilan, Iran, with Legionella pneumophila, Escherichia coli, and Pseudomonas aeruginosa

    Directory of Open Access Journals (Sweden)

    Masoumeh Ahmadi Jalali Moghadam

    2015-01-01

    Full Text Available This study is designed to determine the contamination degree of hospital water supplies with Pseudomonas aeruginosa, Legionella pneumophila, and E. coli in Gilan, Iran. Samples were collected directly into sterile containers and concentrated by centrifuge. Half part of any sample transferred to yeast extract broth and the second part transferred to Trypticase Soy Broth and incubated for 3 days. DNA was extracted by using commercial kit. Four rounds of PCR were performed as follows: multiplex PCR for detecting Pseudomonas aeruginosa, Integron 1, and Metallo-β-lactamases gene; PCR for detecting Legionella pneumophila and mip gene separately; PCR for detecting E. coli; and another PCR for detecting whole bacterial presence. Contamination rates of cold, warm, and incubator water samples with P. aeruginosa, were 16.6%, 37.5%, and 6.8% consequently. Degrees of contamination with L. pneumophila were 3.3%, 9.3%, and 10.9% and with E. coli were zero, 6.2%, and zero. Total bacterial contamination of cold, warm, and incubator water samples was 93.3%, 84.4%, and 89.0% consequently. Metallo-β-lactamases gene was found in 20.0% of all samples. Contamination degree with P. aeruginosa was considerable and with L. pneumophila was moderate. Metallo-β-lactamases gene was found frequently indicating widespread multiple drug resistance bacteria. We suggest using new decontamination method based on nanotechnology.

  14. Invasion of Eukaryotic Cells by Legionella Pneumophila: A Common Strategy for all Hosts?

    Directory of Open Access Journals (Sweden)

    Paul S Hoffman

    1997-01-01

    Full Text Available Legionella pneumophila is an environmental micro-organism capable of producing an acute lobar pneumonia, commonly referred to as Legionnaires’ disease, in susceptible humans. Legionellae are ubiquitous in aquatic environments, where they survive in biofilms or intracellularly in various protozoans. Susceptible humans become infected by breathing aerosols laden with the bacteria. The target cell for human infection is the alveolar macrophage, in which the bacteria abrogate phagolysosomal fusion. The remarkable ability of L pneumophila to infect a wide range of eukaryotic cells suggests a common strategy that exploits very fundamental cellular processes. The bacteria enter host cells via coiling phagocytosis and quickly subvert organelle trafficking events, leading to formation of a replicative phagosome in which the bacteria multiply. Vegetative growth continues for 8 to 10 h, after which the bacteria develop into a short, highly motile form called the ‘mature form’. The mature form exhibits a thickening of the cell wall, stains red with the Gimenez stain, and is between 10 and 100 times more infectious than agar-grown bacteria. Following host cell lysis, the released bacteria infect other host cells, in which the mature form differentiates into a Gimenez-negative vegetative form, and the cycle begins anew. Virulence of L pneumophila is considered to be multifactorial, and there is growing evidence for both stage specific and sequential gene expression. Thus, L pneumophila may be a good model system for dissecting events associated with the host-parasite interactions.

  15. The Legionella pneumophila GIG operon responds to gold and copper in planktonic and biofilm cultures.

    Science.gov (United States)

    Jwanoswki, Kathleen; Wells, Christina; Bruce, Terri; Rutt, Jennifer; Banks, Tabitha; McNealy, Tamara L

    2017-01-01

    Legionella pneumophila contaminates man-made water systems and creates numerous exposure risks for Legionnaires' Disease. Because copper/silver ionization is commonly used to control L. pneumophila, its mechanisms of metal response and detoxification are of significant interest. Here we describe an L. pneumophila operon with significant similarity to the GIG operon of Cupriavidus metallidurans. The Legionella GIG operon is present in a subset of strains and has been acquired as part of the ICE-βox 65-kB integrative conjugative element. We assessed GIG promoter activity following exposure of L. pneumophila to multiple concentrations of HAuCl4, CuSO4 and AgNO3. At 37°C, control stationary phase cultures exhibited GIG promoter activity. This activity increased significantly in response to 20 and 50uM HAuCl4 and CuSO4 but not in response to AgNO3. Conversely, at 26°C, cultures exhibited decreased promoter response to copper. GIG promoter activity was also induced by HAuCl4 or CuSO4 during early biofilm establishment at both temperatures. When an L. pneumophila GIG promoter construct was transformed into E. coli DH5α, cultures showed baseline expression levels that did not increase following metal addition. Analysis of L. pneumophila transcriptional regulatory mutants suggested that GIG up-regulation in the presence of metal ions may be influenced by the stationary phase sigma factor, RpoS.

  16. The Legionella pneumophila GIG operon responds to gold and copper in planktonic and biofilm cultures.

    Directory of Open Access Journals (Sweden)

    Kathleen Jwanoswki

    Full Text Available Legionella pneumophila contaminates man-made water systems and creates numerous exposure risks for Legionnaires' Disease. Because copper/silver ionization is commonly used to control L. pneumophila, its mechanisms of metal response and detoxification are of significant interest. Here we describe an L. pneumophila operon with significant similarity to the GIG operon of Cupriavidus metallidurans. The Legionella GIG operon is present in a subset of strains and has been acquired as part of the ICE-βox 65-kB integrative conjugative element. We assessed GIG promoter activity following exposure of L. pneumophila to multiple concentrations of HAuCl4, CuSO4 and AgNO3. At 37°C, control stationary phase cultures exhibited GIG promoter activity. This activity increased significantly in response to 20 and 50uM HAuCl4 and CuSO4 but not in response to AgNO3. Conversely, at 26°C, cultures exhibited decreased promoter response to copper. GIG promoter activity was also induced by HAuCl4 or CuSO4 during early biofilm establishment at both temperatures. When an L. pneumophila GIG promoter construct was transformed into E. coli DH5α, cultures showed baseline expression levels that did not increase following metal addition. Analysis of L. pneumophila transcriptional regulatory mutants suggested that GIG up-regulation in the presence of metal ions may be influenced by the stationary phase sigma factor, RpoS.

  17. The Legionella pneumophila kai operon is implicated in stress response and confers fitness in competitive environments

    Science.gov (United States)

    Loza-Correa, Maria; Sahr, Tobias; Rolando, Monica; Daniels, Craig; Petit, Pierre; Skarina, Tania; Valero, Laura Gomez; Dervins-Ravault, Delphine; Honoré, Nadine; Savchenko, Aleksey; Buchrieser, Carmen

    2014-01-01

    Summary Legionella pneumophila uses aquatic protozoa as replication niche and protection from harsh environments. Although L. pneumophila is not known to have a circadian clock, it encodes homologues of the KaiBC proteins of Cyanobacteria that regulate circadian gene expression. We show that L. pneumophila kaiB, kaiC and the downstream gene lpp1114, are transcribed as a unit under the control of the stress sigma factor RpoS. KaiC and KaiB of L. pneumophila do not interact as evidenced by yeast and bacterial two-hybrid analyses. Fusion of the C-terminal residues of cyanobacterial KaiB to Legionella KaiB restores their interaction. In contrast, KaiC of L. pneumophila conserved autophosphorylation activity, but KaiB does not trigger the dephosphorylation of KaiC like in Cyanobacteria. The crystal structure of L. pneumophila KaiB suggests that it is an oxidoreductase-like protein with a typical thioredoxin fold. Indeed, mutant analyses revealed that the kai operon-encoded proteins increase fitness of L. pneumophila in competitive environments, and confer higher resistance to oxidative and sodium stress. The phylogenetic analysis indicates that L. pneumophila KaiBC resemble Synechosystis KaiC2B2 and not circadian KaiB1C1. Thus, the L. pneumophila Kai proteins do not encode a circadian clock, but enhance stress resistance and adaption to changes in the environments. PMID:23957615

  18. Subversion of Cell-Autonomous Immunity and Cell Migration by Legionella pneumophila Effectors.

    Science.gov (United States)

    Simon, Sylvia; Hilbi, Hubert

    2015-01-01

    Bacteria trigger host defense and inflammatory processes, such as cytokine production, pyroptosis, and the chemotactic migration of immune cells toward the source of infection. However, a number of pathogens interfere with these immune functions by producing specific so-called "effector" proteins, which are delivered to host cells via dedicated secretion systems. Air-borne Legionella pneumophila bacteria trigger an acute and potential fatal inflammation in the lung termed Legionnaires' disease. The opportunistic pathogen L. pneumophila is a natural parasite of free-living amoebae, but also replicates in alveolar macrophages and accidentally infects humans. The bacteria employ the intracellular multiplication/defective for organelle trafficking (Icm/Dot) type IV secretion system and as many as 300 different effector proteins to govern host-cell interactions and establish in phagocytes an intracellular replication niche, the Legionella-containing vacuole. Some Icm/Dot-translocated effector proteins target cell-autonomous immunity or cell migration, i.e., they interfere with (i) endocytic, secretory, or retrograde vesicle trafficking pathways, (ii) organelle or cell motility, (iii) the inflammasome and programed cell death, or (iv) the transcription factor NF-κB. Here, we review recent mechanistic insights into the subversion of cellular immune functions by L. pneumophila.

  19. Subversion of cell-autonomous immunity and cell migration by Legionella pneumophila effectors

    Directory of Open Access Journals (Sweden)

    Sylvia eSimon

    2015-09-01

    Full Text Available Bacteria trigger host defense and inflammatory processes such as cytokine production, pyroptosis and the chemotactic migration of immune cells towards the source of infection. However, a number of pathogens interfere with these immune functions by producing specific so-called effector proteins, which are delivered to host cells via dedicated secretion systems. Air-borne Legionella pneumophila bacteria trigger an acute and potential fatal inflammation in the lung termed Legionnaires’ disease. The opportunistic pathogen L. pneumophila is a natural parasite of free-living amoebae, but also replicates in alveolar macrophages and accidentally infects humans. The bacteria employ the Icm/Dot type IV secretion system and as many as 300 different effector proteins to govern host cell interactions and establish in phagocytes an intracellular replication niche, the Legionella-containing vacuole. Some Icm/Dot-translocated effector proteins target cell autonomous immunity or cell migration, i.e. they interfere with (i endocytic, secretory or retrograde vesicle trafficking pathways, (ii organelle or cell motility, (iii the inflammasome and programmed cell death, or (iv the transcription factor NF-κB. Here we review recent mechanistic insights into the subversion of cellular immune functions by L. pneumophila.

  20. Pneumonia por Legionella após uso de Infliximabe em paciente com Artrite Reumatoide

    Directory of Open Access Journals (Sweden)

    Karina de Souza Giassi

    2014-10-01

    Full Text Available Os antagonistas do fator de necrose tumoral (anti-TNF têm sido utilizados com sucesso em várias doenças inflamatórias crônicas, como artrite reumatoide (AR, mas alguns estudos observaram a ocorrência de infecções por patógenos intracelulares em pacientes medicados com anti-TNF. Relatamos um caso de paciente mulher com diagnóstico prévio de AR durante 16 anos e que estava sendo medicada com várias drogas antirreumáticas modificadoras de doença (DARMDs, tendo como resultado o insucesso terapêutico, sendo em seguida tratada com infliximab. Depois de transcorridos 15 dias da segunda dose, a paciente foi acome- tida por dor torácica ventilatório-dependente, tosse seca e dispneia. Foi hospitalizada, e o diagnóstico de pneumonia por Legionella pneumophila foi confirmado pela presença do antí- geno de Legionella na urina. TNF é uma citocina inflamatória que também promove inibição do crescimento bacteriano de patógenos intracelulares, e sua inibição parece aumentar a sensibilidade a essas infecções em alguns pacientes.

  1. Legionella jordanis in hematopoietic SCT patients radiographically mimicking invasive mold infection.

    Science.gov (United States)

    Meyer, R; Rappo, U; Glickman, M; Seo, S K; Sepkowitz, K; Eagan, J; Small, T N

    2011-08-01

    Opportunistic pulmonary infections are a major cause of post-transplant morbidity and mortality. Among these infections, Aspergillus is a common cause of fatal pneumonia. Owing to the precarious clinical condition of many patients who acquire invasive mold infections, clinicians often treat them on the basis of radiographic findings, such as the halo sign. However, in patients who do not respond to treatment or who have uncommon presentations, bronchoscopy or lung biopsy looking for other pathogens should be considered. This study describes two cases in which the radiographic halo signs characteristic of Aspergillus were in fact due to Legionella jordanis, a pathogen that has been culture proven only in two patients previously (both of whom had underlying lung pathology) and diagnosed by serologic evidence in several other patients. In immunocompromised patients, Legionella can present as a cavitary lesion. Thus, presumptive treatment for this organism should be considered in post-transplant patients who do not have a classic presentation for invasive fungal infection and/or who fail to respond to conventional treatment. These cases illustrate the importance of obtaining tissue cultures to differentiate among the wide variety of pathogens present in this patient population.

  2. Serotyping, ribotyping, PCR-mediated ribosomal 16S-23S spacer analysis and arbitrarily primed PCR for epidemiological studies on Legionella pneumophila

    NARCIS (Netherlands)

    A.F. van Belkum (Alex); H. Maas (Hugo); H.A. Verbrugh (Henri); N. van Leeuwen (N.)

    1996-01-01

    textabstractFifty clinical and environmental isolates of Legionella pneumophila were typed serologically and by DNA fingerprinting using arbitrarily primed polymerase chain reaction (AP-PCR). Furthermore, variability in and around ribosomal operons was assessed by conventional ribotyping and

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

    Data.gov (United States)

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

  4. Molecular detection of Acanthamoeba spp., Naegleria fowleri and Vermamoeba (Hartmannella) vermiformis as vectors for Legionella spp. in untreated and solar pasteurized harvested rainwater

    OpenAIRE

    Dobrowsky, Penelope H.; Khan, Sehaam; Cloete, Thomas E.; Khan, Wesaal

    2016-01-01

    Background Legionella spp. employ multiple strategies to adapt to stressful environments including the proliferation in protective biofilms and the ability to form associations with free-living amoeba (FLA). The aim of the current study was to identify Legionella spp., Acanthamoeba spp., Vermamoeba (Hartmannella) vermiformis and Naegleria fowleri that persist in a harvested rainwater and solar pasteurization treatment system. Methods Pasteurized (45??C, 65??C, 68??C, 74??C, 84??C and 93??C) a...

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

    Science.gov (United States)

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

    2016-02-01

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

  6. A single Legionella pneumophila genotype in the freshwater system in a ship experiencing three separate outbreaks of legionellosis in 6 years

    Directory of Open Access Journals (Sweden)

    Catrine Ahlen

    2016-08-01

    Full Text Available Background: Recurrent legionella outbreaks at one and the same location are common. We have identified a single Legionella pneumophila genotype associated with recurrent Legionella outbreaks over 6 years. Methods: Field emergency surveys following Legionella outbreaks were performed on a vessel in 2008, 2009 and 2013. Water samples from both the distribution and technical parts of the potable water system were analyzed with respect to L. pneumophila [Real-Time PCR, cultivation, serotyping and genotyping (PFGE] and free-living amoebae, (FLA. Results: Legionella pneumophila serogroup 1 was present in the ship's potable water system during every outbreak. Genotyping of the 2008 survey material showed two separate PFGE genotypes while those in 2009 and 2013 demonstrated the presence of only one of the two genotypes. FLA with intracellular L. pneumophila of the same genotype were also detected. Analyses of the freshwater system on a ship following three separate Legionella outbreaks, for L. pneumophila and FLAs, revealed a single L. pneumophila genotype and FLA (Hartmanella. Conclusions: It is reasonable to assume that the L. pneumophila genotype detected in the freshwater system was the causal agent in the outbreaks onboard. Persistence of an apparently low-pathogenic L. pneumophila genotype and FLA in a potable water system represent a potential risk for recurrent outbreaks.

  7. Rapid quantification of viable Legionella in nuclear cooling tower waters using filter cultivation, fluorescent in situ hybridization and solid-phase cytometry.

    Science.gov (United States)

    Baudart, J; Guillaume, C; Mercier, A; Lebaron, P; Binet, M

    2015-05-01

    To develop a rapid and sensitive method to quantify viable Legionella spp. in cooling tower water samples. A rapid, culture-based method capable of quantifying as few as 600 Legionella microcolonies per litre within 2 days in industrial waters was developed. The method combines a short cultivation step of microcolonies on GVPC agar plate, specific detection of Legionella cells by a fluorescent in situ hybridization (FISH) approach, and a sensitive enumeration using a solid-phase cytometer. Following optimization of the cultivation conditions, the qualitative and quantitative performance of the method was assessed and the method was applied to 262 nuclear power plant cooling water samples. The performance of this method was in accordance with the culture method (NF-T 90-431) for Legionella enumeration. The rapid detection of viable Legionella in water is a major concern to the effective monitoring of this pathogenic bacterium in the main water sources involved in the transmission of legionellosis infection (Legionnaires' disease). The new method proposed here appears to be a robust, efficient and innovative means for rapidly quantifying cultivable Legionella in cooling tower water samples within 48 h. © 2015 The Society for Applied Microbiology.

  8. Applicability assessment of ceramic microbeads coated with hydroxyapatite-binding silver/titanium dioxide ceramic composite earthplus™ to the eradication of Legionella in rainwater storage tanks for household use.

    Science.gov (United States)

    Oana, Kozue; Kobayashi, Michiko; Yamaki, Dai; Sakurada, Tsukasa; Nagano, Noriyuki; Kawakami, Yoshiyuki

    2015-01-01

    Water environments appear to be the habitats of Legionella species. Legionellosis is considered as a preventable illness because bacterial reservoirs can be controlled and removed. Roof-harvested rainwater has attracted significant attention not only as a groundwater recharge but also as a potential alternative source of nonpotable water. We successfully developed ceramic microbeads coated with hydroxyapatite-binding silver/titanium dioxide ceramic composite earthplus™ using the thermal spraying method. The ceramic microbeads were demonstrated to have bactericidal activities against not only Legionella but also coliform and heterotrophic bacteria. Immersing the ceramic microbeads in household rainwater storage tanks was demonstrated to yield the favorable eradication of Legionella organisms. Not only rapid-acting but also long-lasting bactericidal activities of the ceramic microbead were exhibited against Legionella pneumophila. However, time-dependent attenuation of the bactericidal activities against Legionella were also noted in the sustainability appraisal experiment. Therefore, the problems to be overcome surely remain in constantly managing the Legionella-pollution by means of immersing the ceramic microbeads. The results of our investigation apparently indicate that the earthplus™-coated ceramic microbeads would become the favorable tool for Legionella measures in household rainwater storage tanks, which may become the natural reservoir for Legionella species. Our investigation would justify further research and data collection to obtain more reliable procedures to microbiologically regulate the Legionella in rainwater storage tanks.

  9. Brote epidémico de neumonías por Legionella pneumophila en niños cubanos

    Directory of Open Access Journals (Sweden)

    Roberto Razón Behar

    2002-09-01

    Full Text Available La Legionella pneumophila es uno de los patógenos responsable de neumonías atípicas, a través de la inhalación de aerosoles o aspiración de líquidos infectados. Se detectó un brote epidémico de neumonías por Legionella, originado por la aspiración de agua contaminada de una piscina en un grupo de niños cubanos. El agente causal se identificó en 5 de 9 pacientes, por la técnica de inmunofluorescencia indirecta en muestras de sueros pareados. Los síntomas y signos más frecuentes fueron malestar general, anorexia, astenia, fiebre persistente de 39 °C a 40 °C (103 °F a 105 °F, mialgias, cefaleas, náuseas, vómitos, dolor abdominal, diarreas, tos húmeda, dolor torácico y polipnea. Durante el desarrollo de la enfermedad, el tratamiento antibiótico fue empírico (incluyendo los macrólidos, por no tener confirmado el diagnóstico. Todos los pacientes evolucionaron satisfactoriamente. Se reportó un brote epidémico de neumonías por Legionella en niños por primera vez en Cuba, lo cual tiene importancia clínica y epidemiológica.The legionella pneumophila is one of the pathogens responsible for atypic pneumonias by the inhalation of aerosols or aspiration of infected liquids. An epidemic outbreak of pneumonias caused by Legionella was detected among a group of Cuban children. It was originated by the aspiration of contaminated water in a swimming pool. The causal agent was identified in 5 of 9 patients by using the indirect immunofluorescence technique in samples of matched sera. The most frequent symptoms and signs were malaise, anorexia, asthenia, persistent fever from 39°C to 40°C (103° F to 105° F, myalgias, headache, nauseas, vomits, abdominal pain, diarrheas, moist cough, thoracic pain and polypnoea. The antibiotic treatment was empiric (including the macrolides during the development of the disease, since the diagnosis was not confirmed. The patients’ evolution was satisfactory. An epidemic outbreak of pneumonias

  10. 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.

  11. A bacterial protein promotes the recognition of the Legionella pneumophila vacuole by autophagy

    Science.gov (United States)

    Khweek, Arwa Abu; Caution, Kyle; Akhter, Anwari; Abdulrahman, Basant A.; Tazi, Mia; Hassan, Hoda; Majumdar, Neal; Doran, Andrew; Guirado, Evelyn; Schlesinger, Larry S.; Shuman, Howard; Amer, Amal O.

    2013-01-01

    Legionella pneumophila (L. pneumophila) is an intracellular bacterium of human alveolar macrophages that causes Legionnaires' disease. In contrast to humans, most inbred mouse strains are restrictive to L. pneumophila replication. We demonstrate that autophagy targets L. pneumophila vacuoles to lysosomes and that this process requires ubiquitination of L. pneumophila vacuoles and the subsequent binding of the autophagic adaptor p62/SQSTM1 to ubiquitinated vacuoles. The L. pneumophila legA9 encodes for an ankyrin-containing protein with unknown role. We show that the legA9 mutant is the first L. pneumophila mutant to replicate in wild-type (WT) mice and their bone marrow derived macrophages (BMDMs). Less legA9 mutant- containing vacuoles acquired ubiquitin labeling and p62/SQSTM1 staining, evading autophagy uptake and avoiding lysosomal fusion. Thus, we describe a bacterial protein that targets the L. pneumophila -containing vacuole for autophagy uptake. PMID:23420491

  12. Integrated Real-Time PCR for Detection and Monitoring of Legionella pneumophila in Water Systems▿

    Science.gov (United States)

    Yaradou, Diaraf Farba; Hallier-Soulier, Sylvie; Moreau, Sophie; Poty, Florence; Hillion, Yves; Reyrolle, Monique; André, Janine; Festoc, Gabriel; Delabre, Karine; Vandenesch, François; Etienne, Jerome; Jarraud, Sophie

    2007-01-01

    We evaluated a ready-to-use real-time quantitative Legionella pneumophila PCR assay system by testing 136 hot-water-system samples collected from 55 sites as well as 49 cooling tower samples collected from 20 different sites, in parallel with the standard culture method. The PCR assay was reproducible and suitable for routine quantification of L. pneumophila. An acceptable correlation between PCR and culture results was obtained for sanitary hot-water samples but not for cooling tower samples. We also monitored the same L. pneumophila-contaminated cooling tower for 13 months by analyzing 104 serial samples. The culture and PCR results were extremely variable over time, but the curves were similar. The differences between the PCR and culture results did not change over time and were not affected by regular biocide treatment. This ready-to-use PCR assay for L. pneumophila quantification could permit more timely disinfection of cooling towers. PMID:17194840

  13. Effect of Legionella pneumophila sonicate on killing of Listeria monocytogenes by human polymorphonuclear neutrophils and monocytes

    DEFF Research Database (Denmark)

    Rechnitzer, C; Bangsborg, Jette Marie; Shand, G H

    1993-01-01

    polymorphonuclear neutrophils and monocytes. Preincubation of neutrophils with L. pneumophila sonicate did not affect phagocytosis of L. monocytogenes, whereas Listeria killing was significantly inhibited at sonicate concentrations of 1 and 2 mg/ml. The phenol phase of a phenol-water extraction, containing most...... of the lipopolysaccharide (LPS), had no inhibitory effect on the listericidal activity of neutrophils. Killing of Listeria by monocytes was inhibited in a similar manner. The inhibitory activity was mainly recovered in the sonicate fraction above 100 kDa, suggesting that components organized in larger molecular complexes...... are most likely to represent the inhibitory factors. The inhibitory activity of L. pneumophila sonic extract appears to be related to inhibition of killing mechanisms since uptake of Listeria was not affected by the sonicate. Our observations indicate that as Legionella infection progresses, bacterial...

  14. CT features of legionella pneumonia, compared with streptococcal pneumonia. A collaborative study

    International Nuclear Information System (INIS)

    Tokuda, Hitoshi; Sakai, Fumikazu; Goto, Hajime

    2007-01-01

    Legionella pneumophila pneumonia (LPP) is of serious concern for chest physicians for its potential lethality and difficulty of diagnosis. In spite of widespread use of urine antigen detection method, it remains one of the most difficult-to-treat disease among community acquired pneumonia. We investigated CT images of 38 cases of LPP, comparing them with wide spread Streptococcus pneumoniae pneumonia (SPP). We find that sharply demarcated consolidation scattered in ground-glass opacity is the most striking and pathognomonic feature of LPP, which is rarely found in SPP. Bronchiolitis was not found in LPP, while it is a relatively common finding in SPP, which could be the second clue of differentiation of these two diseases. (author)

  15. Neumonía por Legionella pneumophila: Experiencia en un Hospital Universitario de Buenos Aires Neumonia due to Legionella pneumophila. Experience gathered in a University Hospital in Buenos Aires

    Directory of Open Access Journals (Sweden)

    Carlos M. Luna

    2004-04-01

    Full Text Available La enfermedad de los legionarios es una causa de neumonía adquirida en la comunidad (NAC reconocida en todo el mundo. En Latinoamérica su incidencia es desconocida. En este estudio se analizó a 9 pacientes con NAC por Legionella pneumophila atendidos entre 1997 y 2001 en el Hospital de Clínicas José de San Martín de la Universidad de Buenos Aires. Se registraron datos de antecedentes, enfermedad actual, contactos, exposición laboral, examen físico, pruebas de laboratorio y uso previo de antibióticos, y se tomó en cuenta la presencia de criterios de gravedad. Nueve pacientes presentaron diagnóstico de NAC por Legionella, ninguno refirió antecedentes de viajes recientes; cuatro de ellos debieron ser internados en unidades de cuidado intensivo. Siete pacientes tenían antecedentes de tabaquismo, 4 tenían EPOC y un paciente linfoma no-Hodgkin. Nuestra casuística corrobora la baja especificidad de la clínica y estudios complementarios para predecir esta etiología. El aislamiento de Legionella es dificultoso, la seroconversión permite el diagnóstico retrospectivo y requiere plazos prolongados y el antígeno urinario aporta un diagnóstico inmediato. Cuando la legionelosis aparece en casos aislados, como ocurriría en Argentina, si no se piensa en esta etiología no se llegará al diagnóstico. Legionella pneumophila es un patógeno de NAC en nuestro medio, debe buscarse mejor, particularmente en pacientes graves, inmunodeprimidos y en fumadores con enfermedad pulmonar obstructiva crónica (EPOC.Legionnaires’ disease is a well recognized cause of community acquired pneumonia (CAP all around the world. In Latin America its incidence remains unknown. This study analyzed a cohort of 9 patients with CAP due to Legionella pneumophila observed from 1997 to 2001, in the Hospital de Clínicas José de San Martín, University of Buenos Aires. Clinical history included recent illnesses, work exposure, physical exam, prior antibiotic use and

  16. Promotion and Rescue of Intracellular Brucella neotomae Replication during Coinfection with Legionella pneumophila.

    Science.gov (United States)

    Kang, Yoon-Suk; Kirby, James E

    2017-05-01

    We established a new Brucella neotomae in vitro model system for study of type IV secretion system-dependent (T4SS) pathogenesis in the Brucella genus. Importantly, B. neotomae is a rodent pathogen, and unlike B. abortus , B. melitensis , and B. suis , B. neotomae has not been observed to infect humans. It therefore can be handled more facilely using biosafety level 2 practices. More particularly, using a series of novel fluorescent protein and lux operon reporter systems to differentially label pathogens and track intracellular replication, we confirmed T4SS-dependent intracellular growth of B. neotomae in macrophage cell lines. Furthermore, B. neotomae exhibited early endosomal (LAMP-1) and late endoplasmic reticulum (calreticulin)-associated phagosome maturation. These findings recapitulate prior observations for human-pathogenic Brucella spp. In addition, during coinfection experiments with Legionella pneumophila , we found that defective intracellular replication of a B. neotomae T4SS virB4 mutant was rescued and baseline levels of intracellular replication of wild-type B. neotomae were significantly stimulated by coinfection with wild-type but not T4SS mutant L. pneumophila Using confocal microscopy, it was determined that intracellular colocalization of B. neotomae and L. pneumophila was required for rescue and that colocalization came at a cost to L. pneumophila fitness. These findings were not completely expected based on known temporal and qualitative differences in the intracellular life cycles of these two pathogens. Taken together, we have developed a new system for studying in vitro Brucella pathogenesis and found a remarkable T4SS-dependent interplay between Brucella and Legionella during macrophage coinfection. Copyright © 2017 American Society for Microbiology.

  17. Microbiological diagnosis and molecular typing of Legionella strains during an outbreak of legionellosis in Southern Germany.

    Science.gov (United States)

    Essig, Andreas; von Baum, Heike; Gonser, Theodor; Haerter, Georg; Lück, Christian

    2016-02-01

    An explosive outbreak of Legionnaires' disease with 64 reported cases occurred in Ulm/Neu-Ulm in the South of Germany in December 2009/January 2010 caused by Legionella (L.) pneumophila serogroup 1, monoclonal (mAb) subtype Knoxville, sequence type (ST) 62. Here we present the clinical microbiological results from 51 patients who were diagnosed at the University hospital of Ulm, the results of the environmental investigations and of molecular typing of patients and environmental strains. All 50 patients from whom urine specimens were available were positive for L. pneumophila antigen when an enzyme-linked immunosorbent assay (EIA) was used following concentration of those urine samples that tested initially negative. The sensitivity of the BinaxNow rapid immunographic assay (ICA), after 15 min reading and after 60 min reading were 70% and 84%, respectively. Direct typing confirmed the monoclonal subtype Knoxville in 5 out of 8 concentrated urine samples. Real time PCR testing of respiratory tract specimens for L. pneumophila was positive in 15 out of 25 (60%) patients. Direct nested sequence based typing (nSBT) in some of these samples allowed partial confirmation of ST62. L. pneumophila serogroup 1, monoclonal subtype Knoxville ST62, defined as the epidemic strain was isolated from 8 out of 31 outbreak patients (26%) and from one cooling tower confirming it as the most likely source of the outbreak. While rapid detection of Legionella antigenuria was crucial for the recognition and management of the outbreak, culture and molecular typing of the strains from patients and environmental specimens was the clue for the rapid identification of the source of infection. Copyright © 2016 Elsevier GmbH. All rights reserved.

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

    Directory of Open Access Journals (Sweden)

    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

  19. Legionella Bozemanae, a New Cause of Septic Arthritis diagnosed by 16S PCR followed by specific culture

    DEFF Research Database (Denmark)

    Just, Søren Andreas; Bonde Knudsen, John; Skov, Marianne Nielsine

    This is the first report ever to demonstrate that L. Bozemanae can colonize synovial joints leading to infectious arthritis. L. Bozemanae is a rare Legionella species, earlier described as a cause of cavitating lung infections with up to 40% mortality (2). L. Bozemanae is missed by standard cultu...... cultures, however, the case was diagnosed by combining 16S PCR and sequencing followed by culture under specific conditions, a method which may help in the diagnosis of septic arthritis caused by unusual pathogens not detected by standard culture.......This is the first report ever to demonstrate that L. Bozemanae can colonize synovial joints leading to infectious arthritis. L. Bozemanae is a rare Legionella species, earlier described as a cause of cavitating lung infections with up to 40% mortality (2). L. Bozemanae is missed by standard...

  20. Detection of Legionella by quantitative-polymerase chain reaction (qPCR) for monitoring and risk assessment

    DEFF Research Database (Denmark)

    Krøjgaard, Louise H.; Krogfelt, Karen A.; Albrechtsen, Hans-Jorgen

    2011-01-01

    Background: Culture and quantitative polymerase chain reaction (qPCR) assays for the detection of Legionella were compared on samples from a residential area before and after two interventions. A total of 84 samples were collected from shower hoses and taps as first flush samples and at constant...... temperature. Samples were grouped according to the origin of the sample, a) circulation water b) water from empty apartments c) water from shower hoses. The aims were to investigate the usefulness of qPCR compared to culture for monitoring remedial actions for elimination of Legionella bacteria and as a tool...... detection limit (> 10 CFU/L) to 1.6*10(6) CFU/L. In circulating water and in first flush water from shower hoses, culture and qPCR showed the same tendencies. The overall correlation between the bacteria number detected by culture and the two developed qPCR assays (L. spp and L. pneumophila) was relatively...

  1. Effectiveness of different methods to control legionella in the water supply: ten-year experience in an Italian university hospital.

    Science.gov (United States)

    Marchesi, I; Marchegiano, P; Bargellini, A; Cencetti, S; Frezza, G; Miselli, M; Borella, P

    2011-01-01

    We report our ten-year experience of hyperchlorination, thermal shock, chlorine dioxide, monochloramine, boilers and point-of-use filters for controlling legionella contamination in a hospital hot water distribution system. Shock disinfections were associated with a return to pre-treatment contamination levels within one or two months. We found that chlorine dioxide successfully maintained levels at contamination was observed applying point-of-use filters and electric boilers at temperatures of >58°C and no cases of nosocomial legionellosis were detected in the ten-year observation period. Our performance ranking in reducing legionella contamination was filter, boiler, chlorine dioxide, hyperchlorination and thermal shock. Chlorine dioxide was the least expensive procedure followed by thermal shock, hyperchlorination, boiler and filter. We suggest adopting chlorine dioxide and electric boilers in parallel. Copyright © 2010 The Hospital Infection Society. Published by Elsevier Ltd. All rights reserved.

  2. Investigation of Legionella Contamination in Bath Water Samples by Culture, Amoebic Co-Culture, and Real-Time Quantitative PCR Methods.

    Science.gov (United States)

    Edagawa, Akiko; Kimura, Akio; Kawabuchi-Kurata, Takako; Adachi, Shinichi; Furuhata, Katsunori; Miyamoto, Hiroshi

    2015-10-19

    We investigated Legionella contamination in bath water samples, collected from 68 bathing facilities in Japan, by culture, culture with amoebic co-culture, real-time quantitative PCR (qPCR), and real-time qPCR with amoebic co-culture. Using the conventional culture method, Legionella pneumophila was detected in 11 samples (11/68, 16.2%). Contrary to our expectation, the culture method with the amoebic co-culture technique did not increase the detection rate of Legionella (4/68, 5.9%). In contrast, a combination of the amoebic co-culture technique followed by qPCR successfully increased the detection rate (57/68, 83.8%) compared with real-time qPCR alone (46/68, 67.6%). Using real-time qPCR after culture with amoebic co-culture, more than 10-fold higher bacterial numbers were observed in 30 samples (30/68, 44.1%) compared with the same samples without co-culture. On the other hand, higher bacterial numbers were not observed after propagation by amoebae in 32 samples (32/68, 47.1%). Legionella was not detected in the remaining six samples (6/68, 8.8%), irrespective of the method. These results suggest that application of the amoebic co-culture technique prior to real-time qPCR may be useful for the sensitive detection of Legionella from bath water samples. Furthermore, a combination of amoebic co-culture and real-time qPCR might be useful to detect viable and virulent Legionella because their ability to invade and multiply within free-living amoebae is considered to correlate with their pathogenicity for humans. This is the first report evaluating the efficacy of the amoebic co-culture technique for detecting Legionella in bath water samples.

  3. Investigation of Legionella Contamination in Bath Water Samples by Culture, Amoebic Co-Culture, and Real-Time Quantitative PCR Methods

    Directory of Open Access Journals (Sweden)

    Akiko Edagawa

    2015-10-01

    Full Text Available We investigated Legionella contamination in bath water samples, collected from 68 bathing facilities in Japan, by culture, culture with amoebic co-culture, real-time quantitative PCR (qPCR, and real-time qPCR with amoebic co-culture. Using the conventional culture method, Legionella pneumophila was detected in 11 samples (11/68, 16.2%. Contrary to our expectation, the culture method with the amoebic co-culture technique did not increase the detection rate of Legionella (4/68, 5.9%. In contrast, a combination of the amoebic co-culture technique followed by qPCR successfully increased the detection rate (57/68, 83.8% compared with real-time qPCR alone (46/68, 67.6%. Using real-time qPCR after culture with amoebic co-culture, more than 10-fold higher bacterial numbers were observed in 30 samples (30/68, 44.1% compared with the same samples without co-culture. On the other hand, higher bacterial numbers were not observed after propagation by amoebae in 32 samples (32/68, 47.1%. Legionella was not detected in the remaining six samples (6/68, 8.8%, irrespective of the method. These results suggest that application of the amoebic co-culture technique prior to real-time qPCR may be useful for the sensitive detection of Legionella from bath water samples. Furthermore, a combination of amoebic co-culture and real-time qPCR might be useful to detect viable and virulent Legionella because their ability to invade and multiply within free-living amoebae is considered to correlate with their pathogenicity for humans. This is the first report evaluating the efficacy of the amoebic co-culture technique for detecting Legionella in bath water samples.

  4. Effect of temperature and colonization of Legionella pneumophila and Vermamoeba vermiformis on bacterial community composition of copper drinking water biofilms

    OpenAIRE

    Buse, Helen Y.; Ji, Pan; Gomez?Alvarez, Vicente; Pruden, Amy; Edwards, Marc A.; Ashbolt, Nicholas J.

