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Sample records for legionella culture method

  1. 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.

  2. 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.

  3. 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

  4. 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...

  5. 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.

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

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    Carmen Pelaz Antolín

    2006-12-01

    included in the maintenance plans when Legionella prevention is carried out in water installations (Real Decree 865/2003. The comparison of our legislation with other legislations or recommendations adopted in other countries (United Kingdom, France, Australia, America allows to know our degree of demand of some parameters. Bacteria culture is the gold standard method for Legionella detection in water samples, and and there are several normalized assays, such as ISO 11731/98 and 2004 or NF T 90-431/2003 (AFNOR. To help the interpretation of the results, Legionella assays should reflect the standard in which they are based and the limit of detection of the method, that should not be over 100 ufc/L. Moreover, laboratories that carry out these assays should be acredited by our national accreditation body (ENAC. In the last years, fast methods have been developed for Legionella detection based on the amplification of chromosomal DNA in water samples by PCR reactions. PCR assays should not be used alone, but it should be a complement of culture assays, when normative actions are implemented in water installations.

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

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    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.

  8. 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.

  9. 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.

  10. 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...

  11. 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....

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

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    Hsu, B.-M.; Ma, P.-H.; Su, I.-Z.; Chen, N.-S.

    2009-04-01

    Legionella genera are parasites of FLA, and intracellular bacterial replication within the FLA plays a major role in the transmission of disease. At least 13 FLA species—including Acanthamoeba spp., Naegleria spp., and Hartmannella spp.—support intracellular bacterial replication. In the study, Legionellae were detected with microbial culture or by direct DNA extraction and analysis from concentrated water samples or cultured free-living amoebae, combined with molecular methods that allow the taxonomic identification of these pathogens. The water samples were taken from a mud spring recreation area located in a mud-rock-formation area in southern Taiwan. Legionella were detected in 15 of the 34 samples (44.1%). Four of the 34 samples analyzed by Legionella culture were positive for Legionella, five of 34 were positive for Legionella when analyzed by direct DNA extraction and analysis, and 11 of 34 were positive for amoebae-resistant Legionella when analyzed by FLA culture. Ten samples were shown to be positive for Legionella by one analysis method and five samples were shown to be positive by two analysis methods. However, Legionella was detected in no sample by all three analysis methods. This suggests that the three analysis methods should be used together to detect Legionella in aquatic environments. In this study, L. pneumophila serotype 6 coexisted with A. polyphaga, and two uncultured Legionella spp. coexisted with either H. vermiformis or N. australiensis. Of the unnamed Legionella genotypes detected in six FLA culture samples, three were closely related to L. waltersii and the other three were closely related to L. pneumophila serotype 6. Legionella pneumophila serotype 6, L. drancourtii, and L. waltersii are noted endosymbionts of FLA and are categorized as pathogenic bacteria. This is significant for human health because these Legionella exist within FLA and thus come into contact with typically immunocompromised people.

  13. 'Gatekeeper' unit defeats legionella.

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    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.

  14. 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.

  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. 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.

  18. Evaluation of detection methods for Legionella species using ...

    African Journals Online (AJOL)

    In addition, the direct immunofluorescence test and a commercially available latex agglutination test kit were included in the evaluations. The usefulness of treatment with acid or heat prior to culture was also compared. Our results indicated that concentration by membrane filtration using nitro-cellulose filters with a pore size ...

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

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    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.

  20. Survey of Legionella Species Found in Thai Soil

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    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.

  1. [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.

  2. 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.

  3. 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

  4. 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.

  5. 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.

  6. 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.

  7. 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.

  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. 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.

  10. 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.

  11. 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.

  12. 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.

  13. 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.

  14. 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.

  15. 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.

  16. 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...

  17. 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.

  18. 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...

  19. 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

  20. 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.

  1. 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.

  2. 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.

  3. 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.

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

    Directory of Open Access Journals (Sweden)

    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.

  5. 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.

  6. 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.

  7. [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.

  8. A multicenter evaluation of genotypic methods for the epidemiologic typing of Legionella pneumophila serogroup 1

    DEFF Research Database (Denmark)

    Fry, Norman K.; Alexiou-Daniel, Stella; Bangsborg, Jette Marie

    1999-01-01

    length polymorphism analysis, restriction endonuclease analysis, pulsed-field gel electrophoresis (PFGE), PCR using arbitrary/repeat sequence primers (AP-, AP/rep-PCR), and amplified fragment length polymorphism (AFLP) analysis. Results were analyzed visually or using gel analysis software. Each method...... was assessed for its: index of discrimination (D), epidemiologic concordance (E), speed of application and ease of use. In addition, phenotypic analysis was performed in two laboratories using monoclonal antibodies (mAbs). RESULTS: The D of each of the genotypic methods ranged from 0.840 for ribotyping to 0...

  9. A multicenter evaluation of genotypic methods for the epidemiologic typing of Legionella pneumophila serogroup 1

    DEFF Research Database (Denmark)

    Fry, Norman K.; Alexiou-Daniel, Stella; Bangsborg, Jette Marie

    1999-01-01

    was assessed for its: index of discrimination (D), epidemiologic concordance (E), speed of application and ease of use. In addition, phenotypic analysis was performed in two laboratories using monoclonal antibodies (mAbs). RESULTS: The D of each of the genotypic methods ranged from 0.840 for ribotyping to 0.......990 for PFGE using Sfil: E ranged from 0.06 for AP- and AP/rep-PCR to 1.00 for ribotyping using Pstl/EcoRI and AFLP: in general, E was inversely related to D. Although offering only limited discrimination (D=0.838), mAb typing was both rapid and highly epidemiologically concordant (E=1.00). CONCLUSIONS: Two...

  10. 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...

  11. 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

  12. 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.

  13. 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.

  14. 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.

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

    Science.gov (United States)

    Erdoğan, Haluk; Arslan, Hande

    2013-04-01

    samples could not be obtained from the cases, the diagnosis were not confirmed by culture but by urinary antigen test. Besides high antibody titer in single serum sample was accepted as a diagnostic marker. Additionally 26 cases who accommodated in the same hotel and presented with high fever without pneumonia were treated in the outpatient clinics of our hospital. Urinary antigen test was performed in 11 of those patients to confirm the prediagnosis of pontiac fever, however all were found negative. Likewise convalescent phase sera for the confirmation of the diagnosis by seroconversion could not be obtained since they all were foreign tourists. Investigation of water sources of the hotel revealed that the municipal drinking water network had not been connected yet and the hotel supplied water from groundwater sources. The analysis of multiple samples from multiple sites of hotel's water system indicated that the water temperature was between 35-45°C and the iron level was beyond the acceptable limits (245 µg/L) recommended for drinking water in the regulation guides. These properties were considered as the factors that enhanced the growth and survival of Legionella species. Water samples were cultivated on BCYE-_ (Buffered Charcoal Yeast Extract a-Ketoglutarate) and GVPC (Glycine-Vancomycin-Polymyxin-Cycloheximide) agar plates and 11 out of a total 13 samples yielded Legionella spp. growth. All isolates were identified as L.pneumophila serogroup 1 by specific antisera. Legionella decontamination of hotel's water system was managed by implementation of hyperchlorination method as well as superheating (> 60°C) of water. The hotel was not closed during the outbreak and cultures of water samples obtained for one year later did not yield any Legionella spp. growth. This outbreak emphasized that hotel residents are at risk for acquiring LD in the presence of a colonized water system, even in a newly constructed building. In conclusion, effective control and decontamination

  16. 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.

  17. 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

  18. 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.

  19. 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.

  20. 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.

  1. 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.

  2. 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

  3. 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.

  4. 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.

  5. 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.

  6. 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.

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

    Directory of Open Access Journals (Sweden)

    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.

  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. 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.

  10. 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.

  11. 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

  12. 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

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

    Directory of Open Access Journals (Sweden)

    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.

  15. 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.

  16. 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.

  17. 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.

  18. 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.

  19. 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)

  20. 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) ...

  1. 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

  2. The Method of Culture Contrast

    DEFF Research Database (Denmark)

    Hasse, Cathrine; Trentemøller, Stine; Motzkau, Johanna

    2009-01-01

    In this article, we suggest that research is a practical activity building on local category systems belonging specifically to research (etic categories) as well as categories belonging specifically to the national culture of the researcher (emic categories) (Pike 1967). Much cross-cultural resea......In this article, we suggest that research is a practical activity building on local category systems belonging specifically to research (etic categories) as well as categories belonging specifically to the national culture of the researcher (emic categories) (Pike 1967). Much cross......-cultural research can be argued to rest on what has been called implicit comparisons (Nader 1994) of such categorisations. We assume that research of local activities, such as schooling and higher education, is influenced by the researcher's emic and etic categorisations. To get beyond the risk of reproducing......) framework on which the research is conducted. First we present a recent study of European universities as culturally diverse working places and we present an approach in which the researcher's emic and etic categorisations can be challenged when contrasted with each other (Hasse & Trentemøller 2008). Second...

  3. 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.

  4. 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

  5. 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.

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

    Science.gov (United States)

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

    2014-01-01

    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

  7. 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.

  8. 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....

  9. 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

  10. 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.

  11. 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.

  12. 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.

  13. 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

  14. 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.

  15. 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 ...

  16. [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

  17. 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....

  18. 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.

  19. 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

  20. 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.

  1. 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%.

  2. 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...

  3. 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...

  4. 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.

  5. 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.

  6. 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.

  7. 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.

  8. 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.

  9. 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...

  10. 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.

  11. 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

  12. 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

  13. 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.

  14. 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...

  15. 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.

  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. 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.

  18. 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.

  19. 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

  20. 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.

  1. 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.

  2. 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...

  3. 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 ...

  4. 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....

  5. 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.

  6. 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

  7. 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

  8. In vitro culture method of powdery mildew ( Oidium heveae ...

    African Journals Online (AJOL)

    A method for culturing powdery mildew (Oidium heveae) from isolated leaves of Hevea brasiliensis was evaluated, which included three steps: Leaves and fungi selection, nutrient solution and culture dish preparation, fungi inoculation and culture. The culture time and produced conidia number were considered as decision ...

  9. 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.

  10. 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...

  11. 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...

  12. 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.

  13. 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.

  14. 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

  15. 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

  16. 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....

  17. 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...

  18. 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.

  19. 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.

  20. Culture, Interface Design, and Design Methods for Mobile Devices

    Science.gov (United States)

    Lee, Kun-Pyo

    Aesthetic differences and similarities among cultures are obviously one of the very important issues in cultural design. However, ever since products became knowledge-supporting tools, the visible elements of products have become more universal so that the invisible parts of products such as interface and interaction are getting more important. Therefore, the cultural design should be extended to the invisible elements of culture like people's conceptual models beyond material and phenomenal culture. This chapter aims to explain how we address the invisible cultural elements in interface design and design methods by exploring the users' cognitive styles and communication patterns in different cultures. Regarding cultural interface design, we examined users' conceptual models while interacting with mobile phone and website interfaces, and observed cultural difference in performing tasks and viewing patterns, which appeared to agree with cultural cognitive styles known as Holistic thoughts vs. Analytic thoughts. Regarding design methods for culture, we explored how to localize design methods such as focus group interview and generative session for specific cultural groups, and the results of comparative experiments revealed cultural difference on participants' behaviors and performance in each design method and led us to suggest how to conduct them in East Asian culture. Mobile Observation Analyzer and Wi-Pro, user research tools we invented to capture user behaviors and needs especially in their mobile context, were also introduced.

  1. Cultural Impact on Participatory Deisgn Method

    DEFF Research Database (Denmark)

    Jensen, Mika Yasuoka

    2012-01-01

    This is a position paper for Cultural Impact on User Experience Design and Evaluation - One day workshop, NordiCHI, 2012.......This is a position paper for Cultural Impact on User Experience Design and Evaluation - One day workshop, NordiCHI, 2012....

  2. A Method, Computer Program and System for Inferring Relations Between Cultural Specific Concepts in Two Cultures

    DEFF Research Database (Denmark)

    2013-01-01

    The present invention relates to a method, computer program and system for inferring relations between cultural specific concepts (CSC) in two cultures at least comprising the steps of - extracting and listing said cultural specific concepts (CSCs) and features of said CSCs from at least a first...... corpora belonging to a first culture and a second corpora belonging to a second culture, - applying a algorithm to infer relations between said CSCs in the first and the second corpora....

  3. 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...

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

    Directory of Open Access Journals (Sweden)

    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.

  5. 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...

  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. A method for culturing human hair follicle cells.

    Science.gov (United States)

    Weterings, P J; Vermorken, A J; Bloemendal, H

    1981-01-01

    For the first time a method for culturing human hair follicle cells is described. The bovine eye lens capsule, a basement membrane-like structure, is used as the substrate for the cultures. In a culture medium supplemented with hydrocortisone and insulin about 70% of the original follicles will form growing colonies of diploid keratinocytes.

  8. 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.