    2017-01-01

    Summary It is unclear how the water?based pathogen, Legionella pneumophila (Lp), and associated free?living amoeba (FLA) hosts change or are changed by the microbial composition of drinking water (DW) biofilm communities. Thus, this study characterized the bacterial community structure over a 7?month period within mature (>?600?day?old) copper DW biofilms in reactors simulating premise plumbing and assessed the impact of temperature and introduction of Lp and its FLA host, Vermamoeba vermifor...

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

    OpenAIRE

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

    2018-01-01

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

  6. Use of DNA probes for the diagnosis of infections caused by Mycoplasma pneumoniae and legionellae--a review.

    Science.gov (United States)

    Edelstein, P H

    1990-01-01

    Specific DNA probes have been made for both M. pneumoniae and Legionella species. Dot blot methods have been used in research laboratories to test culture isolates of both organisms, and also to test animal tissues with a L. pneumophila-specific probe. Commercial kits are also available for direct specimen testing for these two organisms. The commercial kits are made by a single manufacturer, Gen-Probe, Inc. (San Diego, CA), and use a novel in-solution rapid hybridization assay, using 125I-labeled cDNA to rRNAs of the organisms. The Gen-Probe M. pneumoniae probe appears to be 80% to 100% sensitive, and 97% to 100% specific, based on analysis of two clinical studies using positive culture as the diagnostic criterion. The Gen-Probe legionella probe appears to be 33% to 71% sensitive (mean 57%), and 98.9% to 99.7% specific (mean 99.7%), based on analysis of four prospective clinical studies, using positive culture as the definition of disease, with a total sample size of 3,243 patients, 49 of which were culture-positive. Both Gen-Probe direct tests appear to be clinically useful, although the poor performance of the legionella test in one major university laboratory, and the expense of performing these tests, mandate that thorough evaluations be carried out in each laboratory anticipating using the test. Culture must always be performed for legionella whether or not the DNA probe test is used. It is likely that the use of the M. pneumoniae kit would greatly speed diagnosis, but whether this would alter medical practice or result in lower morbidity and health care costs is unknown.

  7. IFNs Modify the Proteome of Legionella-Containing Vacuoles and Restrict Infection Via IRG1-Derived Itaconic Acid.

    Science.gov (United States)

    Naujoks, Jan; Tabeling, Christoph; Dill, Brian D; Hoffmann, Christine; Brown, Andrew S; Kunze, Mareike; Kempa, Stefan; Peter, Andrea; Mollenkopf, Hans-Joachim; Dorhoi, Anca; Kershaw, Olivia; Gruber, Achim D; Sander, Leif E; Witzenrath, Martin; Herold, Susanne; Nerlich, Andreas; Hocke, Andreas C; van Driel, Ian; Suttorp, Norbert; Bedoui, Sammy; Hilbi, Hubert; Trost, Matthias; Opitz, Bastian

    2016-02-01

    Macrophages can be niches for bacterial pathogens or antibacterial effector cells depending on the pathogen and signals from the immune system. Here we show that type I and II IFNs are master regulators of gene expression during Legionella pneumophila infection, and activators of an alveolar macrophage-intrinsic immune response that restricts bacterial growth during pneumonia. Quantitative mass spectrometry revealed that both IFNs substantially modify Legionella-containing vacuoles, and comparative analyses reveal distinct subsets of transcriptionally and spatially IFN-regulated proteins. Immune-responsive gene (IRG)1 is induced by IFNs in mitochondria that closely associate with Legionella-containing vacuoles, and mediates production of itaconic acid. This metabolite is bactericidal against intravacuolar L. pneumophila as well as extracellular multidrug-resistant Gram-positive and -negative bacteria. Our study explores the overall role IFNs play in inducing substantial remodeling of bacterial vacuoles and in stimulating production of IRG1-derived itaconic acid which targets intravacuolar pathogens. IRG1 or its product itaconic acid might be therapeutically targetable to fight intracellular and drug-resistant bacteria.

  8. Measurements of water temperature in fountains as an indicator of potential secondary water pollution caused by Legionella bacteria

    Directory of Open Access Journals (Sweden)

    Bąk Joanna

    2018-01-01

    Full Text Available At high air temperatures persisting for a long time, water temperature in the fountains may also increase significantly. This can cause a sudden and significant increase in Legionella bacteria, which results in secondary water contamination. This phenomenon with water – air aerosol generated by fountains can be very dangerous for people. During the test, water temperature measurements in fountains in Poland were made. These research tests was conducted in the spring and summer. The research was conducted in order to determine whether there is a possibility of growth of Legionella bacteria. One of the aims of the study was to determine what temperature range occurs in the fountains and how the temperature changes in the basin of the fountain and when the highest temperature occurs. Single temperature measurements were made and also the temperature distribution was measured during daylight hours. The water temperature in most cases was greater than 20°C, but in no case exceed 26°C. The paper presents also the review about the effect of water temperature on the presence and bacterial growth. The study confirmed the existence of the risk of increasing the number of bacteria of the genus Legionella in the water in the fountains.

  9. Measurements of water temperature in fountains as an indicator of potential secondary water pollution caused by Legionella bacteria

    Science.gov (United States)

    Bąk, Joanna

    2018-02-01

    At high air temperatures persisting for a long time, water temperature in the fountains may also increase significantly. This can cause a sudden and significant increase in Legionella bacteria, which results in secondary water contamination. This phenomenon with water - air aerosol generated by fountains can be very dangerous for people. During the test, water temperature measurements in fountains in Poland were made. These research tests was conducted in the spring and summer. The research was conducted in order to determine whether there is a possibility of growth of Legionella bacteria. One of the aims of the study was to determine what temperature range occurs in the fountains and how the temperature changes in the basin of the fountain and when the highest temperature occurs. Single temperature measurements were made and also the temperature distribution was measured during daylight hours. The water temperature in most cases was greater than 20°C, but in no case exceed 26°C. The paper presents also the review about the effect of water temperature on the presence and bacterial growth. The study confirmed the existence of the risk of increasing the number of bacteria of the genus Legionella in the water in the fountains.

  10. Molecular typing of Legionella pneumophila from air-conditioning cooling waters using mip gene, SBT, and FAFLP methods.

    Science.gov (United States)

    Gong, Xiangli; Li, Juntao; Zhang, Ying; Hou, Shuiping; Qu, Pinghua; Yang, Zhicong; Chen, Shouyi

    2017-08-01

    Legionella spp. are important waterborne pathogens. Molecular typing has become an important method for outbreaks investigations and source tracking of Legionnaires. In a survey program conducted by the Guangzhou Center for Disease Control and Prevention, multiple serotypes Legionella pneumophila (L. pneumophila) were isolated from waters in air-conditioning cooling towers in urban Guangzhou region, China between 2008 and 2011. Three genotyping methods, mip (macrophage infectivity potentiator) genotyping, SBT (sequence-based typing), and FAFLP (fluorescent amplified fragment length polymorphism analysis) were used to type these waterborne L. pneumophila isolates. The three methods were capable of typing all the 134 isolates and a reference strain of L. pneumophila (ATCC33153), with discriminatory indices of 0.7034, 0.9218, and 0.9376, for the mip, SBT, and FAFLP methods respectively. Among the 9 serotypes of the 134 isolates, 10, 50, and 34 molecular types were detected by the mip, SBT, and FAFLP methods respectively. The mip genotyping and SBT typing are more feasible for inter-laboratory results sharing and comparison of different types of L. pneumophila. The SBT and FAFLP typing methods were rapid with higher discriminatory abilities. Combinations of two or more of the typing methods enables more accurate typing of Legionella isolates for outbreak investigations and source tracking of Legionnaires. Copyright © 2017 Elsevier B.V. All rights reserved.

  11. Impact of Chlorine and Heat on the Survival of Hartmannella vermiformis and Subsequent Growth of Legionella pneumophila.

    Science.gov (United States)

    Kuchta, J M; Navratil, J S; Shepherd, M E; Wadowsky, R M; Dowling, J N; States, S J; Yee, R B

    1993-12-01

    Hartmannella vermiformis, a common amoebal inhabitant of potable-water systems, supports intracellular multiplication of Legionella pneumophila and is probably important in the transportation and amplification of legionellae within these systems. To provide a practical guide for decontamination of potable-water systems, we assessed the chlorine and heat resistance of H. vermiformis. H. vermiformis cysts and trophozoites were treated independently with chlorine at concentrations of 2.0 to 10.0 ppm for 30 min and then cocultured with L. pneumophila. Both cysts and trophozoites were sensitive to concentrations between 2.0 and 4.0 ppm and above (trophozoites somewhat more so than cysts), and 10.0 ppm was lethal to both forms. Hartmannellae treated with chlorine up to a concentration of 4.0 ppm supported the growth of legionellae. To determine whether heat would be an effective addendum to chlorine treatment of amoebae, hartmannellae were subjected to temperatures of 55 and 60 degrees C for 30 min and alternatively to 50 degrees C followed by treatment with chlorine at a concentration of 2 ppm. Fewer than 0.05% of the amoebae survived treatment at 55 degrees C, and there were no survivors at 60 degrees C. Pretreatment at 50 degrees C appeared to make hartmannella cysts more susceptible to chlorine but did not further reduce the concentration of trophozoites.

  12. ExaSP2

    Energy Technology Data Exchange (ETDEWEB)

    2017-09-08

    ExaSP2 is a reference implementation of typical linear algebra algorithms and workloads for a quantum molecular dynamics (QMD) electronic structure code. The algorithm is based on a recursive second-order Fermi-Operator expansion method (SP2) and is tailored for density functional based tight-binding calculations of material systems.

  13. Growth of Legionella anisa in a model drinking water system to evaluate different shower outlets and the impact of cast iron rust.

    Science.gov (United States)

    van der Lugt, Wilco; Euser, Sjoerd M; Bruin, Jacob P; Den Boer, Jeroen W; Walker, Jimmy T; Crespi, Sebastian

    2017-11-01

    Legionella continues to be a problem in water systems. This study investigated the influence of different shower mixer faucets, and the influence of the presence of cast iron rust from a drinking water system on the growth of Legionella. The research is conducted using a model of a household containing four drinking water systems. All four systems, which contained standard plumbing components including copper pipes and a water heater, were filled with unchlorinated drinking water. Furthermore, all systems had three different shower faucets: (A) a stainless-steel faucet, (B) a brass-ceramic faucet, and (C) a brass thermostatic faucet. System 1 was solely filled with drinking water. System 2 was filled with drinking water, and cast iron rust. System 3 was contaminated with Legionella, and system 4 was contaminated with a Legionella, and cast iron rust. During a period of 34 months, 450 cold water samples were taken from 15 sample points of the four drinking water systems, and tested for Legionella according to the Dutch Standard (NEN 6265). In system 4, with added cast iron rust, the stainless-steel mixer faucet (A) had the highest concentration of Legionella at >4.3log10CFU/l (>20,000CFU/l) and was positive in 46.4% of samples. In contrast, the stainless-steel mixer faucet (A) of system 3 without cast iron rust showed 14.3% positive samples with a maximum concentration of 3.9log10CFU/l (7600CFU/l) Legionella. Additionally, both contaminated systems (3 and 4), with the brass thermostatic faucet (C), tested positive for Legionella. System 3 in 85.7% of the samples, with a maximum concentration of 4.38log10CFU/l (24,200CFU/l), and system 4 in 64.3% of the samples with a maximum concentration of 4.13log10CFU/l (13.400CFU/l). These results suggest that both the type of faucet used in a drinking water system and the presence or absence of cast iron rust influence the growth of Legionella. Copyright © 2017 Elsevier GmbH. All rights reserved.

  14. Métodos analíticos para el estudio de Legionella

    Directory of Open Access Journals (Sweden)

    Carmen Pelaz Antolín

    2006-12-01

    Full Text Available Los ensayos para la determinación de Legionella en muestras de agua son uno de los aspectos contemplados en la legislación española sobre prevención de legionelosis. La periodicidad de estos ensayos en función del tipo de instalación, los laboratorios que los realizan, y las acciones correctoras que derivan de ellos en función de los recuentos bacterianos son acciones incluidas en los planes de mantenimiento preventivo de las instalaciones consideradas de riesgo (Real Decreto 865/2003. La comparación de nuestra legislación con otras legislaciones o recomendaciones adoptadas en otros paises (Reino Unido, Francia, Australia, América permite conocer nuestro grado de exigencia en relación a algunos de los parámetros contemplados. El cultivo de la bacteria es el método de referencia para la detección de Legionella en muestras de agua y existen varios ensayos normalizados, como los estándares ISO 11731/98 y 2004 y NF T 90- 431/2003 (AFNOR. Para ayudar a la interpretación de los resultados, los ensayos deben reflejar el estándar en el que se basan y el límite de detección del método, que no debe ser superior a 100 ufc/L. Además, los laboratorios que realizan estos ensayos deben estar acreditados por nuestra entidad de acreditación ENAC. En los últimos años se han desarrollado métodos rápidos de detección de la bacteria basados en la amplificación de ADN cromosómico en muestras de agua mediante reacciones de PCR. El desarrollo científico de estos métodos va por delante del desarrollo reglamentario, y los ensayos de PCR no deben desplazar a los ensayos de cultivo en cumplimiento de las normativas vigentes, sino que deben complementarlo.

  15. Epidemiology and Ecology of Opportunistic Premise Plumbing Pathogens: Legionella pneumophila, Mycobacterium avium, and Pseudomonas aeruginosa

    Science.gov (United States)

    Hilborn, Elizabeth D.; Arduino, Matthew J.; Pruden, Amy; Edwards, Marc A.

    2015-01-01

    Background Legionella pneumophila, Mycobacterium avium, and Pseudomonas aeruginosa are opportunistic premise plumbing pathogens (OPPPs) that persist and grow in household plumbing, habitats they share with humans. Infections caused by these OPPPs involve individuals with preexisting risk factors and frequently require hospitalization. Objectives The objectives of this report are to alert professionals of the impact of OPPPs, the fact that 30% of the population may be exposed to OPPPs, and the need to develop means to reduce OPPP exposure. We herein present a review of the epidemiology and ecology of these three bacterial OPPPs, specifically to identify common and unique features. Methods A Water Research Foundation–sponsored workshop gathered experts from across the United States to review the characteristics of OPPPs, identify problems, and develop a list of research priorities to address critical knowledge gaps with respect to increasing OPPP-associated disease. Discussion OPPPs share the common characteristics of disinfectant resistance and growth in biofilms in water distribution systems or premise plumbing. Thus, they share a number of habitats with humans (e.g., showers) that can lead to exposure and infection. The frequency of OPPP-infected individuals is rising and will likely continue to rise as the number of at-risk individuals is increasing. Improved reporting of OPPP disease and increased understanding of the genetic, physiologic, and structural characteristics governing the persistence and growth of OPPPs in drinking water distribution systems and premise plumbing is needed. Conclusions Because broadly effective community-level engineering interventions for the control of OPPPs have yet to be identified, and because the number of at-risk individuals will continue to rise, it is likely that OPPP-related infections will continue to increase. However, it is possible that individuals can take measures (e.g., raise hot water heater temperatures and filter

  16. Electric showers as a control measure for Legionella spp. in a renal transplant unit in São Paulo, Brazil. Legionellosis Study Team.

    Science.gov (United States)

    Levin, A S; Gobara, S; Scarpitta, C M; Warschauer, C L; Sinto, S I; Rodrigues, E; Mendes, C M; Sabbaga, E; Boulos, M

    1995-06-01

    After an outbreak of legionnaires' disease Legionella pneumophila serogroup 1 in a renal transplant unit in São Paulo, Brazil, periodic hyperchlorination and flushing of pipes were instituted as control measures. These were only partially effective as every two to five months water cultures turned positive or new cases of the disease occurred. In November 1993 the hot water was disconnected from the unit and small, plastic electric showers were installed in each bathroom. Over a period of 12 months water from showers and taps was cultured for Legionella spp. every two weeks. On only one occasion was a water culture positive for L. pneumophila from a sink tap. No water sample obtained from showers was positive during the study period. No cases of legionnaires' disease occurred. We considered the use of electric showers an inexpensive and effective method of controlling the problem of Legionella spp. in the water system of our renal transplant unit.

  17. [Field study to obtain Legionella-free water from showers and sinks of a transplantation unit by a system of water filters].

    Science.gov (United States)

    Wendt, C; Weist, K; Dietz, E; Schlattmann, P; Rüden, H

    1995-03-01

    The efficiency of filters for elimination of Legionella was examined in a ten month field study. Showers and sinks of twelve patient rooms were equipped with nylon 66-polyamid filters. Before filtration the total number of colonies was very high with an arithmetical mean of 1131 cfu/ml. This could be reduced by filtration at the sinks (mean = 205 cfu/ml). After equipping showers with filters we found a significant increase in the total number of colonies (mean = 1961 cfu/ml). In non-filtered water we found Legionella in nearly 90% of the 648 samples, the concentration extended to 300 cfu/ml (mean = 51 cfu/ml). After filtration the concentration of Legionella was up to 280 cfu/ml in 7.3% of the 665 samples. We calculated a logistic regression model to examine the effects of the age of the filters and the duration of their use since the last sterilization. These parameters were calculated from the time of the break through or retrograde contamination with Legionella. There were clear differences between shower and sink filters. Assuming a 5% probability of break through or retrograde contamination with Legionella, the sink filters should be sterilized every 3 weeks without regards to the age of the filters. In contrast the shower filters may be used for 3 weeks if they are sterilized weekly. If weekly sterilization is not feasible they should be used for 2 weeks only. In our study it was not possible to achieve Legionella-free water by changing either the frequency of sterilization or the age of the filters.

  18. Fuzzy SP-irresolute functions

    International Nuclear Information System (INIS)

    Abbas, S.E.

    2004-01-01

    In this paper, fuzzy SP-irresolute, fuzzy SP-irresolute open and fuzzy SP-irresolute closed functions between fuzzy topological spaces in Sostak sense are defined. Their properties and the relationships between these functions and other functions introduced previously are investigated. Next fuzzy SP-connectedness is introduced and studied with the help of r-fuzzy strongly preopen sets

  19. Hidden Selection of Bacterial Resistance to Fluoroquinolones In Vivo: The Case of Legionella pneumophila and Humans.

    Science.gov (United States)

    Shadoud, Lubana; Almahmoud, Iyad; Jarraud, Sophie; Etienne, Jérôme; Larrat, Sylvie; Schwebel, Carole; Timsit, Jean-François; Schneider, Dominique; Maurin, Max

    2015-09-01

    Infectious diseases are the leading cause of human morbidity and mortality worldwide. One dramatic issue is the emergence of microbial resistance to antibiotics which is a major public health concern. Surprisingly however, such in vivo adaptive ability has not been reported yet for many intracellular human bacterial pathogens such as Legionella pneumophila. We examined 82 unrelated patients with Legionnaire's disease from which 139 respiratory specimens were sampled during hospitalization and antibiotic therapy. We both developed a real time PCR assay and used deep-sequencing approaches to detect antibiotic resistance mutations in L. pneumophila and follow their selection and fate in these samples. We identified the in vivo selection of fluoroquinolone resistance mutations in L. pneumophila in two infected patients treated with these antibiotics. By investigating the mutational dynamics in patients, we showed that antibiotic resistance occurred during hospitalization most likely after fluoroquinolone treatment. In vivo selection of antibiotic resistances in L. pneumophila may be associated with treatment failures and poor prognosis. This hidden resistance must be carefully considered in the therapeutic management of legionellosis patients and in the control of the gradual loss of effectiveness of antibiotics.

  20. The Legionella pneumophila Dps homolog is regulated by iron and involved in multiple stress tolerance.

    Science.gov (United States)

    Yu, Ming-Jia; Ren, Jun; Zeng, Yong-Lun; Zhou, Shi-Ning; Lu, Yong-Jun

    2009-09-01

    Iron homeostasis is essential to almost all organisms. In this study, we identified the putative homolog of the iron-storage protein-encoding gene, dpsL, in the intracellular pathogen Legionella pneumophila and demonstrated its expression under iron-limited conditions and its responses to multiple stresses. Quantitative real-time PCR analysis indicated that the expression of dpsL was enhanced under iron limitation regardless of the growth phase. Compared with the wild-type cells, the cells devoid of dpsL were heat and H(2)O(2)-sensitive. In contrast to the dps mutants of other bacteria, the growth of the dpsL mutant in an iron-deprived medium was delayed but finally reached the same cell density as wild-type cells during the stationary phase of growth. The finding that the dpsL mutant is salt resistant suggested the involvement of DpsL in virulence. Copyright 2009 WILEY-VCH Verlag GmbH & Co. KGaA, Weinheim.

  1. Relationship between Legionella pneumophila and Acanthamoeba polyphaga: physiological status and susceptibility to chemical inactivation.

    Science.gov (United States)

    Barker, J; Brown, M R; Collier, P J; Farrell, I; Gilbert, P

    1992-01-01

    Survival studies were conducted on Legionella pneumophila cells that had been grown intracellularly in Acanthamoeba polyphaga and then exposed to polyhexamethylene biguanide (PHMB), benzisothiazolone (BIT), and 5-chloro-N-methylisothiazolone (CMIT). Susceptibilities were also determined for L. pneumophila grown under iron-sufficient and iron-depleted conditions. BIT was relatively ineffective against cells grown under iron depletion; in contrast, iron-depleted conditions increased the susceptibilities of cells to PHMB and CMIT. The activities of all three biocides were greatly reduced against L. pneumophila grown in amoebae. PHMB (1 x MIC) gave 99.99% reductions in viability for cultures grown in broth within 6 h and no detectable survivors at 24 h but only 90 and 99.9% killing at 6 h and 24 h, respectively, for cells grown in amoebae. The antimicrobial properties of the three biocides against A. polyphaga were also determined. The majority of amoebae recovered from BIT treatment, but few, if any, survived CMIT treatment or exposure to PHMB. This study not only shows the profound effect that intra-amoebal growth has on the physiological status and antimicrobial susceptibility of L. pneumophila but also reveals PHMB to be a potential biocide for effective water treatment. In this respect, PHMB has significant activity, below its recommended use concentrations, against both the host amoeba and L. pneumophila. PMID:1514790

  2. Severe Community-acquired Pneumonia Due to Legionella pneumophila Serogroup 6

    Directory of Open Access Journals (Sweden)

    Chung-Yu Chen

    2006-01-01

    Full Text Available Legionella pneumophila is a common cause of sporadic community-acquired pneumonia, but culture-proven legionellosis is rarely diagnosed. There is no laboratory test for Legionnaires' disease that can detect all patients with the disease. Culture is the standard diagnostic method and should be initiated as soon as possible in suspected cases. We describe a rare case of community-acquired pneumonia caused by L. pneumophila serogroup 6. A 77-year-old man was admitted to a tertiary care hospital because of high fever, productive cough, and progressive dyspnea. Chest radiography showed bilateral pneumonia, which led to respiratory failure necessitating mechanical ventilatory support. Despite antibiotic therapy, his condition continued to deteriorate and acute renal failure also developed. Urine was negative for L. pneumophila. Culture of the sputum yielded L. pneumophila serogroup 6, although there was no elevation of the serum antibody titer. Pneumonia resolved gradually and he was extubated after treatment with levofloxacin followed by erythromycin. L. pneumophila other than serogroup 1 should be included in the differential diagnosis of patients with suspected atypical community-acquired pneumonia.

  3. Detection of Legionella pneumophila from domestic water and their antibiotic resistance profiles

    Directory of Open Access Journals (Sweden)

    Zeki Aras

    2015-04-01

    Full Text Available Objective: To investigate the presence of Legionella pneumophila (L. pneumophila in domestic water in Bitlis province and to determine the in vitro susceptibility of the isolates against several antibiotics. Methods: A total of 320 tap water samples were collected from the urban areas and villages of Bitlis province during the period from May to December 2010. All samples were cultured on plates of buffered charcoal yeast extract agar. L. pneumophila strains were tested for antimicrobial susceptibility by the disk diffusion method. Results: L. pneumophila strains were isolated from six (1.9% domestic water samples. All isolates were typed as L. pneumophila serogroup 1 by latex agglutination test. Four of strains were isolated in July and two of them were detected in August. Antibiotic susceptibility testing was carried out on six L. pneumophila serogroup 1 isolates. Of the six strains, two was resistant to erythromycin and streptomycin, four were resistant to ampicillin and gentamicin, but all were sensitive to chloramphenicol and doxycycline. Conclusions: Our results indicate that L. pneumophila serogroup 1 is the most common type in the domestic water samples and threats public health. This is the first report of L. pneumophila in domestic water samples from Bitlis province.

  4. Protozoa and human macrophages infection by Legionella pneumophila environmental strains belonging to different serogroups.

    Science.gov (United States)

    Messi, Patrizia; Patrizia, Messi; Bargellini, Annalisa; Annalisa, Bargellini; Anacarso, Immacolata; Immacolata, Anacarso; Marchesi, Isabella; Isabella, Marchesi; de Niederhäusern, Simona; Bondi, Moreno; Moreno, Bondi

    2013-02-01

    Three Legionella pneumophila strains isolated from municipal hot tap water during a multicentric Italian survey and belonging to serogroups 1, 6, 9 and the reference strain Philadelphia-1 were studied to determine the intracellular replication capability and the cytopathogenicity in human monocyte cell line U937 and in an Acanthamoeba polyphaga strain. Our results show that both serogroups 1 and Philadelphia-1 were able to multiply into macrophages inducing cytopathogenicity, while serogroup 6 and ever more serogroup 9 were less efficient in leading to death of the infected macrophages. Both serogroups 1 and 6 displayed a quite good capability of intracellular replication in A. polyphaga, although serogroup 1 was less cytopathogenic than serogroup 6. Serogroup 9, like Philadelphia-1 strain, showed a reduced efficiency of infection and replication and a low cytopathogenicity towards the protozoan. Our study suggests that bacterial pathogenesis is linked to the difference in the virulence expression of L. pneumophila serogroups in both hosts, as demonstrated by the fact that only L. pneumophila serogroup 1 shows the contextual expression of the two virulence traits. Serogroup 6 proves to be a good candidate as pathogen since it shows a good capacity for intracellular replication in protozoan.

  5. The many forms of a pleomorphic bacterial pathogen – The developmental network of Legionella pneumophila

    Directory of Open Access Journals (Sweden)

    Peter eRobertson

    2014-12-01

    Full Text Available Legionella pneumophila is a natural intracellular bacterial parasite of free-living freshwater protozoa and an accidental human pathogen that causes Legionnaires’ disease. L. pneumophila differentiates, and does it in style. Recent experimental data on L. pneumophila’s differentiation point at the existence of a complex network that involves many developmental forms. We intend readers to: (i understand the biological relevance of L. pneumophila’s forms found in freshwater and their potential to transmit Legionnaires’ disease, and (ii learn that the common depiction of L. pneumophila’s differentiation as a biphasic developmental cycle that alternates between a replicative and a transmissive form is but an oversimplification of the actual process. Our specific objectives are to provide updates on the molecular factors that regulate L. pneumophila’s differentiation (section 2, and describe the developmental network of L. pneumophila (section 3, which for clarity’s sake we have dissected into five separate developmental cycles. Finally, since each developmental form seems to contribute differently to the human pathogenic process and the transmission of Legionnaires’ disease, readers are presented with a challenge to develop novel methods to detect the various L. pneumophila forms present in water (section 4, as a means to improve our assessment of risk and more effectively prevent legionellosis outbreaks.

  6. Computed tomographic features of 23 sporadic cases with Legionella pneumophila pneumonia

    Energy Technology Data Exchange (ETDEWEB)

    Yu Hui [Department of Respiratory Diseases, Shanghai Pneumology Hospital, Tongji University, Shanghai (China); Higa, Futoshi; Hibiya, Kenji; Furugen, Makoto [Department of Medicine and Therapeutics, Control and Prevention of Infectious Diseases (First Department of Internal Medicine), Faculty of Medicine, University of the Ryukyus, Okinawa (Japan); Sato, Yoko [Tomishiro Chuo Hospital, Okinawa (Japan); Shinzato, Takashi [Nakagami General Hospital, Okinawa (Japan); Haranaga, Shusaku; Yara, Satomi; Tateyama, Masao [Department of Medicine and Therapeutics, Control and Prevention of Infectious Diseases (First Department of Internal Medicine), Faculty of Medicine, University of the Ryukyus, Okinawa (Japan); Fujita, Jiro, E-mail: fujita@med.u-ryukyu.ac.j [Department of Medicine and Therapeutics, Control and Prevention of Infectious Diseases (First Department of Internal Medicine), Faculty of Medicine, University of the Ryukyus, Okinawa (Japan); Li, Huiping [Department of Respiratory Diseases, Shanghai Pneumology Hospital, Tongji University, Shanghai (China)

    2010-06-15

    Objective: To describe the chest computed tomographic (CT) findings of Legionella pneumophila pneumonia. Methods: CT scans obtained from 23 sporadic cases of L. pneumophila pneumonia were retrospectively reviewed. Chest CT findings were analyzed with regard to the patterns and distributions of pulmonary abnormalities. We also analyzed the histopathology of lungs from guinea pigs with experimentally induced L. pneumophila pneumonia. Results: Consolidation and ground-glass opacity (GGO) were the main findings of CT scans in L. pneumophila pneumonia. The distribution of opacities was categorized as non-segmental (n = 20) and segmental (n = 4). Non-segmental distribution may follow an onset of segmental distribution. Pleural effusion was observed in 14 (58.3%) patients, of which 13 were accompanied with non-segmental distribution. Abscess formation was observed in only one immunocompromised patient. In the animal pneumonia model, the lesions comprised of terminal bronchioles, alveolar spaces, and interstitia. Small bacilli were observed to be contained by many macrophages within the alveoli. Conclusion: Non-segmental distribution was significantly more frequent than segmental distribution in L. pneumophila pneumonia. It is possible that L. pneumophila infection initially results in segmental pneumonia, which progresses to typical non-segmental distribution.

  7. Inactivation kinetics and efficiencies of UV-LEDs against Pseudomonas aeruginosa, Legionella pneumophila, and surrogate microorganisms.

    Science.gov (United States)

    Rattanakul, Surapong; Oguma, Kumiko

    2018-03-01

    To demonstrate the effectiveness of UV light-emitting diodes (UV-LEDs) to disinfect water, UV-LEDs at peak emission wavelengths of 265, 280, and 300 nm were adopted to inactivate pathogenic species, including Pseudomonas aeruginosa and Legionella pneumophila, and surrogate species, including Escherichia coli, Bacillus subtilis spores, and bacteriophage Qβ in water, compared to conventional low-pressure UV lamp emitting at 254 nm. The inactivation profiles of each species showed either a linear or sigmoidal survival curve, which both fit well with the Geeraerd's model. Based on the inactivation rate constant, the 265-nm UV-LED showed most effective fluence, except for with E. coli which showed similar inactivation rates at 265 and 254 nm. Electrical energy consumption required for 3-log 10 inactivation (E E,3 ) was lowest for the 280-nm UV-LED for all microbial species tested. Taken together, the findings of this study determined the inactivation profiles and kinetics of both pathogenic bacteria and surrogate species under UV-LED exposure at different wavelengths. We also demonstrated that not only inactivation rate constants, but also energy efficiency should be considered when selecting an emission wavelength for UV-LEDs. Copyright © 2017 Elsevier Ltd. All rights reserved.

  8. Essential roles and regulation of the Legionella pneumophila collagen-like adhesin during biofilm formation.

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    Julia Mallegol

    Full Text Available Legionellosis is mostly caused by Legionella pneumophila (Lp and is defined by a severe respiratory illness with a case fatality rate ranging from 5 to 80%. In a previous study, we showed that a glycosaminoglycan (GAG-binding adhesin of Lp, named Lcl, is produced during legionellosis and is unique to the L. pneumophila species. Importantly, a mutant depleted in Lcl (Δlpg2644 is impaired in adhesion to GAGs and epithelial cells and in biofilm formation. Here, we examine the molecular function(s of Lcl and the transcriptional regulation of its encoding gene during different stages of the biofilm development. We show that the collagen repeats and the C-terminal domains of Lcl are crucial for the production of biofilm. We present evidence that Lcl is involved in the early step of surface attachment but also in intercellular interactions. Furthermore, we address the relationship between Lcl gene regulation during biofilm formation and quorum sensing (QS. In a static biofilm assay, we show that Lcl is differentially regulated during growth phases and biofilm formation. Moreover, we show that the transcriptional regulation of lpg2644, mediated by a prototype of QS signaling homoserine lactone (3OC12-HSL, may play a role during the biofilm development. Thus, transcriptional down-regulation of lpg2644 may facilitate the dispersion of Lp to reinitiate biofilm colonization on a distal surface.

  9. DISTRIBUTION OF LEGIONELLA PNEUMOPHILA SEROGROUPS ISOLATED FROM WATER SYSTEMS OF PUBLIC FACILITIES IN BUSAN, SOUTH KOREA.

    Science.gov (United States)

    Hwang, In-Yeong; Park, Eun-Hee; Park, Yon-Koung; Park, Sun-Hee; Sung, Gyung-Hye; Park, Hye-Young; Lee, Young-Choon

    2016-05-01

    Legionella pneumophila is the major causes of legionellosis worldwide. The distribution of L. pneumophila was investigated in water systems of public facilities in Busan, South Korea during 2007 and 2013-2014. L. pneumophila was isolated from 8.3% of 3,055 samples, of which the highest isolation rate (49%) was from ships and the lowest 4% from fountains. Serogroups of L. pneumophila isolated in 2007 were distributed among serogroups (sgs) 1-7 with the exception of sg 4, while those of isolates during 2013 and 2014 included also 11 sgs ( 1, 2, 3, 4, 5, 6, 7, 8, 12, 13, 15). L. pneumophila sg 1 was predominated among isolates from fountains (75%), hotels (60%), buildings (44%), hospitals (38%), and public baths (37%), whereas sg 3 and sg 7 was the most prevalent from ships (46%) and factories (40%), respectively. The predominated serogroup of L. pneumophila isolates from hot and cooling tower water was sg 1 (35% and 46%, respectively), while from cold water was sg 3 (29%). These results should be useful for epidemiological surveys to identify sources of outbreaks of legionellosis in Busan, South Korea.

  10. Knowledge to Predict Pathogens: Legionella pneumophila Lifecycle Critical Review Part I Uptake into Host Cells

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    Alexis L. Mraz

    2018-01-01

    Full Text Available Legionella pneumophila (L. pneumophila is an infectious disease agent of increasing concern due to its ability to cause Legionnaires’ Disease, a severe community pneumonia, and the difficulty in controlling it within water systems. L. pneumophila thrives within the biofilm of premise plumbing systems, utilizing protozoan hosts for protection from disinfectants and other environmental stressors. While there is a great deal of information regarding how L. pneumophila interacts with protozoa and human macrophages (host for human infection, the ability to use this data in a model to attempt to predict a concentration of L. pneumophila in a water system is not known. The lifecycle of L. pneumophila within host cells involves three processes: uptake, growth, and egression from the host cell. The complexity of these three processes would risk conflation of the concepts; therefore, this review details the available information regarding how L. pneumophila invades host cells (uptake within the context of data needed to model this process, while a second review will focus on growth and egression. The overall intent of both reviews is to detail how the steps in L. pneumophila’s lifecycle in drinking water systems affect human infectivity, as opposed to detailing just its growth and persistence in drinking water systems.

  11. IcmR-regulated membrane insertion and efflux by the Legionella pneumophila IcmQ protein.

    Science.gov (United States)

    Duménil, Guillaume; Montminy, Timothy P; Tang, May; Isberg, Ralph R

    2004-02-06

    Legionella pneumophila proliferates within alveolar macrophages as a central property of Legionnaires' disease. Intracellular growth involves formation of a replicative phagosome, which requires the bacterial Dot/Icm system, a multiprotein secretion apparatus that translocates proteins from the bacterium across the macrophage plasma membrane. Two components of this system, IcmR and IcmQ, are proposed to exhibit a chaperone/substrate relationship similar to that observed in other protein translocation systems. We report here that IcmQ inserts into lipid membranes and forms pores that allow the efflux of the dye calcein but not Dextran 3000. Both membrane insertion and pore formation were inhibited by IcmR. Trypsin digestion mapping demonstrated that IcmQ is subdivided into two functional domains. The N-terminal region of IcmQ was necessary and sufficient for insertion into lipid membranes and calcein efflux. The C-terminal domain was necessary for efficient association of the protein with lipid bilayers. IcmR was found to bind to the N-terminal portion of the protein thus providing a mechanism for its ability to inhibit IcmQ pore-forming activity. Localization of IcmQ on the surface of the L. pneumophila shortly after infection as well as its pore-forming capacities suggest a role for IcmQ in forming a channel that leads translocated effectors out of the bacterium.

  12. The E Block motif is associated with Legionella pneumophila translocated substrates.

    Science.gov (United States)

    Huang, Li; Boyd, Dana; Amyot, Whitney M; Hempstead, Andrew D; Luo, Zhao-Qing; O'Connor, Tamara J; Chen, Cui; Machner, Matthias; Montminy, Timothy; Isberg, Ralph R

    2011-02-01

    Legionella pneumophila promotes intracellular growth by moving bacterial proteins across membranes via the Icm/Dot system. A strategy was devised to identify large numbers of Icm/Dot translocated proteins, and the resulting pool was used to identify common motifs that operate as recognition signals. The 3' end of the sidC gene, which encodes a known translocated substrate, was replaced with DNA encoding 200 codons from the 3' end of 442 potential substrate-encoding genes. The resulting hybrid proteins were then tested in a high throughput assay, in which translocated SidC antigen was detected by indirect immunofluorescence. Among translocated substrates, regions of 6-8 residues called E Blocks were identified that were rich in glutamates. Analysis of SidM/DrrA revealed that loss of three Glu residues, arrayed in a triangle on an α-helical surface, totally eliminated translocation of a reporter protein. Based on this result, a second strategy was employed to identify Icm/Dot substrates having carboxyl terminal glutamates. From the fusion assay and the bioinformatic queries, carboxyl terminal sequences from 49 previously unidentified proteins were shown to promote translocation into target cells. These studies indicate that by analysing subsets of translocated substrates, patterns can be found that allow predictions of important motifs recognized by Icm/Dot. © 2010 Blackwell Publishing Ltd.

  13. Effect of non-Legionellaceae bacteria on the multiplication of Legionella pneumophila in potable water.

    Science.gov (United States)

    Wadowsky, R M; Yee, R B

    1985-05-01

    A naturally occurring suspension of Legionella pneumophila and associated microbiota contained three unidentified non-Legionellaceae bacteria which supported satellite growth of a subculture of L. pneumophila on an L-cysteine-deficient medium and another bacterium which did not support growth of the subculture. Washed suspensions containing 10(3), 10(5), 10(7), or 10(8) CFU of a mixture of isolates of these non-Legionellaceae bacteria failed to support the multiplication of an isolate of agar-grown L. pneumophila which had been washed and seeded into the suspensions. The suspensions which contained 10(3), 10(5), or 10(7) CFU of the non-Legionellaceae bacteria per ml appeared to enhance survival or cryptic growth of agar-grown L. pneumophila. A decline of 1.3 log CFU of L. pneumophila per ml occurred within the first week of incubation in the sample which contained 10(8) CFU of the non-Legionellaceae bacteria per ml. In contrast to these results, naturally occurring L. pneumophila multiplied in the presence of associated microbiota. The necessity to subculture L. pneumophila and the non-Legionellaceae bacteria on artificial medium to obtain pure cultures may have affected the multiplication of L. pneumophila in tap water. Alternatively, other microorganisms may be present in the naturally occurring suspension which support the growth of this bacterium.

  14. Effects of metals on Legionella pneumophila growth in drinking water plumbing systems.

    Science.gov (United States)

    States, S J; Conley, L F; Ceraso, M; Stephenson, T E; Wolford, R S; Wadowsky, R M; McNamara, A M; Yee, R B

    1985-11-01

    An investigation of the chemical environment and growth of Legionella pneumophila in plumbing systems was conducted to gain a better understanding of its ecology in this habitat. Water samples were collected from hospital and institutional hot-water tanks known to have supported L. pneumophila and were analyzed for 23 chemical parameters. The chemical environment of these tanks was found to vary extensively, with the concentrations of certain metals reaching relatively high levels due to corrosion. The effect of various chemical conditions on L. pneumophila growth was then examined by observing its multiplication in the chemically analyzed hot-water tank samples after sterilization and reinoculation with L. pneumophila. L. pneumophila and associated microbiota used in these experiments were obtained from a hot-water tank. These stains were maintained in tap water and had never been passaged on agar. The results of the growth studies indicate that although elevated concentrations of a number of metals are toxic, lower levels of certain metals such as iron, zinc, and potassium enhance growth of naturally occurring L. pneumophila. Parallel observations on accompanying non-Legionellaceae bacteria failed to show the same relationship. These findings suggest that metal plumbing components and associated corrosion products are important factors in the survival and growth of L. pneumophila in plumbing systems and may also be important in related habitats such as cooling towers and air-conditioning systems.

  15. Virulence factor rtx in Legionella pneumophila, evidence suggesting it is a modular multifunctional protein

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    Pelaz Carmen

    2008-01-01

    Full Text Available Abstract Background The repeats in toxin (Rtx are an important pathogenicity factor involved in host cells invasion of Legionella pneumophila and other pathogenic bacteria. Its role in escaping the host immune system and cytotoxic activity is well known. Its repeated motives and modularity make Rtx a multifunctional factor in pathogenicity. Results The comparative analysis of rtx gene among 6 strains of L. pneumophila showed modularity in their structures. Among compared genomes, the N-terminal region of the protein presents highly dissimilar repeats with functionally similar domains. On the contrary, the C-terminal region is maintained with a fashionable modular configuration, which gives support to its proposed role in adhesion and pore formation. Despite the variability of rtx among the considered strains, the flanking genes are maintained in synteny and similarity. Conclusion In contrast to the extracellular bacteria Vibrio cholerae, in which the rtx gene is highly conserved and flanking genes have lost synteny and similarity, the gene region coding for the Rtx toxin in the intracellular pathogen L. pneumophila shows a rapid evolution. Changes in the rtx could play a role in pathogenicity. The interplay of the Rtx toxin with host membranes might lead to the evolution of new variants that are able to escape host cell defences.

  16. Relationship between Legionella pneumophila and Acanthamoeba polyphaga: Physiological status and susceptibility to chemical inactivation

    Energy Technology Data Exchange (ETDEWEB)

    Barker, J.; Farrell, I. (Aston Univ., Aston Triangle, Birmingham (United Kingdom)); Brown, M.R.W.; Collier, P.J.; Gilbert, P. (Univ. of Manchester (United Kingdom))

    1992-08-01

    Survival studies were conducted on Legionella pneumophila cells that had been grown intracellulary in Acanthamoeba polyphaga and then exposed to polyhexamethylene biguanide (PHMB), benzisothiazolone (BIT), and 5-chloro-N-methylisothiazolone (CMIT). Susceptibilities were also determined for L. pneumophila grown under iron-sufficient and iron-depleted conditions. BIT was relatively ineffective against cells to PHMB and CMIT. The activities of all three biocides were greatly reduced against L. pneumophila grown in amoebae. PHMB (1 [times] MIC) gave 99.99% reductions in viability for cultures grown in broth within 6 h and no detectable survivors at 24 h but only 90 and 99.9% killing at 6 h and 24 h, respectively, for cells grown in amoebae. The antimicrobial properties of the three biocides against A. polyphaga were also determined. The majority of amoebae recovered from BIT treatment, but few, if any, survived CMIT treatment or exposure of PHMB. This study not only shows the profound effect that intra-amoebal growth has on the physiological status and antimicrobial susceptibility of L. pneumophila but also reveals PHMB to be a potential biocide for effective water treatment. In this respect, PHMB has significant activity, below its recommended use concentrations, against both the host amoeba and L. pneumophila.

  17. Identification of Conserved ABC Importers Necessary for Intracellular Survival of Legionella pneumophila in Multiple Hosts

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    Amrita Lama

    2017-11-01

    Full Text Available It is established that the human pathogen Legionella pneumophila becomes significantly augmented for infection of macrophages after intracellular growth in amoebae when compared to like-strains cultivated in laboratory media. Based on this observation, we reasoned that the most critical virulence determinants of L.p. are expressed by responding to stimuli generated by the protozoan host specifically; a process we term “protozoan-priming.” We sought to identify L.p. virulence factors that were required for replication in amoebae in order to highlight the genes necessary for production of the most infectious form of the bacterium. Using a transposon mutagenesis screen, we successfully identified 12 insertions that produced bacteria severely attenuated for growth in amoebae, while retaining a functional Dot/Icm type IVb secretion system. Seven of these insertion mutants were found dispensable for growth in macrophages, revealing attractive therapeutic targets that reside upstream of the pathogen-human interface. Two candidates identified, lpg0730 and lpg0122 were required for survival and replication in amoebae and macrophage host cells. Both genes are conserved among numerous important human pathogenic bacteria that can persist or replicate in amoebae. Each gene encodes a component of an ATP binding cassette (ABC transport complex of unknown function. We demonstrate the lpg0730 ortholog in Francisella tularensis subsp. novicida to be essential for colonization of both protozoan and mammalian host cells, highlighting conserved survival mechanisms employed by bacteria that utilize protozoa as an environmental reservoir for replication.

  18. [Legionella pneumonia caused by aspiration of hot spring water after sarin exposure].

    Science.gov (United States)

    Kamimura, M; Katoh, O; Kawata, H; Kudo, K; Yagishita, Y; Niino, H; Saitoh, K; Saitoh, A

    1998-03-01

    A 72-year-old man was exposed to the sarin gas attack in a Tokyo subway on March 20 th, 1995. After exposure, he noticed eye discomfort, chest tightness, headache and weakness of the lower limbs and oropharyngeal muscles. Despite these symptoms, he visited a hot spring on the same day with his family. On March 25 th, his muscle weakness progressed, and a low grade fever appeared. His muscle weakness disappeared 8 days after exposure to sarin, but respiratory failure rapidly developed, necessitating artificial ventilation within four day after hospitalization on March 28th. Chemotherapy with erythromycin, imipenem/cilastatin, and steroid pulse therapy was begu. PCR and culture of sputum collected by bronchofiberscopy were positive for Legionella pneumophila, serogroup I. His respiratory state improved, but subsequent infection with Pseudomonous aeruginosa. Enterobacter cloacae, and Candida tropicalis/glabrata caused his death 71 days after admission. Oropharyngeal muscle weakness caused by sarin-mediated cholinesterase inhibition was strongly suspected as the cause of hot spring water aspiration. Transbronchial lung biopsy revealed organizing pneumonia with fibrosis. Bronchoscopic findings included redness, edema and fragility of all visible areas of the airway, which was thought to be due to bronchitis caused by Legionellosis.

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

    Science.gov (United States)

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

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

    Directory of Open Access Journals (Sweden)

    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

  1. High Prevalence and Genetic Polymorphisms of Legionella in Natural and Man-Made Aquatic Environments in Wenzhou, China

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    Leyi Zhang

    2017-02-01

    Full Text Available Natural and engineered water systems are the main sources of Legionnaires’ disease. It is essential from a public health perspective to survey water environments for the existence of Legionella. To analyze the main serogroups, genotypes and pathogenicity of the pathogen, a stratified sampling method was adopted to collect water samples randomly from shower water, cooling tower water, and local public hot springs in Wenzhou, China. Suspected strains were isolated from concentrated water samples. Serum agglutination assay and real-time PCR (Polymerase chain reaction were used to identify L. pneumophila. Sequence-based typing (SBT and pulsed-field gel electrophoresis (PFGE were used to elucidate the genetic polymorphisms in the collected isolates. The intracellular growth ability of the isolates was determined through their interaction with J774 cells and plating them onto BCYE (Buffered Charcoal Yeast Extract agar plates. Overall, 25.56% (46/180 of water samples were Legionella-positive; fifty-two strains were isolated and two kinds of serogroups were co-detected from six water samples from 2015 to 2016. Bacterial concentrations ranged from 20 CFU/100 mL to 10,720 CFU/100 mL. In detail, the Legionella-positive rates of shower water, cooling tower water and hot springs water were 15.45%, 13.33%, and 62.5%, respectively. The main serogroups were LP1 (30.69% and LP3 (28.85% and all strains carried the dot gene. Among them, 52 isolates and another 10 former isolates were analyzed by PFGE. Nineteen distinct patterns were observed in 52 strains isolated from 2015 to 2016 with three patterns being observed in 10 strains isolated from 2009 to 2014. Seventy-three strains containing 52 from this study and 21 former isolates were selected for SBT analysis and divided into 25 different sequence types in 4 main clonal groups belonging to 4 homomorphic types. Ten strains were chosen to show their abilities to grow and multiply in J744 cells. Taken together

  2. Convective Mixing in Distal Pipes Exacerbates Legionella pneumophila Growth in Hot Water Plumbing

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    William J. Rhoads

    2016-03-01

    Full Text Available Legionella pneumophila is known to proliferate in hot water plumbing systems, but little is known about the specific physicochemical factors that contribute to its regrowth. Here, L. pneumophila trends were examined in controlled, replicated pilot-scale hot water systems with continuous recirculation lines subject to two water heater settings (40 °C and 58 °C and three distal tap water use frequencies (high, medium, and low with two pipe configurations (oriented upward to promote convective mixing with the recirculating line and downward to prevent it. Water heater temperature setting determined where L. pneumophila regrowth occurred in each system, with an increase of up to 4.4 log gene copies/mL in the 40 °C system tank and recirculating line relative to influent water compared to only 2.5 log gene copies/mL regrowth in the 58 °C system. Distal pipes without convective mixing cooled to room temperature (23–24 °C during periods of no water use, but pipes with convective mixing equilibrated to 30.5 °C in the 40 °C system and 38.8 °C in the 58 °C system. Corresponding with known temperature effects on L. pneumophila growth and enhanced delivery of nutrients, distal pipes with convective mixing had on average 0.2 log more gene copies/mL in the 40 °C system and 0.8 log more gene copies/mL in the 58 °C system. Importantly, this work demonstrated the potential for thermal control strategies to be undermined by distal taps in general, and convective mixing in particular.

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

    Science.gov (United States)

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

    2015-01-01

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

  4. Secondary immunity to Legionella pneumophila and Th1 activity are suppressed by delta-9-tetrahydrocannabinol injection.

    Science.gov (United States)

    Newton, C A; Klein, T W; Friedman, H

    1994-01-01

    Resistance to infection with Legionella pneumophila is primarily dependent upon cell-mediated immunity rather than humoral immunity. Recent evidence suggests that activation of cell-mediated immunity depends on Th1 cells and activation of humoral immunity depends on Th2 cells. In this report, delta 9-tetrahydrocannabinol (THC), the major psychoactive cannabinoid of marijuana and an immunomodulator, suppressed development of secondary immunity to L. pneumophila, which correlated with a reduction in Th1 activity. BALB/c mice, infected with a primary sublethal dose of L. pneumophila, developed resistance to a larger challenge infection 3 to 4 weeks later. However, intravenous injection of THC (4 mg/kg of body weight) 1 day prior to primary infection resulted in increased mortality after the challenge infection. The level of anti-L. pneumophila antibodies in serum increased in both THC-treated and control mice; however, in the THC group IgG1 antibodies which are stimulated by Th2 cells were elevated while Th1-regulated, IgG2a antibodies were depressed. Furthermore, cultured splenocytes from THC-treated mice had less L. pneumophila-specific lymphoproliferation, indicating a deficiency in cell-mediated immunity. Normal mouse splenocytes treated in vitro with THC and pokeweed mitogen showed suppressed production of gamma interferon, a cytokine associated with Th1 cells, but increased production of interleukin 4, a cytokine produced by Th2 cells. Splenocytes from THC-treated mice, stimulated in vitro with either pokeweed mitogen or anti-CD3 antibodies, also produced less gamma interferon, indicating less Th1 activity in these mice. These results suggest that THC decreases the development of anti-L. pneumophila immunity by causing a change in the balance of Th1 and Th2 activities. PMID:8063421

  5. [Problems in the serological diagnosis of atypical pneumonia caused by Legionella pneumophila and Mycoplasma pneumoniae].

    Science.gov (United States)

    Śmietańska, Karolina; Chróst, Anna; Rastawicki, Waldemar

    2015-01-01

    The clinical presentation of atypical pneumonia is often similar to the presentation of more typical bacterial pneumonias and the etiological agent must be confirmed by laboratory diagnosis. This article will discuss the problems in the serological diagnosis of atypical pneumonia caused by Legionella pneumophila and Mycoplasma pneumoniae which are the agents most commonly associated with atypical pneumonia. Specifically, seeking the possibility of non-specific response, we evaluated the prevalence of antibodies to M. pneumoniae in serum samples obtained from patients suspected in clinical investigation for legionellosis. The total numbers of 261 serum obtained from patients suspected in clinical investigation for legionellosis, were tested by in-house ELISA with M. pneumoniae sonicated antigen. Some of the positive sera were also re-tested by western-blot with high specific recombinant M. pneumoniae P1 protein. The diagnostic significant level of IgA antibodies to M. pneumoniae were diagnosed by ELISA in 71 (27,2%) of tested serum samples. Some of the IgA-positive sera have also high level of IgG and IgM antibodies to M pneumoniae (respectively 4,2% and 6,5%). Most from the 18 selected positive results obtained by ELISA were also confirmed by western-blot. It was characteristic that IgA antibodies to M pneumoniae were detected more than three times often in serum samples with positive serological tests for Legionnaires' disease than in samples with negative results for L. pneumophila. This study showed the possibility of non-specific reactions in serological diagnosis of atypical pneumonia. However, according to the data of the literature, co-infections of L. pneumophila and M pneumoniae can not be excluded.

  6. Nuclease activity of Legionella pneumophila Cas2 promotes intracellular infection of amoebal host cells.

    Science.gov (United States)

    Gunderson, Felizza F; Mallama, Celeste A; Fairbairn, Stephanie G; Cianciotto, Nicholas P

    2015-03-01

    Legionella pneumophila, the primary agent of Legionnaires' disease, flourishes in both natural and man-made environments by growing in a wide variety of aquatic amoebae. Recently, we determined that the Cas2 protein of L. pneumophila promotes intracellular infection of Acanthamoeba castellanii and Hartmannella vermiformis, the two amoebae most commonly linked to cases of disease. The Cas2 family of proteins is best known for its role in the bacterial and archeal clustered regularly interspaced short palindromic repeat (CRISPR)-CRISPR-associated protein (Cas) system that constitutes a form of adaptive immunity against phage and plasmid. However, the infection event mediated by L. pneumophila Cas2 appeared to be distinct from this function, because cas2 mutants exhibited infectivity defects in the absence of added phage or plasmid and since mutants lacking the CRISPR array or any one of the other cas genes were not impaired in infection ability. We now report that the Cas2 protein of L. pneumophila has both RNase and DNase activities, with the RNase activity being more pronounced. By characterizing a catalytically deficient version of Cas2, we determined that nuclease activity is critical for promoting infection of amoebae. Also, introduction of Cas2, but not its catalytic mutant form, into a strain of L. pneumophila that naturally lacks a CRISPR-Cas locus caused that strain to be 40- to 80-fold more infective for amoebae, unequivocally demonstrating that Cas2 facilitates the infection process independently of any other component encoded within the CRISPR-Cas locus. Finally, a cas2 mutant was impaired for infection of Willaertia magna but not Naegleria lovaniensis, suggesting that Cas2 promotes infection of most but not all amoebal hosts. Copyright © 2015, American Society for Microbiology. All Rights Reserved.

  7. Inhibition of host cell translation elongation by Legionella pneumophila blocks the host cell unfolded protein response.

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    Hempstead, Andrew D; Isberg, Ralph R

    2015-12-08

    Cells of the innate immune system recognize bacterial pathogens by detecting common microbial patterns as well as pathogen-specific activities. One system that responds to these stimuli is the IRE1 branch of the unfolded protein response (UPR), a sensor of endoplasmic reticulum (ER) stress. Activation of IRE1, in the context of Toll-like receptor (TLR) signaling, induces strong proinflammatory cytokine induction. We show here that Legionella pneumophila, an intravacuolar pathogen that replicates in an ER-associated compartment, blocks activation of the IRE1 pathway despite presenting pathogen products that stimulate this response. L. pneumophila TLR ligands induced the splicing of mRNA encoding XBP1s, the main target of IRE1 activity. L. pneumophila was able to inhibit both chemical and bacterial induction of XBP1 splicing via bacterial translocated proteins that interfere with host protein translation. A strain lacking five translocated translation elongation inhibitors was unable to block XBP1 splicing, but this could be rescued by expression of a single such inhibitor, consistent with limitation of the response by translation elongation inhibitors. Chemical inhibition of translation elongation blocked pattern recognition receptor-mediated XBP1 splicing, mimicking the effects of the bacterial translation inhibitors. In contrast, host cell-promoted inhibition of translation initiation in response to the pathogen was ineffective in blocking XBP1 splicing, demonstrating the need for the elongation inhibitors for protection from the UPR. The inhibition of host translation elongation may be a common strategy used by pathogens to limit the innate immune response by interfering with signaling via the UPR.

  8. Active and adaptive Legionella CRISPR-Cas reveals a recurrent challenge to the pathogen.

    Science.gov (United States)

    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.

  9. MultipleLegionella pneumophilaeffector virulence phenotypes revealed through high-throughput analysis of targeted mutant libraries.

    Science.gov (United States)

    Shames, Stephanie R; Liu, Luying; Havey, James C; Schofield, Whitman B; Goodman, Andrew L; Roy, Craig R

    2017-11-28

    Legionella pneumophila is the causative agent of a severe pneumonia called Legionnaires' disease. A single strain of L. pneumophila encodes a repertoire of over 300 different effector proteins that are delivered into host cells by the Dot/Icm type IV secretion system during infection. The large number of L. pneumophila effectors has been a limiting factor in assessing the importance of individual effectors for virulence. Here, a transposon insertion sequencing technology called INSeq was used to analyze replication of a pool of effector mutants in parallel both in a mouse model of infection and in cultured host cells. Loss-of-function mutations in genes encoding effector proteins resulted in host-specific or broad virulence phenotypes. Screen results were validated for several effector mutants displaying different virulence phenotypes using genetic complementation studies and infection assays. Specifically, loss-of-function mutations in the gene encoding LegC4 resulted in enhanced L. pneumophila in the lungs of infected mice but not within cultured host cells, which indicates LegC4 augments bacterial clearance by the host immune system. The effector proteins RavY and Lpg2505 were important for efficient replication within both mammalian and protozoan hosts. Further analysis of Lpg2505 revealed that this protein functions as a metaeffector that counteracts host cytotoxicity displayed by the effector protein SidI. Thus, this study identified a large cohort of effectors that contribute to L. pneumophila virulence positively or negatively and has demonstrated regulation of effector protein activities by cognate metaeffectors as being critical for host pathogenesis.

  10. Antimicrobial activity of solithromycin against clinical isolates of Legionella pneumophila serogroup 1.

    Science.gov (United States)

    Mallegol, Julia; Fernandes, Prabhavathi; Melano, Roberto G; Guyard, Cyril

    2014-01-01

    The activity of solithromycin was evaluated against clinical Legionella pneumophila serogroup 1 (Lp1) isolates (n = 196) collected in Ontario, Canada, from 1980 to 2011. Its in vitro activity was compared to that of azithromycin (AZM) using the broth microdilution method. Solithromycin had a MIC50 of ≤0.015 μg/ml and a MIC90 of 0.031 μg/ml, making its activity at least 8-fold to 32-fold higher than that of AZM (MIC50 and MIC90, 0.125 μg/ml and 1 μg/ml, respectively). Ninety-nine percent of the isolates had MICs for solithromycin ranging from ≤0.015 μg/ml to 0.031 μg/ml, whereas 83.6% of the isolates showed MICs for AZM ranging from 0.062 μg/ml to 0.25 μg/ml. Interestingly, 96.7% (30 out of 31 clinical isolates) identified with higher AZM MICs (0.5 μg/ml to 2 μg/ml) belonged to the clinically prevalent sequence type 1. To investigate the intracellular activity of solithromycin, in vitro invasion assays were also performed against a subset of representative Lp1 isolates internalized within human lung epithelial cells. Solithromycin and AZM both inhibited growth of all intracellular Lp1 isolates at 1× or 8× MICs, displaying bacteriostatic effects, as would be expected with protein synthesis inhibitor rather than bactericidal activity. Solithromycin demonstrated the highest in vitro and intracellular potency against all Lp1 isolates compared to AZM. Given the rapid spread of resistance mechanisms among respiratory pathogens and the reported treatment failures in legionellosis, the development of this new fluoroketolide, already in phase 3 oral clinical studies, constitutes a promising alternative option for the treatment of legionellosis.

  11. Predictive parameters of Legionella pneumophila occurrence in hospital water: HPCs and plumbing system installation age.

    Science.gov (United States)

    Ghanizadeh, Ghader; Mirmohamadlou, Ali; Esmaeli, Davoud

    2016-09-01

    Occurrence of Legionella pneumophila can be relevant to the installation age and the presence of heterotrophic plate counts (HPCs). This research illustrates L. pneumophila contamination of hospital water in accordance with the installation age and the presence of HPCs. One hundred and fifty samples were collected from hot and cold water systems and cultured on R2A and BCYE agar. L. pneumophila identification was done via specific biochemical tests. HPCs and L. pneumophila were detected in 96 and 37.3 % of the samples, respectively. The mean of HPCs density was 947 ± 998 CFU/ml; therefore, 52 % of the samples had higher densities than 500 CFU/ml. High densities of HPCs (>500 CFU/ml) led to colonization of L. pneumophila (≥1000 CFU/ml), mainly observed in cooling systems, gynecological, sonography, and NICU wards. Chi(2) test demonstrated that higher densities (>500 CFU/ml) of HPCs and L. pneumophila contamination in cold water were more frequent than warm water (OR: 2.3 and 1.49, respectively). Univariate regressions implied a significant difference between HPCs density and installation age in positive and negative tests of L. pneumophila (OR = 1.1, p installation age on L. pneumophila occurrences (p installation age (r s  = 0.33, p installation age are relevant; so, plumbing system renovation with appropriate materials and promotion of the effective efforts for hospital's water quality assurance is highly recommended.

  12. 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.

  13. Innate Immune Surveillance in the Central Nervous System in Legionella Pneumophila Infection.

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    Lagana, Pasqualina; Soraci, Luca; Gambuzza, Maria Elsa; Delia, Santi

    2017-11-23

    Innate immune response represents a common event in many neuroinflammatory diseases. The resident immune cells of the central nervous system (CNS) are capable of sensing and reacting to both infections and sterile trauma. Peripheral immune cell migration into CNS is regulated by the blood-brain barrier (BBB), although peripheral immune cells can invade CNS through meninges, choroid plexus, perivascular spaces, and cerebrospinal fluid. Consequently, in the brain immune reactions can be mediated by both resident and peripheral immune cells. Both in the periphery and within the CNS, innate immune response is regulated by a wide array of pattern recognition receptors (PRRs), including Toll-like (TLRs), scavenger (SRs), RIG-1 like (RLRs), and nucleotide-binding oligomerization domains (NOD)-like receptors (NLRs) responsible for inflammasome formation. Inflammasome pathway activation induces pyroptosis, an highly inflammatory form of cell death occurring to remove intracellular pathogens. Legionella pneumophila is an intracellular microorganism responsible for Legionnaires' disease, a lung infection always associated to neurological dysfunctions. Recent studies have been shown that TLRs, NLRs, and RLRs, are activated by L. pneumophila. This flagellated bacterium is capable of replicating in phagocytic cells, including macrophages and microglia, which respond by activating inflammasome pathways, that could be cause of CNS dysfunctions detected in several infected patients. The aim of this review is to bring together the latest findings concerning L. pneumophila infection and innate immune host cell responses. A deeper knowledge of these processes could allow the use of immunomodulatory compounds able to counteract CNS involvement following L. pneumophila infection. Copyright© Bentham Science Publishers; For any queries, please email at epub@benthamscience.org.

  14. Genome-scale identification of Legionella pneumophila effectors using a machine learning approach.

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    Burstein, David; Zusman, Tal; Degtyar, Elena; Viner, Ram; Segal, Gil; Pupko, Tal

    2009-07-01

    A large number of highly pathogenic bacteria utilize secretion systems to translocate effector proteins into host cells. Using these effectors, the bacteria subvert host cell processes during infection. Legionella pneumophila translocates effectors via the Icm/Dot type-IV secretion system and to date, approximately 100 effectors have been identified by various experimental and computational techniques. Effector identification is a critical first step towards the understanding of the pathogenesis system in L. pneumophila as well as in other bacterial pathogens. Here, we formulate the task of effector identification as a classification problem: each L. pneumophila open reading frame (ORF) was classified as either effector or not. We computationally defined a set of features that best distinguish effectors from non-effectors. These features cover a wide range of characteristics including taxonomical dispersion, regulatory data, genomic organization, similarity to eukaryotic proteomes and more. Machine learning algorithms utilizing these features were then applied to classify all the ORFs within the L. pneumophila genome. Using this approach we were able to predict and experimentally validate 40 new effectors, reaching a success rate of above 90%. Increasing the number of validated effectors to around 140, we were able to gain novel insights into their characteristics. Effectors were found to have low G+C content, supporting the hypothesis that a large number of effectors originate via horizontal gene transfer, probably from their protozoan host. In addition, effectors were found to cluster in specific genomic regions. Finally, we were able to provide a novel description of the C-terminal translocation signal required for effector translocation by the Icm/Dot secretion system. To conclude, we have discovered 40 novel L. pneumophila effectors, predicted over a hundred additional highly probable effectors, and shown the applicability of machine learning algorithms for

  15. Genome-scale identification of Legionella pneumophila effectors using a machine learning approach.

    Directory of Open Access Journals (Sweden)

    David Burstein

    2009-07-01

    Full Text Available A large number of highly pathogenic bacteria utilize secretion systems to translocate effector proteins into host cells. Using these effectors, the bacteria subvert host cell processes during infection. Legionella pneumophila translocates effectors via the Icm/Dot type-IV secretion system and to date, approximately 100 effectors have been identified by various experimental and computational techniques. Effector identification is a critical first step towards the understanding of the pathogenesis system in L. pneumophila as well as in other bacterial pathogens. Here, we formulate the task of effector identification as a classification problem: each L. pneumophila open reading frame (ORF was classified as either effector or not. We computationally defined a set of features that best distinguish effectors from non-effectors. These features cover a wide range of characteristics including taxonomical dispersion, regulatory data, genomic organization, similarity to eukaryotic proteomes and more. Machine learning algorithms utilizing these features were then applied to classify all the ORFs within the L. pneumophila genome. Using this approach we were able to predict and experimentally validate 40 new effectors, reaching a success rate of above 90%. Increasing the number of validated effectors to around 140, we were able to gain novel insights into their characteristics. Effectors were found to have low G+C content, supporting the hypothesis that a large number of effectors originate via horizontal gene transfer, probably from their protozoan host. In addition, effectors were found to cluster in specific genomic regions. Finally, we were able to provide a novel description of the C-terminal translocation signal required for effector translocation by the Icm/Dot secretion system. To conclude, we have discovered 40 novel L. pneumophila effectors, predicted over a hundred additional highly probable effectors, and shown the applicability of machine

  16. Autophagy Evasion and Endoplasmic Reticulum Subversion: The Yin and Yang of Legionella Intracellular Infection.

    Science.gov (United States)

    Sherwood, Racquel Kim; Roy, Craig R

    2016-09-08

    The gram-negative bacterial pathogen Legionella pneumophila creates a novel organelle inside of eukaryotic host cells that supports intracellular replication. The L. pneumophila-containing vacuole evades fusion with lysosomes and interacts intimately with the host endoplasmic reticulum (ER). Although the natural hosts for L. pneumophila are free-living protozoa that reside in freshwater environments, the mechanisms that enable this pathogen to replicate intracellularly also function when mammalian macrophages phagocytose aerosolized bacteria, and infection of humans by L. pneumophila can result in a severe pneumonia called Legionnaires' disease. A bacterial type IVB secretion system called Dot/Icm is essential for intracellular replication of L. pneumophila. The Dot/Icm apparatus delivers over 300 different bacterial proteins into host cells during infection. These bacterial proteins have biochemical activities that target evolutionarily conserved host factors that control membrane transport processes, which results in the formation of the ER-derived vacuole that supports L. pneumophila replication. This review highlights research discoveries that have defined interactions between vacuoles containing L. pneumophila and the host ER. These studies reveal how L. pneumophila creates a vacuole that supports intracellular replication by subverting host proteins that control biogenesis and fusion of early secretory vesicles that exit the ER and host proteins that regulate the shape and dynamics of the ER. In addition to recruiting ER-derived membranes for biogenesis of the vacuole in which L. pneumophila replicates, these studies have revealed that this pathogen has a remarkable ability to interfere with the host's cellular process of autophagy, which is an ancient cell autonomous defense pathway that utilizes ER-derived membranes to target intracellular pathogens for destruction. Thus, this intracellular pathogen has evolved multiple mechanisms to control membrane

  17. Non-opsonic phagocytosis of Legionella pneumophila by macrophages is mediated by phosphatidylinositol 3-kinase.

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    Souvenir D Tachado

    Full Text Available BACKGROUND: Legionella pneumophila, is an intracellular pathogen that causes Legionnaires' disease in humans, a potentially lethal pneumonia. L. pneumophila has the ability to enter and replicate in the host and is essential for pathogenesis. METHODOLOGY/PRINCIPAL FINDINGS: Phagocytosis was measured by cell invasion assays. Construction of PI3K mutant by PCR cloning and expression of dominant negative mutant was detected by Western blot. PI3K activity was measured by 32P labeling and detection of phospholipids products by thin layer chromatography. Infection of macrophages with virulent L. pneumophila stimulated the formation of phosphatidylinositol 3-phosphate (PIP3, a phosphorylated lipid product of PI3K whereas two structurally distinct phosphatidylinositol 3 kinase (PI3K inhibitors, wortmannin and LY294002, reduced L. pneumophila entry into macrophages in a dose-dependent fashion. Furthermore, PI3K activation led to Akt stimulation, a serine/threonine kinase, which was also inhibited by wortmannin and LY294002. In contrast, PI3K and protein kinase B (PKB/Akt activities were lower in macrophages infected with an avirulent bacterial strain. Only virulent L. pneumophila increased lipid kinase activity present in immunoprecipitates of the p85alpha subunit of class I PI3K and tyrosine phosphorylated proteins. In addition, macrophages expressing a specific dominant negative mutant of PI3K reduced L. pneumophila entry into these cells. CONCLUSION/SIGNIFICANCE: Entry of L. pneumophila is mediated by PI3K/Akt signaling pathway. These results suggest an important role for PI3K and Akt in the L. pneumophila infection process. They point to possible novel strategies for undermining L. pneumophila host uptake and reducing pathogenesis of Legionnaires' disease.

  18. DIAGNOSTICO SEROLÓGICO DE NEUMONIA POR LEGIONELLA. INCIDENCIA EN UN PERIODO DE TRES AÑOS EN EL AREA SANITARIA OESTE DE VALLADOLID

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    M.A. Mazón

    2005-12-01

    Full Text Available ABSTRACT:Introduction: Legionella species are Gram-negative bacilli. There are described for the first time in 70's associated to the Legionnarie's disease. Presently, more than 34 species of Legionella have been identified, 20 of which have been found pathogenic for the man. The clinical manifestations of Legionella infections are primarily respiratory. The most common presentation is acute pneumonia, which varies in severity from mild illness that does not require hospitalization to fatal pneumonia.Material and Methods: The results corresponding to the determination of antibodies to L. pneumophila (Gull Laboratories by indirect inmunofluorescence (IFI have been analysed and reviewed. This conventional inmunofluorescent test measures antibodies versus a pool of 1, 2, 3, 4, 5, 6, 7 and 8 serogroups. During the 3 years of surveillance period 1249 determinations by IFI were performed in our Hospital. The determination of urinary antigen of Legionella for the qualitative detection of the antigen to L. pneumophila serogroup 1 was made in almost all studied samples by using a fast inmunochromatographical technique (Binax NOW.Results and Discussion: Of the 1249 cases of nosocomial pneumonia (diagnosed by a positive result of IFI Legionella test recorded during the period, 12 cases met the criteria for infection with L. pneumophila serogroup 1. Later, clinical histories were reviewed and it was observed that the ages of the confirmed cases oscillated between the 40 and 87 years, with an average of age of 40, of which 8 were men and 4 women. From the point of view of factors of risk, 6 patients were smokers. Clinically, the displayed symptoms were general malaise, fever high and pattern of pulmonary infiltration in all the cases, migraine in 3 cases, diarrhoea in 5 cases and arthralgias and muscle aches in 3 cases. The analytical parameters in common were high VSG and leukocytosis. Hyponatremia 1/1024, what it demonstrates a clear seroconversion. In

  19. Legionella pneumophila infection presenting as headache, confusion and dysarthria in a human immunodeficiency virus-1 (HIV-1 positive patient: case report

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    Robbins Nathaniel M

    2012-09-01

    Full Text Available Abstract Background Legionella pneumophila is a common cause of community-acquired pneumonia. Central nervous system dysfunction is common, and diagnosis in the absence of pulmonary symptoms can be challenging. Here we describe an atypical clinical presentation of Legionella infection in a patient with HIV who was found to have an unusual neuroradiologic lesion that further served to obscure the diagnosis. This is the first such description in a patient with Legionellosis and HIV coinfection. Case presentation A 43 year-old HIV positive man presented to our hospital with dysarthria, fevers, headache, and altered mental status. Initial work-up revealed pneumonia and a lesion of the splenium of the corpus callosum on magnetic resonance imaging. He was subsequently diagnosed with Legionella pneumonia and treated with complete symptom resolution. Conclusions Neurologic abnormalities are frequent in Legionellosis, but the diagnosis may be difficult in the absence of overt respiratory symptoms and in the presence of HIV coinfection. A high index of suspicion and early initiation of empiric antibiotics is imperative since early treatment may help prevent long-term sequelae. Neuroimaging abnormalities, though rare, can help the physician narrow down the diagnosis and avoid unnecessary invasive testing. Future studies should aim to elucidate the as yet unknown role of neuroimaging in the diagnoses and prognostication of Legionellosis, as well as the interaction between Legionella infection and HIV.

  20. Evaluation of the strategy designed to reduce the incidence of cases produced by the bio contaminant (legionella Sero group I) in the city of Cordoba-Spain

    International Nuclear Information System (INIS)

    Rubio Garcia, A.; Minarro del Moral, R. M.; Guijarro Rodriguez, J. L.

    2009-01-01

    Environmental protection is essential to the quality of lifer for present and future generations. The EU lines of priorities for reucing the adverse health effects associated with certain environmental factors, include. To prevent risks associated with the facilities at risk in the transmission of certain diseases such as Legionella. (Author)

  1. Comments on Whiley Legionella Risk Management and Control in Potable Water Systems: Argument for the Abolishment of Routine Testing. Int. J. Environ. Res. Public Health 2017, 14, 12

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    Samuel Collins

    2017-01-01

    Full Text Available In their recent article, Whiley makes an interesting case for the abolishment of routine testing in Legionella risk management and control plans. Here, we present our views regarding this suggestion, drawing upon our own experiences in the UK. We urge caution against the removal of routine monitoring from guidelines due to the impending public health risks that would result.

  2. Development of a genus-specific next generation sequencing approach for sensitive and quantitative determination of the Legionella microbiome in freshwater systems.