  9. [Research progress of cell co-culture method].

    Science.gov (United States)

    Qin, Yanqin; Chen, Yulong; Li, Jiansheng

    2016-08-01

    Cell culture technology is the most commonly used method in the in vitro experiments at present. However, monolayer cell culture technology has been unable to meet the demand of the researchers. This is because that monolayer cell culture cannot mimic the cellular environment in which multiple cells interact with each other in the body. We cannot discuss the relationship of many cells, because we do not know the relationship between cells through a single kind of cell. So cell co-culture medicine arises at the historic moment for the demand. With the development of research method in recent years, cell co-culture method also has been improved in practice: from direct contact co-cultures to indirect contact co-cultures, from two-dimensional co-cultures to three-dimensional co-cultures. Cell co-culture method is closer to the human body. It is also more advantageous to study the interaction among cells. Nowadays, there are more researchers tend to select this method to study the physiological and pathological in vitro model, tissue engineering, and cell differentiation research. At the same time, it has become the focus of drug research and development, drug analysis, mechanism of drug action, and drug targets. This article will review the studies of cell co-culture method, summarize advantages and disadvantages of various methods, so as to promote improvement of cell culture methods, to build cells co-culture system that more close to human body, and build the in vitro model that simulate internal circulation of human body further.

  10. 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.

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

    Directory of Open Access Journals (Sweden)

    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.

  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. 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

    predicting the presence of Legionella in the cooling towers investigated. We observed that the concentration of TAB was significatively higher in the positive samples for Legionella than in the negative ones, which seems to indicate that TAB is a good tool for evaluating the effectiveness of the Autocontrol Safety Measures of those cooling towers. Our results reinforce the need for controlling the levels of TAB (at 36 ± 1 oC because it is a good indicator of the presence of Legionella spp. despite the fact that the standard established as the action limit should be lowered. Within the study we also reviewed the sampling procedures and provide new data on the incubation period of the disease and on the genotyping methods classically employed in epidemiological investigation.

  15. 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.

  16. 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...

  17. Mouse cell culture - Methods and protocols

    Directory of Open Access Journals (Sweden)

    CarloAlberto Redi

    2010-12-01

    Full Text Available The mouse is, out of any doubt, the experimental animal par excellence for many many colleagues within the scientific community, notably for those working in mammalian biology (in a broad sense, from basic genetic to modeling human diseases, starting at least from 1664 Robert Hooke experiments on air’s propertyn. Not surprising then that mouse cell cultures is a well established field of research itself and that there are several handbooks devoted to this discipline. Here, Andrew Ward and David Tosh provide a necessary update of the protocols currently needed. In fact, nearly half of the book is devoted to stem cells culture protocols, mainly embryonic, from a list of several organs (kidney, lung, oesophagus and intestine, pancreas and liver to mention some........

  18. Early detection of mycobacteria using a novel hydrogel culture method.

    Science.gov (United States)

    Jang, Mi Hee; Kim, Shine Young; Kim, Chang-Ki; Hwang, Sang-Hyun; Park, Byung Kyu; Kim, Sung Soo; Lee, Eun Yup; Chang, Chulhun L

    2014-01-01

    Early laboratory detection of Mycobacterium tuberculosis is crucial for controlling tuberculosis. We developed a hydrogel mycobacterial culture method that retains the advantages of both solid and liquid methods in terms of speed, cost, and efficiency. Mycobacterium bovis bacillus Calmette-Guérin (BCG) suspensions and 200 acid-fast bacilli (AFB)-positive clinical specimens were inoculated in Middlebrook 7H9 liquid media (Becton-Dickinson and Company, USA) and mixed with 75 µL of 9-fluorenylmethoxycarbonyl (Fmoc)-Phe-Phe-OH hydrogel stock solution in an Eppendorf tube just before culture incubation. The mixtures were cultured at 37℃ for as long as 14 days to monitor culture status. The number of M. bovis BCG increased with time. For 200 AFB smear-positive specimens, 155 of 158 conventional culture-positive specimens and 4 culture-negative or contaminated specimens yielded positive cultures within 14 days. For 128 specimens positive with the liquid culture method, the time to positive culture using the hydrogel method (mean, 12.6 days; range, 7 to 14 days) was significantly shorter than that for conventional liquid culture (mean, 16.2 days; range, 6 to 31 days; Pculture system is useful for timely, economical, and efficient detection of mycobacteria in clinical specimens.

  19. The awareness of employees in safety culture through the improved nuclear safety culture evaluation method

    International Nuclear Information System (INIS)

    Kim, Young Ga; Sung, Chan Ho; Jung, Yeon Sub

    2012-01-01

    After the Chernobyl nuclear accident in 1986, nuclear safety culture terminology was at first introduced emphasizing the importance of employees' attitude and organizational safety. The concept of safety culture was spread by INSAG 4 published in 1991. From that time, IAEA had provided the service of ASCOT for the safety culture assessment. However, many people still are thinking that safety culture is abstract and is not clear. It is why the systematic and reliable assessment methodology was not developed. Assessing safety culture is to identify what is the basic assumption for any organization to accept unconsciously. Therefore, it is very difficult to reach a meaningful conclusion by a superficial investigation alone. KHNP had been doing the safety culture assessment which was based on ASCOT methodology every 2 years. And this result had contributed to improving safety culture. But this result could not represent the level of organization's safety culture due to the limitation of method. So, KHNP has improved the safety culture method by benchmarking the over sea assessment techniques in 2011. The effectiveness of this improved methodology was validated through a pilot assessment. In this paper, the level of employees' safety culture awareness was analyzed by the improved method and reviewed what is necessary for the completeness and objectivity of the nuclear safety culture assessment methodology

  20. 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.

  1. 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) ...

  2. 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) ...

  3. 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 ...

  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. 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. An alternative method for Plasmodium culture synchronization.

    Science.gov (United States)

    Lelièvre, J; Berry, A; Benoit-Vical, F

    2005-03-01

    Since the synchronization of Plasmodium falciparum has become an essential tool in research, we have investigated the use of a commercial gelatine solution, Plasmion, to replace Plasmagel, which is now difficult to obtain. This method also avoids the use of techniques based on Percoll-glucose gradients. The Plasmion-based technique proved to be a good method and could become an alternative to Plasmagel.

  7. 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.

  8. Rapid method for culturing embryonic neuron-glial cell cocultures

    DEFF Research Database (Denmark)

    Svenningsen, Åsa Fex; Shan, Wei-Song; Colman, David R

    2003-01-01

    A streamlined, simple technique for primary cell culture from E17 rat tissue is presented. In an attempt to standardize culturing methods for all neuronal cell types in the embryo, we evaluated a commercial medium without serum and used similar times for trypsinization and tested different surfaces...

  9. Tissue-Culture Method of Cloning Rubber Plants

    Science.gov (United States)

    Ball, E. A.

    1983-01-01

    Guayule plant, a high-yield rubber plant cloned by tissue-culture method to produce multiple new plants that mature quickly. By adjusting culture medium, excised shoot tip produces up to 50 identical guayule plants. Varying concentration of cytokinin, single excised tip produces either 1 or several (up to 50) new plants.

  10. 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

  11. The effect of Montessori Method on teaching cultural and creative ...

    African Journals Online (AJOL)

    The Effect of the Montessori Method on teaching was investigated among children to discover their artistic development in Zaria, Kaduna State. The problem of the study is that the Montessori Method on teaching cultural and creative arts is not adequately explored in the primary schools, while other teaching methods used, ...

  12. Intercultural and Cross-Cultural Communication Research: Some Reflections about Culture and Qualitative Methods

    Directory of Open Access Journals (Sweden)

    Maria Assumpta Aneas

    2009-01-01

    Full Text Available This article attempts to offer a response, from a general perspective, to the question of how culture reveals itself in the application of qualitative research methods in intercultural communication. When we use the term "culture" it is important to bear in mind that culturally attributed social interaction processes are themselves the result of socially constructed processes. They are part of an individual-collective dialectic with multiple potential meanings, which are emergent and in constant reformulation from a wide variety of social and cultural perspectives. Much of the recent research in intercultural communication has been directed towards the study of these systems of culturally related meanings. The literature we review offers perspectives from a variety of disciplines and insights into the role of culture in communication processes. URN: urn:nbn:de:0114-fqs0901519

  13. 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.

  14. 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.

  15. 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.

  16. 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.

  17. 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).

  18. Method and system of culturing an algal mat

    Science.gov (United States)

    Das, Keshav C; Cannon, Benjamin R; Bhatnagar, Ashish; Chinnasamy, Senthil

    2014-05-13

    A system and method for culturing algae are presented. The system and method utilize a fog of growth medium that is delivered to an algal mat generator along with a stream of CO.sub.2 to promote growth of algal cells contained in the generator.

  19. 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.

  20. Improved Cell Culture Method for Growing Contracting Skeletal Muscle Models

    Science.gov (United States)

    Marquette, Michele L.; Sognier, Marguerite A.

    2013-01-01

    An improved method for culturing immature muscle cells (myoblasts) into a mature skeletal muscle overcomes some of the notable limitations of prior culture methods. The development of the method is a major advance in tissue engineering in that, for the first time, a cell-based model spontaneously fuses and differentiates into masses of highly aligned, contracting myotubes. This method enables (1) the construction of improved two-dimensional (monolayer) skeletal muscle test beds; (2) development of contracting three-dimensional tissue models; and (3) improved transplantable tissues for biomedical and regenerative medicine applications. With adaptation, this method also offers potential application for production of other tissue types (i.e., bone and cardiac) from corresponding precursor cells.

  1. 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...

  2. 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.

  3. 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.

  4. 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.

  5. Comparison of Nested-PCR technique and culture method in ...

    African Journals Online (AJOL)

    USER

    2010-04-05

    Apr 5, 2010 ... tuberculosis (GUTB) compared with acid fast staining and culture method. In total ... tuberculosis; PCR, polymerase chain reaction; MTB,. Mycobacterium tuberculosis;. EPTB, extrapulmonary tuberculosis; AFB, acid-fast bacilli; SDS, sodium ..... Dingtoumda B, Diallo B, Defer MC, Sombié I, Zanetti S, Sechi LA.

  6. Effect of integration of cultural, botanical, and chemical methods of ...

    African Journals Online (AJOL)

    A field experiment was conducted from November 2011 to June 2013 to evaluate the effects of botanical, cultural, and chemical methods on termite colony survival, crop and wooden damage, and other biological activities in Ghimbi district of western Ethiopia. The termite mounds were dug and the following treatments were ...

  7. Methods of epithelial tissue culture in albino rabbit skin

    Directory of Open Access Journals (Sweden)

    Anarluki J

    1998-05-01

    Full Text Available With the intention of research of various methods of epithelial tissue culture we've studied five French Albino rabbits with an average of 8 weeks. In order to evaluate and control growth and proliferation of autologus cultured tissue samples were obtained on 1st, 5th and 8th days. After fixation of these samples and passing them through various processes, histologic sections were prepared. These sections were stained with H-E and studied by light microscope, we succeeded in developing the original donor surface by 18 times.

  8. 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.

  9. 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.

  10. 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.

  11. 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.

  12. Red porgy Pagrus pagrus:ontogeny and culture methods

    OpenAIRE

    Andrade, Carlos Alberto Pestana, 1962-

    2012-01-01

    Tese de doutoramento, Biologia (Biologia Marinha e Aquacultura), Universidade de Lisboa, Faculdade de Ciências, 2012 Red porgy is a candidate species for the diversification of aquaculture production (Chapter 1). Different larvae ontogenetic stages and associated culture methods are critical to larvae development and survival, being considered as hatchery production bottlenecks and investigated in the course of the thesis. Larvae rearing trials were conducted using mesocosm semi-intensive ...

  13. Methods to Improve Cultural Communication Skills in Special Operations Forces

    National Research Council Canada - National Science Library

    Wise, J

    1998-01-01

    .... Three culture-specific templates were developed, each describing a culture and identifying culture-specific behaviors that were prioritized through focus groups of cultural informants and clustering analysis...

  14. Socio-cultural Foundations of the Development of the Education System. The Socio-cultural Situation Method

    OpenAIRE

    Anatoliy Tsirul'nikov

    2009-01-01

    This article deals with the method of Socio-cultural Situation (SCS) Analysis. This method is used to design development strategies and models of school education in districts and cities. Four classic types of SCS are determined: a school in a cultural center, a school in a former cultural center, a school in a potential cultural center, a school in a cultural desert. A number of intermediate complex SCS are described: a socio-cultural break, an unidentifi ed pedagogical object, a rural schoo...

  15. 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

  16. 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.

  17. 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.

  18. 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 ...

  19. 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...

  20. Diversity of endophytic bacteria of Dendrobium officinale based on culture-dependent and culture-independent methods

    Directory of Open Access Journals (Sweden)

    Cong Pei

    2017-01-01

    Full Text Available Culture-dependent and culture-independent methods were compared and evaluated in the study of the endophytic diversity of Dendrobium officinale. Culture-independent methods consisted of polymerase chain reaction–denaturing gradient gel electrophoresis (PCR-DGGE and metagenome methods. According to the results, differences were found between the three methods. Three phyla, namely Firmicutes, Proteobacteria, and Actinobacteria, were detected using the culture-dependent method, and two phyla, Firmicutes and Proteobacteria, were detected by the DGGE method. Using the metagenome method, four major phyla were determined, including Proteobacteria (76.54%, Actinobacteria (18.56%, Firmicutes (2.27%, and Bacteroidetes (1.56%. A distinct trend was obtained at the genus level in terms of the method and the corresponding number of genera determined. There were 449 genera and 16 genera obtained from the metagenome and DGGE methods, respectively, and only 7 genera were obtained through the culture-dependent method. By comparison, all the genera from the culture-dependent and DGGE methods were contained in the members determined using the metagenome method. Overall, culture-dependent methods are limited to ‘finding’ endophytic bacteria in plants. DGGE is an alternative to investigating primary diversity patterns; however, the metagenome method is still the best choice for determining the endophytic profile in plants. It is essential to use multiphasic approaches to study cultured and uncultured microbes.

  1. Electron microscopy methods in studies of cultural heritage sites

    Energy Technology Data Exchange (ETDEWEB)

    Vasiliev, A. L., E-mail: a.vasiliev56@gmail.com; Kovalchuk, M. V.; Yatsishina, E. B. [National Research Centre “Kurchatov Institute” (Russian Federation)

    2016-11-15

    The history of the development and application of scanning electron microscopy (SEM), transmission electron microscopy (TEM), and energy-dispersive X-ray microanalysis (EDXMA) in studies of cultural heritage sites is considered. In fact, investigations based on these methods began when electron microscopes became a commercial product. Currently, these methods, being developed and improved, help solve many historical enigmas. To date, electron microscopy combined with microanalysis makes it possible to investigate any object, from parchment and wooden articles to pigments, tools, and objects of art. Studies by these methods have revealed that some articles were made by ancient masters using ancient “nanotechnologies”; hence, their comprehensive analysis calls for the latest achievements in the corresponding instrumental methods and sample preparation techniques.

  2. Electron microscopy methods in studies of cultural heritage sites

    International Nuclear Information System (INIS)

    Vasiliev, A. L.; Kovalchuk, M. V.; Yatsishina, E. B.

    2016-01-01

    The history of the development and application of scanning electron microscopy (SEM), transmission electron microscopy (TEM), and energy-dispersive X-ray microanalysis (EDXMA) in studies of cultural heritage sites is considered. In fact, investigations based on these methods began when electron microscopes became a commercial product. Currently, these methods, being developed and improved, help solve many historical enigmas. To date, electron microscopy combined with microanalysis makes it possible to investigate any object, from parchment and wooden articles to pigments, tools, and objects of art. Studies by these methods have revealed that some articles were made by ancient masters using ancient “nanotechnologies”; hence, their comprehensive analysis calls for the latest achievements in the corresponding instrumental methods and sample preparation techniques.

  3. 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.

  4. Culture-independent methods for identifying microbial communities in cheese.

    Science.gov (United States)

    Jany, Jean-Luc; Barbier, Georges

    2008-10-01

    This review focuses on the culture-independent methods available for the description of both bacterial and fungal communities in cheese. Important steps of the culture-independent strategy, which relies on bulk DNA extraction from cheese and polymerase chain reaction (PCR) amplification of selected sequences, are discussed. We critically evaluate the identification techniques already used for monitoring microbial communities in cheese, including PCR-denaturing gradient gel electrophoresis (PCR-DGGE), PCR-temporal temperature gradient gel electrophoresis (PCR-TTGE) or single-strand conformation polymorphism-PCR (SSCP-PCR) as well as some other techniques that remain to be adapted to the study of cheese communities. Further, our analysis draws attention to the lack of data available on suitable DNA sequences for identifying fungal communities in cheese and proposes some potential DNA targets.

  5. Comparison of PCR method with the culture method for identification of gonococci from endocervical swabs

    Directory of Open Access Journals (Sweden)

    Alam A

    2002-01-01

    Full Text Available Gonococcal infection remains still a major cause of morbidity among sexually active individuals. Diagnosis of the infection in a female case is more difficult than that in a male. This was a prospective study among 269 female commercial sex workers (CSWs to screen them for gonococcal infection, comparing the rapid method of identification of gonococci by polymerase chain reaction (PCR with the selective culture method. A total of 92 (34.2% CSWs were identified positive for Neisseria gonorrhoeae by combination of the two methods. The PCR method identified 87 of the specimens to harbour cppB gene of N. gonorrhoeae, whereas culture method identified 83 specimens showing colonies of gonococci. Taking into consideration of the total positive cases (92, the PCR method showed a sensitivity of 94.57%, whereas sensitivity of culture method was 90.22%. The selective culture method appears to be the most applicable in the identification of gonococci from clinical specimens, particularly in the less resourceful countries like Bangladesh.

  6. 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

    to monitor all Legionella species in qualitative and quantitative terms if a spiked-in genome standard is used to calibrate the method. Overall, the genus-specific NGS approach opens up a new avenue to massive parallel diagnostics in a quantitative, specific and sensitive way.

  7. Understanding Global / Local Cultural Leadership : Issues and Methods

    NARCIS (Netherlands)

    Kolsteeg, Johan

    2017-01-01

    Cultural leaders sail between the Scylla and Charibdis of aggregated trans- and supranational cultural-political discourses and the cultural needs of local communities. How do these dynamics influence the work of cultural leaders? How can we understand the work of cultural leaders to connect

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

    Directory of Open Access Journals (Sweden)

    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

  9. A new method to culture sweetpotato in space farming

    Science.gov (United States)

    Tsuyuki, I.; Ishii, Y.; Oda, M.; Kitaya, Y.; Mori, G.

    Sweetpotato production in space has many advantages over that of other crops; the plant has a higher growth rate and a higher yield with less fertilizer and less water, and functions as an efficient CO_2/O_2 converter. In a limited space in space farming, however, it is not favorable that sweetpotato shoots develop vigorously while the roots have not enlarged yet, because the sweetpotato organ of interest is not the shoot but the tuberous root. Cuttings of sweetpotato (Ipomoea batatas Lam. "Beniazuma") were used in this study. Each cutting was cut off from the 2nd - 10th nodes from the apices of mother branches and consisted of one expanded leaf, one node and five cm long stem. The cuttings were cultured suboptimally on a mixed soil (peat-moss:vermiculite=1:1 in volume) in a greenhouse under sunlight. Growth characteristics of the cuttings removed axillary buds were compared with cuttings with axillary buds in the first experiment. The cuttings without axillary buds started tuberous root bulking about 30 days after the onset of the experiment. The harvest index (tuberous root dry mass/total dry mass) was 0.5 after 70 days. Whereas, the control plant with an axillary bud developed a lateral shoot and formed no tuberous root during 70 days in the experiment. It was necessary to remove the axillary buds in order to form the tuberous roots in this method. To evaluate the effect of light intensity on tuberous root formation, cuttings without axillary buds were shaded with cheesecloth having 43% of light transmittance in the second experiment. The tuberous root formation was retarded 50 days in shaded cuttings compared with control cuttings. The tuberous roots were quickly formed and the large harvest index was ensured in this method with cuttings without axillary buds. Therefore the method is expected to be advantageous to culture sweetpotato at a high density with rapid turn over in a limited culture space in space farming.

  10. 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.

  11. 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.

  12. [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.

  13. Participatory Methods and UCA Project: understanding technologies as culture

    Directory of Open Access Journals (Sweden)

    Magda Pischetola

    2015-12-01

    Full Text Available Abstract In the complex and changing context of digital culture, the media become an important space of relation, as they have the crucial role of articulating new cultural logics that lead to disruptions in the school environment. To understand this change, new methods of analysis and research have been created, the so-called Participatory Methodologies. They are action research strategies aimed at intervening in a given social situation. In the analysis proposed here, such methodologies will help us to address the challenge of involving digital technologies in school culture, through the participation of different individuals involved. Two qualitative case studies about the project Um Computador por Aluno – the Brazilian One Laptop per Child -, carried out in 2012 in the schools of Santa Catarina and Bahia, are the first of two phases of the research presented. The results concern a "vertical" form of technology insertion in schools, which led to frustration and de-motivation at several levels. Starting from these considerations, the second stage of research proposes a pedagogical intervention in one of four schools in the field. The methodologies of participatory video and photography are chosen as possibilities of action-reflection-action on the sociocultural reality of students through the experience of sharing. The results show the importance of carrying out creative activities, appropriate to a social conception of learning, as well as the centrality of children and youth as agency and a broader need to redefine the relationship between teacher and student, in a more "horizontal" perspective process of teaching and learning. Keywords: Projeto UCA. Participatory Research Method. Innovative teaching-learning.

  14. 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.

  15. 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...

  16. 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.

  17. 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 ...

  18. 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....

  19. 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.

  20. 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.

  1. 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.

  2. 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.

  3. 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.

  4. Legionella pneumophila serogroup 3 pneumonia in a patient with low-grade 4 non-Hodgkin lymphoma: a case report

    Directory of Open Access Journals (Sweden)

    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.

  5. 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.

  6. 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.

  7. 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.

  8. 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.

  9. 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...

  10. 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

  11. 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 ...

  12. 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%.

  13. 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%.

  14. 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...

  15. Two years' performance of an in-house ELISA for diagnosis of Legionnaires' disease: detection of specific IgM and IgG antibodies against Legionella pneumophila serogroup 1, 3 and 6 in human serum.

    Science.gov (United States)

    Elverdal, P L; Jørgensen, C S; Krogfelt, K A; Uldum, S A

    2013-08-01

    The aim of this study was to evaluate the performance of an in-house ELISA for the diagnosis of Legionnaires' disease (LD) by detection of IgM and IgG antibodies against Legionella (L.) pneumophila serogroups (sg) 1, 3 and 6. The evaluation was done throughout a two-year period in a diagnostic routine laboratory. Furthermore, the sensitivity of four different methods, the in-house L. pneumophila antibody test (ELISA), the urinary antigen test (Binax® EIA), an in-house PCR and culture, both alone and in combination was evaluated. From 2008 to 2010, 12,158 serum samples from 10,503 patients were analysed. During the same period, 361 cases of laboratory-confirmed LD cases were recorded in Denmark, but of these only 113 had a serum sample examined. The positive predictive value of the in-house ELISA was calculated to be 12.8 and the negative predictive value was 99.6, using only the confirmed LD cases as true positives. The sensitivity of the in-house ELISA for the detection of IgM and IgG antibodies in the confirmed LD cases was 61% and 36%, respectively. By combining the two ELISA assays the sensitivity increased to 66%. The sensitivity of the Legionella urinary antigen test (Binax® EIA) was 63%, of the in-house PCR 87% and of culture 69%. When all the different methods were combined, a higher sensitivity was calculated--for in-house ELISA (IgM+IgG) and Binax® EIA 91%, in-house ELISA (IgM+IgG) and in-house PCR 93%, in-house ELISA (IgM+IgG) and culture 93%, Binax® EIA and in-house PCR 79%, Binax® EIA and culture 68% and in-house PCR and culture 94%. This study confirms that the detection of IgG and IgM antibodies by ELISA is an important diagnostic tool, also during the initial phase of the disease. Furthermore, we showed that LD in Denmark with or without serum samples collected exhibits the same age and sex distribution and epidemiology, as in the rest of Europe, i.e., mostly men are infected, infections are mostly community acquired, followed by infection from

  16. Methods for the Organogenesis of Skeletal Muscle in Tissue Culture

    Science.gov (United States)

    Vandenburgh, Herman; Shansky, Janet; DelTatto, Michael; Chromiak, Joseph

    1997-01-01

    Skeletal muscle structure is regulated by many factors, including nutrition, hormones, electrical activity, and tension. The muscle cells are subjected to both passive and active mechanical forces at all stages of development and these forces play important but poorly understood roles in regulating muscle organogenesis and growth. For example, during embryogenesis, the rapidly growing skeleton places large passive mechanical forces on the attached muscle tissue. These forces not only help to organize the proliferating mononucleated myoblasts into the oriented, multinucleated myofibers of a functional muscle but also tightly couple the growth rate of muscle to that of bone. Postnatally, the actively contracting, innervated muscle fibers are subjected to different patterns of active and passive tensions which regulate longitudinal and cross sectional myofiber growth. These mechanically-induced organogenic processes have been difficult to study under normal tissue culture conditions, resulting in the development of numerous methods and specialized equipment to simulate the in vivo mechanical environment.These techniques have led to the "engineering" of bioartificial muscles (organoids) which display many of the characteristics of in vivo muscle including parallel arrays of postmitotic fibers organized into fascicle-like structures with tendon-like ends. They are contractile, express adult isoforms of contractile proteins, perform directed work, and can be maintained in culture for long periods. The in vivo-like characteristics and durability of these muscle organoids make them useful for long term in vitro studies on mechanotransduction mechanisms and on muscle atrophy induced by decreased tension. In this report, we described a simple method for generating muscle organoids from either primary embrionic avain or neonatal rodent myoblasts.