    Science.gov (United States)

    Pereira, Rui P A; Peplies, Jörg; Brettar, Ingrid; Höfle, Manfred G

    2017-03-31

    Next Generation Sequencing (NGS) has revolutionized the analysis of natural and man-made microbial communities by using universal primers for bacteria in a PCR based approach targeting the 16S rRNA gene. In our study we narrowed primer specificity to a single, monophyletic genus because for many questions in microbiology only a specific part of the whole microbiome is of interest. We have chosen the genus Legionella, comprising more than 20 pathogenic species, due to its high relevance for water-based respiratory infections. A new NGS-based approach was designed by sequencing 16S rRNA gene amplicons specific for the genus Legionella using the Illumina MiSeq technology. This approach was validated and applied to a set of representative freshwater samples. Our results revealed that the generated libraries presented a low average raw error rate per base (approach also showed high in situ specificity (>95%) and very good repeatability. Only in samples in which the gammabacterial clade SAR86 was present more than 1% non-Legionella sequences were observed. Next-generation sequencing read counts did not reveal considerable amplification/sequencing biases and showed a sensitive as well as precise quantification of L. pneumophila along a dilution range using a spiked-in, certified genome standard. The genome standard and a mock community consisting of six different Legionella species demonstrated that the developed NGS approach was quantitative and specific at the level of individual species, including L. pneumophila. The sensitivity of our genus-specific approach was at least one order of magnitude higher compared to the universal NGS approach. Comparison of quantification by real-time PCR showed consistency with the NGS data. Overall, our NGS approach can determine the quantitative abundances of Legionella species, i. e. the complete Legionella microbiome, without the need for species-specific primers. The developed NGS approach provides a new molecular surveillance tool

  3. Legionella pneumophila serogroup 3 pneumonia in a patient with low-grade 4 non-Hodgkin lymphoma: a case report

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    Bistoni Francesco

    2011-08-01

    Full Text Available Abstract Introduction Nosocomial legionellosis has generally been described in immunodepressed patients, but Legionella pneumophila serogroup 3 has rarely been identified as the causative agent. Case presentation We report the case of nosocomial L. pneumophila serogroup 3 pneumonia in a 70-year-old Caucasian man with non-Hodgkin lymphoma. Diagnosis was carried out by culture and real-time polymerase chain reaction of bronchoalveolar lavage fluid. The results of a urinary antigen test were negative. A hospital environmental investigation revealed that the hospital water system was highly colonized by L. pneumophila serogroups 3, 4, and 8. The hospital team involved in the prevention of infections was informed, long-term control measures to reduce the environmental bacterial load were adopted, and clinical monitoring of legionellosis occurrence in high-risk patients was performed. No further cases of Legionella pneumonia have been observed so far. Conclusions In this report, we describe a case of legionellosis caused by L. pneumophila serogroup 3, which is not usually a causative agent of nosocomial infection. Our research confirms the importance of carrying out cultures of respiratory secretions to diagnose legionellosis and highlights the limited value of the urinary antigen test for hospital infections, especially in immunocompromised patients. It also indicates that, to reduce the bacterial load and prevent nosocomial legionellosis, appropriate control measures should be implemented with systematic monitoring of hospital water systems.

  4. Comparison of pulsed corona plasma and pulsed electric fields for the decontamination of water containing Legionella pneumophila as model organism.

    Science.gov (United States)

    Banaschik, Robert; Burchhardt, Gerhard; Zocher, Katja; Hammerschmidt, Sven; Kolb, Juergen F; Weltmann, Klaus-Dieter

    2016-12-01

    Pulsed corona plasma and pulsed electric fields were assessed for their capacity to kill Legionella pneumophila in water. Electrical parameters such as in particular dissipated energy were equal for both treatments. This was accomplished by changing the polarity of the applied high voltage pulses in a coaxial electrode geometry resulting in the generation of corona plasma or an electric field. For corona plasma, generated by high voltage pulses with peak voltages of +80kV, Legionella were completely killed, corresponding to a log-reduction of 5.4 (CFU/ml) after a treatment time of 12.5min. For the application of pulsed electric fields from peak voltages of -80kV a survival of log 2.54 (CFU/ml) was still detectable after this treatment time. Scanning electron microscopy images of L. pneumophila showed rupture of cells after plasma treatment. In contrast, the morphology of bacteria seems to be intact after application of pulsed electric fields. The more efficient killing for the same energy input observed for pulsed corona plasma is likely due to induced chemical processes and the generation of reactive species as indicated by the evolution of hydrogen peroxide. This suggests that the higher efficacy and efficiency of pulsed corona plasma is primarily associated with the combined effect of the applied electric fields and the promoted reaction chemistry. Copyright © 2016 Elsevier B.V. All rights reserved.

  5. [Rate and level of contamination of potentially dangerous water supply facilities in Moscow region by Legionella pneumophila].

    Science.gov (United States)

    Tartakovskiĭ, I S; Novokshonova, I V; Sadretdinova, O V; Gruzdeva, O A; Aliapkina, Iu S; Karpova, T I; Dronina, Iu E; Fokina, V G; Shustrova, N M

    2010-01-01

    To assess rate and level of contamination by Legionella pneumophila of cooling water systems in industrial facilities as well as hot water supply systems of administrative buildings in Moscow region. Cooling water systems of 8 industrial facilities and hot water supply systems of 12 administrative buildings or complexes located in Moscow or Moscow region were examined. Samples of water, washes and biofilms were studied by bacteriologic methods and RT-PCR. Results. Significant level of contamination of water systems by L. pneumophila was revealed in examined objects. Rate of contamination of cooling water systems in industrial facilities was 70%. The agent was detected in stagnant, end-capped, and rarely used segments of all hot water supply systems during decrease of water temperature to 36-52 degrees C. Visual detection of natural biofilms on the object correlated with high concentration of L. pneumophila in water samples. In some cases, associations of L. pneumophila with Pseudomonas aeruginosa were detected, including water samples from supply systems of 2 healthcare facilities. Obtained results confirm the importance of implementation of modem concept of legionellosis prevention in our country, based on regular quantitative monitoring for Legionella in potentially dangerous water objects and conduction of preventative measures if contamination exceeds acceptable level.

  6. 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.

  7. Cryptosporidium sp. in lizards

    Czech Academy of Sciences Publication Activity Database

    Koudela, Břetislav; Modrý, D.

    1998-01-01

    Roč. 45, č. 1 (1998), s. 8 ISSN 1066-5234. [ Cryptosporidium sp. in lazards. 01.01.1998-02.01.1998, Praha] R&D Projects: GA ČR GA508/95/0273; GA AV ČR IPP2020702 Subject RIV: fp - Other Medical Disciplines

  8. Criterios microbiológicos y de muestreo establecidos en la legislación vigente para el control de Legionella

    Directory of Open Access Journals (Sweden)

    Isabel Inza Rojas

    2006-12-01

    Full Text Available Se ha investigado retrospectivamente la relación existente entre las bacterias aerobias totales (BAT y la presencia de Legionella en un total de 576 muestras de agua procedentes de torres de refrigeración industriales. Los resultados revelan un menor porcentaje de muestras positivas para Legionella (19,8% por encima del nivel de 104 UFC/ml BAT establecido como límite de acción, que por debajo de este (80,2%, lo que indica que este limite no cumple su función en las instalaciones investigadas. Niveles de BAT superiores 100 y 1000 UFC/ml parecen ser más útiles para predecir la presencia de Legionella. Hemos observado una concentración de BAT significativamente más elevada en las muestras positivas para Legionella que en las negativas. Esto indica que las BAT son una buena herramienta para la validación de la eficacia del Plan de Autocontol (PA establecido en estas instalaciones. Estos resultados apoyan la importancia de seguir manteniendo el control de las BAT (a 36 ± 1 oC como un indicador de riego de presencia de Legionella y evidencian la necesidad de introducir un límite de acción más bajo para las BAT. Por otra parte se discuten algunos aspectos vinculados a la toma de muestras y a su transporte, y algunos tópicos vinculados al periodo de incubación de la enfermedad y a los métodos de tipado epidemiológico.

  9. Tsukamurella inchonensis sp. nov.

    Science.gov (United States)

    Yassin, A F; Rainey, F A; Brzezinka, H; Burghardt, J; Lee, H J; Schaal, K P

    1995-07-01

    Chemotaxonomic and genomic 16S ribosomal DNA sequence analyses of two isolates obtained from two different clinical materials clearly delineated a new species of the genus Tsukamurella. This new species can be identified by its 16S ribosomal DNA similarity values, as well as its physiological characteristics. The name Tsukamurella inchonensis sp. nov. is proposed for these isolates, which are represented by strain IMMIB D-771T (= DSM 44067T) (T = type strain). This strain exhibits only 45% DNA relatedness to Tsukamurella paurometabola.

  10. Biofilm Composition and Threshold Concentration for Growth of Legionella pneumophila on Surfaces Exposed to Flowing Warm Tap Water without Disinfectant

    Science.gov (United States)

    Bakker, Geo L.; Italiaander, Ronald; Veenendaal, Harm R.; Wullings, Bart A.

    2017-01-01

    ABSTRACT Legionella pneumophila in potable water installations poses a potential health risk, but quantitative information about its replication in biofilms in relation to water quality is scarce. Therefore, biofilm formation on the surfaces of glass and chlorinated polyvinyl chloride (CPVC) in contact with tap water at 34 to 39°C was investigated under controlled hydraulic conditions in a model system inoculated with biofilm-grown L. pneumophila. The biofilm on glass (average steady-state concentration, 23 ± 9 pg ATP cm−2) exposed to treated aerobic groundwater (0.3 mg C liter−1; 1 μg assimilable organic carbon [AOC] liter−1) did not support growth of the organism, which also disappeared from the biofilm on CPVC (49 ± 9 pg ATP cm−2) after initial growth. L. pneumophila attained a level of 4.3 log CFU cm−2 in the biofilms on glass (1,055 ± 225 pg ATP cm−2) and CPVC (2,755 ± 460 pg ATP cm−2) exposed to treated anaerobic groundwater (7.9 mg C liter−1; 10 μg AOC liter−1). An elevated biofilm concentration and growth of L. pneumophila were also observed with tap water from the laboratory. The Betaproteobacteria Piscinibacter and Methyloversatilis and amoeba-resisting Alphaproteobacteria predominated in the clones and isolates retrieved from the biofilms. In the biofilms, the Legionella colony count correlated significantly with the total cell count (TCC), heterotrophic plate count, ATP concentration, and presence of Vermamoeba vermiformis. This amoeba was rarely detected at biofilm concentrations of TCC = 1 × 106 to 2 × 106 cells cm−2) was derived for growth of L. pneumophila in biofilms. IMPORTANCE Legionella pneumophila is the etiologic agent in more than 10,000 cases of Legionnaires' disease that are reported annually worldwide and in most of the drinking water-associated disease outbreaks reported in the United States. The organism proliferates in biofilms on surfaces exposed to warm water in engineered freshwater installations. An

  11. SP-100 surety evaluation

    International Nuclear Information System (INIS)

    1985-06-01

    This report describes surety evaluations conducted during GFY 1985 in support of the General Electric design for a Space Nuclear Power System - SP-100. Those surety evaluations address both safety and safeguards requirements, which are derived from OSNP-1 and supporting documents. The report includes results of neutronics (criticality) calculations performed by Los Alamos. The results have been benchmarked against independent calculations performed by General Electric with different codes. These comparisons show close agreement, and are summarized. Los Alamos has also provided specifications of explosion and fire environments, which have been used in evaluation of the GE SP-100 concept. Following the summary of key results, surety requirements are given and recommendations toward specification of requirements for later SP-100 project phases are presented. A conceptual design summary is presented. To establish a comprehensive background for surety evaluations, a reference mission profile and potential accidents for each phase of the mission are identified. The main body of the report addresses surety of the General Electric Thermoelectric Conversion design. GE has also developed a Stirling Engine concept, and performed comprehensive surety evaluations for it. These evaluations are reported

  12. Legionella risk assessment in water installations Algoritmo de evaluación del riesgo de transmisión de 'Legionella' en instalaciones de transporte y tratamiento de masa de agua

    Directory of Open Access Journals (Sweden)

    Paulino Pastor Pérez

    2005-12-01

    Full Text Available Legionelose is an illness produced as a consequence of the inhalation of bacteria of the specie Legionella, which occasionally may colonize some of the water installations of the buildings.This article describes a Legionella risk assessment algorithm particularized for every type of installation covering all the specific characteristics of every piece of equipment, but keeping a common similar structure that allows easiness of use and homogeneous results. This algorithm has been developed by the Experts Commission of the Ministry of Health.The main use of these kind of tools is to minimize as much as possible the subjectivity of the inspectors in the identification and assessment of risks associated to the installations. These types of algorithms are applicable in risks depending on a relative low number of parameters that are somehow easy to control, as it happens in the water installations in the buildings.It has been chosen to develop a global index rated from 0 to 100 (meaning 0 minimum risk and 100 maximum risk distributed in term in three sub indexes, which define the main risk aspects to be taken into account.Structural Index: design.Maintenance Index: maintenance quality.Operational Index: performance of the equipment.Covering all the different aspects of the life of the installations.La legionelosis es una enfermedad producida por la inhalación de bacterias de la especie Legionella, que ocasionalmente pueden colonizar algunas instalaciones de la edificación que utilizan agua para su funcionamiento.El presente articulo describe un algoritmo de evaluación del riesgo de transmisión de legionelosis particularizado para cada tipo de instalación de riesgo a fin de cubrir las características especificas de cada equipo, pero con una estructura común similar, para asegurar la sencillez de uso y la homogeneidad de resultados. Este algoritmo ha sido desarrollado en el seno de la Comisión de Expertos de Legionella del Ministerio de Sanidad

  13. UV-A photocatalytic treatment of Legionella pneumophila bacteria contaminated airflows through three-dimensional solid foam structured photocatalytic reactors

    Energy Technology Data Exchange (ETDEWEB)

    Josset, Sebastien; Hajiesmaili, Shabnam; Begin, Dominique; Edouard, David; Pham-Huu, Cuong [Laboratoire des Materiaux, Surfaces et Procedes pour la Catalyse (LMSPC), European Laboratory for Catalysis and Surface Sciences (ELCASS), CNRS, Strasbourg University, 25 rue Becquerel 67087 Strasbourg (France); Lett, Marie-Claire [Laboratoire de Genetique Moleculaire, Genomique, Microbiologie, CNRS, Strasbourg University, 28, rue Goethe 67083 Strasbourg Cedex (France); Keller, Nicolas, E-mail: nkeller@chimie.u-strasbg.fr [Laboratoire des Materiaux, Surfaces et Procedes pour la Catalyse (LMSPC), European Laboratory for Catalysis and Surface Sciences (ELCASS), CNRS, Strasbourg University, 25 rue Becquerel 67087 Strasbourg (France); Keller, Valerie [Laboratoire des Materiaux, Surfaces et Procedes pour la Catalyse (LMSPC), European Laboratory for Catalysis and Surface Sciences (ELCASS), CNRS, Strasbourg University, 25 rue Becquerel 67087 Strasbourg (France)

    2010-03-15

    A 3D-structured photocatalytic media was designed for allowing a tubular reactor to work in a traversing-flow mode at low pressure drops with a strong increase in the surface area-to-volume ratio inside the reactor. A protective polysiloxane coating was performed for protecting a structured polyurethane foam and anchoring the active TiO{sub 2} particles. Filled with the 3D-structured solid foam supporting TiO{sub 2} photocatalyst, the reactor could thus take advantages from the static mixer effect and from the low pressure drop resulting from the reticulated foam support. Very efficient decontamination levels towards airborne Legionella pneumophila bacteria were reached in a single-pass test mode.

  14. UV-A photocatalytic treatment of Legionella pneumophila bacteria contaminated airflows through three-dimensional solid foam structured photocatalytic reactors.

    Science.gov (United States)

    Josset, Sébastien; Hajiesmaili, Shabnam; Begin, Dominique; Edouard, David; Pham-Huu, Cuong; Lett, Marie-Claire; Keller, Nicolas; Keller, Valérie

    2010-03-15

    A 3D-structured photocatalytic media was designed for allowing a tubular reactor to work in a traversing-flow mode at low pressure drops with a strong increase in the surface area-to-volume ratio inside the reactor. A protective polysiloxane coating was performed for protecting a structured polyurethane foam and anchoring the active TiO(2) particles. Filled with the 3D-structured solid foam supporting TiO(2) photocatalyst, the reactor could thus take advantages from the static mixer effect and from the low pressure drop resulting from the reticulated foam support. Very efficient decontamination levels towards airborne Legionella pneumophila bacteria were reached in a single-pass test mode. (c) 2009 Elsevier B.V. All rights reserved.

  15. Planktonic replication is essential for biofilm formation by Legionella pneumophila in a complex medium under static and dynamic flow conditions

    DEFF Research Database (Denmark)

    Mampel, J.; Spirig, T.; Weber, S.S.

    2006-01-01

    Legionella pneumophila persists for a long time in aquatic habitats, where the bacteria associate with biofilms and replicate within protozoan predators. While L. pneumophila serves as a paradigm for intracellular growth within protozoa, it is less clear whether the bacteria form or replicate......A mutant lacking the alternative sigma factor sigma(28) was reduced, which demonstrated that bacterial factors are required. Accumulation of biomass coincided with an increase in the optical density at 600 nm and ceased when the bacteria reached the stationary growth phase. L. pneumophila neither grew nor......, and no sizeable patches of clonally growing bacteria were observed. Our findings indicate that biofilm formation by L. pneumophila in a rich medium is due to growth of planktonic bacteria rather than to growth of sessile bacteria. In agreement with this conclusion, GFP-labeled L. pneumophila initially adhered...

  16. Supply of domestic hot Water at comfortable temperatures by low-temperature district heating without risk of Legionella

    DEFF Research Database (Denmark)

    Yang, Xiaochen

    . According to the Danish standard, the supply for DHW should be able to reach 45 °C for the kitchen use and 40 °C for other uses for comfort. Regarding to the hygiene requirements, large DHW system with DHW storage tank and circulation has to use high temperature regime to get rid of Legionella. The storage...... the sterilization methods and optimized DHW system design methods are included. For the sterilization methods, we selected the most widely used treatments, and analyzed the feasibility of applying them in the LTDH scenario, the installation and operation difficulty, and the economy based on the review...... of substantial documents and relevant standards. In terms of the DHW system design methods, the optimal system configurations and operation methods were designed and evaluated with respect to the different DH scenarios and building typologies. Model studies were built to simulate the performances of the proposed...

  17. Alternative solutions for inhibiting Legionella in domestic hot water systems based on low-temperature district heating

    DEFF Research Database (Denmark)

    Yang, Xiaochen; Li, Hongwei; Svendsen, Svend

    2015-01-01

    , and includes a comprehensive overview of their efficiency, installation, operation and costs, so as to give adequate information for selecting appropriate solutions. In addition to sterilization methods, alternative system design implemented with new technologies can also help prevent Legionella in hot water...... systems. They have the additional benefit of reducing the heat loss of the hot water system. The alternative design solutions both enrich our options for water sanitation and improve the energy efficiency of our energy systems....... methods, thermal treatment, ionization, chlorine, chlorine dioxide, ultraviolet light, photocatalysis and filtration are discussed as the most frequently used methods in hot water systems. The characteristics, efficacy and operation methods of LTDH using the solutions investigated are documented...

  18. Multifaceted effects of synthetic TLR2 ligand and Legionella pneumophilia on Treg-mediated suppression of T cell activation

    Directory of Open Access Journals (Sweden)

    Sutmuller Roger PM

    2011-03-01

    Full Text Available Abstract Background Regulatory T cells (Treg play a crucial role in maintaining immune homeostasis and self-tolerance. The immune suppressive effects of Tregs should however be limited in case effective immunity is required against pathogens or cancer cells. We previously found that the Toll-like receptor 2 (TLR2 agonist, Pam3CysSK4, directly stimulated Tregs to expand and temporarily abrogate their suppressive capabilities. In this study, we evaluate the effect of Pam3CysSK4 and Legionella pneumophila, a natural TLR2 containing infectious agent, on effector T (Teff cells and dendritic cells (DCs individually and in co-cultures with Tregs. Results TLR2 agonists can directly provide a co-stimulatory signal inducing enhanced proliferation and cytokine production of naive CD4+ Teff cells. With respect to cytokine production, DCs appear to be most sensitive to low amounts of TLR agonists. Using wild type and TLR2-deficient cells in Treg suppression assays, we accordingly show that all cells (e.g. Treg, Teff cells and DCs contributed to overcome Treg-mediated suppression of Teff cell proliferation. Furthermore, while TLR2-stimulated Tregs readily lost their ability to suppress Teff cell proliferation, cytokine production by Teff cells was still suppressed. Similar results were obtained upon stimulation with TLR2 ligand containing bacteria, Legionella pneumophila. Conclusions These findings indicate that both synthetic and natural TLR2 agonists affect DCs, Teff cells and Treg directly, resulting in multi-modal modulation of Treg-mediated suppression of Teff cells. Moreover, Treg-mediated suppression of Teff cell proliferation is functionally distinct from suppression of cytokine secretion.

  19. 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.

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

    Directory of Open Access Journals (Sweden)

    Barbara A Weissenmayer

    2011-03-01

    Full Text Available 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.

  1. Enhanced chlorine resistance of tap water-adapted Legionella pneumophila as compared with agar medium-passaged strains.

    Science.gov (United States)

    Kuchta, J M; States, S J; McGlaughlin, J E; Overmeyer, J H; Wadowsky, R M; McNamara, A M; Wolford, R S; Yee, R B

    1985-07-01

    Previous studies have shown that bacteria maintained in a low-nutrient "natural" environment such as swimming pool water are much more resistant to disinfection by various chemical agents than strains maintained on rich media. In the present study a comparison was made of the chlorine (Cl2) susceptibility of hot-water tank isolates of Legionella pneumophila maintained in tap water and strains passaged on either nonselective buffered charcoal-yeast extract or selective differential glycine-vancomycin-polymyxin agar medium. Our earlier work has shown that environmental and clinical isolates of L. pneumophila maintained on agar medium are much more resistant to Cl2 than coliforms are. Under the present experimental conditions (21 degrees C, pH 7.6 to 8.0, and 0.25 mg of free residual Cl2 per liter, we found the tap water-maintained L. pneumophila strains to be even more resistant than the agar-passaged isolates. Under these conditions, 99% kill of tap water-maintained strains of L. pneumophila was usually achieved within 60 to 90 min compared with 10 min for agar-passaged strains. Samples from plumbing fixtures in a hospital yielded legionellae which were "super"-chlorine resistant when assayed under natural conditions. After one agar passage their resistance dropped to levels of comparable strains which had not been previously exposed to additional chlorination. These studies more closely approximate natural conditions than our previous work and show that tap water-maintained L. pneumophila is even more resistant to Cl2 than its already resistant agar medium-passaged counterpart.

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

    Directory of Open Access Journals (Sweden)

    Carlos Méndez

    2012-09-01

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

  3. Tsukamurella pulmonis sp. nov.

    Science.gov (United States)

    Yassin, A F; Rainey, F A; Brzezinka, H; Burghardt, J; Rifai, M; Seifert, P; Feldmann, K; Schaal, K P

    1996-04-01

    Chemotaxonomic and 16S ribosomal DNA sequence analyses of an isolate from the sputum of a patient with a mycobacterial lung infection clearly delineated a new species of the genus Tsukamurella. This new species can be defined on the basis of genotypic and phenotypic data. The name Tsukamurella pulmonis sp. nov. is proposed for this organism; the type strain is IMMIB D-1321T (= DSM 44142T). This isolate shows 44.2 and 36.2% DNA relatedness to Tsukamurella paurometabola DSM 20162T (T = type strain) and Tsukamurella inchonensis DSM 44067T, respectively.

  4. Origin and prevention of infection with Legionella pneumophila through cooling towers and evaporative cooling towers; Entstehung und Vermeidung von Legionelleninfektionen durch Kuehltuerme und Rueckkuehlwerke

    Energy Technology Data Exchange (ETDEWEB)

    Schulze-Roebbecke, R. [Bonn Univ. (Germany). Hygiene-Institut; Richter, M. [Staatshochbauamt, Bonn (Germany)

    1994-04-01

    Evaporative cooling towers and industrial ventilator cooling towers have repeatedly been described as the origin of Legionnaires` disease. This article describes the design and function of cooling towers and evaporative cooling towers, sums up knowledge on the colonization of such systems with Legionella pneumophila, and describes conditions permitting the transmission of Legionella. Furthermore, design, maintenance, cleaning and disinfection measures are indicated which are believed to reduce the risk of infection through industrial and evaporative cooling towers. (orig.) [Deutsch] Rueckkuehlwerke klimatechnischer Anlagen und industrielle Ventilator-Kuehltuerme sind mehrfach als Ausgangspunkt von Legionellose-Epidemien beschrieben worden. In der vorliegenden Arbeit werden Aufbau und Funktion von Kuehltuermen und Rueckkuehlwerken erlaeutert, Erkenntnisse ueber die Besiedlung dieser Anlagen mit Legionellen zusammengefassten und die Situationen geschildert, in denen es zur Uebertragung von Legionellen kam. Darueber hinaus werden Konstruktions-, Wartungs-, Reinigungs-, und Desinfektionsmassnahmen aufgefuehrt, die zur Verminderung des Infektionsrisikos durch Kuehltuerme und Rueckkuehlwerke als wirksam angesehen werden. (orig.)

  5. The use of heteroduplex analysis of polymerase chain reaction products to support the possible transmission of Legionella pneumophila from a malfunctioning automobile air conditioner.

    Science.gov (United States)

    Pinar, Ahmet; Ramirez, Julio A; Schindler, Laura L; Miller, Richard D; Summersgill, James T

    2002-03-01

    Air conditioner condensates have not been previously associated with cases of Legionnaires' disease. We report the possible transmission of Legionella pneumophila serogroup 1 from a malfunctioning automobile air conditioning system's leaking water onto the floorboard of a car driven for a long distance by the patient. Heteroduplex analysis of polymerase chain reaction products was used to help establish an epidemiologic link between the water specimen and the patient.

  6. Field testing hot water temperature reduction as an energy-saving measure--does the Legionella presence change in a clinic's plumbing system?

    Science.gov (United States)

    Völker, Sebastian; Kistemann, Thomas

    2015-01-01

    Legionella spp. represent a significant health risk for humans. To ensure hygienically safe drinking water, technical guidelines recommend a central potable water hot (PWH) supply temperature of at least 60°C at the calorifier. In a clinic building we monitored whether slightly lowered temperatures in the PWH system led to a systemic change in the growth of these pathogens. In four separate phases we tested different scenarios concerning PWH supply temperatures and disinfection with chlorine dioxide (ClO2). In each phase, we took 5 sets of samples at 17 representative sampling points in the building's drinking water plumbing system. In total we collected 476 samples from the PWH system. All samples were tested (culture-based) for Legionella spp. and serogroups. Additionally, quantitative parameters at each sampling point were collected, which could possibly be associated with the presence of Legionella spp. (Pseudomonas aeruginsoa, heterotrophic plate count at 20°C and 36°C, temperatures, time until constant temperatures were reached, and chlorine dioxide concentration). The presence of Legionella spp. showed no significant reactions after reducing the PWH supply temperature from 63°C to 60°C and 57°C, as long as disinfection with ClO2 was maintained. After omitting the disinfectant, the PWH system showed statistically significant growth rates at 57°C. PWH temperatures which are permanently lowered to less than recommended values should be carefully accompanied by frequent testing, a thorough evaluation of the building's drinking water plumbing system, and hygiene expertise.

  7. Qualitative detection of Legionella species in bronchoalveolar lavages and induced sputa by automated DNA extraction and real-time polymerase chain reaction.

    Science.gov (United States)

    Raggam, R B; Leitner, E; Mühlbauer, G; Berg, J; Stöcher, M; Grisold, A J; Marth, E; Kessler, H H

    2002-10-01

    Molecular assays for qualitative detection of Legionella spp. in clinical specimens were evaluated. DNA extraction was done either with a fully automated DNA extraction protocol on the MagNA Pure LC System or with manual DNA extraction. Amplification and detection were done by real-time polymerase chain reaction (PCR) on the LightCycler (LC) instrument. Oligonucleotides were derived from the 16S rRNA gene of Legionella spp. The assays included a specially designed DNA fragment as Legionella-specific internal control. For both molecular assays, the detection limit was determined to be 5 CFU per LC PCR run. Sixty-one clinical specimens were tested with the molecular assays. Results were compared to culture. Five samples were found to be positive with the molecular assays. Three of them were positive in culture. No inhibition was found throughout the whole study. In conclusion, the molecular assays described may lead to safe and early diagnosis of Legionnaires' disease. They proved to be suitable for the routine molecular diagnostics laboratory.

  8. De aanwezigheid van Legionella species in door drinkwater gevoede watersystemen in Nederland. Deel I. 48 pp. (ook los verkrijgbaar). Idem, Deel II (ADDENDUM) Chemische en bacteriologische analyse resultaten per onderzocht object. Technische informatie van de onderzochte systemen per object. (Alleen samen met deel I verkrijgbaar)

    NARCIS (Netherlands)

    van Kranen HJ

    1988-01-01

    Legionella's werden geisoleerd uit warm watervoorzieningen, water van koeltorens, water van luchtbevochtigers en incidenteel uit drinkwater zelf. In 32% van de onderzochte objecten werden, uit water met een temperawtuur varierend van 6 graden C tot en met 59 graden C, legionella's

  9. Acetobacter intermedius, sp. nov.

    Science.gov (United States)

    Boesch, C; Trcek, J; Sievers, M; Teuber, M

    1998-03-01

    Strains of a new species in the genus Acetobacter, for which we propose the name A. intermedius sp. nov., were isolated and characterized in pure culture from different sources (Kombucha beverage, cider vinegar, spirit vinegar) and different countries (Switzerland, Slovenia). The isolated strains grow in media with 3% acetic acid and 3% ethanol as does A. europaeus, do, however, not require acetic acid for growth. These characteristics phenotypically position A. intermedius between A. europaeus and A. xylinus, DNA-DNA hybridizations of A. intermedius-DNA with DNA of the type strains of Acetobacter europaeus, A. xylinus, A. aceti, A. hansenii, A. liquefaciens, A. methanolicus, A. pasteurianus, A. diazotrophicus, Gluconobacter oxydans and Escherichia coli HB 101 indicated less than 60% DNA similarity. The important features of the new species are described. Acetobacter intermedius strain TF2 (DSM11804) isolated from the liquid phase of a tea fungus beverage (Kombucha) is the type strain.

  10. Tsukamurella tyrosinosolvens sp. nov.

    Science.gov (United States)

    Yassin, A F; Rainey, F A; Burghardt, J; Brzezinka, H; Schmitt, S; Seifert, P; Zimmermann, O; Mauch, H; Gierth, D; Lux, I; Schaal, K P

    1997-07-01

    Chemotaxonomic and 16S ribosomal DNA sequence analyses of four bacterial isolates from blood cultures from patients with cardiac pacemaker implants and sputa of patients with chronic lung infections clearly demonstrated that these bacteria belong to the genus Tsukamurella. DNA-DNA hybridization data, as well as the physiological characteristics of the isolates, indicate that they are closely related and belong to a single species that differs from previously described members of the genus Tsukamurella. The name Tsukamurella tyrosinosolvens sp. nov. is proposed for these isolates, and the new species is represented by strain IMMIB D-1397T (= DSM 44234T). Strain IMMIB D-1397T exhibits 53.4, 53.5, and 54.7% DNA-DNA relatedness to Tsukamurella paurometabola DSM 20162T, Tsukamurella inchonensis DSM 44067T, and Tsukamurella pulmonis DSM 44142T, respectively.

  11. Temperature-Dependent Growth Modeling of Environmental and Clinical Legionella pneumophila Multilocus Variable-Number Tandem-Repeat Analysis (MLVA) Genotypes.

    Science.gov (United States)

    Sharaby, Yehonatan; Rodríguez-Martínez, Sarah; Oks, Olga; Pecellin, Marina; Mizrahi, Hila; Peretz, Avi; Brettar, Ingrid; Höfle, Manfred G; Halpern, Malka

    2017-04-15

    Legionella pneumophila causes waterborne infections resulting in severe pneumonia. High-resolution genotyping of L. pneumophila isolates can be achieved by multiple-locus variable-number tandem-repeat analysis (MLVA). Recently, we found that different MLVA genotypes of L. pneumophila dominated different sites in a small drinking-water network, with a genotype-related temperature and abundance regime. The present study focuses on understanding the temperature-dependent growth kinetics of the genotypes that dominated the water network. Our aim was to model mathematically the influence of temperature on the growth kinetics of different environmental and clinical L. pneumophila genotypes and to compare it with the influence of their ecological niches. Environmental strains showed a distinct temperature preference, with significant differences among the growth kinetics of the three studied genotypes (Gt4, Gt6, and Gt15). Gt4 strains exhibited superior growth at lower temperatures (25 and 30°C), while Gt15 strains appeared to be best adapted to relatively higher temperatures (42 and 45°C). The temperature-dependent growth traits of the environmental genotypes were consistent with their distribution and temperature preferences in the water network. Clinical isolates exhibited significantly higher growth rates and reached higher maximal cell densities at 37°C and 42°C than the environmental strains. Further research on the growth preferences of L. pneumophila clinical and environmental genotypes will result in a better understanding of their ecological niches in drinking-water systems as well as in the human body. IMPORTANCE Legionella pneumophila is a waterborne pathogen that threatens humans in developed countries. The bacteria inhabit natural and man-made freshwater environments. Here we demonstrate that different environmental L. pneumophila genotypes have different temperature-dependent growth kinetics. Moreover, Legionella strains that belong to the same species

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

    Directory of Open Access Journals (Sweden)

    Pasqualina Laganà

    2014-09-01

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

  13. Bakteri Legionella pneumophila Terdeteksi pada Air Kolam Renang di Kota Surabaya dengan Nested Polymerase Chain Reaction (LEGIONELLA PNEUMOPHILA BACTERIADETECTED IN SWIMMING POOL WATER OF SURABAYA BY USING NESTED POLYMERASE CHAIN REACTION

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    Eduardus Bimo Aksono

    2017-06-01

    Full Text Available Legionella pneumophila is a Gram-negative bacillus that causes nosocomial and community-acquired pneumonia. The aim of this research was to detect the presence of bacteria of L. pneumophila species in the swimming pools water of Surabaya city by using nested Polymerase Chain Reaction (PCR assay of a specific gene for L. pneumophila (mip gene. This study used purposive sampling method. A total of 10 water samples were collected from five swimming pools consisting of 200 mL water for each swimming pool. The results showed that of 10 samples tested by nested PCR, one sample was positive for L. pneumophila, and nine samples were negative. L. pneumophila were found in pool water samples with a higher temperature (>30ºC.Serogrouping analysis of positive sample that L. pneumophila bacteria detected in the water sample of swimming pool in Surabaya was L. pneumophila serogroup 9 (98% and serogroup 10 (98%. L. pneumophila detection of bacteria is expected to raise the awareness of physician and microbiologists about the transmission of L. pneumophila and will also be useful for controlling the agents. ABSTRAK Legionella pneumophila adalah bakteri Gram-negatif berbentuk batang yang dapat menyebabkan penyakit nosokomial dan pneumonia. Tujuan penelitian ini adalah untuk mendeteksi keberadaan bakteri L. pneumophila pada air kolam renang di Kota Surabaya dengan menggunakan nested Polymerase Chain Reaction (PCR berbasis gen spesifik L. pneumophila (mip gene. Penelitian ini menggunakan metode purposive sampling. Sebanyak sepuluh sampel diambil dari lima kolam renang. Sampel diambil sebanyak 200 mL dari air kolam renang di setiap lokasi. Hasil dari 10 sampel yang diuji menggunakan nested PCR, satu sampel menunjukkan hasil positif untuk L.pneumophila, dan sembilan sampel menunjukkan hasil negatif. Bakteri L. pneumophila ditemukan pada sampel air kolam dengan suhu yang lebih tinggi (>30ºC. Satu sampel positip tersebut ketika dilanjutkan terhadap analisis serogrup

  14. Biofilm Composition and Threshold Concentration for Growth of Legionella pneumophila on Surfaces Exposed to Flowing Warm Tap Water without Disinfectant.

    Science.gov (United States)

    van der Kooij, Dick; Bakker, Geo L; Italiaander, Ronald; Veenendaal, Harm R; Wullings, Bart A

    2017-03-01

    Legionella pneumophila in potable water installations poses a potential health risk, but quantitative information about its replication in biofilms in relation to water quality is scarce. Therefore, biofilm formation on the surfaces of glass and chlorinated polyvinyl chloride (CPVC) in contact with tap water at 34 to 39°C was investigated under controlled hydraulic conditions in a model system inoculated with biofilm-grown L. pneumophila The biofilm on glass (average steady-state concentration, 23 ± 9 pg ATP cm -2 ) exposed to treated aerobic groundwater (0.3 mg C liter -1 ; 1 μg assimilable organic carbon [AOC] liter -1 ) did not support growth of the organism, which also disappeared from the biofilm on CPVC (49 ± 9 pg ATP cm -2 ) after initial growth. L. pneumophila attained a level of 4.3 log CFU cm -2 in the biofilms on glass (1,055 ± 225 pg ATP cm -2 ) and CPVC (2,755 ± 460 pg ATP cm -2 ) exposed to treated anaerobic groundwater (7.9 mg C liter -1 ; 10 μg AOC liter -1 ). An elevated biofilm concentration and growth of L. pneumophila were also observed with tap water from the laboratory. The Betaproteobacteria Piscinibacter and Methyloversatilis and amoeba-resisting Alphaproteobacteria predominated in the clones and isolates retrieved from the biofilms. In the biofilms, the Legionella colony count correlated significantly with the total cell count (TCC), heterotrophic plate count, ATP concentration, and presence of Vermamoeba vermiformis This amoeba was rarely detected at biofilm concentrations of <100 pg ATP cm -2 A threshold concentration of approximately 50 pg ATP cm -2 (TCC = 1 × 10 6 to 2 × 10 6 cells cm -2 ) was derived for growth of L. pneumophila in biofilms. IMPORTANCE Legionella pneumophila is the etiologic agent in more than 10,000 cases of Legionnaires' disease that are reported annually worldwide and in most of the drinking water-associated disease outbreaks reported in the United States. The organism proliferates in biofilms on surfaces

  15. Water heater temperature set point and water use patterns influence Legionella pneumophila and associated microorganisms at the tap.