  17. 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.

  18. 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)

  19. 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.

  20. 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.

  1. 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 .

  2. Utilizing the Project Method for Teaching Culture and Intercultural Competence

    Science.gov (United States)

    Euler, Sasha S.

    2017-01-01

    This article presents a detailed methodological outline for teaching culture through project work. It is argued that because project work makes it possible to gain transferrable and applicable knowledge and insight, it is the ideal tool for teaching culture with the aim of achieving real intercultural communicative competence (ICC). Preceding the…

  3. Research on fuzzy comprehensive assessment method of nuclear power plant safety culture

    International Nuclear Information System (INIS)

    Xiang Yuanyuan; Chen Xukun; Xu Rongbin

    2012-01-01

    Considering the traits of safety culture in nuclear plant, 38 safety culture assessment indexes are established from 4 aspects such as safety values, safety institution, safety behavior and safety sub- stances. Based on it, a comprehensive assessment method for nuclear power plant safety culture is constructed by using AHP (Analytic Hierarchy Process) approach and fuzzy mathematics. The comprehensive assessment method has the quality of high precision and high operability, which can support the decision making of safety culture development. (authors)

  4. 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.

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

    Directory of Open Access Journals (Sweden)

    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

  6. Transnational Cultural Leadership as a Situated Practice : Dilemmas and Methods

    NARCIS (Netherlands)

    Kolsteeg, Johan

    2017-01-01

    In a longitudinal transnational research project, a network of European research institutions and field organisations aims to understand how cultural managers mediate global and local pressures concerning creative autonomy, economy and ideology. Among the research questions are which variables

  7. 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.

  8. Individualism-Collectivism and Power Distance Cultural Dimensions: How Each Influences Parental Disciplinary Methods

    Science.gov (United States)

    Schwab, Karen Walker

    2013-01-01

    This paper is a literature review using the Douglas-Widavasky Grid/Group theory as a framework to examine, from a cross cultural perspective, preferred parental disciplinary methods. The four rival cultures defined in the Grid/Group theory mirror the cultural dimensions of individualism-collectivism and power distance described by Geert Hofstede.…

  9. Multiple Contexts, Multiple Methods: A Study of Academic and Cultural Identity among Children of Immigrant Parents

    Science.gov (United States)

    Urdan, Tim; Munoz, Chantico

    2012-01-01

    Multiple methods were used to examine the academic motivation and cultural identity of a sample of college undergraduates. The children of immigrant parents (CIPs, n = 52) and the children of non-immigrant parents (non-CIPs, n = 42) completed surveys assessing core cultural identity, valuing of cultural accomplishments, academic self-concept,…

  10. Methods and Techniques of Sampling, Culturing and Identifying of Subsurface Bacteria

    International Nuclear Information System (INIS)

    Lee, Seung Yeop; Baik, Min Hoon

    2010-11-01

    This report described sampling, culturing and identifying of KURT underground bacteria, which existed as iron-, manganese-, and sulfate-reducing bacteria. The methods of culturing and media preparation were different by bacteria species affecting bacteria growth-rates. It will be possible for the cultured bacteria to be used for various applied experiments and researches in the future

  11. Cultural Continuity in an Educational Institution: A Case Study of the Suzuki Method of Music Instruction.

    Science.gov (United States)

    Taniuchi, Lois

    As the Suzuki method of music instruction has spread from Japan to other countries, its methods have been modified to adapt to the culture of those countries. In this paper the Japanese cultural background, and the principles and methods developed in Japan are discussed and compared with the adaptations made in the United States. The Suzuki method…

  12. 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.

  13. 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

  14. 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...

  15. 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...

  16. 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.

  17. 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...

  18. 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

  19. Development of an Evaluation Method for Team Safety Culture Competencies using Social Network Analysis

    International Nuclear Information System (INIS)

    Han, Sang Min; Kim, Ar Ryum; Seong, Poong Hyun

    2016-01-01

    In this study, team safety culture competency of a team was estimated through SNA, as a team safety culture index. To overcome the limit of existing safety culture evaluation methods, the concept of competency and SNA were adopted. To estimate team safety culture competency, we defined the definition, range and goal of team safety culture competencies. Derivation of core team safety culture competencies is performed and its behavioral characteristics were derived for each safety culture competency, from the procedures used in NPPs and existing criteria to assess safety culture. Then observation was chosen as a method to provide the input data for the SNA matrix of team members versus insufficient team safety culture competencies. Then through matrix operation, the matrix was converted into the two meaningful values, which are density of team members and degree centralities of each team safety culture competency. Density of tem members and degree centrality of each team safety culture competency represent the team safety culture index and the priority of team safety culture competency to be improved

  20. A Method to Preclude Moisture Condensation in Plated Tissue Cultures

    Science.gov (United States)

    Alex M. Diner

    1992-01-01

    Excessive condensate normally accumulates in in vitro-illuminated petri dishes containing plant tissue cultures, causing avariety of problems. A dark-colored rubber net-mesh placed over the petri dishes prevented such condensation, even when charcoal-supplemented media are used under high light intensity in a growth chamber.

  1. 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.

  2. [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.

  3. 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...

  4. 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...

  5. The method of rodent whole embryo culture using the rotator-type bottle culture system.

    Science.gov (United States)

    Takahashi, Masanori; Osumi, Noriko

    2010-08-28

    Whole embryo culture (WEC) technique has been developed in 1950's by New and his colleagues, and applied for developmental biology (1). Although development and growth of mammalian embryos are critically dependent on the function of the placenta, WEC technique allows us to culture mouse and rat embryos ex vivo condition during limited periods corresponding to midgestation stages during embryonic day (E) 6.5-E12.5 in the mouse or E8.5-E14.5 in the rat (2, 3, 4). In WEC, we can directly target desired areas of embryos using fine glass capillaries because embryos can be manipulated under the microscope. Therefore, rodent WEC is very useful technique when we want to study dynamic developmental processes of postimplanted mammalian embryos. Up to date, several types of WEC systems have been developed (1). Among those, the rotator-type bottle culture system is most popular and suitable for long-term culture of embryos at midgestation, i.e., after E9.5 and E11.5 in the mouse and rat, respectively (1). In this video protocol, we demonstrate our standard procedures of rat WEC after E12.5 using a refined model of the original rotator system, which was designed by New and Cockroft (5, 6), and introduce various applications of WEC technique for studies in mammalian developmental biology.

  6. 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

  7. 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.

  8. 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.

  9. [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.

  10. Using case methods to study cultural diversity within the development of telematic systems

    DEFF Research Database (Denmark)

    Nøhr, Christian; Bertelsen, Pernille Scholdan; Brender, Jytte

    The report " Using case methods to study cultural diversity within the development of telematic systems" discusses a case study method which is an extension of work orginallly done in Babel, a 5th framework EU project. The report contributes to the discussion identifying operational cultural...

  11. Characterization of microbes in prosthetic joint specimens by culture-independent molecular methods

    DEFF Research Database (Denmark)

    Xu, Yijuan; Rudkjøbing, Vibeke Børsholt; Simonsen, Ole

    in situ hybridization (FISH). Concomitant samples were cultured by standard methods. Overall, the results of culture-based and molecular methods showed concordant results for 13 patients and discrepant results for 6 patients. In the remaining cases, culture methods identified one species or a group......, and polymicrobial communities were found in 9 patients. Culture-based methods detected bacteria in 7 patients, and multiple species were only isolated from one of them. Additionally, quantification of Propionibacterium and Staphylococcus aureus using specific qPCR assays confirmed the findings by clone library...... was to investigate the microbial diversity in surgical samples (eg. synovial fluid, periprosthetic tissue, removed prosthesis) from 22 prosthetic patients using a range of culture-independent molecular methods including broad range 16S rRNA gene PCR, cloning, phylogeny, quantitative PCR (qPCR), and fluorescence...

  12. 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...

  13. 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.

  14. Survey of blood cultures methods in Italy in 2010

    Directory of Open Access Journals (Sweden)

    Antonio Goglio

    2011-09-01

    Full Text Available Sepsis is a serious clinical condition, associated with high mortality despite advanced modern medical treatment. Traditionally, the detection and identification of bacteria and fungi circulating in the blood-stream is based on blood cultures. A number of factors influence the yield of blood culture, most of them concerning the microbiologist skill and the laboratory organization. In order to collect information about the practices and procedures used for the detection of microrganisms in blood cultures in the italian laboratory (lab, an e-mail with the invitation to participate in the survey was sent to 2000 members of the Italian Association of Clinical Microbiology. Responses were received from 100 lab, located from all over the country (in 18/20 italian regions. The results presented hereby concern specimen collection, culture techniques, rapid identification and susceptibility testing, laboratory organization, relationships with physicians. In summary, most lab use automated systems (96%, the bottles are incubated immediately during public holidays in 72/96 lab (75% and in 49/97 lab at night (50.5%, the lenght of incubation was 5 or 7 days in 93% of the lab, although it is common to extend the incubation period when brucellosis (74 lab, endocarditis (49 lab, systemic mycosis (33 lab is suspected. A wide variety of media are employed for subcultures. All lab process the positive bottles at least once a day, while only in 42 of 81 (51.9% lab the positive blood are processed on holiday. Communication between clinicians and microbiologist include: distribution of specimen collection guidelines (96/100 lab, availability to microbiologist of patients’ clinical situation (77/96 lab, 80.2%, and adding to report the microbiologist’ suggestion (75/98 lab, 76.5%. The results, compared with those collected with a similar questionnaire in 2001, show a greater adherence to guidelines: the number of bottles examined by lab yearly is almost doubled

  15. Comparison of Standard Culture-Based Method to Culture-Independent Method for Evaluation of Hygiene Effects on the Hand Microbiome.

    Science.gov (United States)

    Zapka, C; Leff, J; Henley, J; Tittl, J; De Nardo, E; Butler, M; Griggs, R; Fierer, N; Edmonds-Wilson, S

    2017-03-28

    Hands play a critical role in the transmission of microbiota on one's own body, between individuals, and on environmental surfaces. Effectively measuring the composition of the hand microbiome is important to hand hygiene science, which has implications for human health. Hand hygiene products are evaluated using standard culture-based methods, but standard test methods for culture-independent microbiome characterization are lacking. We sampled the hands of 50 participants using swab-based and glove-based methods prior to and following four hand hygiene treatments (using a nonantimicrobial hand wash, alcohol-based hand sanitizer [ABHS], a 70% ethanol solution, or tap water). We compared results among culture plate counts, 16S rRNA gene sequencing of DNA extracted directly from hands, and sequencing of DNA extracted from culture plates. Glove-based sampling yielded higher numbers of unique operational taxonomic units (OTUs) but had less diversity in bacterial community composition than swab-based sampling. We detected treatment-induced changes in diversity only by using swab-based samples ( P microbiome studies. On the basis of our results and previously published studies, we propose recommendations for best practices in hand microbiome research. IMPORTANCE The hand microbiome is a critical area of research for diverse fields, such as public health and forensics. The suitability of culture-independent methods for assessing effects of hygiene products on microbiota has not been demonstrated. This is the first controlled laboratory clinical hand study to have compared traditional hand hygiene test methods with newer culture-independent characterization methods typically used by skin microbiologists. This study resulted in recommendations for hand hygiene product testing, development of methods, and future hand skin microbiome research. It also demonstrated the importance of inclusion of skin physiological metadata in skin microbiome research, which is atypical for skin

  16. A method for isolating identifying and culturing of rat trachea-bronchia epithelial cells

    International Nuclear Information System (INIS)

    Cui Fengmei; Su Shibiao; Nie Jihua; Li Bingyan; Tong Jian

    2005-01-01

    Objective: To explore a method for isolating identifying and culturing the rat trachea-bronchia epithelial cells. Methods: The rat trachea-bronchia epithelial cells were isolated by digestion with pronase and brushing with cell brush, identified using confocul and cultured in entire F12 media with no serum. Results: With this method, cells in high purity and high viability could be obtained, and about 10 6 cells per rat. The cells grow well in entire F12 media with no serum. Conclusion: The method is useful for isolating rate trachea-bronchia epithelial cells and the entire F12 media with no serum is effective for culturing. (authors)

  17. 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

  18. Triangulation and the importance of establishing valid methods for food safety culture evaluation.

    Science.gov (United States)

    Jespersen, Lone; Wallace, Carol A

    2017-10-01

    The research evaluates maturity of food safety culture in five multi-national food companies using method triangulation, specifically self-assessment scale, performance documents, and semi-structured interviews. Weaknesses associated with each individual method are known but there are few studies in food safety where a method triangulation approach is used for both data collection and data analysis. Significantly, this research shows that individual results taken in isolation can lead to wrong conclusions, resulting in potentially failing tactics and wasted investments. However, by applying method triangulation and reviewing results from a range of culture measurement tools it is possible to better direct investments and interventions. The findings add to the food safety culture paradigm beyond a single evaluation of food safety culture using generic culture surveys. Copyright © 2017. Published by Elsevier Ltd.