    Science.gov (United States)

    Rhoads, William J; Ji, Pan; Pruden, Amy; Edwards, Marc A

    2015-12-01

    Lowering water heater temperature set points and using less drinking water are common approaches to conserving water and energy; yet, there are discrepancies in past literature regarding the effects of water heater temperature and water use patterns on the occurrence of opportunistic pathogens, in particular Legionella pneumophila. Our objective was to conduct a controlled, replicated pilot-scale investigation to address this knowledge gap using continuously recirculating water heaters to examine five water heater set points (39-58 °C) under three water use conditions. We hypothesized that L. pneumophila levels at the tap depend on the collective influence of water heater temperature, flow frequency, and the resident plumbing ecology. We confirmed temperature setting to be a critical factor in suppressing L. pneumophila growth both in continuously recirculating hot water lines and at distal taps. For example, at 51 °C, planktonic L. pneumophila in recirculating lines was reduced by a factor of 28.7 compared to 39 °C and was prevented from re-colonizing biofilm. However, L. pneumophila still persisted up to 58 °C, with evidence that it was growing under the conditions of this study. Further, exposure to 51 °C water in a low-use tap appeared to optimally select for L. pneumophila (e.g., 125 times greater numbers than in high-use taps). We subsequently explored relationships among L. pneumophila and other ecologically relevant microbes, noting that elevated temperature did not have a general disinfecting effect in terms of total bacterial numbers. We documented the relationship between L. pneumophila and Legionella spp., and noted several instances of correlations with Vermamoeba vermiformis, and generally found that there is a dynamic relationship with this amoeba host over the range of temperatures and water use frequencies examined. Our study provides a new window of understanding into the microbial ecology of potable hot water systems and helps to resolve

  16. Interactive effects of temperature, organic carbon, and pipe material on microbiota composition and Legionella pneumophila in hot water plumbing systems.

    Science.gov (United States)

    Proctor, Caitlin R; Dai, Dongjuan; Edwards, Marc A; Pruden, Amy

    2017-10-04

    Several biotic and abiotic factors have been reported to influence the proliferation of microbes, including Legionella pneumophila, in hot water premise plumbing systems, but their combined effects have not been systematically evaluated. Here, we utilize simulated household water heaters to examine the effects of stepwise increases in temperature (32-53 °C), pipe material (copper vs. cross-linked polyethylene (PEX)), and influent assimilable organic carbon (0-700 μg/L) on opportunistic pathogen gene copy numbers and the microbiota composition, as determined by quantitative polymerase chain reaction and 16S rRNA gene amplicon sequencing. Temperature had an overarching influence on both the microbiota composition and L. pneumophila numbers. L. pneumophila peaked at 41 °C in the presence of PEX (1.58 × 10 5 gene copies/mL). At 53 °C, L. pneumophila was not detected. Several operational taxonomic units (OTUs) persisted across all conditions, accounting for 50% of the microbiota composition from 32 to 49 °C and 20% at 53 °C. Pipe material most strongly influenced microbiota composition at lower temperatures, driven by five to six OTUs enriched with each material. Copper pipes supported less L. pneumophila than PEX pipes (mean 2.5 log 10 lower) at temperatures ≤ 41 °C, but showed no difference in total bacterial numbers. Differences between pipe materials diminished with elevated temperature, probably resulting from decreased release of copper ions. At temperatures ≤ 45 °C, influent assimilable organic carbon correlated well with total bacterial numbers, but not with L. pneumophila numbers. At 53 °C, PEX pipes leached organic carbon, reducing the importance of dosed organic carbon. L. pneumophila numbers correlated with a Legionella OTU and a Methylophilus OTU identified by amplicon sequencing. Temperature was the most effective factor for the control of L. pneumophila, while microbiota composition shifted with each stepwise temperature

  17. Molecular mimicry by an F-box effector of Legionella pneumophila hijacks a conserved polyubiquitination machinery within macrophages and protozoa.

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    Christopher T Price

    2009-12-01

    Full Text Available The ability of Legionella pneumophila to proliferate within various protozoa in the aquatic environment and in macrophages indicates a remarkable evolution and microbial exploitation of evolutionarily conserved eukaryotic processes. Ankyrin B (AnkB of L. pneumophila is a non-canonical F-box-containing protein, and is the only known Dot/Icm-translocated effector of L. pneumophila essential for intra-vacuolar proliferation within both macrophages and protozoan hosts. We show that the F-box domain of AnkB and the (9L(10P conserved residues are essential for intracellular bacterial proliferation and for rapid acquisition of polyubiquitinated proteins by the Legionella-containing vacuole (LCV within macrophages, Dictyostelium discoideum, and Acanthamoeba. Interestingly, translocation of AnkB and recruitment of polyubiquitinated proteins in macrophages and Acanthamoeba is rapidly triggered by extracellular bacteria within 5 min of bacterial attachment. Ectopically expressed AnkB within mammalian cells is localized to the periphery of the cell where it co-localizes with host SKP1 and recruits polyubiquitinated proteins, which results in restoration of intracellular growth to the ankB mutant similar to the parental strain. While an ectopically expressed AnkB-(9L(10P/AA variant is localized to the cell periphery, it does not recruit polyubiquitinated proteins and fails to trans-rescue the ankB mutant intracellular growth defect. Direct in vivo interaction of AnkB but not the AnkB-(9L(10P/AA variant with the host SKP1 is demonstrated. Importantly, RNAi-mediated silencing of expression of SKP1 renders the cells non-permissive for intracellular proliferation of L. pneumophila. The role of AnkB in exploitation of the polyubiquitination machinery is essential for intrapulmonary bacterial proliferation in the mouse model of Legionnaires' disease. Therefore, AnkB exhibits a novel molecular and functional mimicry of eukaryotic F-box proteins that exploits

  18. Bradysia sp. em morangueiro Bradysia sp. in strawberry

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    Bernadete Radin

    2009-04-01

    Full Text Available No trabalho, relatam-se os primeiros registros de Bradysia sp. (Insecta: Diptera: Sciaridae em morangueiro (Fragaria x ananassa Duch., cultivado no Município de Eldorado do Sul, RS. O cultivo foi realizado em sacolas com três metros de comprimento, preenchidas com substrato composto de casca de arroz e turfa, dispostas horizontalmente sobre bancadas de madeira, em ambiente protegido. A presença de Bradysia sp. foi observada na segunda quinzena de agosto de 2005. Neste trabalho, estão descritos os sintomas apresentados no morangueiro pela praga, prováveis conseqüências sobre o aparecimento de doenças e uma breve descrição morfológica da Bradysia sp., adulto e fase larval.This paper describes the first record of Bradysia sp. (Insecta; Diptera; Sciaridae in strawberry (Fragaria x ananassa, cultivated in the city of Eldorado do Sul, RS, Brazil. Strawberry was planted in plastic bags filled with a mixture of burnt rice hulls and peat and cultivated in a greenhouse. The presence of Bradysia sp was noticed in the second fortnight of August, 2005. The symptoms in strawberry and the probable consequences in terms of disease arising were described in the present study, as well as the morphological characterization of Bradysia sp. and its illustrations.

  19. SP-100 reactor design

    International Nuclear Information System (INIS)

    Armijo, J.S.; Atwell, J.; Pluta, P.R.; Smith, M.A.; Solorzano, E.R.

    1987-01-01

    The SP-100 space reactor power system is being designed and developed as part of the Ground Engineering System (GES) contract between General Electric Company as the system developer and the Department of Energy. Other key participants in the GES program include Westinghouse Hanford Company (site operator), Los Alamos National Laboratory (fuel development and production), Oak Ridge National Laboratory (materials), and Westinghouse, Advanced Energy Systems Division (shield, HTS equipment). The GES Program includes two major elements. First, the development of a Reference Flight System design at 100 kWe output to the user, and second the validation of the Reference Flight System design by analysis and by testing. Development of key technologies along with component and system testing is an essential part of the validation program. The nuclear subsystem validation includes the design, manufacture, assembly and operational testing of a Ground Reactor Test Assembly. The subject of this paper is the reactor design for the Reference Flight System. The reference flight design is in the preliminary design stage and will evolve over the next year

  20. Structural basis for the recruitment and activation of the Legionella phospholipase VipD by the host GTPase Rab5

    Science.gov (United States)

    Lucas, María; Gaspar, Andrew H.; Pallara, Chiara; Rojas, Adriana Lucely; Fernández-Recio, Juan; Machner, Matthias P.; Hierro, Aitor

    2014-01-01

    A challenge for microbial pathogens is to assure that their translocated effector proteins target only the correct host cell compartment during infection. The Legionella pneumophila effector vacuolar protein sorting inhibitor protein D (VipD) localizes to early endosomal membranes and alters their lipid and protein composition, thereby protecting the pathogen from endosomal fusion. This process requires the phospholipase A1 (PLA1) activity of VipD that is triggered specifically on VipD binding to the host cell GTPase Rab5, a key regulator of endosomes. Here, we present the crystal structure of VipD in complex with constitutively active Rab5 and reveal the molecular mechanism underlying PLA1 activation. An active site-obstructing loop that originates from the C-terminal domain of VipD is repositioned on Rab5 binding, thereby exposing the catalytic pocket within the N-terminal PLA1 domain. Substitution of amino acid residues located within the VipD–Rab5 interface prevented Rab5 binding and PLA1 activation and caused a failure of VipD mutant proteins to target to Rab5-enriched endosomal structures within cells. Experimental and computational analyses confirmed an extended VipD-binding interface on Rab5, explaining why this L. pneumophila effector can compete with cellular ligands for Rab5 binding. Together, our data explain how the catalytic activity of a microbial effector can be precisely linked to its subcellular localization. PMID:25114243

  1. Interaction of Legionella pneumophila with Acanthamoeba castellanii: uptake by coiling phagocytosis and inhibition of phagosome-lysosome fusion.

    Science.gov (United States)

    Bozue, J A; Johnson, W

    1996-01-01

    Legionella pneumophila is a facultative intracellular parasite able to survive within both human monocytes and amoebae. We have demonstrated that processing of L. pneumophila by the free-living amoeba Acanthamoeba castellanii shows many similarities to the processing of L. pneumophila by monocytes. These similarities include uptake of L. pneumophila by coiling phagocytosis and the subsequent confinement of L. pneumophila in a ribosome-studded phagosome. In addition, as in monocytes, inhibition of lysosomal fusion with phagosomes containing L. pneumophila was detected in amoebae. With all clinical isolates, inhibition of phagosomes-lysosome fusion correlated with virulence. However, with one of the environmental isolates tested, no significant difference in phagosome-lysosome fusion was observed between the virulent and avirulent forms. These results indicate that the avirulent form of this isolate differed from the virulent form in some other respect critical to intracellular survival. Therefore, intracellular multiplication of L. pneumophila within A. castellanii may not be solely dependent upon the inhibition of lysosomal fusion. PMID:8550225

  2. Natural transformation occurs independently of the essential actin-like MreB cytoskeleton in Legionella pneumophila.

    Science.gov (United States)

    Juan, Pierre-Alexandre; Attaiech, Laetitia; Charpentier, Xavier

    2015-11-03

    Natural transformation is the process by which bacteria can actively take up and integrate exogenous DNA thereby providing a source of genetic diversity. Under specific growth conditions the coordinated expression of several genes--a situation referred to as "competence"--allows bacteria to assemble a highly processive and dedicated system that can import high molecular weight DNA. Within the cell these large imported DNA molecules are protected from degradation and brought to the chromosome for recombination. Here, we report elevated expression of mreB during competence in the Gram-negative pathogen Legionella pneumophila. Interestingly a similar observation had previously been reported in the distantly-related Gram-positive organism Bacillus subtilis. MreB is often viewed as the bacterial actin homolog contributing to bacterial morphogenesis by coordinating peptidoglycan-synthesising complexes. In addition MreB is increasingly found to be involved in a growing number of processes including chromosome segregation and motor-driven motility. Using genetic and pharmacological approaches, we examined the possible role of MreB during natural transformation in L. pneumophila. Our data show that natural transformation does not require MreB dynamics and exclude a direct role of MreB filaments in the transport of foreign DNA and its recombination in the chromosome.

  3. Inflammasome-mediated cell death in response to bacterial pathogens that access the host cell cytosol: lessons from Legionella pneumophila

    Directory of Open Access Journals (Sweden)

    Cierra Nichole Casson

    2013-12-01

    Full Text Available Cell death can be critical for host defense against intracellular pathogens because it eliminates a crucial replicative niche, and pro-inflammatory cell death can alert neighboring cells to the presence of pathogenic organisms and enhance downstream immune responses. Pyroptosis is a pro-inflammatory form of cell death triggered by the inflammasome, a multi-protein complex that assembles in the cytosol to activate caspase-1. Inflammasome activation by pathogens hinges upon violation of the host cell cytosol by activities such as the use of pore-forming toxins, the use of specialized secretion systems, or the cytosolic presence of the pathogen itself. Recently, a non-canonical inflammasome has been described that activates caspase-11 and also leads to pro-inflammatory cell death. Caspase-11 is activated rapidly and robustly in response to violation of the cytosol by bacterial pathogens as well. In this mini-review, we describe the canonical and non-canonical inflammasome pathways that are critical for host defense against a model intracellular bacterial pathogen that accesses the host cytosol—Legionella pneumophila.

  4. Identification of host cytosolic sensors and bacterial factors regulating the type I interferon response to Legionella pneumophila.

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    Kathryn M Monroe

    2009-11-01

    Full Text Available Legionella pneumophila is a gram-negative bacterial pathogen that replicates in host macrophages and causes a severe pneumonia called Legionnaires' Disease. The innate immune response to L. pneumophila remains poorly understood. Here we focused on identifying host and bacterial factors involved in the production of type I interferons (IFN in response to L. pneumophila. It was previously suggested that the delivery of L. pneumophila DNA to the host cell cytosol is the primary signal that induces the type I IFN response. However, our data are not easily reconciled with this model. We provide genetic evidence that two RNA-sensing proteins, RIG-I and MDA5, participate in the IFN response to L. pneumophila. Importantly, these sensors do not seem to be required for the IFN response to L. pneumophila DNA, whereas we found that RIG-I was required for the response to L. pneumophila RNA. Thus, we hypothesize that bacterial RNA, or perhaps an induced host RNA, is the primary stimulus inducing the IFN response to L. pneumophila. Our study also identified a secreted effector protein, SdhA, as a key suppressor of the IFN response to L. pneumophila. Although viral suppressors of cytosolic RNA-sensing pathways have been previously identified, analogous bacterial factors have not been described. Thus, our results provide new insights into the molecular mechanisms by which an intracellular bacterial pathogen activates and also represses innate immune responses.

  5. Neutrophil and Alveolar Macrophage-Mediated Innate Immune Control of Legionella pneumophila Lung Infection via TNF and ROS.

    Directory of Open Access Journals (Sweden)

    Pascal Ziltener

    2016-04-01

    Full Text Available Legionella pneumophila is a facultative intracellular bacterium that lives in aquatic environments where it parasitizes amoeba. However, upon inhalation of contaminated aerosols it can infect and replicate in human alveolar macrophages, which can result in Legionnaires' disease, a severe form of pneumonia. Upon experimental airway infection of mice, L. pneumophila is rapidly controlled by innate immune mechanisms. Here we identified, on a cell-type specific level, the key innate effector functions responsible for rapid control of infection. In addition to the well-characterized NLRC4-NAIP5 flagellin recognition pathway, tumor necrosis factor (TNF and reactive oxygen species (ROS are also essential for effective innate immune control of L. pneumophila. While ROS are essential for the bactericidal activity of neutrophils, alveolar macrophages (AM rely on neutrophil and monocyte-derived TNF signaling via TNFR1 to restrict bacterial replication. This TNF-mediated antibacterial mechanism depends on the acidification of lysosomes and their fusion with L. pneumophila containing vacuoles (LCVs, as well as caspases with a minor contribution from cysteine-type cathepsins or calpains, and is independent of NLRC4, caspase-1, caspase-11 and NOX2. This study highlights the differential utilization of innate effector pathways to curtail intracellular bacterial replication in specific host cells upon L. pneumophila airway infection.

  6. [Accreditation of a hygiene hospital laboratory for sampling and analysis activities for the detection and counting of Legionella in water].

    Science.gov (United States)

    Lecointe, Didier; Noël, Coralie; Beauvais, Raphaëlle; Descaves, Carole; Gouot, Armelle; Bourgeois, Sandrine; Koutcherenko, Stéphane; Kassidi, Noura

    2015-01-01

    Since January 1(st) 2012, detection and counting of Legionella bacteria have been obligatory in France and must be carried out by COFRAC-accredited laboratories. In our establishment, sampling and analysis were outsourced and our hospital was scheduled to move to a new site. We aimed to develop both these activities in-house and to obtain COFRAC accreditation, whilst organizing the move to the new site. We set up a quality assurance system bringing together staff from the hygiene laboratory and institutional resource managers. We set up sampling and analysis activities in-house 13 months before requesting accreditation. The initial evaluation took place before we moved and identified 17 areas of deficiency, six of which were considered critical. After we had moved, a subsequent evaluation considered 14 of these deficiencies to have been corrected, included the six initially identified as critical. We were therefore awarded accreditation. The quality assurance system established during the year before our request was submitted led to accreditation two and a half years after the transfer in-house of sampling and analysis activities, despite our hospital moving during this period.

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

    Directory of Open Access Journals (Sweden)

    Zhang Qin-fen

    2010-02-01

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

  8. Structure and Function of Interacting IcmR-IcmQ Domains from a Type IVb Secretion System in Legionella pneumophila

    Energy Technology Data Exchange (ETDEWEB)

    Raychaudhury, S.; Farelli, J; Montminy, T; Matthews, M; Menetret, J; Dumenil, G; Roy, C; Head, J; Isberg, R; Akey, C

    2009-01-01

    During infection, Legionella pneumophila creates a replication vacuole within eukaryotic cells and this requires a Type IVb secretion system (T4bSS). IcmQ plays a critical role in the translocase and associates with IcmR. In this paper, we show that the N-terminal domain of IcmQ (Qn) mediates self-dimerization, whereas the C-terminal domain with a basic linker promotes membrane association. In addition, the binding of IcmR to IcmQ prevents self-dimerization and also blocks membrane permeabilization. However, IcmR does not completely block membrane binding by IcmQ. We then determined crystal structures of Qn with the interacting region of IcmR. In this complex, each protein forms an ?-helical hairpin within a parallel four-helix bundle. The amphipathic nature of helices in Qn suggests two possible models for membrane permeabilization by IcmQ. The Rm-Qn structure also suggests how IcmR-like proteins in other L. pneumophila species may interact with their IcmQ partners.

  9. Structure and function of interacting IcmR-IcmQ domains from a type IVb secretion system in Legionella pneumophila.

    Science.gov (United States)

    Raychaudhury, Suchismita; Farelli, Jeremiah D; Montminy, Timothy P; Matthews, Miguelina; Ménétret, Jean-François; Duménil, Guillaume; Roy, Craig R; Head, James F; Isberg, Ralph R; Akey, Christopher W

    2009-04-15

    During infection, Legionella pneumophila creates a replication vacuole within eukaryotic cells and this requires a Type IVb secretion system (T4bSS). IcmQ plays a critical role in the translocase and associates with IcmR. In this paper, we show that the N-terminal domain of IcmQ (Qn) mediates self-dimerization, whereas the C-terminal domain with a basic linker promotes membrane association. In addition, the binding of IcmR to IcmQ prevents self-dimerization and also blocks membrane permeabilization. However, IcmR does not completely block membrane binding by IcmQ. We then determined crystal structures of Qn with the interacting region of IcmR. In this complex, each protein forms an alpha-helical hairpin within a parallel four-helix bundle. The amphipathic nature of helices in Qn suggests two possible models for membrane permeabilization by IcmQ. The Rm-Qn structure also suggests how IcmR-like proteins in other L. pneumophila species may interact with their IcmQ partners.

  10. Neutrophil and Alveolar Macrophage-Mediated Innate Immune Control of Legionella pneumophila Lung Infection via TNF and ROS

    Science.gov (United States)

    Ziltener, Pascal; Reinheckel, Thomas; Oxenius, Annette

    2016-01-01

    Legionella pneumophila is a facultative intracellular bacterium that lives in aquatic environments where it parasitizes amoeba. However, upon inhalation of contaminated aerosols it can infect and replicate in human alveolar macrophages, which can result in Legionnaires’ disease, a severe form of pneumonia. Upon experimental airway infection of mice, L. pneumophila is rapidly controlled by innate immune mechanisms. Here we identified, on a cell-type specific level, the key innate effector functions responsible for rapid control of infection. In addition to the well-characterized NLRC4-NAIP5 flagellin recognition pathway, tumor necrosis factor (TNF) and reactive oxygen species (ROS) are also essential for effective innate immune control of L. pneumophila. While ROS are essential for the bactericidal activity of neutrophils, alveolar macrophages (AM) rely on neutrophil and monocyte-derived TNF signaling via TNFR1 to restrict bacterial replication. This TNF-mediated antibacterial mechanism depends on the acidification of lysosomes and their fusion with L. pneumophila containing vacuoles (LCVs), as well as caspases with a minor contribution from cysteine-type cathepsins or calpains, and is independent of NLRC4, caspase-1, caspase-11 and NOX2. This study highlights the differential utilization of innate effector pathways to curtail intracellular bacterial replication in specific host cells upon L. pneumophila airway infection. PMID:27105352

  11. Effect of temperature, pH, and oxygen level on the multiplication of naturally occurring Legionella pneumophila in potable water.

    Science.gov (United States)

    Wadowsky, R M; Wolford, R; McNamara, A M; Yee, R B

    1985-05-01

    A water culture containing naturally occurring Legionella pneumophila and associated microbiota was maintained in the laboratory by serially transferring the culture in tap water which had been sterilized by membrane filtration. Successful maintenance of the water culture depended upon transferring the culture when the growth of L. pneumophila was in the late-exponential to early-stationary phase. The water culture was used as a source of naturally occurring bacteria to determine some of the parameters which affect the multiplication of L. pneumophila in tap water. Naturally occurring L. pneumophila multiplied at a temperature between 25 and 37 degrees C, at pH levels of 5.5 to 9.2, and at concentrations of dissolved oxygen of 6.0 to 6.7 mg/liter. Multiplication did not occur in tap water which contained less than 2.2 mg of dissolved oxygen per liter. An association was observed between the multiplication of L. pneumophila and the non-Legionellaceae bacteria which were also present in the water culture. The method of preserving naturally occurring L. pneumophila and associated microbiota may facilitate studies on the symbiosis of L. pneumophila with other microorganisms.

  12. Genotypic diversity of Legionella pneumophila in environmental and clinical strains assessed by Sequence-Based Typing, in association with retrospective clinical surveillance in Northern Italy.

    Science.gov (United States)

    Bianchi, Annalisa; Pregliasco, Fabrizio Ernesto; Consonni, Michela; Tesauro, Marina

    2016-06-02

    The study aimed to evidence previous cases of legionellosis or proven or suspected pneumonia in seven hospital facilities for the mentally disabled in Northern Italy, where no clinical surveillance had been previously carried out. An additional aim was to highlight the occurrence of strains of Legionella pneumophila of clinical and environmental origin by Sequence-Based Typing (SBT), comparing them to world surveillance. A clinical survey was perfomed from 2003-2012, analyzing 615 medical records for hospital- and community-acquired pneumonia, with particular attention to legionellosis. Clinical (n=4) and environmental (n=25) isolates of Legionella pneumophila, isolated in the same period (2003-2012), were characterized by SBT and the Sequence Types (STs) compared with the European Working Group for Legionella Infections (EWGLI) database. Surveillance revealed that there were seven detected cases of legionellosis; most pneumonia cases could not be confirmed by diagnostic tests because of the disabilities of the patients and their lack of cooperation. The same ST was found in two of the clinical strains and also in a corresponding environment, i.e. ST685 and ST16, and two clinical strains belonging to the same ST (ST1). The other environmental strains were isolated in department with confirmed/suspected clinical cases. Five other STs found in this study were new to the database: ST685 was isolated both from a patient and from water; ST694, ST1181, ST1370 and ST1371 have not been described previously. The study confirmed that the routine collection and analysis of environmental strains may be an important strategy for preventing sporadic and epidemic cases of legionellosis, in association with clinical surveillance.

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

    Science.gov (United States)

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

    2014-12-01

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

  14. Penicillium araracuarense sp. nov., Penicillium elleniae sp. nov., Penicillium penarojense sp. nov., Penicillium vanderhammenii sp. nov. and Penicillium wotroi sp. nov., isolated from leaf litter

    DEFF Research Database (Denmark)

    Houbraken, Jos; López-Quintero, Carlos A.; Frisvad, Jens Christian

    2011-01-01

    Several species of the genus Penicillium were isolated during a survey of the mycobiota of leaf litter and soil in Colombian Amazon forest. Five species, Penicillium penarojense sp. nov. (type strain CBS 113178T = IBT 23262T), Penicillium wotroi sp. nov. (type strain CBS 118171T = IBT 23253T......), Penicillium araracuarense sp. nov. (type strain CBS 113149T = IBT 23247T), Penicillium elleniae sp. nov. (type strain CBS 118135T = IBT 23229T) and Penicillium vanderhammenii sp. nov. (type strain CBS 126216T = IBT 23203T) are described here as novel species. Their taxonomic novelty was determined using......, andrastin A, pulvilloric acid, paxillin, paspaline and janthitrem, were commonly produced by these phylogenetically related species. The novel species had a high growth rate on agar media, but could be distinguished from each other by several macro- and microscopical characteristics....

  15. Uji Fitokimia, Kandungan Total Fenol dan Aktivitas Antioksidan Mikroalga Spirulina sp., Chlorella sp., dan Nannochloropsis sp.

    Directory of Open Access Journals (Sweden)

    Diini Fithriani

    2015-12-01

    Full Text Available Penelitian ini bertujuan untuk mengetahui kandungan fitokimia, total fenol, dan aktivitas antioksidan dari mikroalga Spirulina sp., Nannochloropsis sp., dan Chlorella sp. Mikroalga diekstrak dengan ekstraksi tunggal menggunakan etanol. Skrining fitokimia dilakukan secara kualitatif. Analisis total fenol dilakukan secara spektrofotometri dengan metode Folin-Ciocalteu. Analisis antioksidan dilakukan dengan metode 2,2-diphenyl-1-picrylhydrazyl (DPPH dan Ferric Reducing Antioxidant Power (FRAP. Skrining fitokimia menunjukkan keberadaan tanin, flavonoid, steroid, glikosida, dan alkaloid pada ekstrak etanol ketiga jenis mikroalga, sedangkan saponin hanya terdeteksi pada ekstrak etanol Spirulina sp. dan Chlorella sp., adapun triterpenoid tidak terdeteksi pada ekstrak etanol ketiga jenis mikroalga. Hasil penelitian menunjukkan bahwa kandungan total fenol, aktivitas antioksidan (IC50, dan kapasitas antioksidan (FRAP tertinggi diperoleh pada ekstrak etanol Spirulina sp. dengan nilai berturut turut sebesar 0,32 ± 0,025 mg GAE g-1D.W., 518,94 ppm, dan  49,95 ± 2,02 (mmol Fe2+ eq.g-1D.W. Dalam penelitian ini diketahui bahwa kandungan fenol total memiliki korelasi  yang kuat  dengan kapasitas antioksidan metode FRAP (R2= 0,84, dan aktivitas antioksidan metode DPPH (R2= 0,79.

  16. Uji Fitokimia, Kandungan Total Fenol Dan Aktivitas Antioksidan Mikroalga Spirulina Sp., Chlorella Sp., dan Nannochloropsis Sp.

    Directory of Open Access Journals (Sweden)

    Diini Fithriani

    2015-12-01

    Full Text Available Penelitian ini bertujuan untuk mengetahui kandungan fitokimia, total fenol, dan aktivitas antioksidan dari mikroalga Spirulina sp., Nannochloropsis sp., dan Chlorella sp. Mikroalga diekstrak dengan ekstraksi tunggal menggunakan etanol. Skrining fitokimia dilakukan secara kualitatif. Analisis total fenol dilakukan secara spektrofotometri dengan metode Folin-Ciocalteu. Analisis antioksidan dilakukan dengan metode 2,2-diphenyl-1-picrylhydrazyl (DPPH dan Ferric Reducing Antioxidant Power (FRAP. Skrining fitokimia menunjukkan keberadaan tanin, flavonoid, steroid, glikosida, dan alkaloid pada ekstrak etanol ketiga jenis mikroalga, sedangkan saponin hanya terdeteksi pada ekstrak etanol Spirulina sp. dan Chlorella sp., adapun triterpenoid tidak terdeteksi pada ekstrak etanol ketiga jenis mikroalga. Hasil penelitian menunjukkan bahwa kandungan total fenol, aktivitas antioksidan (IC50, dan kapasitas antioksidan (FRAP tertinggi diperoleh pada ekstrak etanol Spirulina sp. dengan nilai berturut turut sebesar 0,32 ± 0,025 mg GAE g-1D.W., 518,94 ppm, dan  49,95 ± 2,02 (mmol Fe2+ eq.g-1D.W. Dalam penelitian ini diketahui bahwa kandungan fenol total memiliki korelasi  yang kuat  dengan kapasitas antioksidan metode FRAP (R2= 0,84, dan aktivitas antioksidan metode DPPH (R2= 0,79.

  17. Distribution of Legionella pneumophila serogroups, monoclonal antibody subgroups and DNA sequence types in recent clinical and environmental isolates from England and Wales (2000-2008).

    Science.gov (United States)

    Harrison, T G; Afshar, B; Doshi, N; Fry, N K; Lee, J V

    2009-07-01

    Clinical isolates of Legionella pneumophila, obtained from 167 patients, who acquired their illness in the community in England and Wales between January 2000 and March 2008, were compared with 276 environmental isolates of L. pneumophila obtained over the same period as part of the routine sampling of 'managed' water systems. The 443 isolates were typed by monoclonal antibody (mAb) subgrouping and the internationally standardised, seven-gene loci, sequence-based typing (SBT) scheme of the European Working Group for Legionella Infections (EWGLI). Of the clinical isolates, 97.6% were L. pneumophila serogroup (sgp) 1, compared with only 55.8% of environmental isolates (P = 0.0002); 91.6% were subgrouped as mAb3/1+ve, compared with only 8.3% of environmental isolates (P accounting for 25.7% and three (ST47, ST37 and ST62) accounting for 46.1% of all isolates. Eighty-two STs were identified among the environmental isolates, with two (ST1 and ST79) accounting for 34.1% of these. Comparison of the STs seen among clinical and environmental isolates showed that there was very little overlap between the two populations (P accounted for 22.8% of cases, but was only found once in the environment. Conversely, mAb 'Oxford/OLDA' ST1 was the most common environmental strain (17.0%), but only caused two infections. A review of the published data shows that mAb 'Allentown' ST47 is also an important cause of infection in France and possibly in the Netherlands. However, it was not found in a large study of German clinical isolates. This study confirms previous work showing that just a few strains of L. pneumophila cause the majority of community-acquired Legionella infection in England and Wales, and that these clinically significant strains are only rarely found in managed water systems. These data suggest that knowing which particular strain is present in an environment might be at least as important as knowing the quantity in which legionellae are present.

  18. Differences in Virulence Between Legionella pneumophila Isolates From Human and Non-human Sources Determined in Galleria mellonella Infection Model

    Directory of Open Access Journals (Sweden)

    Patrícia S. Sousa

    2018-04-01

    Full Text Available Legionella pneumophila is a ubiquitous bacterium in freshwater environments and in many man-made water systems capable of inducing pneumonia in humans. Despite its ubiquitous character most studies on L. pneumophila virulence focused on clinical strains and isolates from man-made environments, so little is known about the nature and extent of virulence variation in strains isolated from natural environments. It has been established that clinical isolates are less diverse than man-made and natural environmental strains, suggesting that only a subset of environmental isolates is specially adapted to infect humans. In this work we intended to determine if unrelated L. pneumophila strains, isolated from different environments and with distinct virulence-related genetic backgrounds, displayed differences in virulence, using the Wax Moth Galleria mellonella infection model. We found that all tested strains were pathogenic in G. mellonella, regardless of their origin. Indeed, a panoply of virulence-related phenotypes was observed sustaining the existence of significant differences on the ability of L. pneumophila strains to induce disease. Taken together our results suggest that the occurrence of human infection is not related with the increased capability of some strains to induce disease since we also found a concentration threshold above which L. pneumophila strains are equally able to cause disease. In addition, no link could be established between the sequence-type (ST and L. pneumophila pathogenicity. We envision that in man-made water distribution systems environmental filtering selection and biotic competition acts structuring L. pneumophila populations by selecting more resilient and adapted strains that can rise to high concentration if no control measures are implemented. Therefore, public health strategies based on the sequence based typing (STB scheme analysis should take into account that the major disease-associated clones of L

  19. RavN is a member of a previously unrecognized group of Legionella pneumophila E3 ubiquitin ligases

    Science.gov (United States)

    Lin, Yi-Han; Evans, Timothy R.; Doms, Alexandra G.; Beauchene, Nicole A.; Hierro, Aitor

    2018-01-01

    The eukaryotic ubiquitylation machinery catalyzes the covalent attachment of the small protein modifier ubiquitin to cellular target proteins in order to alter their fate. Microbial pathogens exploit this post-translational modification process by encoding molecular mimics of E3 ubiquitin ligases, eukaryotic enzymes that catalyze the final step in the ubiquitylation cascade. Here, we show that the Legionella pneumophila effector protein RavN belongs to a growing class of bacterial proteins that mimic host cell E3 ligases to exploit the ubiquitylation pathway. The E3 ligase activity of RavN was located within its N-terminal region and was dependent upon interaction with a defined subset of E2 ubiquitin-conjugating enzymes. The crystal structure of the N-terminal region of RavN revealed a U-box-like motif that was only remotely similar to other U-box domains, indicating that RavN is an E3 ligase relic that has undergone significant evolutionary alteration. Substitution of residues within the predicted E2 binding interface rendered RavN inactive, indicating that, despite significant structural changes, the mode of E2 recognition has remained conserved. Using hidden Markov model-based secondary structure analyses, we identified and experimentally validated four additional L. pneumophila effectors that were not previously recognized to possess E3 ligase activity, including Lpg2452/SdcB, a new paralog of SidC. Our study provides strong evidence that L. pneumophila is dedicating a considerable fraction of its effector arsenal to the manipulation of the host ubiquitylation pathway. PMID:29415051

  20. A Legionella pneumophila effector protein encoded in a region of genomic plasticity binds to Dot/Icm-modified vacuoles.

    Directory of Open Access Journals (Sweden)

    Shira Ninio

    2009-01-01

    Full Text Available Legionella pneumophila is an opportunistic pathogen that can cause a severe pneumonia called Legionnaires' disease. In the environment, L. pneumophila is found in fresh water reservoirs in a large spectrum of environmental conditions, where the bacteria are able to replicate within a variety of protozoan hosts. To survive within eukaryotic cells, L. pneumophila require a type IV secretion system, designated Dot/Icm, that delivers bacterial effector proteins into the host cell cytoplasm. In recent years, a number of Dot/Icm substrate proteins have been identified; however, the function of most of these proteins remains unknown, and it is unclear why the bacterium maintains such a large repertoire of effectors to promote its survival. Here we investigate a region of the L. pneumophila chromosome that displays a high degree of plasticity among four sequenced L. pneumophila strains. Analysis of GC content suggests that several genes encoded in this region were acquired through horizontal gene transfer. Protein translocation studies establish that this region of genomic plasticity encodes for multiple Dot/Icm effectors. Ectopic expression studies in mammalian cells indicate that one of these substrates, a protein called PieA, has unique effector activities. PieA is an effector that can alter lysosome morphology and associates specifically with vacuoles that support L. pneumophila replication. It was determined that the association of PieA with vacuoles containing L. pneumophila requires modifications to the vacuole mediated by other Dot/Icm effectors. Thus, the localization properties of PieA reveal that the Dot/Icm system has the ability to spatially and temporally control the association of an effector with vacuoles containing L. pneumophila through activities mediated by other effector proteins.

  1. The Influence of Programmed Cell Death in Myeloid Cells on Host Resilience to Infection with Legionella pneumophila or Streptococcus pyogenes.

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    Pia Gamradt

    2016-12-01

    Full Text Available Pathogen clearance and host resilience/tolerance to infection are both important factors in surviving an infection. Cells of the myeloid lineage play important roles in both of these processes. Neutrophils, monocytes, macrophages, and dendritic cells all have important roles in initiation of the immune response and clearance of bacterial pathogens. If these cells are not properly regulated they can result in excessive inflammation and immunopathology leading to decreased host resilience. Programmed cell death (PCD is one possible mechanism that myeloid cells may use to prevent excessive inflammation. Myeloid cell subsets play roles in tissue repair, immune response resolution, and maintenance of homeostasis, so excessive PCD may also influence host resilience in this way. In addition, myeloid cell death is one mechanism used to control pathogen replication and dissemination. Many of these functions for PCD have been well defined in vitro, but the role in vivo is less well understood. We created a mouse that constitutively expresses the pro-survival B-cell lymphoma (bcl-2 protein in myeloid cells (CD68(bcl2tg, thus decreasing PCD specifically in myeloid cells. Using this mouse model we explored the impact that decreased cell death of these cells has on infection with two different bacterial pathogens, Legionella pneumophila and Streptococcus pyogenes. Both of these pathogens target multiple cell death pathways in myeloid cells, and the expression of bcl2 resulted in decreased PCD after infection. We examined both pathogen clearance and host resilience and found that myeloid cell death was crucial for host resilience. Surprisingly, the decreased myeloid PCD had minimal impact on pathogen clearance. These data indicate that the most important role of PCD during infection with these bacteria is to minimize inflammation and increase host resilience, not to aid in the clearance or prevent the spread of the pathogen.