  19. 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.

  20. The Story-Presenting Method: a Method for Constructing Multiple Viewpoints to Understand Different Cultures.

    Science.gov (United States)

    Watanabe, Tadaharu

    2017-09-01

    This study will show the results of four dialogical cultural exchange classes, which were held between Japanese and Chinese high school students, and examine the shifts in students' viewpoints and changes in cultural understandings that occurred during those classes. In the first cultural exchange class, students of both countries read a story which described an older student who carelessly wore a T-shirt inside out, and younger students passed by without greeting him. Students of both countries were then asked to write their comments about it. From the second to the fourth class, students discussed the story with each other through exchanging their comments. By presenting another story, which introduced the viewpoint of a third person, and asking them questions that allowed them to reflect on their lives, students also experienced four different viewpoints during these cultural exchange classes. At the beginning of the cultural exchange, students of both countries tended to focus on the similarities in each other's comments, which led to the closing down of the discussion. However, through discussions and experiencing the four different viewpoints, they found there are some essential differences between them around 'ways of greeting' and 'hierarchical relationships between older and younger students', which motivated them to understand their counterparts' culture. Moreover, in the last comments of these cultural exchange classes, it was found that they acquired the viewpoints of cultural others. Given the results of these classes, it is shown that it is effective to present various stories to stimulate cultural understanding.

  1. 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.

  2. Perceptions of Organizational Culture of a Multi-Campus Community College District: Mixed Methods in Concert

    Science.gov (United States)

    Kuster Dale, Kimberly

    2012-01-01

    This concurrent, mixed-methods case study analyzed perceptions of current and preferred organizational culture within a rural, multi-campus community college district. This phenomenon was examined by analyzing and comparing data collected by surveying all full-time employees utilizing the Organizational Culture Assessment Instrument (OCAI) and…

  3. Choice of Appropriate Multimedia Technology and Teaching Methods for Different Culture Groups

    Science.gov (United States)

    Taratoukhina, Julia

    2014-01-01

    This paper describes the prerequisites for development in the area of cross-cultural multimedia didactics. This approach is based on research studies of differences between mentalities, ways of working with educational information, culturally-specific teaching methods and teaching techniques that determine differentiated approaches to the choice…

  4. 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

  5. An Evaluation Method for Team Competencies to Enhance Nuclear Safety Culture

    International Nuclear Information System (INIS)

    Hang, S. M.; Seong, P. H.; Kim, A. R.

    2016-01-01

    Safety culture has received attention in safety-critical industries, including nuclear power plants (NPPs), due to various prominent accidents such as concealment of a Station Blackout (SBO) of Kori NPP unit 1 in 2012, the Sewol ferry accident in 2014, and the Chernobyl accident in 1986. Analysis reports have pointed out that one of the major contributors to the cause of the accidents is ‘the lack of safety culture’. The term, nuclear safety culture, was firstly defined after the Chernobyl accident by the IAEA in INSAG report no. 4, as follows “Safety culture is that assembly of characteristics and attitudes in organizations and individuals which establishes that, as an overriding priority, nuclear plant safety issues receive the attention warranted their significance.” Afterwards, a wide consensus grew among researchers and nuclear-related organizations, that safety culture should be evaluated and managed in a certain manner. Consequently, each nuclear-related organization defined and developed their own safety culture definitions and assessment methods. However, none of these methods provides a way for an individual or a team to enhance the safety culture of an organization. Especially for a team, which is the smallest working unit in NPPs, team members easily overlook their required practices to improve nuclear safety culture. Therefore in this study, we suggested a method to estimate nuclear safety culture of a team, by approaching with the ‘competency’ point of view. The competency is commonly focused on individuals, and defined as, “underlying characteristics of an individual that are causally related to effective or superior performance in a job.” Similar to safety culture, the definition of competency focuses on characteristics and attitudes of individuals. Thus, we defined ‘safety culture competency’ as “underlying characteristics and outward attitudes of individuals that are causally related to a healthy and strong nuclear safety

  6. Towards a culturally independent participatory design method: Fusing game elements into the design process

    DEFF Research Database (Denmark)

    Jensen, Mika Yasuoka; Nakatani, Momoko; Ohno, Takehiko

    2013-01-01

    Historically, Participatory Design (PD) was introduced and applied in the Scandinavian and American context as a practical design method for collective creativity and stakeholder involvement. In this paper, by fusing game elements into PD, we suggest a first step towards a culturally independent PD...... and the created Persona characteristics, probably due to cultural differences. Of more interestingly, despite these process differences, the game elements of our PD variant were found to promote the key values of PD in culturally diverse settings by stimulating idea creation and participation. Our experiments...... imply that the introduction of game elements allows PD to be effectively utilized in culturally diverse settings....

  7. 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.

  8. 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.

  9. 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.

  10. 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.

  11. 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.

  12. Method for organotypic tissue culture in the aged animal

    Directory of Open Access Journals (Sweden)

    Jared Schommer

    2017-01-01

    • Our method permits slices from mice as old as 16 months and rabbits as old as years of age to survive ex vivo up to 8 weeks [6–9]. Such a slice system may be relevant to investigating age-related brain diseases.

  13. Preservation of live cultures of basidiomycetes - Recent methods

    Czech Academy of Sciences Publication Activity Database

    Homolka, Ladislav

    2014-01-01

    Roč. 118, č. 2 (2014), s. 107-125 ISSN 1878-6146 R&D Projects: GA ČR GAP504/12/0709 Institutional support: RVO:61388971 Keywords : Fungi * Maintenance methods * Long-term preservation Subject RIV: EE - Microbiology, Virology Impact factor: 2.342, year: 2014

  14. QUANTIFICATION OF LISTERIA MONOCYTOGENES IN MILK BY MPN-PCR AND MPN-CULTURE METHODS

    Directory of Open Access Journals (Sweden)

    Mahzad Hosseini

    2014-10-01

    Full Text Available The aim of this study was to compare the MPN-PCR (Most Probable Number- Polymerase Chain Reaction and MPN-Culture methods in enumerating of Listeria monocytogenes in milk. In order to compare the accuracy of these methods, 103 cell/ml Listeria monocytogenes and different background bacteria which may be present in raw milk, were inoculated in sterilized milk. After preparing serial dilutions, three replicates per dilution were inoculated in tubes containing listeria enrichment broth. After 48 hours of incubation, for MPN-Culture three inoculated replicates were subcultured on Oxford agar and suspected colonies were confirmed by performing by biochemical tests. For MPN-PCR assay, the DNA extraction was performed from the three inoculated replicates which were already used for MPN-Culture and PCR assay was performed using primers specific for Listeria monocytogenes. The experiment was repeated three times and the average of enumerated bacteria was calculated by each method separately. Statistical analysis using one sample Wilcoxon signed rank test showed that enumeration by MPN-PCR method was more accurate than enumeration by MPN-Culture method. The result of this study showed that MPN-PCR method in comparision with MPN-Culture even in the presence of different background microorganisms is more rapid and reliable. It is concluded that MPN-PCR method facilitates the enumeration of Listeria monocytogenes without excessive work and could be considered as an alternative to MPN-Culture technique.

  15. Tourismological valorization of intangible cultural heritage of Serbia according to the Hilary du Cros method

    Directory of Open Access Journals (Sweden)

    Željko Bjeljac

    2016-02-01

    Full Text Available The folk artistry of Serbia is rich in spiritual values tied to customs, celebrations, music, song, dance, games, stories and legends, and this kind of cultural heritage is presented through numerous festivals, events and tourist manifestations. In 2012, the network for the safeguarding of intangible cultural heritage was formed, comprised of the National committee for intangible cultural heritage, the Commission for admission into the registry of intangible cultural heritage, a network of coordinators and the Center for intangible cultural heritage of Serbia. These institutions have chosen 6 elements of intangible cultural heritage, out of 27 suggestions: the slava, the Đurđevdan ritual, the kolo dance, singing accompanied by gusle, Slovakian naive painting, the custom of making and lighting farmers’ candles, Pirot carpet weaving, and Zlakusa pottery as elements of cultural heritage which reflect the national and cultural identity of the Serbian people, and Slavic minorities. These elements of intangible cultural heritage have a certain tourism potential and can represent an important factor in the forming of the tourist brand of Serbia. In order to determine the importance of the 27 suggestions of intangible cultural elements of Serbia, an analysis was conducted, using an adapted form of the Hilary du Cros method of tourist valorization.

  16. MODIFIED SIMULATION LEARNING METHOD ON KNOWLEDGE AND ATTITUDE OF NURSING STUDEN T’S CULTURAL AWARENESS AT UNIVERSITAS INDONESIA

    Directory of Open Access Journals (Sweden)

    Enie Novieastari

    2012-06-01

    Full Text Available Nursing students should be prepared to be culturally competent nurses. Cultural awareness is believed as the most important elements of cultural competence. The purpose of this article is to describe the effectiveness of Modified Simulation Learning Methods on cultural awareness as one atribute of cultural competence. Aquasi-experimental (control group design was used to explore the relationship between variabels among 98 first year nursing studentattending Basic Nursing Concept I course at Faculty of Nursing Universitas Indonesia. The knowledge of cultural awareness was found statistically different between participants in Modified Simulation Methods group (intervention and participants using the regular method. However, there were no statistical differences in attitude of cultural awareness between intervention and control groups. It could be concluded that Modified Simulation Learning methods is an effective learning method for increasing cultural knowledge of nusing student to be a competent nurse. Further research should be developed in continuing the improvement of cultural competence such as cultural skills.

  17. Microbial Diversity of a Camembert-Type Cheese Using Freeze-Dried Tibetan Kefir Coculture as Starter Culture by Culture-Dependent and Culture-Independent Methods

    Science.gov (United States)

    Mei, Jun; Guo, Qizhen; Wu, Yan; Li, Yunfei

    2014-01-01

    The biochemical changes occurring during cheese ripening are directly and indirectly dependent on the microbial associations of starter cultures. Freeze-dried Tibetan kefir coculture was used as a starter culture in the Camembert-type cheese production for the first time. Therefore, it's necessary to elucidate the stability, organization and identification of the dominant microbiota presented in the cheese. Bacteria and yeasts were subjected to culture-dependent on selective media and culture-independent polymerase chain reaction (PCR)-denaturing gradient gel electrophoresis (DGGE) analysis and sequencing of dominant bands to assess the microbial structure and dynamics through ripening. In further studies, kefir grains were observed using scanning electron microscopy (SEM) methods. A total of 147 bacteria and 129 yeasts were obtained from the cheese during ripening. Lactobacillus paracasei represents the most commonly identified lactic acid bacteria isolates, with 59 of a total of 147 isolates, followed by Lactococcus lactis (29 isolates). Meanwhile, Kazachstania servazzii (51 isolates) represented the mainly identified yeast isolate, followed by Saccharomyces cerevisiae (40 isolates). However, some lactic acid bacteria detected by sequence analysis of DGGE bands were not recovered by plating. The yeast S. cerevisiae and K. servazzii are described for the first time with kefir starter culture. SEM showed that the microbiota were dominated by a variety of lactobacilli (long and curved) cells growing in close association with a few yeasts in the inner portion of the grain and the short lactobacilli were observed along with yeast cells on the exterior portion. Results indicated that conventional culture method and PCR-DGGE should be combined to describe in maximal detail the microbiological composition in the cheese during ripening. The data could help in the selection of appropriate commercial starters for Camembert-type cheese. PMID:25360757

  18. Microbial diversity of a Camembert-type cheese using freeze-dried Tibetan kefir coculture as starter culture by culture-dependent and culture-independent methods.

    Directory of Open Access Journals (Sweden)

    Jun Mei

    Full Text Available The biochemical changes occurring during cheese ripening are directly and indirectly dependent on the microbial associations of starter cultures. Freeze-dried Tibetan kefir coculture was used as a starter culture in the Camembert-type cheese production for the first time. Therefore, it's necessary to elucidate the stability, organization and identification of the dominant microbiota presented in the cheese. Bacteria and yeasts were subjected to culture-dependent on selective media and culture-independent polymerase chain reaction (PCR-denaturing gradient gel electrophoresis (DGGE analysis and sequencing of dominant bands to assess the microbial structure and dynamics through ripening. In further studies, kefir grains were observed using scanning electron microscopy (SEM methods. A total of 147 bacteria and 129 yeasts were obtained from the cheese during ripening. Lactobacillus paracasei represents the most commonly identified lactic acid bacteria isolates, with 59 of a total of 147 isolates, followed by Lactococcus lactis (29 isolates. Meanwhile, Kazachstania servazzii (51 isolates represented the mainly identified yeast isolate, followed by Saccharomyces cerevisiae (40 isolates. However, some lactic acid bacteria detected by sequence analysis of DGGE bands were not recovered by plating. The yeast S. cerevisiae and K. servazzii are described for the first time with kefir starter culture. SEM showed that the microbiota were dominated by a variety of lactobacilli (long and curved cells growing in close association with a few yeasts in the inner portion of the grain and the short lactobacilli were observed along with yeast cells on the exterior portion. Results indicated that conventional culture method and PCR-DGGE should be combined to describe in maximal detail the microbiological composition in the cheese during ripening. The data could help in the selection of appropriate commercial starters for Camembert-type cheese.