  2. Glucose Metabolism in Legionella pneumophila: Dependence on the Entner-Doudoroff Pathway and Connection with Intracellular Bacterial Growth† ▿

    Science.gov (United States)

    Harada, Eiji; Iida, Ken-Ichiro; Shiota, Susumu; Nakayama, Hiroaki; Yoshida, Shin-Ichi

    2010-01-01

    Glucose metabolism in Legionella pneumophila was studied by focusing on the Entner-Doudoroff (ED) pathway with a combined genetic and biochemical approach. The bacterium utilized exogenous glucose for synthesis of acid-insoluble cell components but manifested no discernible increase in the growth rate. Assays with permeabilized cell preparations revealed the activities of three enzymes involved in the pathway, i.e., glucokinase, phosphogluconate dehydratase, and 2-dehydro-3-deoxy-phosphogluconate aldolase, presumed to be encoded by the glk, edd, and eda genes, respectively. Gene-disrupted mutants for the three genes and the ywtG gene encoding a putative sugar transporter were devoid of the ability to metabolize exogenous glucose, indicating that the pathway is almost exclusively responsible for glucose metabolism and that the ywtG gene product is the glucose transporter. It was also established that these four genes formed part of an operon in which the gene order was edd-glk-eda-ywtG, as predicted by genomic information. Intriguingly, while the mutants exhibited no appreciable change in growth characteristics in vitro, they were defective in multiplication within eukaryotic cells, strongly indicating that the ED pathway must be functional for the intracellular growth of the bacterium to occur. Curiously, while the deficient glucose metabolism of the ywtG mutant was successfully complemented by the ywtG+ gene supplied in trans via plasmid, its defect in intracellular growth was not. However, the latter defect was also manifested in wild-type cells when a plasmid carrying the mutant ywtG gene was introduced. This phenomenon, resembling so-called dominant negativity, awaits further investigation. PMID:20363943

  3. Energy Conservation and the Promotion of Legionella pneumophila Growth: The Probable Role of Heat Exchangers in a Nosocomial Outbreak.

    Science.gov (United States)

    Bédard, Emilie; Lévesque, Simon; Martin, Philippe; Pinsonneault, Linda; Paranjape, Kiran; Lalancette, Cindy; Dolcé, Charles-Éric; Villion, Manuela; Valiquette, Louis; Faucher, Sébastien P; Prévost, Michèle

    2016-12-01

    OBJECTIVE To determine the source of a Legionella pneumophila serogroup 5 nosocomial outbreak and the role of the heat exchanger installed on the hot water system within the previous year. SETTING A 400-bed tertiary care university hospital in Sherbrooke, Canada. METHODS Hot water samples were collected and cultured for L. pneumophila from 25 taps (baths and sinks) within wing A and 9 taps in wing B. Biofilm (5) and 2 L water samples (3) were collected within the heat exchangers for L. pneumophila culture and detection of protists. Sequence-based typing was performed on strain DNA extracts and pulsed-field gel electrophoresis patterns were analyzed. RESULTS Following 2 cases of hospital-acquired legionellosis, the hot water system investigation revealed a large proportion of L. pneumophila serogroup 5 positive taps (22/25 in wing A and 5/9 in wing B). High positivity was also detected in the heat exchanger of wing A in water samples (3/3) and swabs from the heat exchanger (4/5). The outbreak genotyping investigation identified the hot water system as the source of infections. Genotyping results revealed that all isolated environmental strains harbored the same related pulsed-field gel electrophoresis pattern and sequence-based type. CONCLUSIONS Two cases of hospital-acquired legionellosis occurred in the year following the installation of a heat exchanger to preheat hospital hot water. No cases were reported previously, although the same L. pneumophila strain was isolated from the hot water system in 1995. The heat exchanger promoted L. pneumophila growth and may have contributed to confirmed clinical cases. Infect. Control Hosp. Epidemiol. 2016;1475-1480.

  4. Efficacy of copper and silver ions and reduced levels of free chlorine in inactivation of Legionella pneumophila

    Energy Technology Data Exchange (ETDEWEB)

    Landeen, L.K.; Yahya, M.T.; Gerba, C.P. (Univ. of Arizona, Tucson (USA))

    1989-12-01

    Water disinfection systems utilizing electrolytically generated copper and silver ions (200 and 20, 400 and 40, or 800 and 80 {mu}g/liter) and low levels of free chlorine (0.1 to 0.4 mg/liter) were evaluated at room (21 to 23{degree}C) and elevated (39 to 40{degree}C) temperatures in filtered well water (pH 7.3) for their efficacy in inactivating Legionella pneumophila (ATCC 33155). At room temperature, a contact time of at least 24 h was necessary for copper and silver (400 and 40 {mu}g/liter) to achieve a 3-log{sub 10} reduction in bacterial numbers. As the copper and silver concentration increased to 800 and 80 {mu}g/liter, the inactivation rate significantly increased. In water systems with and without copper and silver (400 and 40 {mu}g/liter), to inactivation rates significantly increased as the free chlorine concentration increased from 0.1 mg/liter to 0.4 mg/liter. Compared to room temperature, no significant differences were observed when 0.2 mg of free chlorine per liter with and without 400 and 40 {mu}g of copper and silver per liter was tested at 39 to 40{degree}C. All disinfection systems, regardless of temperature or free chlorine concentration, showed increased inactivation rates when 400 and 40 {mu}g of copper and silver per liter was added; however, this trend was significant only at 0.4 mg of free chlorine per liter.

  5. Enzymatic properties of an ecto-nucleoside triphosphate diphosphohydrolase from Legionella pneumophila: substrate specificity and requirement for virulence.

    Science.gov (United States)

    Sansom, Fiona M; Riedmaier, Patrice; Newton, Hayley J; Dunstone, Michelle A; Müller, Christa E; Stephan, Holger; Byres, Emma; Beddoe, Travis; Rossjohn, Jamie; Cowan, Peter J; d'Apice, Anthony J F; Robson, Simon C; Hartland, Elizabeth L

    2008-05-09

    Legionella pneumophila is the predominant cause of Legionnaires disease, a severe and potentially fatal form of pneumonia. Recently, we identified an ecto-nucleoside triphosphate diphosphohydrolase (NTPDase) from L. pneumophila, termed Lpg1905, which enhances intracellular replication of L. pneumophila in eukaryotic cells. Lpg1905 is the first prokaryotic member of the CD39/NTPDase1 family of enzymes, which are characterized by the presence of five apyrase conserved regions and the ability to hydrolyze nucleoside tri- and diphosphates. Here we examined the substrate specificity of Lpg1905 and showed that apart from ATP and ADP, the enzyme catalyzed the hydrolysis of GTP and GDP but had limited activity against CTP, CDP, UTP, and UDP. Based on amino acid residues conserved in the apyrase conserved regions of eukaryotic NTPDases, we generated five site-directed mutants, Lpg1905E159A, R122A, N168A, Q193A, and W384A. Although the mutations E159A, R122A, Q193A, and W384A abrogated activity completely, N168A resulted in decreased activity caused by reduced affinity for nucleotides. When introduced into the lpg1905 mutant strain of L. pneumophila, only N168A partially restored the ability of L. pneumophila to replicate in THP-1 macrophages. Following intratracheal inoculation of A/J mice, none of the Lpg1905 mutants was able to restore virulence to an lpg1905 mutant during lung infection, thereby demonstrating the importance of NTPDase activity to L. pneumophila infection. Overall, the kinetic studies undertaken here demonstrated important differences to mammalian NTPDases and different sensitivities to NTPDase inhibitors that may reflect underlying structural variations.

  6. Tratamientos y diseños alternativos de las instalaciones de riesgo de proliferación de Legionella neumophila

    Directory of Open Access Journals (Sweden)

    José Macias Macias

    2006-12-01

    Full Text Available La dilatada experiencia del autor en el campo de la ingeniería de mantenimiento hospitalario, ha servido de base para sugerir distintas alternativas para prevenir la aparición de la bacteria Legionella neumophila en las instalaciones de riesgo. Lo que se pretende con las recomendaciones propuestas de diseño, uso y tratamiento de dichas instalaciones, es crear en ellas entornos hostiles para la vida microbiana y especialmente para la legionela, combinando métodos físicos y químicos. De esta forma lograremos disminuir las poblaciones de la bacteria, por debajo de los niveles que a la luz de los conocimientos actuales, resultan infectivos.Se describen opciones diferentes a los métodos de tratamiento tradicionales, que permiten obtener los mismos resultados minimizando los riesgos, tanto para las personas como para el medio ambiente. También se exponen los resultados de varios experimentos, mediante los cuales se ha intentado conocer como se comportan las instalaciones y los materiales que las componen, frente a la agresión que producen los métodos biocidas. Y por último, se comparan los distintos tipos de instalaciones centralizadas de producción y almacenamiento de agua caliente y se estudian las ventajas e inconvenientes de cada una de ellas.Las propuestas que se formulan pretenden tres cosas y por este orden: controlar las poblaciones de legionela evitando que originen la enfermedad en las personas, proteger las instalaciones en las que puede desarrollarse la bacteria, y en la medida de lo posible, evitar daños al medio ambiente derivados del vertido de sustancias peligrosas.

  7. The machinery at endoplasmic reticulum-plasma membrane contact sites contributes to spatial regulation of multiple Legionella effector proteins.

    Directory of Open Access Journals (Sweden)

    Andree Hubber

    2014-07-01

    Full Text Available The Dot/Icm system of the intracellular pathogen Legionella pneumophila has the capacity to deliver over 270 effector proteins into host cells during infection. Important questions remain as to spatial and temporal mechanisms used to regulate such a large array of virulence determinants after they have been delivered into host cells. Here we investigated several L. pneumophila effector proteins that contain a conserved phosphatidylinositol-4-phosphate (PI4P-binding domain first described in the effector DrrA (SidM. This PI4P binding domain was essential for the localization of effectors to the early L. pneumophila-containing vacuole (LCV, and DrrA-mediated recruitment of Rab1 to the LCV required PI4P-binding activity. It was found that the host cell machinery that regulates sites of contact between the plasma membrane (PM and the endoplasmic reticulum (ER modulates PI4P dynamics on the LCV to control localization of these effectors. Specifically, phosphatidylinositol-4-kinase IIIα (PI4KIIIα was important for generating a PI4P signature that enabled L. pneumophila effectors to localize to the PM-derived vacuole, and the ER-associated phosphatase Sac1 was involved in metabolizing the PI4P on the vacuole to promote the dissociation of effectors. A defect in L. pneumophila replication in macrophages deficient in PI4KIIIα was observed, highlighting that a PM-derived PI4P signature is critical for biogenesis of a vacuole that supports intracellular multiplication of L. pneumophila. These data indicate that PI4P metabolism by enzymes controlling PM-ER contact sites regulate the association of L. pneumophila effectors to coordinate early stages of vacuole biogenesis.

  8. 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

  9. Phenylalanine hydroxylase from Legionella pneumophila is a thermostable enzyme with a major functional role in pyomelanin synthesis.

    Directory of Open Access Journals (Sweden)

    Marte I Flydal

    Full Text Available Legionella pneumophila is a pathogenic bacterium that can cause Legionnaires' disease and other non-pneumonic infections in humans. This bacterium produces a pyomelanin pigment, a potential virulence factor with ferric reductase activity. In this work, we have investigated the role of phenylalanine hydroxylase from L. pneumophila (lpPAH, the product of the phhA gene, in the synthesis of the pyomelanin pigment and the growth of the bacterium in defined compositions.Comparative studies of wild-type and phhA mutant corroborate that lpPAH provides the excess tyrosine for pigment synthesis. phhA and letA (gacA appear transcriptionally linked when bacteria were grown in buffered yeast extract medium at 37°C. phhA is expressed in L. pneumophila growing in macrophages. We also cloned and characterized lpPAH, which showed many characteristics of other PAHs studied so far, including Fe(II requirement for activity. However, it also showed many particular properties such as dimerization, a high conformational thermal stability, with a midpoint denaturation temperature (T(m = 79 ± 0.5°C, a high specific activity at 37°C (10.2 ± 0.3 µmol L-Tyr/mg/min and low affinity for the substrate (K(m (L-Phe = 735 ± 50 µM.lpPAH has a major functional role in the synthesis of pyomelanin and promotes growth in low-tyrosine media. The high thermal stability of lpPAH might reflect the adaptation of the enzyme to withstand relatively high survival temperatures.

  10. The Influence of Programmed Cell Death in Myeloid Cells on Host Resilience to Infection with Legionella pneumophila or Streptococcus pyogenes

    Science.gov (United States)

    Gamradt, Pia; Xu, Yun; Gratz, Nina; Duncan, Kellyanne; Kobzik, Lester; Högler, Sandra; Decker, Thomas

    2016-01-01

    Pathogen clearance and host resilience/tolerance to infection are both important factors in surviving an infection. Cells of the myeloid lineage play important roles in both of these processes. Neutrophils, monocytes, macrophages, and dendritic cells all have important roles in initiation of the immune response and clearance of bacterial pathogens. If these cells are not properly regulated they can result in excessive inflammation and immunopathology leading to decreased host resilience. Programmed cell death (PCD) is one possible mechanism that myeloid cells may use to prevent excessive inflammation. Myeloid cell subsets play roles in tissue repair, immune response resolution, and maintenance of homeostasis, so excessive PCD may also influence host resilience in this way. In addition, myeloid cell death is one mechanism used to control pathogen replication and dissemination. Many of these functions for PCD have been well defined in vitro, but the role in vivo is less well understood. We created a mouse that constitutively expresses the pro-survival B-cell lymphoma (bcl)-2 protein in myeloid cells (CD68(bcl2tg), thus decreasing PCD specifically in myeloid cells. Using this mouse model we explored the impact that decreased cell death of these cells has on infection with two different bacterial pathogens, Legionella pneumophila and Streptococcus pyogenes. Both of these pathogens target multiple cell death pathways in myeloid cells, and the expression of bcl2 resulted in decreased PCD after infection. We examined both pathogen clearance and host resilience and found that myeloid cell death was crucial for host resilience. Surprisingly, the decreased myeloid PCD had minimal impact on pathogen clearance. These data indicate that the most important role of PCD during infection with these bacteria is to minimize inflammation and increase host resilience, not to aid in the clearance or prevent the spread of the pathogen. PMID:27973535

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

    Science.gov (United States)

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

    1988-11-01

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

  12. Legionella bacteria and legionnaires illness - Directives from Germany and their use in technical installations; Legionellen und Legionellose - Richtlinien aus Deutschland und deren technische Umsetzung

    Energy Technology Data Exchange (ETDEWEB)

    Schaefer, B.

    2003-07-01

    This comprehensive article presents directives, regulations and technical guidelines in force in Germany in what regards protection against legionnaires illness. These documents formulate the requirements applicable to domestic hot water installations, bathing pools and ventilation/air conditioning systems, and clarify liability issues. They have been worked out after casualties have been recorded and a European directive on water quality for human usage issued. Whereas the water utility is responsible for water quality up to the main water inlet into the building, the building owner is responsible for in-house water installations. Regulations do not apply to small systems and single-family houses. Planners and installers have to follow recommendations and prescriptions, and regional authorities have to check the regulations' enforcement. Periodic water quality checks are mandatory. Education and training of operation and maintenance staff is prescribed. The documents include the procedures to be followed in the case of the detection of a high legionella concentration in a part of the checked installation: immediate measures, modification of the installation, re-checks of water quality. It is emphasized that the bacteria are always present in water installations and that only their proliferation shall be prevented, especially by avoiding standing hot water in distribution pipework, through appropriate design, installation and water flow balancing, as well as by avoiding critical water temperatures favourable to legionella proliferation.

  13. Cytochrome c4 is required for siderophore expression by Legionella pneumophila, whereas cytochromes c1 and c5 promote intracellular infection.

    Science.gov (United States)

    Yip, Emily S; Burnside, Denise M; Cianciotto, Nicholas P

    2011-03-01

    A panel of cytochrome c maturation (ccm) mutants of Legionella pneumophila displayed a loss of siderophore (legiobactin) expression, as measured by both the chrome azurol S assay and a Legionella-specific bioassay. These data, coupled with the finding that ccm transcripts are expressed by wild-type bacteria grown in deferrated medium, indicate that the Ccm system promotes siderophore expression by L. pneumophila. To determine the basis of this newfound role for Ccm, we constructed and tested a set of mutants specifically lacking individual c-type cytochromes. Whereas ubiquinol-cytochrome c reductase (petC) mutants specifically lacking cytochrome c(1) and cycB mutants lacking cytochrome c(5) had normal siderophore expression, cyc4 mutants defective for cytochrome c(4) completely lacked legiobactin. These data, along with the expression pattern of cyc4 mRNA, indicate that cytochrome c(4) in particular promotes siderophore expression. In intracellular infection assays, petC mutants and cycB mutants, but not cyc4 mutants, had a reduced ability to infect both amoebae and macrophage hosts. Like ccm mutants, the cycB mutants were completely unable to grow in amoebae, highlighting a major role for cytochrome c(5) in intracellular infection. To our knowledge, these data represent both the first direct documentation of the importance of a c-type cytochrome in expression of a biologically active siderophore and the first insight into the relative importance of c-type cytochromes in intracellular infection events.

  14. Effects of oakmoss and its components on Acanthamoeba castellanii ATCC 30234 and the uptake of Legionella pneumophila JCM 7571 (ATCC 33152) into A. castellanii.

    Science.gov (United States)

    Nomura, Harue; Isshiki, Yasunori; Sakuda, Keisuke; Sakuma, Katsuya; Kondo, Seiichi

    2015-01-01

    Acanthamoeba castellanii, a ubiquitous organism in water environments, is pathogenic toward humans and also is a host for bacteria of the genus Legionella, a causative agent of legionellosis. Oakmoss, a natural fragrance ingredient, and its components are antibacterial agents specifically against the genus Legionella. In the present study, oakmoss and its components were investigated for their amoebicidal activity against A. castellanii ATCC 30234 and the inhibitory effect on the uptake of L. pneumophila JCM 7571 (ATCC 33152) into A. castellanii. The oakmoss and its components 3-hydroxy-5-methylphenyl 2,4-dihydroxy-6-methylbenzoate(5), and 6,8-dihydroxy-3-pentyl-1H-isochromen-1-one (12) exhibited high amoebicidal activity (IC50 values; 10.5 ± 2.3, 16.3 ± 4.0 and 17.5 ± 2.8 μg/mL, respectively) after 48 h of treatment, which were equivalent to that of the reference compound, chlorhexidine gluconate. Pretreatment of L. pneumophila with sub-minimal inhibitory concentration of oakmoss, compound 5, 3-hydroxy-5-methylphenyl 2-hydroxy-4-methoxy-6-methylbenzoate (10) and 8-(2,4-dihydroxy-6-pentylphenoxy)-6-hydroxy-3-pentyl-1H-isochromen-1-one (14) obviously reduced the uptake of L. pneumophila into A.castellanii (p castellanii.

  15. Lactobacillus apinorum sp. nov., Lactobacillus mellifer sp. nov., Lactobacillus mellis sp. nov., Lactobacillus melliventris sp. nov., Lactobacillus kimbladii sp. nov., Lactobacillus helsingborgensis sp. nov. and Lactobacillus kullabergensis sp. nov., isolated from the honey stomach of the honeybee Apis mellifera

    Science.gov (United States)

    Alsterfjord, Magnus; Nilson, Bo; Butler, Èile; Vásquez, Alejandra

    2014-01-01

    We previously discovered a symbiotic lactic acid bacterial (LAB) microbiota in the honey stomach of the honeybee Apis mellifera. The microbiota was composed of several phylotypes of Bifidobacterium and Lactobacillus. 16S rRNA gene sequence analyses and phenotypic and genetic characteristics revealed that the phylotypes isolated represent seven novel species. One grouped with Lactobacillus kunkeei and the others belong to the Lactobacillus buchneri and Lactobacillus delbrueckiisubgroups of Lactobacillus. We propose the names Lactobacillus apinorum sp. nov., Lactobacillus mellifer sp. nov., Lactobacillus mellis sp. nov., Lactobacillus melliventris sp. nov., Lactobacillus kimbladii sp. nov., Lactobacillus helsingborgensis sp. nov. and Lactobacillus kullabergensis sp. nov. for these novel species, with the respective type strains being Fhon13NT ( = DSM 26257T = CCUG 63287T), Bin4NT ( = DSM 26254T = CCUG 63291T), Hon2NT ( = DSM 26255T = CCUG 63289T), Hma8NT ( = DSM 26256T = CCUG 63629T), Hma2NT ( = DSM 26263T = CCUG 63633T), Bma5NT ( = DSM 26265T = CCUG 63301T) and Biut2NT ( = DSM 26262T = CCUG 63631T). PMID:24944337

  16. Ecdysteroids from a Zoanthus sp.

    Science.gov (United States)

    Suksamrarn, Apichart; Jankam, Aroon; Tarnchompoo, Bongkoch; Putchakarn, Sumaitt

    2002-08-01

    A new ecdysteroid, zoanthusterone, has been isolated from a marine zoanthid, Zoanthus sp. Ten known ecdysteroids, ponasterone A, 20-hydroxyecdysone 2-acetate, viticosterone E, integristerone A 25-acetate, 2-deoxy-20-hydroxyecdysone, ecdysone, ajugasterone C, dacryhainansterone, inokosterone, and 20-hydroxyecdysone, have also been isolated. This is the first report of ecdysteroids in a Zoanthus species.

  17. Sporulation of Cyclospora sp. oocysts.

    OpenAIRE

    Smith, H V; Paton, C A; Mitambo, M M; Girdwood, R W

    1997-01-01

    Cyclospora sp. oocysts sporulated maximally at 22 and 30 degrees C for 14 days retarded sporulation. Up to 12% of human- and baboon-derived oocysts previously stored at 4 degrees C for 1 to 2 months sporulated when stored for 6 to 7 days at 30 degrees C.

  18. De aanwezigheid van Legionella species in door drinkwater gevoede watersystemen in Nederland. Deel I. 48 pp. (ook los verkrijgbaar). Idem, Deel II (ADDENDUM) Chemische en bacteriologische analyse resultaten per onderzocht object. Technische informatie van de onderzochte systemen per object. (Alleen samen met deel I verkrijgbaar)

    OpenAIRE

    van Kranen HJ

    1988-01-01

    Legionella's werden geisoleerd uit warm watervoorzieningen, water van koeltorens, water van luchtbevochtigers en incidenteel uit drinkwater zelf. In 32% van de onderzochte objecten werden, uit water met een temperawtuur varierend van 6 graden C tot en met 59 graden C, legionella's geisoleerd overwegend (75%) in concentraties kleiner dan 10 c.f.u./ml. In deze objecten werden nergens gevallen van legionellose onderkend. De in dit onderzoek voor het eerst geconstateerde besmettingen va...

  19. An Sp1/Sp3 binding polymorphism confers methylation protection.

    Directory of Open Access Journals (Sweden)

    Yanis A Boumber

    2008-08-01

    Full Text Available Hundreds of genes show aberrant DNA hypermethylation in cancer, yet little is known about the causes of this hypermethylation. We identified RIL as a frequent methylation target in cancer. In search for factors that influence RIL hypermethylation, we found a 12-bp polymorphic sequence around its transcription start site that creates a long allele. Pyrosequencing of homozygous tumors revealed a 2.1-fold higher methylation for the short alleles (P<0.001. Bisulfite sequencing of cancers heterozygous for RIL showed that the short alleles are 3.1-fold more methylated than the long (P<0.001. The comparison of expression levels between unmethylated long and short EBV-transformed cell lines showed no difference in expression in vivo. Electrophorectic mobility shift assay showed that the inserted region of the long allele binds Sp1 and Sp3 transcription factors, a binding that is absent in the short allele. Transient transfection of RIL allele-specific transgenes showed no effects of the additional Sp1 site on transcription early on. However, stable transfection of methylation-seeded constructs showed gradually decreasing transcription levels from the short allele with eventual spreading of de novo methylation. In contrast, the long allele showed stable levels of expression over time as measured by luciferase and approximately 2-3-fold lower levels of methylation by bisulfite sequencing (P<0.001, suggesting that the polymorphic Sp1 site protects against time-dependent silencing. Our finding demonstrates that, in some genes, hypermethylation in cancer is dictated by protein-DNA interactions at the promoters and provides a novel mechanism by which genetic polymorphisms can influence an epigenetic state.

  20. Antioxidant capacity, polyphenol content and iron bioavailability from algae (Ulva sp., Sargassum sp. and Porphyra sp.) in human subjects.

    Science.gov (United States)

    García-Casal, Maria N; Ramírez, José; Leets, Irene; Pereira, Ana C; Quiroga, Maria F

    2009-01-01

    Marine algae are easily produced and are good sources of Fe. If this Fe is bioavailable, algae consumption could help to combat Fe deficiency and anaemia worldwide. The objective of the present study was to evaluate Fe bioavailability, polyphenol content and antioxidant capacity from three species of marine algae distributed worldwide. A total of eighty-three subjects received maize- or wheat-based meals containing marine algae (Ulva sp., Sargassum sp. and Porphyra sp.) in different proportions (2.5, 5.0 and 7.5 g) added to the water to prepare the dough. All meals administered contained radioactive Fe. Absorption was evaluated calculating radioactive Fe incorporation in subjects' blood. The three species of marine algae were analysed for polyphenol content and reducing power. Algae significantly increased Fe absorption in maize- or wheat-based meals, especially Sargassum sp., due to its high Fe content. Increases in absorption were dose-dependent and higher in wheat- than in maize-based meals. Total polyphenol content was 10.84, 18.43 and 80.39 gallic acid equivalents/g for Ulva sp., Porphyra sp. and Sargassum sp., respectively. The antioxidant capacity was also significantly higher in Sargassum sp. compared with the other two species analysed. Ulva sp., Sargassum sp. and Porphyra sp. are good sources of bioavailable Fe. Sargassum sp. resulted in the highest Fe intake due to its high Fe content, and a bread containing 7.5 g Sargassum sp. covers daily Fe needs. The high polyphenol content found in Sargassum sp. could be partly responsible for the antioxidant power reported here, and apparently did not affect Fe absorption.

  1. Identification of Variable-Number Tandem-Repeat (VNTR) Sequences in Legionella pneumophila and Development of an Optimized Multiple-Locus VNTR Analysis Typing Scheme▿

    Science.gov (United States)

    Pourcel, Christine; Visca, Paolo; Afshar, Baharak; D'Arezzo, Silvia; Vergnaud, Gilles; Fry, Norman K.

    2007-01-01

    The utility of a genotypic typing assay for Legionella pneumophila was investigated. A multiple-locus variable number of tandem repeats (VNTR) analysis (MLVA) scheme using PCR and agarose gel electrophoresis is proposed based on eight minisatellite markers. Panels of well-characterized strains were examined in a multicenter analysis to validate the assay and to compare its performance to that of other genotyping assays. Excellent typeability, reproducibility, stability, and epidemiological concordance were observed. The MLVA type or profile is composed of a string of allele numbers, corresponding to the number of repeats at each VNTR locus, separated by commas, in a predetermined order. A database containing information from 99 L. pneumophila serogroup 1 strains and four strains of other serogroups and their MLVA profiles, which can be queried online, is available from http://bacterial-genotyping.igmors.u-psud.fr/. PMID:17251393

  2. Pantoea rodasii sp. nov., Pantoea rwandensis sp. nov. and Pantoea wallisii sp. nov., isolated from Eucalyptus.

    Science.gov (United States)

    Brady, Carrie L; Cleenwerck, Ilse; van der Westhuizen, Lorinda; Venter, Stephanus N; Coutinho, Teresa A; De Vos, Paul

    2012-07-01

    Several Gram-negative-staining, facultatively anaerobic bacterial isolates were obtained from Eucalyptus seedlings showing symptoms of bacterial blight and dieback in Colombia, Rwanda and South Africa. Partial 16S rRNA gene sequencing, together with partial gyrB sequencing, placed the isolates in the genus Pantoea and indicated that they constituted three novel species. Multilocus sequence analysis (MLSA) based on partial sequences of gyrB, rpoB, infB and atpD revealed Pantoea dispersa, Pantoea eucrina and Pantoea cypripedii as their closest phylogenetic relatives. DNA-DNA hybridization studies confirmed the classification of the new isolates as three novel species and phenotypic tests allowed them to be differentiated from their closest phylogenetic neighbours. The names Pantoea rodasii sp. nov. [type strain LMG 26273(T)=BD 943(T) (deposited with the Plant Pathogenic and Plant Protecting Bacteria Collection, South Africa)=BCC 581(T) (deposited with the Bacterial Culture Collection, Forestry and Agricultural Institute, South Africa)], Pantoea rwandensis sp. nov. (type strain LMG 26275(T)=BD 944(T)=BCC 571(T)) and Pantoea wallisii sp. nov. (type strain LMG 26277(T)=BD 946(T)=BCC 682(T)) are proposed.

  3. Comparative Proteomics of Purified Pathogen Vacuoles Correlates Intracellular Replication of Legionella pneumophila with the Small GTPase Ras-related protein 1 (Rap1).

    Science.gov (United States)

    Schmölders, Johanna; Manske, Christian; Otto, Andreas; Hoffmann, Christine; Steiner, Bernhard; Welin, Amanda; Becher, Dörte; Hilbi, Hubert

    2017-04-01

    Legionella pneumophila is an opportunistic bacterial pathogen that causes a severe lung infection termed "Legionnaires' disease." The pathogen replicates in environmental protozoa as well as in macrophages within a unique membrane-bound compartment, the Legionella -containing-vacuole (LCV). LCV formation requires the bacterial Icm/Dot type IV secretion system, which translocates ca. 300 "effector proteins" into host cells, where they target distinct host factors. The L. pneumophila "pentuple" mutant (Δpentuple) lacks 5 gene clusters (31% of the effector proteins) and replicates in macrophages but not in Dictyostelium discoideum amoeba. To elucidate the host factors defining a replication-permissive compartment, we compare here the proteomes of intact LCVs isolated from D. discoideum or macrophages infected with Δpentuple or the parental strain Lp02. This analysis revealed that the majority of host proteins are shared in D. discoideum or macrophage LCVs containing the mutant or the parental strain, respectively, whereas some proteins preferentially localize to distinct LCVs. The small GTPase Rap1 was identified on D. discoideum LCVs containing strain Lp02 but not the Δpentuple mutant and on macrophage LCVs containing either strain. The localization pattern of active Rap1 on D. discoideum or macrophage LCVs was confirmed by fluorescence microscopy and imaging flow cytometry, and the depletion of Rap1 by RNA interference significantly reduced the intracellular growth of L. pneumophila Thus, comparative proteomics identified Rap1 as a novel LCV host component implicated in intracellular replication of L. pneumophila . © 2017 by The American Society for Biochemistry and Molecular Biology, Inc.

  4. Development of a new CARD-FISH protocol for quantification of Legionella pneumophila and its application in two hospital cooling towers.

    Science.gov (United States)

    Kirschner, A K T; Rameder, A; Schrammel, B; Indra, A; Farnleitner, A H; Sommer, R

    2012-06-01

    Open cooling towers are frequent sources of infections with Legionella pneumophila. The gold standard for the detection of Leg. pneumophila is based on cultivation lasting up to 10 days and detecting only culturable cells. Alternative fluorescence in situ hybridization (FISH) protocols have been proposed, but they result in faint fluorescence signals and lack specificity because of cross-hybridization with other Legionella species. Our aim was thus to develop a new FISH protocol for rapid and specific detection of Leg. pneumophila in water samples. A novel catalysed reporter deposition FISH (CARD-FISH) protocol for the detection of Leg. pneumophila was developed, which significantly enhanced signal intensity as well as specificity of the probe through the use of a novel competitor probe. The developed protocol was compared with the culture method for monitoring the seasonal development of culturable and nonculturable Leg. pneumophila in two hospital cooling tower systems. Seasonal fluctuations of Leg. pneumophila concentrations detected via CARD-FISH were related to the development of the total bacterial community in both cooling towers, with temperature and biocide as the main factors controlling this development. Our results clearly showed that the majority of the Leg. pneumophila cells were in a nonculturable state. Thus, detection of Leg. pneumophila with culture methods may underestimate the total numbers of Leg. pneumophila present. Rapid, sensitive and specific detection and quantification of Leg. pneumophila in water systems is prerequisite for reliable risk estimation. The new protocol significantly improves current methodology and can be used to monitor and screen for Leg. pneumophila concentrations in cooling towers or other water systems. © 2012 The Authors. Journal of Applied Microbiology © 2012 The Society for Applied Microbiology.

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

    Science.gov (United States)

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

    2013-05-01

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

  6. Inter-kingdom Signaling by the Legionella Quorum Sensing Molecule LAI-1 Modulates Cell Migration through an IQGAP1-Cdc42-ARHGEF9-Dependent Pathway.

    Directory of Open Access Journals (Sweden)

    Sylvia Simon

    2015-12-01

    Full Text Available Small molecule signaling promotes the communication between bacteria as well as between bacteria and eukaryotes. The opportunistic pathogenic bacterium Legionella pneumophila employs LAI-1 (3-hydroxypentadecane-4-one for bacterial cell-cell communication. LAI-1 is produced and detected by the Lqs (Legionella quorum sensing system, which regulates a variety of processes including natural competence for DNA uptake and pathogen-host cell interactions. In this study, we analyze the role of LAI-1 in inter-kingdom signaling. L. pneumophila lacking the autoinducer synthase LqsA no longer impeded the migration of infected cells, and the defect was complemented by plasmid-borne lqsA. Synthetic LAI-1 dose-dependently inhibited cell migration, without affecting bacterial uptake or cytotoxicity. The forward migration index but not the velocity of LAI-1-treated cells was reduced, and the cell cytoskeleton appeared destabilized. LAI-1-dependent inhibition of cell migration involved the scaffold protein IQGAP1, the small GTPase Cdc42 as well as the Cdc42-specific guanine nucleotide exchange factor ARHGEF9, but not other modulators of Cdc42, or RhoA, Rac1 or Ran GTPase. Upon treatment with LAI-1, Cdc42 was inactivated and IQGAP1 redistributed to the cell cortex regardless of whether Cdc42 was present or not. Furthermore, LAI-1 reversed the inhibition of cell migration by L. pneumophila, suggesting that the compound and the bacteria antagonistically target host signaling pathway(s. Collectively, the results indicate that the L. pneumophila quorum sensing compound LAI-1 modulates migration of eukaryotic cells through a signaling pathway involving IQGAP1, Cdc42 and ARHGEF9.

  7. DISTRIBUSI Solen sp DI PERAIRAN KABUPATEN BANGKALAN

    Directory of Open Access Journals (Sweden)

    Eva Ari Wahyuni

    2016-03-01

    Full Text Available DISTRIBUTION OF Solen sp IN BANGKALAN WATERSSolen sp potential needs to be developed on the island of Madura, particularly in Bangkalan. Solen sp utilization has increased which has the potential to overfishing. Therefore, this study aims to determine the density of Solen sp and their ecology in the waters Modung village, Modung District, Bangkalan. The experiment was conducted in April 2015 using the descriptive method. The materials used include Solen sp and physico-chemical parameters of the environment (temperature, salinity, pH, and substrate. The analyzes were conducted at the Laboratory of Marine Science, Department of Marine Sciences, Trunojoyo University of Madura by using the tool grabsampler, sieveshaker, and pipetting with gravimetric method. The analysis shows the range of values of temperature between 29-300C, salinity between 31-32 ppt, pH were 7.9-8.0 and the type of substrate in the form of sandy mud, as well as the density of Solen sp from 8-10 individuals/m2. All measurement results indicate normal conditions and in accordance with the sea water quality standard for marine life, which can be a suitable habitat for the growth and development of Solen sp. This condition is thought to affect the density of Solen sp.Keywords: Bangkalan, density, distribution, Solen sp, substrate.ABSTRAKPotensi Solen sp perlu dikembangkan di pulau Madura, khususnya di Kabupaten Bangkalan. Pemanfaatan Solen sp mengalami peningkatan sehingga berpotensi overfishing. Untuk itu, penelitian ini bertujuan untuk mengetahui kepadatan Solen sp dan ekologinya di perairan desa Modung, Kecamatan Modung, Kabupaten Bangkalan. Penelitian dilaksanakan pada bulan April 2015 dengan metode deskriptif. Materi dan bahan yang digunakan diantaranya Solen sp dan parameter fisika-kimia lingkungan (suhu, salinitas, pH, dan substrat. Analisa dilakukan di Laboratorium Ilmu Kelautan, Program studi/Jurusan Ilmu Kelautan Universitas Trunojoyo Madura dengan menggunakan alat

  8. Risk assessment and monitoring of Legionella by culture and q-PCR in a newly built block of flats associated with a small outbreak of legionnaires´ disease

    DEFF Research Database (Denmark)

    Krøjgaard, Louise Hjelmar; Krogfelt, Karen A.; Albrechtsen, Hans-Jørgen

    2010-01-01

    ) and permanent changes of the flow and temperature were conducted to overcome the high concentrations of Legionella in the water. Water samples (104 in total, both cold and warm water) from taps and shower hoses were collected and cultivated. The samples consisted of: A samples: the first one litre, B samples......: one litre collected after flushing until constant temperature and water from shower hoses at a temperature of 38ºC. The survey pointed at important risk factors 1) low temperature of the hot water in some of the most distant taps 2) low flow of the water circulating system because of too small pipe...... dimensions and too low pump capacity 3) high numbers of Legionella in the shower hoses 4) stagnancy of water before residents move in and in unoccupied apartments and 5) lack of or inappropriate control measures. Following cultivation, DNA from all water samples was extracted by a simple method with Chelex...

  9. Especificidad del hongo micorrizico (Rhizoctonia sp.) en Phalaenopsis sp., Cymbidium sp., Trichoceros antenifer, Oncidium excavatum, y Cyrtochilum sp.

    OpenAIRE

    Ordoñez, Silvia L.; Pillacela Zhunio, Dora Priscila; Salazar, Jazmín M.; Peña Tapia, Denisse Fabiola

    2016-01-01

    Las orquídeas producen abundantes semillas pequeñas, careciendo de endospermo, cotiledones y sustancias de reserva para llevar a cabo su germinación. Es por esto que estratégicamente las semillas establecen una relación simbiótica con un hongo micorrízico que favorece a su germinación y desarrollo. El objetivo de este estudio fue determinar la especificidad del hongo micorrízico (Rhizoctonia sp.) en la germinación de cinco géneros de orquídeas. Se usaron dos medios de cultivo: 1) PhytamaxTM y...

  10. Rodentibacter gen. nov including Rodentibacter pneumotropicus comb. nov., Rodentibacter heylii sp nov., Rodentibacter myodis sp nov., Rodentibacter ratti sp nov., Rodentibacter heidelbergensis sp nov., Rodentibacter trehalosifermentans sp nov., Rodentibacter rarus sp nov., Rodentibacter mrazii

    DEFF Research Database (Denmark)

    Adhikary, Sadhana; Nicklas, Werner; Bisgaard, Magne

    2017-01-01

    genomic comparison allowed the estimation of DNA–DNA renaturation. Rodentibacter heylii sp. nov. was proposed for a group that included the biovar Heyl of [ Pasteurella ] pneumotropica with the type strain ATCC 12555T (=CCUG 998T). A group was proposed as Rodentibacter ratti sp. nov., which included...

  11. Approches moléculaires de l'épidémiologie de la légionellose et de la résistance aux antibiotiques chez Legionella pneumophila

    OpenAIRE

    Shadoud, Lubana

    2014-01-01

    Legionella pneumophila is a Gram- negative, facultative intracellular bacterium responsible for legionellosis (or Legionnaires' disease ) in humans. The fluoroquinolones and the macrolides are used as first-line antibiotic treatment of this disease. However, treatment failures are common, and the mortality rates remain high (10-15 % of cases, more than 30% in immunocompromised patients). Although L. pneumophila strain resistant to these antibiotics have never been isolated, treatment failures...

  12. Xylanolytic enzyme systems in Arthrobacter sp MTCC 5214 and Lactobacillus sp.

    Digital Repository Service at National Institute of Oceanography (India)

    Khandeparker, R.; Jalal, T.

    The production of extracellular xylanolytic enzymes such as xylanase, alfa-L-arabinofuranosidase (alfa-l-AFase), and acetyl xylan esterase (Axe) by marine Arthrobacter sp and Lactobacillus sp was investigated using different carbon sources Induction...

  13. Evaluation of antioxidant and cytotoxic properties of Cynobacteria, Limnothrix sp. and Leptolyngbya sp. from Arabian sea

    Digital Repository Service at National Institute of Oceanography (India)

    Anas, A.; Vinothkumar, S.; Gupta, S.; Jasmin, C.; Joseph, V.; Parameswaran, P.S.; Nair, S.

    The hexane fractions of the marine cyanobacteria: Leptolyngbya sp. and Limnothrix sp., collected from Arabian Sea were found to display promising antioxidant properties than their ethyl acetate fraction during radical scavenging ABTS/DPPH assays (IC...

  14. First report of Anisakis sp. in Epinephelus sp. in East Indonesia

    OpenAIRE

    Annytha Ina Rohi Detha; Diana Agustiani Wuri; Julianty Almet; Yuni Riwu; Christin Melky

    2018-01-01

    Objective: The present research was conducted to identify the prevalence of Anisakis sp. as fish-borne zoonoses in Epinephelus sp. in territorial waters of East Nusa Tenggara, Indonesia. Materials and methods: A total of 50 fish (Epinephelus sp.) were collected from Kupang Fish Market in East Nusa Tenggara. Identification of Anisakis sp. was performed based on morphological observations considering shape of ventriculus, boring tooth, and mucron using binocular microscope. Results: Prev...

  15. Specialised predation by Palpimanus sp. (Araneae: Palpimanidae ...

    African Journals Online (AJOL)

    This is the first detailed report on the natural prey and the prey-capture tactics of a Palpimanus sp. from Entebbe (Uganda). Although this species fed occasionally on insects, its dominant prey in the field was other spiders, especially jumping spiders (Salticidae) and their eggs. Encounters between Palpimanus sp. and ...

  16. Guide til gode spørgeskemaer

    DEFF Research Database (Denmark)

    Olsen, Henning

    Spørgeskemaundersøgelser bliver ofte brugt til at dokumentere forskellige forhold og begrunde politiske beslutninger. Men resultaterne kan være forbundet med stor usikkerhed. Det kræver omhu og stor sproglig formåen at udarbejde spørgeskemaer. Seniorforsker Henning Olsen har i flere år arbejdet med...... viden om, hvordan folk forstår sproglige meddelelser og genkalder sig informationer. I guiden behandles emner som fx styrende problemstillinger og spørgsmåls fokus og neutralitet, formulering af åbne eller lukkede spørgsmål og svarkategorier, tematiske spørgeforløb, aflastning af svarpersoners...

  17. Listeria floridensis sp. nov., Listeria aquatica sp. nov., Listeria cornellensis sp. nov., Listeria riparia sp. nov. and Listeria grandensis sp. nov., from agricultural and natural environments.

    Science.gov (United States)

    den Bakker, Henk C; Warchocki, Steven; Wright, Emily M; Allred, Adam F; Ahlstrom, Christina; Manuel, Clyde S; Stasiewicz, Matthew J; Burrell, Angela; Roof, Sherry; Strawn, Laura K; Fortes, Esther; Nightingale, Kendra K; Kephart, Daniel; Wiedmann, Martin

    2014-06-01

    Sampling of agricultural and natural environments in two US states (Colorado and Florida) yielded 18 Listeria-like isolates that could not be assigned to previously described species using traditional methods. Using whole-genome sequencing and traditional phenotypic methods, we identified five novel species, each with a genome-wide average BLAST nucleotide identity (ANIb) of less than 85% to currently described species. Phylogenetic analysis based on 16S rRNA gene sequences and amino acid sequences of 31 conserved loci showed the existence of four well-supported clades within the genus Listeria; (i) a clade representing Listeria monocytogenes, L. marthii, L. innocua, L. welshimeri, L. seeligeri and L. ivanovii, which we refer to as Listeria sensu stricto, (ii) a clade consisting of Listeria fleischmannii and two newly described species, Listeria aquatica sp. nov. (type strain FSL S10-1188(T) = DSM 26686(T) = LMG 28120(T) = BEI NR-42633(T)) and Listeria floridensis sp. nov. (type strain FSL S10-1187(T) = DSM 26687(T) = LMG 28121(T) = BEI NR-42632(T)), (iii) a clade consisting of Listeria rocourtiae, L. weihenstephanensis and three novel species, Listeria cornellensis sp. nov. (type strain TTU A1-0210(T) = FSL F6-0969(T) = DSM 26689(T) = LMG 28123(T) = BEI NR-42630(T)), Listeria grandensis sp. nov. (type strain TTU A1-0212(T) = FSL F6-0971(T) = DSM 26688(T) = LMG 28122(T) = BEI NR-42631(T)) and Listeria riparia sp. nov. (type strain FSL S10-1204(T) = DSM 26685(T) = LMG 28119(T) = BEI NR- 42634(T)) and (iv) a clade containing Listeria grayi. Genomic and phenotypic data suggest that the novel species are non-pathogenic. © 2014 IUMS.

  18. Predictive value of Sp1/Sp3/FLIP signature for prostate cancer recurrence.

    Directory of Open Access Journals (Sweden)

    Roble G Bedolla

    Full Text Available Prediction of prostate cancer prognosis is challenging and predictive biomarkers of recurrence remain elusive. Although prostate specific antigen (PSA has high sensitivity (90% at a PSA level of 4.0 ng/mL, its low specificity leads to many false positive results and considerable overtreatment of patients and its performance at lower ranges is poor. Given the histopathological and molecular heterogeneity of prostate cancer, we propose that a panel of markers will be a better tool than a single marker. We tested a panel of markers composed of the anti-apoptotic protein FLIP and its transcriptional regulators Sp1 and Sp3 using prostate tissues from 64 patients with recurrent and non-recurrent cancer who underwent radical prostatectomy as primary treatment for prostate cancer and were followed with PSA measurements for at least 5 years. Immunohistochemical staining for Sp1, Sp3, and FLIP was performed on these tissues and scored based on the proportion and intensity of staining. The predictive value of the FLIP/Sp1/Sp3 signature for clinical outcome (recurrence vs. non-recurrence was explored with logistic regression, and combinations of FLIP/Sp1/Sp3 and Gleason score were analyzed with a stepwise (backward and forward logistic model. The discrimination of the markers was identified by sensitivity-specificity analysis and the diagnostic value of FLIP/Sp1/Sp3 was determined using area under the curve (AUC for receiver operator characteristic curves. The AUCs for FLIP, Sp1, Sp3, and Gleason score for predicting PSA failure and non-failure were 0.71, 0.66, 0.68, and 0.76, respectively. However, this increased to 0.93 when combined. Thus, the "biomarker signature" of FLIP/Sp1/Sp3 combined with Gleason score predicted disease recurrence and stratified patients who are likely to benefit from more aggressive treatment.

  19. Antibacterial Actions and Potential Phototoxic Effects of Volatile oils of Foeniculum sp. (fennel, Salvia sp. (sage, Vitis sp. (grape, Lavandula sp. (lavender

    Directory of Open Access Journals (Sweden)

    Elif Ayse Erdogan Eliuz

    2016-09-01

    Full Text Available In the present study, the volatile compounds of essential oil of Foeniculum vulgare (fennel, Salvia officinalis (sage, Vitis vinifera (grape, Lavandula angustifolia (lavender were analysed by gas chromatography-mass spectrometry (GC-MS using the Nist and Willey libraries. It was determined that the main components of Foeniculum sp. were anethole (41.11%, carvacrol (9.18%. whereas main components of Salvia sp were 1.8 cineole (34.09%, caryophyllene (10.95%, camphor (9.44%, α-pinene (8.42%. Vitis sp. contained linoleic acid (36.98%, 2,4-decadienal (30.79%. Finally, volatile component of Lavandula sp. was linalool (33.57%, linalyl acetate (30.74%. Photoxic antibacterial activity of volatile oil of those plants against Escherichia coli (ATCC 25293, Klebsiella pneumoniae (10031, Salmonella thyphimurium, Bacillus subtilis (ATCC 6633, Staphylococcus aureus (ATCC 25925, Enterococcus feacalis (ATCC 29212 were examined by using disc diffusion method. We demonstrated that volatile oil effectively can be activated by a standard LED light. In vitro, significant phototoxicity was demonstrated by volatile oil of Foeniculum sp. and Vitis sp. (P < 0.05, while minor phototoxicity was induced by Lavandula sp. Therefore, volatile oil of plant can be considered as a potential photosensitizer in the photochemical therapy.

  20. TurboSP and the Topological Trigger

    CERN Document Server

    Belavin, Vladislav

    2016-01-01

    TurboSP was originally proposed as an alternative to Full stream in LHCb data flow. TurboSP is a data flow strategy which not only selects events that should be preserved, like in Full stream, but also provides selective persistence. This is achieved by saving candidates and subset of the reconstruction. During this summer project we investigated the physics viability of using TurboSP with the topological lines and found out a possibility to reduce the number of kept tracks per event by two times while keeping a ratio of fully picked up interesting decay modes on $\\sim 97 \\%$ level.

  1. Amphibian (Xenopus sp.) iodothyronine deiodinase ...

    Science.gov (United States)

    The U.S. EPA-MED amphibian thyroid group is currently screening chemicals for inhibition of human iodothyronine deiodinase activity as components of the thyroid system important in human development. Amphibians are a bellwether taxonomic group to gauge toxicity of chemicals in the environment. Amphibian thyroid function is not only important in development but also metamorphosis. Xenopus sp. have been used extensively as model organisms and are well characterized genetically. We propose to screen a list of chemicals (selected from the human DIO screening results) to test for inhibition of Xenopus deiodinases. Large quantities of the enzymes will be produced using an adenovirus system. Our preliminary results show that there may be catalytic differences between human and Xenopus deiodinases. The Twin Ports Early Career Scientists is a new group formed within the Duluth-Superior scientific community. This presentation will provide a basic introduction to my research and our mission at EPA, and help to establish networking and collaboration relationships across disciplines and institutions.

  2. Detection of Salmonella sp., Vibrio sp. and total plate count bacteria on blood cockle (Anadara granosa)

    Science.gov (United States)

    Ekawati, ER; Yusmiati, S. N. H.

    2018-01-01

    Blood cockle (Anadara granosa) has high level of zinc and protein, which is beneficial for therapeutic function for malnourished particularly stunting case in children. Zinc in animal foods is more absorbable than that from vegetable food. Blood cockle (Anadara granosa) is rich in nutrient and an excellent environment for the growth of microorganisms. This research aimed to identify the contamination of Salmonella sp., Vibrio sp. and total plate count bacteria on blood cockle (Anadara granosa). This was observation research with laboratory analysis. Salmonella sp. and Vibrio sp. were detected from blood cockle. Total plate count was determine of the total amount of the bacteria. Results detected from 20 samples of blood cockle showed that all samples were negative of Salmonella sp. and 1 sample positive Vibrio sp. The result of total plate count bacteria was sample.

  3. Identification of a gene that affects the efficiency of host cell infection by Legionella pneumophila in a temperature-dependent fashion.

    Science.gov (United States)

    Ridenour, Dennis A; Cirillo, Suat L G; Feng, Sheng; Samrakandi, Mustapha M; Cirillo, Jeffrey D

    2003-11-01

    The ability to infect host cells is critical for the survival and replication of intracellular pathogens in humans. We previously found that many genes involved in the ability of Legionella pneumophila to infect macrophages are not expressed efficiently under standard laboratory growth conditions. We have developed an approach using expression of L. pneumophila genes from an exogenous constitutive promoter on a low-copy-number vector that allows identification of genes involved in host cell infection. Through the use of this strategy, we found that expression of a gene, lvhB2, enhances the efficiency of L. pneumophila infection of mammalian cells. The putative protein encoded by lvhB2 has similarity to structural pilin subunits of type IV secretion systems. We confirmed that this gene plays a role in host cell infection by the construction of an in-frame deletion in the L. pneumophila lvhB2 gene and complementation of this mutant with the wild-type gene. The lvhB2 mutant does not display a very obvious defect in interactions with host cells when the bacteria are grown at 37 degrees C, but it has an approximately 100-fold effect on entry and intracellular replication when grown at 30 degrees C. These data suggest that lvhB2 plays an important role in the efficiency of host cell infection by L. pneumophila grown at lower temperatures.

  4. Recombinant flagellin-PAL fusion protein of Legionella pneumophila induced cell-mediated and protective immunity against bacteremia in BALB/c mice.

    Science.gov (United States)

    Mohabati Mobarez, Ashraf; Ahmadrajabi, Roya; Khoramabadi, Nima; Salmanian, Ali Hatef

    2017-09-08

    We report a new recombinant fusion protein composed of full-length Legionella pneumophila flagellin A and peptidoglycan-associated lipoprotein (PAL), rFLA-PAL, capable of inducing protective immunity against L. pneumophila. The recombinant protein was over expressed in Escherichia coli strain BL21 (DE3) using pET-28a (+) expression vector (pET28a-flaA-pal) and purified by Ni 2+ exchange chromatography. Immunological properties of rFLA-PAL were assessed in a mouse model. Female BALB/c mice, immunized with rFLA-PAL, exhibited a rapid increase in serum antibody concentration against each of its protein portions. Furthermore, a strong activation of both innate and adaptive cell-mediated immunity was observed as indicated by antigen-specific splenocyte proliferation, IFN-γ and IL-12 production, and early production of TNF-α in the serum and in splenocyte cultures which were separately assessed against PAL and FLA. BALB/c mice were challenged with a lethal dose of L. pneumophila intravenously. In a 10-days follow-up after intravenous lethal challenge with L. pneumophila, a 100% survival rate was observed for mice immunized with rFLA-PAL, same as for those immunized with a sublethal dose of L. pneumophila. Based on the potent immune responses observed in mice immunized with rFLA-PAL, this recombinant fusion protein could be a potential vaccine candidate against the intracellular pathogen L. pneumophila.

  5. Legionella pneumophila in bronchoalveolar lavage samples of patients suffering from severe respiratory infections: Role of age, sex and history of smoking in the prevalence of bacterium

    Directory of Open Access Journals (Sweden)

    Faradonbeh Fatemeh Alaei

    2015-01-01

    Full Text Available Introduction. Legionella pneumophila is the most commonly detected cause of legionellosis, which is an acute respiratory tract infection with high morbidity and mortality rates. Objective. The aim of this study was to investigate the prevalence rate of L. pneumophila in bronchoalveolar lavages and study the role of sex, age and history of smoking as risk factors for susceptibility to the bacterium. Methods. One hundred bronchoalveolar lavage samples were collected from the Iranian health centers and immediately transferred to laboratory. The samples were cultured and those that were L. pneumophila positive were subjected to PCR method with respect to the 16S rRNA gene. Results. Twelve out of 100 samples were positive for L. pneumophila (12%. Patients older than 70 years had the highest incidence of L. pneumophila (17.77%. Prevalence of L. pneumophila in male and female patients was 14.81% and 8.69%, respectively. Total incidence of L. pneumophila in patients with and without history of smoking was 18% and 6%, respectively. There were significant differences in the incidence of bacterium between groups of our study. Conclusion. Sex, age and history of smoking are predominant risk factors for the occurrence of L. pneumophila. However, more studies should be undertaken to confirm these results.

  6. Multiplex PMA-qPCR Assay with Internal Amplification Control for Simultaneous Detection of Viable Legionella pneumophila, Salmonella typhimurium, and Staphylococcus aureus in Environmental Waters.

    Science.gov (United States)

    Li, Haiyan; Xin, Hongyi; Li, Sam Fong Yau

    2015-12-15

    Pathogenic microorganisms are responsible for many infectious diseases, and pathogen monitoring is important and necessary for water quality control. This study for the first time explored a multiplex quantitative real-time PCR (qPCR) technique combined with propidium monoazide (PMA) to simultaneously detect viable Legionella pneumophila, Salmonella typhimurium, and Staphylococcus aureus in one reaction from water samples. Sodium lauroyl sarcosinate (sarkosyl) was applied to enhance the dead bacterial permeability of PMA. The sensitivity of the multiplex PMA-qPCR assay achieved two colony-forming units (CFU) per reaction for L. pneumophila and three CFU per reaction for S. typhimurium and S. aureus. No PCR products were amplified from all nontarget control samples. Significantly, with comparable specificity and sensitivity, this newly invented multiplex PMA-qPCR assay took a much shorter time than did conventional culture assays when testing water samples with spiked bacteria and simulated environmental water treatment. The viable multiplex PMA-qPCR assay was further successfully applied to pathogen detection from rivers, canals, and tap water samples after simple water pretreatment.

  7. Long-term persistence of a single Legionella pneumophila strain possessing the mip gene in a municipal shower despite repeated cycles of chlorination.

    Science.gov (United States)

    Cooper, I R; White, J; Mahenthiralingam, E; Hanlon, G W

    2008-10-01

    The ability of Legionella pneumophila to colonise domestic water systems is a primary cause of outbreaks of Legionnaire's disease in humans. World Health Organization guidelines recommend that drinking water is chlorinated to between 0.2 and 1mg/L [Chlorine in drinking-water. Guidelines for drinking-water quality, 2nd edn. Geneva: World Health Organization; 1996], but L. pneumophila is repeatedly isolated from chlorinated water systems, indicating that this treatment is not effective at preventing colonisation. Current UK guidelines recommend a one-off treatment of 20-50mg/L of free chlorine to remove the bacteria. In this study we report on the persistence of L. pneumophila serogroup 1 in a domestic shower system despite repeated cycles of chlorination at 50mg/L for 1h exposure time, over the course of two and a half years. Persisting isolates were subjected to in-vitro phenotypic analyses and polymerase chain reaction analysis for the toxin-encoding mip gene. Random amplified polymorphic DNA typing was also performed to determine whether the isolates recovered on different occasions were the same strain. We found that seven isolates of L. pneumophila recovered over a two-and-a-half year period are the same genetically defined strain, indicating that the bacteria can persist despite repeated cycles of chlorination after each successive isolation.

  8. Legionella dumoffii Tex-KL Mutated in an Operon Homologous to traC-traD is Defective in Epithelial Cell Invasion.

    Science.gov (United States)

    Qin, Tian; Ken-Ichiro, Iida; Ren, Hong Yu; Zhou, Hai Jian; Yoshida, Shin-Ichi

    2016-06-01

    To understand the mechanism of invasion by Legionella dumoffii. The L. dumoffii strain Tex-KL was mutated using the Tn903 derivative, Tn903dIIlacZ. After screening 799 transposon insertion mutants, we isolated one defective mutant. We then constructed the gene-disrupted mutant, KL16, and studied its invasion of and intracellular growth in HeLa and A549 cells, and in A/J mice survival experiments. The structure of traC-traD operon was analyzed by RT-PCR. The transposon insertion was in a gene homologous to Salmonella typhi traC, which is required for the assembly of F pilin into the mature F pilus structure and for conjugal DNA transmission. Results from RT-PCR suggested that the traC-traD region formed an operon. We found that when the traC gene was disrupted, invasion and intracellular growth of L. dumoffii Tex-KL were impaired in human epithelial cells. When mice were infected by intranasal inoculation with a traC deficient mutant, their survival significantly increased when compared to mice infected with the wild-type strain.. Our results indicated that the traC-traD operon is required for the invasion and intracellular growth abilities of L. dumoffii Tex-KL in epithelial cells. Copyright © 2016 The Editorial Board of Biomedical and Environmental Sciences. Published by China CDC. All rights reserved.

  9. The htpAB operon of Legionella pneumophila cannot be deleted in the presence of the groE chaperonin operon of Escherichia coli.

    Science.gov (United States)

    Nasrallah, Gheyath K; Gagnon, Elizabeth; Orton, Dennis J; Garduño, Rafael A

    2011-11-01

    HtpB, the chaperonin of the intracellular bacterial pathogen Legionella pneumophila , displays several virulence-related functions in vitro. To confirm HtpB's role in vivo, host infections with an htpB deletion mutant would be required. However, we previously reported that the htpAB operon (encoding co-chaperonin and chaperonin) is essential. We attempted here to delete htpAB in a L. pneumophila strain carrying the groE operon (encoding the Escherichia coli co-chaperonin and chaperonin). The groE operon was inserted into the chromosome of L. pneumophila Lp02, and then allelic replacement of htpAB with a gentamicin resistance cassette was attempted. Although numerous potential postallelic replacement transformants showed a correct selection phenotype, we still detected htpAB by PCR and full-size HtpB by immunoblot. Southern blot and PCR analysis indicated that the gentamicin resistance cassette had apparently integrated in a duplicated htpAB region. However, we showed by Southern blot that strain Lp02, and the Lp02 derivative carrying the groE operon, have only one copy of htpAB. These results confirmed that the htpAB operon cannot be deleted, not even in the presence of the groE operon, and suggested that attempts to delete htpAB under strong phenotypic selection result in aberrant genetic recombinations that could involve duplication of the htpAB locus.

  10. Molecular diversity and high virulence of Legionella pneumophila strains isolated from biofilms developed within a warm spring of a thermal spa

    Directory of Open Access Journals (Sweden)

    Chaabna Zineddine

    2013-01-01

    Full Text Available Abstract Background Several cases of legionellosis have been diagnosed in the same French thermal spa in 1986, 1994 and 1997. L. pneumophila serogroup 1 (Lp1 strains have been isolated from several patients, but the source of contamination was not identified despite the presence of different Lp1 in water samples of the three natural springs feeding the spa at this period. Results Our strategy was to investigate L. pneumophila (Lp strains from natural biofilms developed in a sulphur-rich warm spring of this contaminated site. Biofilm analysis revealed the presence of three Lp serogroups (Lp1, Lp10 and Lp12. Surprisingly, Lp10 and Lp12 were not reported in the previous described studies from water samples. Besides, the new seven Lp1 we isolated exhibit a high molecular diversity and have been differentiated in five classes according to their DNA genome patterns obtained by PFGE and mip sequences. It must be noted that these DNA patterns are original and unknown in databases. Interestingly, the 27 Lp environmental strains we isolated display a higher cytotoxicity and virulence towards the amoeba Acanthamoeba castellanii than those of known Lp1 epidemic strains. Conclusion The characteristics of Legionella pneumophila Lp1 strains isolated from the warm spring are in agreement with their presence in biofilms and their probable long-term persistence in this ecosystem.

  11. Efficacy of thermal treatment and copper-silver ionization for controlling Legionella pneumophila in high-volume hot water plumbing systems in hospitals.

    Science.gov (United States)

    Mietzner, S; Schwille, R C; Farley, A; Wald, E R; Ge, J H; States, S J; Libert, T; Wadowsky, R M; Miuetzner, S

    1997-12-01

    Thermal treatment and copper-silver ionization are often used for controlling Legionella pneumophila in high-volume hospital plumbing systems, although the comparative efficacies of these measures in high-volume systems are unknown. Thermal treatment of a hot water circuit was accomplished by flushing hot water (> 60 degrees C) through distal fixtures for 10 minutes. Copper-silver ionization was conducted in three circuits by installing units into return lines immediately upstream from hot water tanks. Recovery rates of L. pneumophila were monitored by culturing swab samples from faucets. Concentrations of copper and silver in water samples were determined by atomic absorption spectrophotometry. Four heat-flush treatments failed to provide long-term control of L. pneumophila. In contrast, ionization treatment reduced the rate of recovery of L. pneumophila from 108 faucets from 72% to 2% within 1 month and maintained effective control for at least 22 months. Only three samples (1.9%) of hot water from faucets exceeded Environmental Protection Agency standards for silver, and none exceeded the standards for copper. Of 24 samples obtained from hot water tanks, 42% and 50% exceeded the silver and copper standards, respectively. Copper-silver ionization effectively controls L. pneumophila in high-volume plumbing systems and is superior to thermal treatment; however, high concentrations of copper and silver can accumulate at the bottom of hot water tanks.

  12. Gentamicin-Containing Peptone-Yeast Extract Medium for Cocultivation of Hartmannella vermiformis ATCC 50256 and Virulent Strains of Legionella pneumophila.

    Science.gov (United States)

    Wadowsky, R M; Wang, L; Laus, S; Dowling, J N; Kuchta, J M; States, S J; Yee, R B

    1995-12-01

    We evaluated the use of peptone-yeast extract (PY) medium, different strains of Hartmannella vermiformis, and gentamicin in a coculture system to improve the discrimination of virulent and avirulent strains of Legionella pneumophila. H. vermiformis ATCC 50256 was unique among four strains of H. vermiformis, in that it multiplied equally well in Medium 1034 and PY medium (Medium 1034 without fetal calf serum, folic acid, hemin, and yeast nucleic acid and with a 50% reduction of peptone). However, both a virulent strain of L. pneumophila and its avirulent derivative strain multiplied in cocultures when PY medium was used. The multiplication of this avirulent strain was greatly reduced by incorporating gentamicin (1 (mu)g/ml) into the cocultivation system. Five virulent-avirulent sets of L. pneumophila strains were then tested for multiplication in cocultures with H. vermiformis ATCC 50256 and the gentamicin-containing PY medium. Only the virulent strains multiplied. The modified cocultivation system can discriminate between virulent and avirulent strains of L. pneumophila.

  13. [Legionella pneumophila eukaryotic-like effector LegK3 inhibits growth of Saccharomyces cerevisiae and modulates its vesicle trafficking pathway].

    Science.gov (United States)

    Wang, Jiaming; Li, Xianghui; Chen, Aifeng; Lu, Yongjun

    2014-04-04

    To study biochemical functions of the Legionella pneumophila eukaryotic-like effector protein LegK3, the budding yeast Saccharomyces cerevisiae was used as an alternative host in which growth defect induced by the ectopic expression of LegK3 was assessed. Using genomic DNA of the L. pneumophila strain Lp02 as template, we respectively amplified and inserted the ORF sequences of legK3, ralF or lidA into the plasmid pESC-HK to yield the ectopic-expression plasmids. Then, the recombination plasmids were transformed into the yeast strain W301-1A. With 2% -galactose induction, growth defect and carboxypeptidase Y (CPY) delay were determined simultaneously. In parallel, total yeast proteins before or after induction were extracted and subjected to Immunoblot assay. For detecting the expression of effector proteins or determining CPY delay, anti-c-myc or anti-PGK/anti-CPY antibodies were utilized respectively. The expression of LegK3 resulted in visible growth defect in yeast cells, together with obvious retard in CPY processing. L. pneumophila eukaryotic-like effector LegK3 might target and interfere with the vesicle-trafficking pathways, thereby to inhibit the growth and division of host cells.

  14. Prevalence of [i]Legionella pneumophila[/i] in water distribution systems in hospitals and public buildings of the Lublin region of eastern Poland

    Directory of Open Access Journals (Sweden)

    Agnieszka Sikora

    2015-05-01

    Full Text Available Objective. The aim of this study was to determine the prevalence of [i]L. pneumophila[/i] in water supply systems, hospitals and public buildings in the Lublin region of eastern Poland. Material and methods. The study was carried out in 26 different objects in the Lublin region. The number of [i]Legionella[/i] bacteria in water samples was determined by the membrane filtration method and/or by surface inoculation in accordance with the standards. Results. The study showed the presence of[i] L. pneumophila[/i] in 166 hot water samples (74.77%. In 34.33% (n=57 of water samples the count of tested bacteria exceeded the acceptable level of >100 CFU/100 ml. Of the samples where an acceptable level of bacteria was exceeded, 49 samples had an average level of [i]L. pneumophila[/i] (100–1,000 CFU/100 ml, and the level in 8 samples was high (>1,000 CFU/100 ml. Conclusions. The water samples collected form the hot water supply system of hospitals and public buildings showed exceeded counts of[i] L. pneumophila[/i], indicating the risk of infection. The constant monitoring of water distribution systems is an important element of the control of infections caused by these organisms.

  15. Human health risks for Legionella and Mycobacterium avium complex (MAC) from potable and non-potable uses of roof-harvested rainwater.

    Science.gov (United States)

    Hamilton, Kerry A; Ahmed, Warish; Toze, Simon; Haas, Charles N

    2017-08-01

    A quantitative microbial risk assessment (QMRA) of opportunistic pathogens Legionella pneumophila (LP) and Mycobacterium avium complex (MAC) was undertaken for various uses of roof-harvested rainwater (RHRW) reported in Queensland, Australia to identify appropriate usages and guide risk management practices. Risks from inhalation of aerosols due to showering, swimming in pools topped up with RHRW, use of a garden hose, car washing, and toilet flushing with RHRW were considered for LP while both ingestion (drinking, produce consumption, and accidental ingestion from various activities) and inhalation risks were considered for MAC. The drinking water route of exposure presented the greatest risks due to cervical lymphadenitis and disseminated infection health endpoints for children and immune-compromised populations, respectively. It is therefore not recommended that these populations consume untreated rainwater. LP risks were up to 6 orders of magnitude higher than MAC risks for the inhalation route of exposure for all scenarios. Both inhalation and ingestion QMRA simulations support that while drinking, showering, and garden hosing with RHRW may present the highest risks, car washing and clothes washing could constitute appropriate uses of RHRW for all populations, and toilet flushing and consumption of lettuce irrigation with RHRW would be appropriate for non- immune-compromised populations. Copyright © 2017 Elsevier Ltd. All rights reserved.

  16. Helicotylenchus stylocercus n. sp. and Rotylenchus phaliurus n. sp. (Nematoda: Hoplolaimidae) from Costa Rica.

    Science.gov (United States)

    Siddiqi, M R; Pinochet, J

    1979-10-01

    Two new species of plant-parasitic nematodes from Costa Rica are described. Helicotygenchus styloeercus n. sp., from soil around roots of banana at Coto, is distinguished hy the female tail, which bears a large pillarlike ventral projection. Rotylenchus phaliurus n. sp., from soil artmnd roots of Dioscoroea sp. at Sixaola, differs from R. caudaphasmidius in having the conus equal to or more than half the spear length, and large terminal annules on the female tail.

  17. SP (4,R) symmetry in light nuclei

    International Nuclear Information System (INIS)

    Peterson, D.R.

    1979-01-01

    A classification of nuclear states according to the noncompact sympletic Lie algebras sp(2n,R), n = 1, 2, 3, is investigated. Such a classification has recently been shown to be physically meaningful. This classification scheme is the appropriate generalization fo Elliott's SU 3 model of rotational states in deformed light nuclei to include core excitations. A restricted classification according to the Lie algebra, sp(4,R), is motivated. Truncation of the model space to a single sp(4,R) irreducible representation allows the inclusion of states possessing very high excitation energy. An sp(4,R) model study is performed on S = T = 0 positive-parity rotational bands in the deformed light nuclei 16 O and 24 Mg. States are included in the model space that possess up to 10h ω in excitation energy. Results for the B(E2) transition rates compare favorable with experiment, without resort to effective charges

  18. EOP Gold Coral (Gerardia sp.) Growth Measurements

    Data.gov (United States)

    National Oceanic and Atmospheric Administration, Department of Commerce — Gold coral (Gerardia sp.) trees that were inspected years earlier on Pisces submersible dives were revisited and their change in size measured. The fishery for...

  19. Silencing the SpMPK1, SpMPK2, and SpMPK3 Genes in Tomato Reduces Abscisic Acid—Mediated Drought Tolerance

    Directory of Open Access Journals (Sweden)

    Yan Liang

    2013-11-01

    Full Text Available Drought is a major threat to agriculture production worldwide. Mitogen-activated protein kinases (MAPKs play a pivotal role in sensing and converting stress signals into appropriate responses so that plants can adapt and survive. To examine the function of MAPKs in the drought tolerance of tomato plants, we silenced the SpMPK1, SpMPK2, and SpMPK3 genes in wild-type plants using the virus-induced gene silencing (VIGS method. The results indicate that silencing the individual genes or co-silencing SpMPK1, SpMPK2, and SpMPK3 reduced the drought tolerance of tomato plants by varying degrees. Co-silencing SpMPK1 and SpMPK2 impaired abscisic acid (ABA-induced and hydrogen peroxide (H2O2-induced stomatal closure and enhanced ABA-induced H2O2 production. Similar results were observed when silencing SpMPK3 alone, but not when SpMPK1 and SpMPK2 were individually silenced. These data suggest that the functions of SpMPK1 and SpMPK2 are redundant, and they overlap with that of SpMPK3 in drought stress signaling pathways. In addition, we found that SpMPK3 may regulate H2O2 levels by mediating the expression of CAT1. Hence, SpMPK1, SpMPK2, and SpMPK3 may play crucial roles in enhancing tomato plants’ drought tolerance by influencing stomatal activity and H2O2 production via the ABA-H2O2 pathway.

  20. Estudo comparativo da apresentação clínica da pneumonia a Legionella e outras pneumonias adquiridas na comuuidade

    Directory of Open Access Journals (Sweden)

    Nieves Sopena

    1998-11-01

    Full Text Available RESUMO: A Legionella pneumophila (LP é considerada em muitos estudos como uma das três causas mais comuns de pneumonia adquirida na comunidade (2,4 e a segunda causa em termos de gravidade.O principal objectivo do presente estudo realizado no Hospital da Universidade Autónoma de Barcelona, foi comparar os diferentes aspectos clínicos, biológicos e radiológicos da Pneumonia a Legionella pneumophilla e das outras Pneumonias adquiridas oa comunidade (PAC, de modo a auxiliar o diagnóstico precoce da pneumonia a LP.Foi realizado um estudo prospectivo de 392 doentes com PAC. Procedeu-se à análise comparativa dos aspectos epidemiológicos (hospitalizações ou viagens recentes, residência próximo de escavações ou trabalhos de constrção, contacto com animais, aves, demográficos (idade e sexo, clinicos (febre, tosse, expectoração, toracalgia, dispneia, cefaleias, confusão mental, dor aboominal, náuseas, vómitos, diarreia, artromialgias, dias de evolução, antibioterapia prévia, patologiaassociada, analiticos (leucocitose, natrémia­Na, creatina Kinase-CK, aspartato aminotransferase­AST e radiológicos. Estes aspectos foram estudados em 48 doentes com PAC por LP e 125 doentes com PAC de outra etiologia (68 por Streptococcus pneumoniae, 41 por Clamydia pneumoniae, 5 por Mycoplasma pneumoniae, 4 por Coxiella burnetii, 3 por Pseudomonas aeruginosa, 2 por Haemophilus injluenzae e 2 por Nocardia.A análise unifactorial mostrou que a PAC por LP foi mais frequente hos doentes de meia idade, do sexo masculino, em saúde aparente, mas com hábitos alcoó1icos acentuados, relativamente aos doentes com PAC de outra etioçogia. Também a auseocia de resposta aos fármacos beta-lactamicos prévios, cefaleias, diarreia, hiponatrémia grave e a elevação dos níveis de creatina Kinase sérica (CK foram mais frequentes na PAC por LP, enquanto que a tosse, expectoração e a toracalgia., foram mais frequeotes na pneumonia bacteriana de outra

  1. Penggunaan Jamur Antagonis Trichoderma SP. Dan Gliocladium SP. Untuk Mengendalikan Penyakit Layu Fusarium Pada Tanaman Bawang Merah (Allium Ascalonicum L.)

    OpenAIRE

    Arie Ramadhina, Arie Ramadhina; Lisnawita, Lisnawita; Lubis, Lahmuddin

    2013-01-01

    The use of antagonism fungus of Trichoderma sp. and Gliocladium sp. for controlling wilt(Fusarium oxysporum) in red onion plants. The aim of the research was to know the effectiviness ofantagonism fungus of Trichoderma sp. and Gliocladium sp. in controlling wilt in red onion plants.The research used non-factorial RAK (random group design) with eight treatments: control, 10grams of F. oxysporum, 12 grams of Trichoderma sp., 18 grams of Trichoderma sp., 24 grams ofTrichoderma sp., and 12 grams ...