  19. Comparative evaluation of the teratogenicity of genistein and genistin using rat whole embryo culture and limbud micromass culture methods.

    Science.gov (United States)

    Zou, Peng; Xing, Lina; Tang, Qiuqiong; Liu, Ran; Hao, Weidong

    2012-08-01

    Genistein (GEN) is one kind of phytoestrogen. Several studies have demonstrated the teratogenic potential of GEN in vitro by postimplantation rat whole embryo culture (WEC) assay, but GEN showed no teratogenic effects in vivo even at a dose up to 1000 mg/kg bw/day. The mechanism of such discrepancy is still unclear. Because more than 80% of total genistein (free plus glycoside form) in circulation is its glycoside metabolite, genistin (GIN), we thus hypothesize that genistin is non-teratogenic. To prove this hypothesis, rat whole embryo culture (WEC) and limbud micromass culture methods were applied to compare the teratogenic effects of GEN and GIN on developing embryos in vitro. In WEC assay, we found that the development of embryos was affected by GEN treatment dose-dependently, while GIN-treated embryos displayed slight developmental defects only at the highest dose (222 μM). In micromass culture assay, the IC50 of cell proliferation and differentiation for GEN were 15.6 and 37.2 μM, respectively, while neither was influenced by GIN treatment up to 111 μM. Collectively, our study indicated that GEN showed no teratogenic effects in vivo probably due to its transformation to the non-teratogenic metabolite, GIN. Copyright © 2012 Elsevier Ltd. All rights reserved.

  20. Direct microscopical versus cultural method in screening for candidiasis among non gravid nigerian women.

    Science.gov (United States)

    Elegbe, I A; Botu, M

    1982-09-17

    The incidence of candidiasis in 125 symptomless, non-gravid women in Ile-Ife State Hospital, Ile-Ife, Oyo-State of Nigeria was determined by either microscopic observation or cultural methods. Among the three techniques investigated, the cultural method had the highest recovery rate with 60% of the cultures from 125 patients positive. Statistical analysis of the data (X2) demonstrated a significant difference between the recovery rates at the P less than 0.01 level. The implications of these findings were discussed in the text.

  1. Bacterial diversity determination using culture-dependent and culture-independent methods

    Directory of Open Access Journals (Sweden)

    M. Ghiasian

    2017-04-01

    Full Text Available Mud volcanoes are taken into consideration by geologists and oil industry experts have given their association with oil and gas reserves and methane greenhouse gas production in hydrosphere and atmosphere. Gomishan mud volcano phenomenon in the southeastern edge of the Caspian Sea, given its oil and gas resources, has been studied by some geologists in terms of geology and tectonics but not in terms of microbiology. Accordingly, it seems necessary to study this phenomenon from the perspective of microbiology in order to identify prokaryotes living in this area. Prokaryotes diversity in Mud volcano has been studied by cultivation techniques, fluorescence in situ hybridization, and denaturing gradient gel electrophoresis of PCR-amplified fragments of 16S rRNA genes. Total cell abundance in the mud volcano from 1×101-6×101per milliliter was determined by 4', 6-diamidino-2-phenylindole direct count. The detectable proportion of Archaea to Bacteria in the community by FISH was one to five. High viable counts (1 – 3 × 106 were obtained in culture media. A total of 122 isolates were obtained, 46 colonies were selected based on primarily morphological and physiological traits, and their 16S rRNA sequences were determined. The isolated genera included Halomonas (20%, Arthrobacter (5%, Kocuria (5%, Thalassobacillus (5%, Marinobacter (20%, Paracoccus (5%, Roseovarius (5%, Jeotgalicoccus (5%, Bacillus (15%, and Staphylococcus (15%. Regarding DGGE analysis, selected bands were obtained from the gels, reamplified and sequenced. Overall, 75% of the bacterial sequences were related to Rahnella and 25% related to Serratia.

  2. 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.

  3. 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.

  4. Plant management in natural areas: balancing chemical, mechanical, and cultural control methods

    Science.gov (United States)

    Steven Manning; James. Miller

    2011-01-01

    After determining the best course of action for control of an invasive plant population, it is important to understand the variety of methods available to the integrated pest management professional. A variety of methods are now widely used in managing invasive plants in natural areas, including chemical, mechanical, and cultural control methods. Once the preferred...

  5. METHODS OF EDUCATION DEVELOPMENT MANAGEMENT IN THE FIELD OF PHYSICAL CULTURE AND SPORTS OF POLAND

    Directory of Open Access Journals (Sweden)

    Oleksandr Mozolev

    2016-05-01

    Full Text Available The author researches the content of management activities to develop education in the field of physical culture and sports in Poland. The analysis of revealing the term "method" by Polish scientists has been made. The concept of "management method" is defined as a way to implement administrative functions by organizing preliminary impact of management subject on object of management with the purpose of implementation of its public functions or a set of controls that are used in an appropriate manner that can most effectively achieve their managerial objectives defining the qualitative aspect of system administration. The concept "management methods for developing education in the field of physical culture and sports in Poland" is formulated. The content of management methods for developing education in the field of physical culture and sports in Poland in which the author includes: economic; social and political; administrative; ethical methods; methods of cognitive activity; forecasting; planning; management decision-making and control.

  6. Microbiota of high-pressure-processed Serrano ham investigated by culture-dependent and culture-independent methods.

    Science.gov (United States)

    Martínez-Onandi, N; Castioni, A; San Martín, E; Rivas-Cañedo, A; Nuñez, M; Torriani, S; Picon, A

    2017-01-16

    The microbiota of Serrano dry-cured ham of different chemical composition, subjected or not to high-pressure processing (HPP), was investigated using culture-dependent and culture-independent methods. Microbial counts were submitted to analysis of variance with physicochemical parameters (a w , NaCl concentration, salt-in-lean ratio and intramuscular fat content) or HPP as main effects. In untreated hams, physicochemical parameters significantly affected counts of aerobic mesophiles, psychrotrophs, and moulds and yeasts. NaCl concentration and fat content influenced the levels of four and three of the five studied microbial groups, respectively, whereas no influence of a w was stated. The HPP treatment had a significant effect on counts of all investigated microbial groups. Culture-independent methods showed the presence of bacteria such as Staphylococcus equorum, Staphylococcus succinus, Bacillus subtilis and Cellulosimicrobium sp., moulds like Penicillium commune, Aspergillus fumigatus, Sclerotinia sclerotiorum, Eurotium athecium and Moniliella mellis, and yeasts like Debaryomyces hansenii and Candida glucosophila. Absence of B. subtilis bands and weaker bands of E. athecium were recorded for HPP-treated hams. The higher microbial levels found in lean ham might result in a quicker deterioration. HPP treatment confirmed its suitability as a procedure to control spoilage microorganisms. DGGE did not seem to be sensitive enough to highlight changes caused by HPP treatment in the microbiota of ham, but contributed to the detection of microbial species not previously found in ham. Copyright © 2016 Elsevier B.V. All rights reserved.

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

    Science.gov (United States)

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

    2015-08-01

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

  8. Diversity of Termitomyces associated with fungus-farming termites assessed by cultural and culture-independent methods.

    Science.gov (United States)

    Makonde, Huxley M; Boga, Hamadi I; Osiemo, Zipporah; Mwirichia, Romano; Stielow, J Benjamin; Göker, Markus; Klenk, Hans-Peter

    2013-01-01

    Fungus-cultivating termites make use of an obligate mutualism with fungi from the genus Termitomyces, which are acquired through either vertical transmission via reproductive alates or horizontally transmitted during the formation of new mounds. Termitomyces taxonomy, and thus estimating diversity and host specificity of these fungi, is challenging because fruiting bodies are rarely found. Molecular techniques can be applied but need not necessarily yield the same outcome than morphological identification. Culture-dependent and culture-independent methods were used to comprehensively assess host specificity and gut fungal diversity. Termites were identified using mitochondrial cytochrome oxidase II (COII) genes. Twenty-three Termitomyces cultures were isolated from fungal combs. Internal transcribed spacer (ITS) clone libraries were constructed from termite guts. Presence of Termitomyces was confirmed using specific and universal primers. Termitomyces species boundaries were estimated by cross-comparison of macromorphological and sequence features, and ITS clustering parameters accordingly optimized. The overall trends in coverage of Termitomyces diversity and host associations were estimated using Genbank data. Results indicate a monoculture of Termitomyces in the guts as well as the isolation sources (fungal combs). However, cases of more than one Termitomyces strains per mound were observed since mounds can contain different termite colonies. The newly found cultures, as well as the clustering analysis of GenBank data indicate that there are on average between one and two host genera per Termitomyces species. Saturation does not appear to have been reached, neither for the total number of known Termitomyces species nor for the number of Termitomyces species per host taxon, nor for the number of known hosts per Termitomyces species. Considering the rarity of Termitomyces fruiting bodies, it is suggested to base the future taxonomy of the group mainly on well

  9. Diversity of Termitomyces associated with fungus-farming termites assessed by cultural and culture-independent methods.

    Directory of Open Access Journals (Sweden)

    Huxley M Makonde

    Full Text Available BACKGROUND: Fungus-cultivating termites make use of an obligate mutualism with fungi from the genus Termitomyces, which are acquired through either vertical transmission via reproductive alates or horizontally transmitted during the formation of new mounds. Termitomyces taxonomy, and thus estimating diversity and host specificity of these fungi, is challenging because fruiting bodies are rarely found. Molecular techniques can be applied but need not necessarily yield the same outcome than morphological identification. METHODOLOGY: Culture-dependent and culture-independent methods were used to comprehensively assess host specificity and gut fungal diversity. Termites were identified using mitochondrial cytochrome oxidase II (COII genes. Twenty-three Termitomyces cultures were isolated from fungal combs. Internal transcribed spacer (ITS clone libraries were constructed from termite guts. Presence of Termitomyces was confirmed using specific and universal primers. Termitomyces species boundaries were estimated by cross-comparison of macromorphological and sequence features, and ITS clustering parameters accordingly optimized. The overall trends in coverage of Termitomyces diversity and host associations were estimated using Genbank data. RESULTS AND CONCLUSION: Results indicate a monoculture of Termitomyces in the guts as well as the isolation sources (fungal combs. However, cases of more than one Termitomyces strains per mound were observed since mounds can contain different termite colonies. The newly found cultures, as well as the clustering analysis of GenBank data indicate that there are on average between one and two host genera per Termitomyces species. Saturation does not appear to have been reached, neither for the total number of known Termitomyces species nor for the number of Termitomyces species per host taxon, nor for the number of known hosts per Termitomyces species. Considering the rarity of Termitomyces fruiting bodies, it is

  10. 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.

  11. 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

  12. 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

  13. 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.

  14. Comparison of Standard Culture-Based Method to Culture-Independent Method for Evaluation of Hygiene Effects on the Hand Microbiome

    Directory of Open Access Journals (Sweden)

    C. Zapka

    2017-03-01

    Full Text Available Hands play a critical role in the transmission of microbiota on one’s own body, between individuals, and on environmental surfaces. Effectively measuring the composition of the hand microbiome is important to hand hygiene science, which has implications for human health. Hand hygiene products are evaluated using standard culture-based methods, but standard test methods for culture-independent microbiome characterization are lacking. We sampled the hands of 50 participants using swab-based and glove-based methods prior to and following four hand hygiene treatments (using a nonantimicrobial hand wash, alcohol-based hand sanitizer [ABHS], a 70% ethanol solution, or tap water. We compared results among culture plate counts, 16S rRNA gene sequencing of DNA extracted directly from hands, and sequencing of DNA extracted from culture plates. Glove-based sampling yielded higher numbers of unique operational taxonomic units (OTUs but had less diversity in bacterial community composition than swab-based sampling. We detected treatment-induced changes in diversity only by using swab-based samples (P < 0.001; we were unable to detect changes with glove-based samples. Bacterial cell counts significantly decreased with use of the ABHS (P < 0.05 and ethanol control (P < 0.05. Skin hydration at baseline correlated with bacterial abundances, bacterial community composition, pH, and redness across subjects. The importance of the method choice was substantial. These findings are important to ensure improvement of hand hygiene industry methods and for future hand microbiome studies. On the basis of our results and previously published studies, we propose recommendations for best practices in hand microbiome research.

  15. 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.

  16. 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

  17. 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.