  2. Chaenorhinum semispeluncarum sp. nov. and C. yildirimlii sp. nov. (Scrophulariaceae) from east Anatolia, Turkey

    DEFF Research Database (Denmark)

    Yildirim, Hasan; Tan, Kit; Senol, Serdar

    2010-01-01

    Chaenorhinum semispeluncarum H. Yildirim, Kit Tan, S. Senol & A. Pirhan sp. nov. and C. yildirimlii Kit Tan, H. Yildirim, S. Senol & A. Pirhan sp. nov. (Scrophulariaceae, C. sect. Microrrhinum) from east Anatolia are described and illustrated. They are both narrow endemics related to the rare C...

  3. Metallaphotoredox-catalysed sp3-sp3 cross-coupling of carboxylic acids with alkyl halides

    Science.gov (United States)

    Johnston, Craig P.; Smith, Russell T.; Allmendinger, Simon; MacMillan, David W. C.

    2016-08-01

    In the past 50 years, cross-coupling reactions mediated by transition metals have changed the way in which complex organic molecules are synthesized. The predictable and chemoselective nature of these transformations has led to their widespread adoption across many areas of chemical research. However, the construction of a bond between two sp3-hybridized carbon atoms, a fundamental unit of organic chemistry, remains an important yet elusive objective for engineering cross-coupling reactions. In comparison to related procedures with sp2-hybridized species, the development of methods for sp3-sp3 bond formation via transition metal catalysis has been hampered historically by deleterious side-reactions, such as β-hydride elimination with palladium catalysis or the reluctance of alkyl halides to undergo oxidative addition. To address this issue, nickel-catalysed cross-coupling processes can be used to form sp3-sp3 bonds that utilize organometallic nucleophiles and alkyl electrophiles. In particular, the coupling of alkyl halides with pre-generated organozinc, Grignard and organoborane species has been used to furnish diverse molecular structures. However, the manipulations required to produce these activated structures is inefficient, leading to poor step- and atom-economies. Moreover, the operational difficulties associated with making and using these reactive coupling partners, and preserving them through a synthetic sequence, has hindered their widespread adoption. A generically useful sp3-sp3 coupling technology that uses bench-stable, native organic functional groups, without the need for pre-functionalization or substrate derivatization, would therefore be valuable. Here we demonstrate that the synergistic merger of photoredox and nickel catalysis enables the direct formation of sp3-sp3 bonds using only simple carboxylic acids and alkyl halides as the nucleophilic and electrophilic coupling partners, respectively. This metallaphotoredox protocol is suitable for

  4. Five novel Wickerhamomyces- and Metschnikowia-related yeast species, Wickerhamomyces chaumierensis sp. nov., Candida pseudoflosculorum sp. nov., Candida danieliae sp. nov., Candida robnettiae sp. nov. and Candida eppingiae sp. nov., isolated from plants

    NARCIS (Netherlands)

    Groenewald, Marizeth; Robert, Vincent; Smith, Maudy Th

    On the basis of nucleotide divergences in the D1/D2 domain of the 26S rRNA gene and the internal transcribed spacers (ITS) domain of the rRNA gene, five novel yeast species, Wickerhamomyces chaumierensis sp. nov. (CBS 8565(T)  = JCM 17246(T)), Candida pseudoflosculorum sp. nov. (CBS 8584(T)  = JCM

  5. Crystal structure of Legionella DotD: insights into the relationship between type IVB and type II/III secretion systems.

    Directory of Open Access Journals (Sweden)

    Noboru Nakano

    2010-10-01

    Full Text Available The Dot/Icm type IVB secretion system (T4BSS is a pivotal determinant of Legionella pneumophila pathogenesis. L. pneumophila translocate more than 100 effector proteins into host cytoplasm using Dot/Icm T4BSS, modulating host cellular functions to establish a replicative niche within host cells. The T4BSS core complex spanning the inner and outer membranes is thought to be made up of at least five proteins: DotC, DotD, DotF, DotG and DotH. DotH is the outer membrane protein; its targeting depends on lipoproteins DotC and DotD. However, the core complex structure and assembly mechanism are still unknown. Here, we report the crystal structure of DotD at 2.0 Å resolution. The structure of DotD is distinct from that of VirB7, the outer membrane lipoprotein of the type IVA secretion system. In contrast, the C-terminal domain of DotD is remarkably similar to the N-terminal subdomain of secretins, the integral outer membrane proteins that form substrate conduits for the type II and the type III secretion systems (T2SS and T3SS. A short β-segment in the otherwise disordered N-terminal region, located on the hydrophobic cleft of the C-terminal domain, is essential for outer membrane targeting of DotH and Dot/Icm T4BSS core complex formation. These findings uncover an intriguing link between T4BSS and T2SS/T3SS.

  6. A Unique cis-Encoded Small Noncoding RNA Is Regulating Legionella pneumophila Hfq Expression in a Life Cycle-Dependent Manner

    Directory of Open Access Journals (Sweden)

    Giulia Oliva

    2017-01-01

    Full Text Available Legionella pneumophila is an environmental bacterium that parasitizes protozoa, but it may also infect humans, thereby causing a severe pneumonia called Legionnaires’ disease. To cycle between the environment and a eukaryotic host, L. pneumophila is regulating the expression of virulence factors in a life cycle-dependent manner: replicating bacteria do not express virulence factors, whereas transmissive bacteria are highly motile and infective. Here we show that Hfq is an important regulator in this network. Hfq is highly expressed in transmissive bacteria but is expressed at very low levels in replicating bacteria. A L. pneumophila hfq deletion mutant exhibits reduced abilities to infect and multiply in Acanthamoeba castellanii at environmental temperatures. The life cycle-dependent regulation of Hfq expression depends on a unique cis-encoded small RNA named Anti-hfq that is transcribed antisense of the hfq transcript and overlaps its 5′ untranslated region. The Anti-hfq sRNA is highly expressed only in replicating L. pneumophila where it regulates hfq expression through binding to the complementary regions of the hfq transcripts. This results in reduced Hfq protein levels in exponentially growing cells. Both the small noncoding RNA (sRNA and hfq mRNA are bound and stabilized by the Hfq protein, likely leading to the cleavage of the RNA duplex by the endoribonuclease RNase III. In contrast, after the switch to transmissive bacteria, the sRNA is not expressed, allowing now an efficient expression of the hfq gene and consequently Hfq. Our results place Hfq and its newly identified sRNA anti-hfq in the center of the regulatory network governing L. pneumophila differentiation from nonvirulent to virulent bacteria.

  7. SP-100/Brayton power system concepts

    International Nuclear Information System (INIS)

    Owen, D.F.

    1989-01-01

    Use of closed Brayton cycle (CBC) power conversion technology has been investigated for use with SP-100 reactors for space power systems. The CBC power conversion technology is being developed by Rockwell International under the Dynamic Isotype Power System (DIPS) and Space Station Freedom solar dynamic power system programs to provide highly efficient power conversion with radioisotype and solar collector heat sources. Characteristics including mass, radiator area, thermal power, and operating temperatures for systems utilizing SP-100 reactor and CBC power conversion technology were determined for systems in the 10-to 100-kWe power range. Possible SP-100 reactor/CBC power system configurations are presented. Advantages of CBC power conversion technology with regard to reactor thermal power, operating temperature, and development status are discussed

  8. Professional training for hygienic and sanitation maintenance of systems and equipments with risk of legionelosis Capacitación del personal que realiza las operaciones de mantenimiento higiénico-sanitario de las instalaciones de riesgo frente a Legionella

    Directory of Open Access Journals (Sweden)

    Milagros Fernández de Lezeta Sáez de Jáuregui

    2006-12-01

    Full Text Available Training for technicians in charge of maintenance, cleaning and disinfection of the equipment and installations that could be amplify and spread Legionella need to be checked and updated. This revision must to be done keeping with the kind of tasks performed and requested knowledges for technicians.In the following lines there is an analysis about the present training and ANECPLA proposes different levels based on its training experience and skills need by technicians to prevent and control legionnaires’ disease.La formación del personal técnico dedicado a las tareas de mantenimiento, limpieza y desinfección de equipos e instalaciones de riesgo frente a Legionella requiere una revisión y actualización que debe ser realizada teniendo en cuenta el tipo de tareas desempeñadas y conocimientos exigidos para su desarrollo.En este artículo se hace un análisis sobre la formación actual y se proponen diferentes niveles de capacitación en base a los criterios y experiencia que ANECPLA posee en materia de formación y conocimiento de las habilidades que se tienen que desarrollar en el trabajo de prevención y control de legionelosis.

  9. Ovos de Toxocara sp. e larvas de Ancylostoma sp. em praça pública de Lavras, MG Toxocara sp. eggs and Ancylostoma sp. larva in public parks, Brazil

    Directory of Open Access Journals (Sweden)

    Antônio Marcos Guimarães

    2005-04-01

    Full Text Available Larva migrans visceral e cutânea são zoonoses parasitárias causadas pela infecção da larva de Toxocara sp. e Ancylostoma sp., respectivamente. O objetivo do estudo foi verificar a contaminação por ovos de Toxocara sp. e ovos e larvas de Ancylostoma sp. em amostras de solos coletadas de praças públicas e de áreas de recreação infantil de Lavras, Estado de Minas Gerais, por meio da técnica de centrífugo-flutuação e do método de Baermann. A ocorrência de ovos de Toxocara sp. e, ovos e larvas de Ancylostoma sp. foi observada em 69,6% (16/23 das amostras de solo coletadas de praças públicas. A contaminação somente por ovos de Ancylostoma sp. em amostras de solo coletadas em escolas/creches foi de 22,2% (4/18. A percentagem de amostras de areia coletadas de escolas/creches contaminadas somente com larvas de Ancylostoma sp. foi de 11,1% (2/18. Praças públicas são as áreas com maior risco potencial de infecção por Toxocara sp. e Ancylostoma sp. Exame coproparasitológico realizado em 174 amostras de fezes de cães observou 58% e 23%, respectivamente, com ovos de Ancylostoma sp. e Toxocara sp.Visceral and cutaneous larva migrans are parasitic zoonoses caused by the infection of larval nematodes Toxocara sp. and Ancylostoma sp. respectively. The objective of this study was to investigate the contamination by Toxocara sp. eggs and Ancylostoma sp. eggs and larva of soil samples collected from public parks and children's playground areas in state of Minas Gerais, Brazil, using both Baermann's method and centrifugal flotation technique. Toxocara sp. and Ancylostoma sp. eggs were observed in soil samples collected from public squares in 17.4% (4/23 and 69.6 (16/23 respectively. In schools and child day care settings the contamination by Ancylostoma sp. larva in sand samples was 11.1% (2/18. Public parks are settings of more potential risk of Toxocara sp. eggs and Ancylostoma sp. infection. Stool parasitology testing of 174 stool

  10. Nitrogen fixation in Asaia sp. (family Acetobacteraceae).

    Science.gov (United States)

    Samaddar, Neeloy; Paul, Arundhati; Chakravorty, Somnath; Chakraborty, Writachit; Mukherjee, Joydeep; Chowdhuri, Debarati; Gachhui, Ratan

    2011-08-01

    The genus Asaia (family Acetobacteraceae) was first introduced with a single species-Asaia bogorensis and later six more species were described namely A. siamensis, A. krungthepensis, A. lannaensis, A. platycodi, A. prunellae, and A. astilbes. Acetobacteraceae family has been divided into ten genera but, only three of them include nitrogen fixing species: Gluconacetobacter, Acetobacter, and Swaminathania. This article originated from our study primarily aimed to isolate new endosymbiotic nitrogen fixer among Acetobacteraceae during which we have isolated, for the first time in India, four different strains of Asaia sp. from three different sources: Michalia champaca flower, Anopheles mosquito, and ant Tetraponera rufonigra. All the endosymbiotic strains isolated possess the ability to fix nitrogen. Evidence for both nitrogenase activity and the presence of nifH gene in isolated Asaia sp. is presented. Asaia bogorensis (MTCC 4041(T)) and A. siamensis (MTCC 4042(T)), two of the validated type strains available from the repository, were tested positive for the presence of functional nitrogenase. The nifH gene sequences from these type strains were also confirmed and compared with other nitrogen fixing members of the family Acetobacteraceae. Our result corroborate with the previous reports that Asaia sp. are indeed widely distributed in nature but this is the first time demonstration of their functional nitrogenase activity. This study shows Asaia sp. as fourth genera of nitrogen fixing bacteria in the family Acetobacteraceae.

  11. Molecular characterization of Trichoderma sp. isolated from ...

    African Journals Online (AJOL)

    The genetic relatedness among 15 isolates of Trichoderma sp. was analyzed with six micro-satellite primers. ISSR profiles showed 83.7% genetic diversity among the isolates with the formation of four clusters. Analysis of dendrogram revealed that similarity coefficient ranged from 0.27 to 0.95. ITS-PCR of rDNA region with ...

  12. Som man spørger

    DEFF Research Database (Denmark)

    Bjerg, Lars

    Indhold: Input og outputinterview ; Tre interviewtyper ; Tre grundprincipper ; De syv dødssynder ; Åbne, simple og neutrale spørgsmål ; Skuffespørgsmål ; Fokus ; Aktiv lytning ; Mål ; Strategi ; Abstrakt og konkret ; Det kritiske interview ; Kilder med et budskab ; Oldfruens fif ; Resumé i punktform...

  13. POTENSI MELANOTUS SP. DALAM MENDEGRADASI LIGNIN

    Directory of Open Access Journals (Sweden)

    NUNIK SULISTINAH

    2008-06-01

    Full Text Available Ten isolates of fungus were isolated from oil palm stem at oil palm plantation in Medan All of them were tested its abilities to degrade lignin. The results showed that one of them was able to grow on ligninase media and the fungi has the ability to degrade ligin. The fungi is identified as Melanotus sp.

  14. Biosorption of uranium by Azolla, SP, Brazil

    Energy Technology Data Exchange (ETDEWEB)

    Vieira, Ludmila C.; Alves, Eliakim G.; Marumo, Julio T., E-mail: lcvieira@ipen.br [Instituto de Pesquisas Energeticas e Nucleares (IPEN/CNEN-SP), Sao Paulo, SP (Brazil); Ferreira, Rafael V. de P., E-mail: rafael@itatijuca.com [Itatijuca Biotech, Sao Paulo, SP (Brazil); Canevesi, Rafael L.S.; Silva, Edson A., E-mail: edson.silva2@unioeste.br [Universidade Estadual do Oeste Parana (UNIOESTE), Toledo, PR (Brazil)

    2015-07-01

    Radioactive liquid waste needs special attention and requires suitable treatment before deposition. Among the potential technologies under development for the treatment of liquid radioactive wastes the biosorption has been highlighted by being an efficient and low cost technique. Biosorption process involves the exchange of ions contained in the biomass matrix by others present in solution. There are many biomasses that could be applied in treatment of radioactive wastes, for example, agricultural residues and macrophyte. The aim of this study is evaluate the ability of the Azolla sp., a floating aquatic plant, to absorb uranium in solution. Azolla sp. is a macrophyte that has been used to treat effluents containing heavy metals. The biosorption capacity of uranium by Azolla sp. was experimentally determined and modeled by isotherms. Experiments were performed to determine metal uptake, and then the solutions were analyzed by inductively coupled plasma optical emission spectrometry (ICP-OES). The isotherms applied to model the data was Langmuir, Freundlich, Sips Toth, Redlich Peternson, Two-Site-Langmuir, Radke Prausnitz to develop a technique for the treatment of radioactive liquid waste generated at the Nuclear and Energy Research Institute (IPEN-CNEN/SP), Brazil. (author)

  15. Biosorption of uranium by Azolla, SP, Brazil

    International Nuclear Information System (INIS)

    Vieira, Ludmila C.; Alves, Eliakim G.; Marumo, Julio T.; Ferreira, Rafael V. de P.; Canevesi, Rafael L.S.; Silva, Edson A.

    2015-01-01

    Radioactive liquid waste needs special attention and requires suitable treatment before deposition. Among the potential technologies under development for the treatment of liquid radioactive wastes the biosorption has been highlighted by being an efficient and low cost technique. Biosorption process involves the exchange of ions contained in the biomass matrix by others present in solution. There are many biomasses that could be applied in treatment of radioactive wastes, for example, agricultural residues and macrophyte. The aim of this study is evaluate the ability of the Azolla sp., a floating aquatic plant, to absorb uranium in solution. Azolla sp. is a macrophyte that has been used to treat effluents containing heavy metals. The biosorption capacity of uranium by Azolla sp. was experimentally determined and modeled by isotherms. Experiments were performed to determine metal uptake, and then the solutions were analyzed by inductively coupled plasma optical emission spectrometry (ICP-OES). The isotherms applied to model the data was Langmuir, Freundlich, Sips Toth, Redlich Peternson, Two-Site-Langmuir, Radke Prausnitz to develop a technique for the treatment of radioactive liquid waste generated at the Nuclear and Energy Research Institute (IPEN-CNEN/SP), Brazil. (author)

  16. Carbon sp chains in graphene nanoholes

    DEFF Research Database (Denmark)

    Castelli, Ivano Eligio; Ferri, Nicola; Onida, Giovanni

    2012-01-01

    Nowadays sp carbon chains terminated by graphene or graphitic-like carbon are synthesized routinely in several nanotech labs. We propose an ab initio study of such carbon-only materials, by computing their structure and stability, as well as their electronic, vibrational and magnetic properties. We...

  17. Lipid contents of the sponge Haliclona sp.

    Digital Repository Service at National Institute of Oceanography (India)

    Parameswaran, P.S.; Das, B.; Kamat, S.Y.

    Several fatty acids, sterols, batyl alcohol and its analogs and an N-acylated sphingosine (ceramide) have been isolated from the lipid fraction of the extract of the sponge Haliclona sp. The major sterol is found to be cholesterol (54%), followed...

  18. Test af spørgeskema

    DEFF Research Database (Denmark)

    Hermansen, Jonathan

    2017-01-01

    Denne bog gennemgår alle surveyarbejdets faser og giver en praktisk indføring i •design af undersøgelsen og udvælgelse af stikprøver, •formulering af spørgeskemaer samt indsamling og kodning af data, •metoder til at analysere resultaterne....

  19. Botrallin from the endophytic fungus Hyalodendriella sp ...

    African Journals Online (AJOL)

    use

    2011-12-12

    Dec 12, 2011 ... Bioassay-guided fractionation of the crude methanol extract of the mycelia from the endophytic fungus. Hyalodendriella sp. Ponipodef12, associated with the hybrid 'Neva' of Populus deltoides Marsh × P. nigra L., led to the isolation of one compound coded as P12-1 which was identified as botrallin (1,7-.

  20. Gamma-proteobacteria Aquicella lusitana gen. nov., sp. nov., and Aquicella siphonis sp. nov. infect protozoa and require activated charcoal for growth in laboratory media.

    Science.gov (United States)

    Santos, Paula; Pinhal, Isabel; Rainey, Fred A; Empadinhas, Nuno; Costa, Joana; Fields, Barry; Benson, Robert; Veríssimo, António; Da Costa, Milton S

    2003-11-01

    Several isolates, belonging to two new species of the same novel genus of gamma-proteobacteria, were recovered from drilled well (borehole) and spa water at São Gemil in central Portugal. These organisms are phylogenetically most closely related to the strictly intracellular uncultured species of the genus Rickettsiella, which cause disease in arthropods, and to the facultatively intracellular species of the genus Legionella, some of which cause Legionnaires' disease and Pontiac fever. The São Gemil strains grew only on media containing charcoal, as is also true of the species of the genus LEGIONELLA: Unlike the vast majority of Legionella isolates, the new isolates did not require L-cysteine or ferric pyrophosphate for growth but like the legionellae had an absolute requirement for alpha-ketoglutarate. Strains SGT-39(T) and SGT-56 grew consistently between 30 and 43 degrees C, while strains SGT-108(T) and SGT-109 grew between 30 and 40 degrees C. The pH ranges for growth of these organisms were surprisingly narrow: strains SGT-39(T) and SGT-56 grew between pH 6.3 and 7.3, while strains SGT-108(T) and SGT-109 grew between pH 6.3 and 7.0. Both organisms proliferated in the amoeba Hartmannella vermiformis but did not grow in U937 human cells. Based on 16S rRNA gene sequence analysis and physiological, biochemical, and chemical analysis we describe two new species of one novel genus; one species is represented by strain SGT-39(T), for which we propose the name Aquicella lusitana, while strain SGT-108(T) represents a second species of the same genus, for which we propose the name Aquicella siphonis.

  1. C (sp2)–C (sp2) cross coupling reaction catalyzed by a palladacycle ...

    Indian Academy of Sciences (India)

    (sp2) cross coupling reaction catalyzed by a palladacycle phosphine complex: A simple and sustainable protocol in aqueous media. Seyyed Javad Sabounchei Marjan Hosseinzadeh. Articles Volume 127 Issue 11 November 2015 pp 1919- ...

  2. Nanocrystalline sp{sup 2} and sp{sup 3} carbons: CVD synthesis and applications

    Energy Technology Data Exchange (ETDEWEB)

    Terranova, M. L. [Università degli Studi di Roma “Tor Vergata,” via Della Ricerca Scientifica, Dipartimento di Scienze e Tecnologie Chimiche—MinimaLab (Italy); Rossi, M. [Università degli Studi di Roma “Sapienza,” via A. Scarpa, Dipartimento di Scienze di Base e Applicate per l’Ingegneria and Centro di Ricerca per le Nanotecnologie Applicate all’Ingegneria (CNIS) (Italy); Tamburri, E., E-mail: emanuela.tamburri@uniroma2.it [Università degli Studi di Roma “Tor Vergata,” via Della Ricerca Scientifica, Dipartimento di Scienze e Tecnologie Chimiche—MinimaLab (Italy)

    2016-11-15

    The design and production of innovative materials based on nanocrystalline sp{sup 2}- and sp{sup 3}-coordinated carbons is presently a focus of the scientific community. We present a review of the nanostructures obtained in our labs using a series of synthetic routes, which make use of chemical vapor deposition (CVD) techniques for the selective production of non-planar graphitic nanostructures, nanocrystalline diamonds, and hybrid two-phase nanostructures.

  3. Analisis Fosfor pada Cacing Tanah (Megascolex sp. dan Fridericia sp.) Secara Spektrofotometri Sinar Tampak

    OpenAIRE

    Safira, Cut Shafa

    2015-01-01

    Earthworm is natural resource which can be used for medication due to its highly amount of minerals. One of these minerals is phosphorus. The aim of this research are to identify, determine and know the difference content of phosphorus in Megascolex sp. and Fridericia sp. Qualitative analysis shows positive results with addition of ammonium molybdate 4% and BaCl2 5%. Quantitative analysis was done using visible spectrophotometer with ascorbic acid method, measuring blu-colored molybdenum ...

  4. Differences in nutrient uptake capacity of the benthic filamentous algae Cladophora sp., Klebsormidium sp. and Pseudanabaena sp. under varying N/P conditions.

    Science.gov (United States)

    Liu, Junzhuo; Vyverman, Wim

    2015-03-01

    The N/P ratio of wastewater can vary greatly and directly affect algal growth and nutrient removal process. Three benthic filamentous algae species Cladophora sp., Klebsormidium sp. and Pseudanabaena sp. were isolated from a periphyton bioreactor and cultured under laboratory conditions on varying N/P ratios to determine their ability to remove nitrate and phosphorus. The N/P ratio significantly influenced the algal growth and phosphorus uptake process. Appropriate N/P ratios for nitrogen and phosphorus removal were 5-15, 7-10 and 7-20 for Cladophora sp., Klebsormidium sp. and Pseudanabaena sp., respectively. Within these respective ranges, Cladophora sp. had the highest biomass production, while Pseudanabaena sp. had the highest nitrogen and phosphorus contents. This study indicated that Cladophora sp. had a high capacity of removing phosphorus from wastewaters of low N/P ratio, and Pseudanabaena sp. was highly suitable for removing nitrogen from wastewaters with high N/P ratio. Copyright © 2014 Elsevier Ltd. All rights reserved.

  5. Anomalous self potential (sp) log signatures observed in a water ...

    African Journals Online (AJOL)

    Geophysical logging was done after drilling had been completed in a water well at Okwudor, South Eastern Nigeria. Three electric logs were run viz: Self Potential (SP), Resistivity N16″ and N64″ logs. An anomaly was observed in the SP log. The SP results from this well show some deviation from the standard norm.

  6. Biosorption of chromium by mangrove-derived Aplanochytrium sp ...

    African Journals Online (AJOL)

    The microbial dried biomass of Thraustochytrids is used as bioadsorbent for the removal of the chromium in aqueous solution. In this investigation, three species of Thraustochydrids namely Aplanochytrium sp., Thraustochytrium sp. and Schizochytrium sp. were tested for the efficiency of chromium accumulation by culturing ...

  7. Babesia peircei sp. nov. from the jackass penguin

    African Journals Online (AJOL)

    1992-01-09

    Jan 9, 1992 ... An avian piroplasm, Babesia peircei sp. nov. is described from the jackass penguin Spheniscus demersus. Morphological differences between Babesia peircei sp. nov. and the other valid Babesia spp. are discussed together with the possible vectors. 'n Voal-piroplasma, Babesia peircei sp. nov. afkomstig ...

  8. Occurrence of Aspergillus sp., Fusarium sp., and aflatoxins in corn hybrids with different systems of storage

    Directory of Open Access Journals (Sweden)

    Adriana Sbardelotto Di Domenico

    2016-01-01

    Full Text Available The aim of this study was to assess alternatives for viable corn storage for small rural properties in two annual storage experiments. A 4×5 factorial design was used with four types of storage (conventional bags, hermetic bags, metal silos and corncobs and five periods of storage (0, 3, 6, 9 and 12 months. We used corn hybrids 2B688RR and 30K73Hx cultivated in winter 2012 and summer 2012/2013 in the city of Dois Vizinhos, Paraná, Brazil. The moisture contents, counts of Aspergillus sp. and Fusarium sp., and the occurrence of aflatoxins (B1, B2, G1 and G2 were assessed. The kernels stored in hermetic bags had lower moisture contents. Aspergillus sp. and Fusarium sp. were observed in 20.37 and 86.11% of winter storage samples, respectively, and in 83.3 and 91.6% of summer storage samples, respectively. The storage system and time of storage had no influence on the occurrence of Aspergillus sp. and aflatoxins in the winter crop samples. The corncobs from the summer crop samples had the lowest counts of Aspergillus sp. and did not have aflatoxins. We detected aflatoxins at concentrations of 2.8-14.5 and 3-197.5 µg kg-1 in the winter and summer crop samples, respectively.

  9. Two new species of suctorians, Acineta satyanandani sp. nov. and Paracineta karunakarani sp. nov. epizoic on ostracods

    Digital Repository Service at National Institute of Oceanography (India)

    Santhakumari, V.

    Two, species of protozoic suctorians, Acineta satyanandani sp. nov. and Paracineta karunakarani sp. nov., are described. These were found attached on the body of the marine ostracod, Cypridina dentata (Muller), collected from the shelf and slope...

  10. The family Carabodidae (Acari, Oribatida) VIII. The genus Machadocepheus (first part) Machadocepheusleoneae sp. n. and Machadocepheusrachii sp. n. from Gabon.

    Science.gov (United States)

    Fernandez, Nestor; Theron, Pieter; Rollard, Christine; Leiva, Sergio

    2014-01-01

    The genus Machadocepheus, being one of the more complex genera of the Carabodidae family, is briefly outlined to demonstrate this complexity. Descriptions of two new species from Gabon, Machadocepheusleoneae sp. n. and Machadocepheusrachii sp. n. are given.

  11. First report of Giardia sp. and Cryptosporidium sp. in fecal samples in blue macaw (Anodorhynchus hyacinthinus) in southern Brazil

    OpenAIRE

    Matheus Hillard Farret; Vinicius da Rosa Fanfa; Luisa Ragagnin; Aleksandro Schafer da Silva; Silvia Gonzalez Monteiro

    2010-01-01

    This study reports the gastrointestinal parasitism by Giardia sp. and Cryptosporidium sp. in blue macaw (Anodorhynchus hyacinthinus) in the southern region of Brazil. Fecal samples of two species kept in captivity in the state of Rio Grande do Sul were analyzed by the direct smear method, centrifugal flotation technique with zinc sulfate and Kinyoun staining technique for research of parasites. Mixed infection by eggs of Capillaria, cysts of Giardia sp. and oocysts of Cryptosporidium sp. was ...

  12. Agromyces italicus sp. nov., Agromyces humatus sp. nov. and Agromyces lapidis sp. nov., isolated from Roman catacombs.

    Science.gov (United States)

    Jurado, Valme; Groth, Ingrid; Gonzalez, Juan M; Laiz, Leonila; Schuetze, Barbara; Saiz-Jimenez, Cesareo

    2005-03-01

    A polyphasic study was carried out to clarify the taxonomic positions of three Gram-positive isolates from the Catacombs of Domitilla, Rome (Italy). 16S rRNA gene sequence comparisons placed these strains within the genus Agromyces. The morphological and chemotaxonomic characteristics of these isolates were consistent with the description of the genus Agromyces. The three isolates could be readily distinguished from one another and from representatives of all Agromyces species with validly published names by a broad range of phenotypic characteristics and DNA-DNA relatedness studies. Therefore, these isolates are proposed to represent three novel species of the genus Agromyces, Agromyces italicus sp. nov. (type strain CD1(T)=HKI 0325(T)=DSM 16388(T)=NCIMB 14011(T)), Agromyces humatus sp. nov. (type strain CD5(T)=HKI 0327(T)=DSM 16389(T)=NCIMB 14012(T)) and Agromyces lapidis sp. nov. (type strain CD55(T)=HKI 0324(T)=DSM 16390(T)=NCIMB 14013(T)).

  13. Cryptotrichosporon argae sp. nov., Cryptotrichosporon brontae sp. nov. and Cryptotrichosporon steropae sp. nov., isolated from forest soils.

    Science.gov (United States)

    Pontes, Ana; Röhl, Oliver; Maldonado, Cristina; Yurkov, Andrey M; Sampaio, José Paulo

    2017-09-01

    Yeast strains belonging to the basidiomycetous genus Cryptotrichosporon were isolated from forest soils in Serra da Arrábida Natural Park in Portugal. Similar to the already-known representatives of this genus, the new isolates formed pigmented colonies of a distinctive pale orange colour. Phylogenetic analyses employing concatenated sequences of the D1/D2 domains of the 26S (large subunit) rRNA gene and the internal transcribed spacer (ITS) region supported the recognition of three novel species: Cryptotrichosporon argae sp. nov. (type strain CM 19T=CBS 14376T=PYCC 7010T=DSM 104550T; MycoBank accession number MB 817168), Cryptotrichosporon brontae sp. nov. (type strain CM 1562T=CBS 14303T=PYCC 7011T=DSM 104551T; MycoBank accession number MB 817077) and Cryptotrichosporon steropae sp. nov. (type strain OR 395T=CBS 14302T=PYCC 7012T=DSM 104552T; MycoBank accession number MB 817078).

  14. Fitoremediasi limbah budidaya sidat menggunakan filamentous algae (Spirogyra sp.

    Directory of Open Access Journals (Sweden)

    Tri Apriadi

    2014-09-01

    Full Text Available Penelitian ini bertujuan untuk mengetahui potensi dari filamentous algae (Spirogyra sp. sebagai agen bioremediasi dalam mereduksi kandungan bahan organik limbah budidaya sidat. Penelitian menggunakan rancangan acak lengkap dengan perlakuan perbedaan dosis limbah (25 %, 50 %, 75 %, 100%. Wadah penelitian berupa akuarium resirkulasi menggunakan sistem carrousel. Dilakukan pengukuran secara rutin terhadap beberapa parameter kualitas air serta perubahan bobot Spirogyra sp. selama dua minggu retensi. Diperoleh hasil bahwa penurunan konsentrasi bahan organik menggunakan Spirogyra sp. berlangsung efektif hingga hari keenam. Spirogyra sp. mampu mentolelir limbah budidaya sidat pada dosis limbah 25% dan 50%. Spirogyra sp. pada perlakuan dosis limbah 50% memiliki kemampuan yang lebih baik dalam menurunkan bahan organik limbah budidaya sidat.

  15. A superhard sp3 microporous carbon with direct bandgap

    Science.gov (United States)

    Pan, Yilong; Xie, Chenlong; Xiong, Mei; Ma, Mengdong; Liu, Lingyu; Li, Zihe; Zhang, Shuangshuang; Gao, Guoying; Zhao, Zhisheng; Tian, Yongjun; Xu, Bo; He, Julong

    2017-12-01

    Carbon allotropes with distinct sp, sp2, and sp3 hybridization possess various different properties. Here, a novel all-sp3 hybridized tetragonal carbon, namely the P carbon, was predicted by the evolutionary particle swarm structural search. It demonstrated a low density among all-sp3 carbons, due to the corresponding distinctive microporous structure. P carbon is thermodynamically stable than the known C60 and could be formed through the single-walled carbon nanotubes (SWCNTs) compression. P carbon is a direct bandgap semiconductor displaying a strong and superhard nature. The unique combination of electrical and mechanical properties constitutes P carbon a potential superhard material for semiconductor industrial fields.

  16. Sp6 and Sp8 Transcription Factors Control AER Formation and Dorsal-Ventral Patterning in Limb Development

    Science.gov (United States)

    Haro, Endika; Delgado, Irene; Junco, Marisa; Yamada, Yoshihiko; Mansouri, Ahmed; Oberg, Kerby C.; Ros, Marian A.

    2014-01-01

    The formation and maintenance of the apical ectodermal ridge (AER) is critical for the outgrowth and patterning of the vertebrate limb. The induction of the AER is a complex process that relies on integrated interactions among the Fgf, Wnt, and Bmp signaling pathways that operate within the ectoderm and between the ectoderm and the mesoderm of the early limb bud. The transcription factors Sp6 and Sp8 are expressed in the limb ectoderm and AER during limb development. Sp6 mutant mice display a mild syndactyly phenotype while Sp8 mutants exhibit severe limb truncations. Both mutants show defects in AER maturation and in dorsal-ventral patterning. To gain further insights into the role Sp6 and Sp8 play in limb development, we have produced mice lacking both Sp6 and Sp8 activity in the limb ectoderm. Remarkably, the elimination or significant reduction in Sp6;Sp8 gene dosage leads to tetra-amelia; initial budding occurs, but neither Fgf8 nor En1 are activated. Mutants bearing a single functional allele of Sp8 (Sp6−/−;Sp8+/−) exhibit a split-hand/foot malformation phenotype with double dorsal digit tips probably due to an irregular and immature AER that is not maintained in the center of the bud and on the abnormal expansion of Wnt7a expression to the ventral ectoderm. Our data are compatible with Sp6 and Sp8 working together and in a dose-dependent manner as indispensable mediators of Wnt/βcatenin and Bmp signaling in the limb ectoderm. We suggest that the function of these factors links proximal-distal and dorsal-ventral patterning. PMID:25166858

  17. Bioakumulasi logam berat Cu oleh Bacillus sp

    Directory of Open Access Journals (Sweden)

    Riesta Primaharinastiti

    2012-02-01

    Full Text Available The research was conducted to investigate the ability of Bacillus sp in accumulating Cu and how much it can be acumulated. Themedium used to growth the bacterium was Nutrient Broth and Atomic Absorption Spectrophotometry methods was used to assay theCu, both in the cells and medium. The result of this study showed that Bacillus sp incubated in the Nutrient Broth medium containing10 ppm of Cu, with continuous stirring in the room temperature was able to reduce Cu in the medium 8.912–12.623% and accumulateCu in the cell 0.1149–0.1400 %/mg cells. Based on this result, it is necessary to develop more studies to find out what factors thatinfluence the accumulation process and to optimize the bioprocess.

  18. Analysis list: SP2 [Chip-atlas[Archive

    Lifescience Database Archive (English)

    Full Text Available SP2 Blood,Liver,Pluripotent stem cell + hg19 http://dbarchive.biosciencedbc.jp/kyus...hu-u/hg19/target/SP2.1.tsv http://dbarchive.biosciencedbc.jp/kyushu-u/hg19/target/SP2.5.tsv http://dbarchive.bioscience...dbc.jp/kyushu-u/hg19/target/SP2.10.tsv http://dbarchive.biosciencedbc.jp/kyushu-u/hg19/colo/SP2.B...lood.tsv,http://dbarchive.biosciencedbc.jp/kyushu-u/hg19/colo/SP2.Liver.tsv,http://dbarchive.bioscience...dbc.jp/kyushu-u/hg19/colo/SP2.Pluripotent_stem_cell.tsv http://dbarchive.biosciencedbc

  19. Superfield quantization in Sp(2) covariant formalism

    CERN Document Server

    Lavrov, P M

    2001-01-01

    The rules of the superfield Sp(2) covariant quantization of the arbitrary gauge theories for the case of the introduction of the gauging with the derivative equations for the gauge functional are generalized. The possibilities of realization of the expanded anti-brackets are considered and it is shown, that only one of the realizations is compatible with the transformations of the expanded BRST-symmetry in the form of super translations along the Grassmann superspace coordinates

  20. Carbon sp chains in graphene nanoholes

    Science.gov (United States)

    Castelli, Ivano E.; Ferri, Nicola; Onida, Giovanni; Manini, Nicola

    2012-03-01

    Nowadays sp carbon chains terminated by graphene or graphitic-like carbon are synthesized routinely in several nanotech labs. We propose an ab initio study of such carbon-only materials, by computing their structure and stability, as well as their electronic, vibrational and magnetic properties. We adopt a fair compromise of microscopic realism with a certain level of idealization in the model configurations, and predict a number of properties susceptible to comparison with experiment.