  18. 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

  19. 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.

  20. Cultural cognition in the thinking-aloud method for usability evaluation

    DEFF Research Database (Denmark)

    Clemmensen, Torkil; Hertzum, Morten; Hornbæk, Kasper

    2008-01-01

    and evaluator. In conclusion, we point to the importance of matching the task presentation to users' cultural background, the different effects of thinking aloud on task performance between Easterners and Westerners, the differences in nonverbal behaviour that affect usability problem detection, and, finally......We discuss the impact of cultural differences on usability evaluations that are based on the thinking-aloud method (TA). The term ‘cultural differences' helps distinguish differences in the perception and thinking of Westerners (people from Western Europe and US citizens with European origins......) and Easterners (people from China and the countries heavily influenced by its culture). We illustrate the impact of cultural cognition on four central elements of TA: (1) instructions and tasks, (2) the user's verbalizations, (3) the evaluator's reading of the user, and (4) the overall relationship between user...

  1. Cultural cognition in the thinking-aloud method for usability evaluation

    DEFF Research Database (Denmark)

    Clemmensen, Torkil; Hertzum, Morten; Hornbæk, Kasper

    2008-01-01

    We discuss the impact of cultural differences on usability evaluations that are based on the thinking-aloud method (TA). The term ‘cultural differences' helps distinguish differences in the perception and thinking of Westerners (people from Western Europe and US citizens with European origins......) and Easterners (people from China and the countries heavily influenced by its culture). We illustrate the impact of cultural cognition on four central elements of TA: (1) instructions and tasks, (2) the user's verbalizations, (3) the evaluator's reading of the user, and (4) the overall relationship between user...... and evaluator. In conclusion, we point to the importance of matching the task presentation to users' cultural background, the different effects of thinking aloud on task performance between Easterners and Westerners, the differences in nonverbal behaviour that affect usability problem detection, and, finally...

  2. A fluorescence anisotropy method for measuring protein concentration in complex cell culture media.

    Science.gov (United States)

    Groza, Radu Constantin; Calvet, Amandine; Ryder, Alan G

    2014-04-22

    The rapid, quantitative analysis of the complex cell culture media used in biopharmaceutical manufacturing is of critical importance. Requirements for cell culture media composition profiling, or changes in specific analyte concentrations (e.g. amino acids in the media or product protein in the bioprocess broth) often necessitate the use of complicated analytical methods and extensive sample handling. Rapid spectroscopic methods like multi-dimensional fluorescence (MDF) spectroscopy have been successfully applied for the routine determination of compositional changes in cell culture media and bioprocess broths. Quantifying macromolecules in cell culture media is a specific challenge as there is a need to implement measurements rapidly on the prepared media. However, the use of standard fluorescence spectroscopy is complicated by the emission overlap from many media components. Here, we demonstrate how combining anisotropy measurements with standard total synchronous fluorescence spectroscopy (TSFS) provides a rapid, accurate quantitation method for cell culture media. Anisotropy provides emission resolution between large and small fluorophores while TSFS provides a robust measurement space. Model cell culture media was prepared using yeastolate (2.5 mg mL(-1)) spiked with bovine serum albumin (0 to 5 mg mL(-1)). Using this method, protein emission is clearly discriminated from background yeastolate emission, allowing for accurate bovine serum albumin (BSA) quantification over a 0.1 to 4.0 mg mL(-1) range with a limit of detection (LOD) of 13.8 μg mL(-1). Copyright © 2014. Published by Elsevier B.V.

  3. Bayesian analysis of culture and PCR methods for detection of Campylobacter spp. in broiler caecal samples.

    Science.gov (United States)

    Arnold, M E; Jones, E M; Lawes, J R; Vidal, A B; Clifton-Hadley, F A; Rodgers, J D; Powell, L F

    2015-01-01

    The objective of this study was to estimate the sensitivity and specificity of a culture method and a polymerase chain reaction (PCR) method for detection of two Campylobacter species: C. jejuni and C. coli. Data were collected during a 3-year survey of UK broiler flocks, and consisted of parallel sampling of caeca from 436 batches of birds by both PCR and culture. Batches were stratified by season (summer/non-summer) and whether they were the first depopulation of the flock, resulting in four sub-populations. A Bayesian approach in the absence of a gold standard was adopted, and the sensitivity and specificity of the PCR and culture for each Campylobacter subtype was estimated, along with the true C. jejuni and C. coli prevalence in each sub-population. Results indicated that the sensitivity of the culture method was higher than that of PCR in detecting both species when the samples were derived from populations infected with at most one species of Campylobacter. However, from a mixed population, the sensitivity of culture for detecting both C. jejuni or C. coli is reduced while PCR is potentially able to detect both species, although the total probability of correctly identifying at least one species by PCR is similar to that of the culture method.

  4. [The comparison of different bronchial aspirate culturing methods in patients with ventilator-associated pneumonia (VAP)].

    Science.gov (United States)

    Kowalczyk, Wojciech; Rybicki, Zbigniew; Tomaszewski, Dariusz; Truszczyński, Andrzej; Guzek, Aneta

    2011-01-01

    Although broncho-alveolar lavage (BAL) culture and protected specimen brush (PSB) are regarded as the most effective methods in the diagnosis of VAP, a simple endotracheal aspiration (EA) is frequently performed during routine care, because of its simplicity and low cost. We compared the effectiveness of EA with BAL and PSB in VAP patients. Sixty-one adult VAP patients, ventilated for longer than 48 h, were cultured with all three methods. Positive cultures were obtained from 63.9% of patients, with Acinetobacter baumannii being the most common pathogen. There was a high positive correlation between simple aspirates and BAL (k 0.817, CI 0.664-0.840, p aspirates and PSB (k 0.667, CI 0.483-0.871, p aspirate culturing, compared to BAL and PSB, it can be used successfully in most cases.

  5. A method for establishing human primary gastric epithelial cell culture from fresh surgical gastric tissues.

    Science.gov (United States)

    Aziz, Faisal; Yang, Xuesong; Wen, Qingping; Yan, Qiu

    2015-08-01

    At present, biopsy specimens, cancer cell lines and tissues obtained by gastric surgery are used in the study and analysis of gastric cancer, including the molecular mechanisms and proteomics. However, fibroblasts and other tissue components may interfere with these techniques. Therefore, the present study aimed to develop a procedure for the isolation of viable human gastric epithelial cells from gastric surgical tissues. A method was developed to culture human gastric epithelial cells using fresh, surgically excised tissues and was evaluated using immunocytochemistry, periodic acid-Schiff (PAS) staining and cell viability assays. Low cell growth was observed surrounding the gastric tissue on the seventh day of tissue explant culture. Cell growth subsequently increased, and at 12 days post-explant a high number of pure epithelial cells were detected. The gastric cancer cells exhibited rapid growth with a doubling time of 13-52 h, as compared to normal cells, which had a doubling time of 20-53 h. Immunocytochemical analyses of primary gastric cells revealed positive staining for cytokeratin 18 and 19, which indicated that the culture was comprised of pure epithelial cells and contained no fibroblasts. Furthermore, PAS staining demonstrated that the cultured gastric cells produced neutral mucin. Granulin and carbohydrate antigen 724 staining confirmed the purity of gastric cancer and normal cells in culture. This method of cell culture indicated that the gastric cells in primary culture consisted of mucin-secreting gastric epithelial cells, which may be useful for the study of gastric infection with Helicobacter pylori and gastric cancer.

  6. Urine culture collected from gel-based diapers: developing a novel experimental laboratory method.

    Science.gov (United States)

    Shvartzman, Pesach; Nasri, Yussuf

    2004-01-01

    Urinary tract infection diagnosis is based on urine culture, taken from a midstream collection. Obtaining samples in this manner is difficult in elderly patients suffering from incontinence and in infants. (1) Develop a method for urine collection using gel-based diapers and (2) compare culture results from gel-based diapers with those of the same urine, examined by accepted methods. Urine was collected and cultured by standard bacteriologic techniques at the microbiology laboratory of Soroka Medical Center, Beer-Sheva, Israel. Gel-based diapers were manually dampened with the same urine samples, and samples from diapers were cultured 0 to 3 hours after dampening. Comparison showed good correlation between direct urine cultures and cultures obtained from gel based diapers (R = 1.000). Total sensitivity was 100% and specificity 97%. (1) Urine can be cultured from gel-based diapers and (2) a larger sample of in vivo trials will be needed to make this technique clinically applicable in an outpatient setting.

  7. The bacteriology of chronic venous leg ulcer examined by culture-independent molecular methods

    DEFF Research Database (Denmark)

    Thomsen, Trine R; Aasholm, Martin S; Rudkjøbing, Vibeke B

    2010-01-01

    The bacterial microbiota plays an important role in the prolonged healing of chronic venous leg ulcers. The present study compared the bacterial diversity within ulcer material from 14 skin graft operations of chronic venous leg ulcers using culture-based methods and molecular biological methods...... was different. All the wounds contained Staphylococcus aureus, whereas Pseudomonas aeruginosa was in six out of 14 wounds. Molecular methods detected anaerobic pathogens in four ulcers that were not detected with anaerobic culture methods. Quantitative polymerase chain reaction was used to compare the abundance...... of S. aureus and P. aeruginosa at different locations in the ulcers and their numbers varied greatly between samples taken at different locations in the same ulcer. This should be considered when ulcers are investigated in routine clinical care. The differences between the results obtained with culture...

  8. Conversion of primordial germ cells to pluripotent stem cells: methods for cell tracking and culture conditions.

    Science.gov (United States)

    Nagamatsu, Go; Suda, Toshio

    2013-01-01

    Primordial germ cells (PGCs) are unipotent cells committed to germ lineage: PGCs can only differentiate into gametes in vivo. However, upon fertilization, germ cells acquire the capacity to differentiate into all cell types in the body, including germ cells. Therefore, germ cells are thought to have the potential for pluripotency. PGCs can convert to pluripotent stem cells in vitro when cultured under specific conditions that include bFGF, LIF, and the membrane-bound form of SCF (mSCF). Here, the culture conditions which efficiently convert PGCs to pluripotent embryonic germ (EG) cells are described, as well as methods used for identifying pluripotent candidate cells during culture.

  9. Optimization of chemically defined cell culture media - Replacing fetal bovine serum in mammalian in vitro methods

    DEFF Research Database (Denmark)

    van der Valk, J; Brunner, D; De Smet, K

    2010-01-01

    Quality assurance is becoming increasingly important. Good laboratory practice (GLP) and good manufacturing practice (GMP) are now established standards. The biomedical field aims at an increasing reliance on the use of in vitro methods. Cell and tissue culture methods are generally fast, cheap, ...

  10. A novel culture method for high level production of heterologous protein in Saccharomyces cerevisiae.

    Science.gov (United States)

    Nagashima, T; Yamamoto, Y; Gomi, K; Kitamoto, K; Kumagai, C

    1994-07-01

    A high level production system for heterologous protein by cold culture of yeast transformants at 15 degrees C was developed. The yeast transformants, carrying a plasmid containing cDNA for Aspergillus oryzae alpha-amylase (Taka-amylase A) or human lysozyme synthetic DNA, were cultivated in a selective medium for 1 or 2 days until full growth at 30 degrees C. The yeast cells were harvested by centrifugation from the culture fluid and then were transferred to YPD medium. These inoculated broths were incubated for 2 days at 15 degrees C and then for another 2 days at 30 degrees C. By the cold culture method described above, higher amounts of Taka-amylase A (28.6 mg/liter) and human lysozyme (6.1 mg/liter) were produced by the yeast transformants compared to those by conventional methods. Heterologous protein productions using YEp, YCp, and YIp types of yeast expression vectors with ADH1 or GAPDH promoter by the cold culture method showed effective productivity of about 2-fold compared to those by the conventional method of culture at 30 degrees C. The high level production of heterologous protein by this method was not specific to the S. cerevisiae strains examined.

  11. Culture methods of allograft musculoskeletal tissue samples in Australian bacteriology laboratories.

    Science.gov (United States)

    Varettas, Kerry

    2013-12-01

    Samples of allograft musculoskeletal tissue are cultured by bacteriology laboratories to determine the presence of bacteria and fungi. In Australia, this testing is performed by 6 TGA-licensed clinical bacteriology laboratories with samples received from 10 tissue banks. Culture methods of swab and tissue samples employ a combination of solid agar and/or broth media to enhance micro-organism growth and maximise recovery. All six Australian laboratories receive Amies transport swabs and, except for one laboratory, a corresponding biopsy sample for testing. Three of the 6 laboratories culture at least one allograft sample directly onto solid agar. Only one laboratory did not use a broth culture for any sample received. An international literature review found that a similar combination of musculoskeletal tissue samples were cultured onto solid agar and/or broth media. Although variations of allograft musculoskeletal tissue samples, culture media and methods are used in Australian and international bacteriology laboratories, validation studies and method evaluations have challenged and supported their use in recovering fungi and aerobic and anaerobic bacteria.

  12. Searching for Innovations and Methods of Using the Cultural Heritage on the Example of Upper Silesia

    Science.gov (United States)

    Wagner, Tomasz

    2017-10-01

    The basic subject of this paper is historical and cultural heritage of some parts of Upper Silesia, bind by common history and similar problems at present days. The paper presents some selected historical phenomena that have influenced contemporary space, mentioned above, and contemporary issues of heritage protection in Upper Silesia. The Silesian architecture interpretation, since 1989, is strongly covered with some ideological and national ideas. The last 25 years are the next level of development which contains rapidly transformation of the space what is caused by another economical transformations. In this period, we can observe landscape transformations, liquidation of objects and historical structures, loos of regional features, spontaneous adaptation processes of objects and many methods of implementation forms of protection, and using of cultural resources. Some upheaval linked to the state borders changes, system, economy and ethnic transformation caused that former Upper Silesia border area focuses phenomena that exists in some other similar European areas which are abutments of cultures and traditions. The latest period in the history of Upper Silesia gives us time to reflect the character of changes in architecture and city planning of the area and appraisal of efficiency these practices which are connected to cultural heritage perseveration. The phenomena of the last decades are: decrement of regional features, elimination of objects, which were a key feature of the regional cultural heritage, deformation of these forms that were shaped in the history and some trials of using these elements of cultural heritage, which are widely recognized as cultural values. In this situation, it is important to seek creative solutions that will neutralize bad processes resulting from bad law and practice. The most important phenomena of temporary space is searching of innovative fields and methods and use of cultural resources. An important part of the article is

  13. An efficient method for the establishment of cell suspension cultures in potato (Solanum tuberosum L.)

    International Nuclear Information System (INIS)

    Sajid, Z.A.

    2016-01-01

    Cell suspension cultures offers an In vitro system that can be used as a tool for various studies involving mutant selection, mass propagation, protoplast isolation, gene transfer and selection of cell-lines which are resistant to various biotic or abiotic stresses. Research work on the development of cell suspension cultures was carried out to establish the most efficient method in Potato (cv. Desiree). Healthy, well-proliferating tissues from different types of callus cultures (compact, friable, embryogenic or non-embryogenic) were inoculated on various media combinations, i.e., MS, MS2 or AA liquid medium containing 18.09 micro M 2, 4-D. A fixed quantity (0.5-1.0 g) of callus tissue from 60-day-old callus cultures was transferred to 10-25 ml of liquid medium in 100 ml Erlenmeyer flask. Cultures were placed on an orbital shaker and agitated at different speeds (75, 100 or 125 rpm) under 16-h photoperiod at 25 ± 2 degree C. Medium was changed after every 3 days and fractionated tissue was filtered after every 6 days through sterile mesh (100-800 micro m) to develop a cell-line by transferring resulting suspension to fresh medium under the same conditions. Results indicated that eight-week-old translucent, friable, off-white callus cultures were an excellent starting material for the initiation of homogeneous cell suspension cultures as compared to other tested sources. Of the three tested media (MS, MS2 or AA medium containing 18.09 micro M 2, 4-D), MS2 was found to be a better medium for the initiation of cell suspension cultures. Cell suspension cultures, placed in 16-h photoperiod at 25 ± 2 degree C and agitated at 120 rpm using a gyratory shaker showed excellent results. Several other factors influencing quick establishment of cell suspension cultures in this cultivar are also discussed in this communication. (author)

  14. 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.

  15. Characterization of microbes in prosthetic joint specimens by culture-independent molecular methods

    DEFF Research Database (Denmark)

    Xu, Yijuan; Rudkjøbing, Vibeke Børsholt; Simonsen, Ole

    , and polymicrobial communities were found in 9 patients. Culture-based methods detected bacteria in 7 patients, and multiple species were only isolated from one of them. Additionally, quantification of Propionibacterium and Staphylococcus aureus using specific qPCR assays confirmed the findings by clone library...... was to investigate the microbial diversity in surgical samples (eg. synovial fluid, periprosthetic tissue, removed prosthesis) from 22 prosthetic patients using a range of culture-independent molecular methods including broad range 16S rRNA gene PCR, cloning, phylogeny, quantitative PCR (qPCR), and fluorescence...... of bacteria (e.g., coryneform rods), while molecular methods detected several distinct species including the species identified by culture. Broad range 16S rRNA gene PCR and cloning detected 39 bacterial species from 11 patients, including known pathogens and species not previously reported in PJIs...

  16. Establishment of primary bovine intestinal epithelial cell culture and clone method.

    Science.gov (United States)

    Zhan, Kang; Lin, Miao; Liu, Ming-Mei; Sui, Yang-Nan; Zhao, Guo-Qi

    2017-01-01

    The aim of this study was to establish bovine intestinal epithelial cell (BIEC) line and provide a novel clone cell method. Although various strategies of bovine cell culture and clone techniques have been reported, these methods remain not established. Here, we culture successfully primary BIECs and establish a novel clone cell method. Our result showed that BIECs could be successfully cultured and passaged about generation 5. These cellular aggregates and clusters were adherent loosely at day 2 of culture. Cell aggregates and clusters start to proliferate after approximately 4 d. The BIECs showed positive reaction against cytokeratin 18, E-cadherin, and characteristics of epithelial-like morphology. In addition, the fatty acid-binding proteins (FABPs), villin, and intestinal peptidase (IP) band were positive in BIECs. Our results suggest that the establishment of culturing and clone BIEC methods will apply to isolate and clone other primary cells. These BIECs could therefore contribute to the study of bovine intestinal nutrient absorption and regulation, immune regulation, and the pathogenesis of the bovine intestinal disease, which will provide intestinal cell model in vitro.

  17. 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.

  18. 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.

  19. Cultural

    Science.gov (United States)

    Wilbur F. LaPage

    1971-01-01

    A critical look at outdoor recreation research and some underlying premises. The author focuses on the concept of culture as communication and how it influences our perception of problems and our search for solutions. Both outdoor recreation and science are viewed as subcultures that have their own bodies of mythology, making recreation problems more difficult to...

  20. 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 ...

  1. 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.

  2. 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...

  3. 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...

  4. 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

  5. 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

  6. 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.

  7. 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.

  8. 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.

  9. Detection of food hazards in foods: comparison of real time polymerase chain reaction and cultural methods

    Directory of Open Access Journals (Sweden)

    Paolo Bonilauri

    2016-01-01

    Full Text Available Foodstuffs should not contain microorganisms or their toxins or metabolites in quantities suggesting an unacceptable risk for human health. The detection of food hazards in foods is performed by several tests that produce results dependent on the analytical method used: an analytical reference method, defined as standard, is associated with each microbiological criterion laid down in Regulation 2073/2005/EC, but, analytical methods other than the reference ones, in particular more rapid methods, could be used. Combined screening methods performed by real time-polymerase chain reaction (RT-PCR are currently validated as alternative methods according to the ISO 16140:2003 and certified by the Association Française de Normalisation. However, the positive results obtained with these alternative methods, the investigated molecular relations that resulted positive have to be confirmed with cultural methods using the same enrichment media in which the molecular screening was performed. Since it is necessary to assess if these testing schemes provide equivalent guarantees of food safety, the aim of this retrospective study is to analyse the data collected, from 2012 to 2014 by Emilia Romagna Region in the field of Piano Regionale Alimenti (Food Regional Plan during official controls monitoring food samples of animal and other than animal origin. Records performed by combined methods of molecular screening of Salmonella spp., Listeria monocytogenes and thermophilic Campylobacter and cultural confirmation results were gathered together and the results were compared in order to assess the sensitivity of the methods. A total of 10,604 food samples were considered in this study: the comparison of the data revealed that the RT-PCR method detected Salmonella, L. monocytogenes, and thermophilic Campylobacter in 2.18, 3.85 and 3.73% of the samples, respectively, whereas by using cultural method these pathogens were isolated in 0.43, 1.57 and 1.57% of samples

  10. Detection of Food Hazards in Foods: Comparison of Real Time Polymerase Chain Reaction and Cultural Methods.

    Science.gov (United States)

    Bonilauri, Paolo; Bardasi, Lia; Leonelli, Roberto; Ramini, Mattia; Luppi, Andrea; Giacometti, Federica; Merialdi, Giuseppe

    2016-01-18

    Foodstuffs should not contain microorganisms or their toxins or metabolites in quantities suggesting an unacceptable risk for human health. The detection of food hazards in foods is performed by several tests that produce results dependent on the analytical method used: an analytical reference method, defined as standard, is associated with each microbiological criterion laid down in Regulation 2073/2005/EC, but, analytical methods other than the reference ones, in particular more rapid methods, could be used. Combined screening methods performed by real time-polymerase chain reaction (RT-PCR) are currently validated as alternative methods according to the ISO 16140:2003 and certified by the Association Française de Normalisatio n. However, the positive results obtained with these alternative methods, the investigated molecular relations that resulted positive have to be confirmed with cultural methods using the same enrichment media in which the molecular screening was performed. Since it is necessary to assess if these testing schemes provide equivalent guarantees of food safety, the aim of this retrospective study is to analyse the data collected, from 2012 to 2014 by Emilia Romagna Region in the field of Piano Regionale Alimenti (Food Regional Plan) during official controls monitoring food samples of animal and other than animal origin. Records performed by combined methods of molecular screening of Salmonella spp., Listeria monocytogenes and thermophilic Campylobacter and cultural confirmation results were gathered together and the results were compared in order to assess the sensitivity of the methods. A total of 10,604 food samples were considered in this study: the comparison of the data revealed that the RT-PCR method detected Salmonella, L. monocytogenes , and thermophilic Campylobacter in 2.18, 3.85 and 3.73% of the samples, respectively, whereas by using cultural method these pathogens were isolated in 0.43, 1.57 and 1.57% of samples, respectively. In

  11. Enhanced growth medium and method for culturing human mammary epithelial cells

    Science.gov (United States)

    Stampfer, Martha R.; Smith, Helene S.; Hackett, Adeline J.

    1983-01-01

    Methods are disclosed for isolating and culturing human mammary epithelial cells of both normal and malignant origin. Tissue samples are digested with a mixture including the enzymes collagenase and hyaluronidase to produce clumps of cells substantially free from stroma and other undesired cellular material. Growing the clumps of cells in mass culture in an enriched medium containing particular growth factors allows for active cell proliferation and subculture. Clonal culture having plating efficiencies of up to 40% or greater may be obtained using individual cells derived from the mass culture by plating the cells on appropriate substrates in the enriched media. The clonal growth of cells so obtained is suitable for a quantitative assessment of the cytotoxicity of particular treatment. An exemplary assay for assessing the cytotoxicity of the drug adriamycin is presented.

  12. Method and Apparatus for a Miniature Bioreactor System for Long-Term Cell Culture

    Science.gov (United States)

    Kleis, Stanley J. (Inventor); Geffert, Sandra K. (Inventor); Gonda, Steve R. (Inventor)

    2015-01-01

    A bioreactor and method that permits continuous and simultaneous short, moderate, or long term cell culturing of one or more cell types or tissue in a laminar flow configuration is disclosed, where the bioreactor supports at least two laminar flow zones, which are isolated by laminar flow without the need for physical barriers between the zones. The bioreactors of this invention are ideally suited for studying short, moderate and long term studies of cell cultures and the response of cell cultures to one or more stressors such as pharmaceuticals, hypoxia, pathogens, or any other stressor. The bioreactors of this invention are also ideally suited for short, moderate or long term cell culturing with periodic cell harvesting and/or medium processing for secreted cellular components.

  13. A Simple Method to Reduce both Lactic Acid and Ammonium Production in Industrial Animal Cell Culture.

    Science.gov (United States)

    Freund, Nathaniel W; Croughan, Matthew S

    2018-01-28

    Fed-batch animal cell culture is the most common method for commercial production of recombinant proteins. However, higher cell densities in these platforms are still limited due to factors such as excessive ammonium production, lactic acid production, nutrient limitation, and/or hyperosmotic stress related to nutrient feeds and base additions to control pH. To partly overcome these factors, we investigated a simple method to reduce both ammonium and lactic acid production-termed Lactate Supplementation and Adaptation (LSA) technology-through the use of CHO cells adapted to a lactate-supplemented medium. Using this simple method, we achieved a reduction of nearly 100% in lactic acid production with a simultaneous 50% reduction in ammonium production in batch shaker flasks cultures. In subsequent fed-batch bioreactor cultures, lactic acid production and base addition were both reduced eight-fold. Viable cell densities of 35 million cells per mL and integral viable cell days of 273 million cell-days per mL were achieved, both among the highest currently reported for a fed-batch animal cell culture. Investigating the benefits of LSA technology in animal cell culture is worthy of further consideration and may lead to process conditions more favorable for advanced industrial applications.

  14. Improving Evaluation of Dental Hygiene Students' Cultural Competence with a Mixed-Methods Approach.

    Science.gov (United States)

    Flynn, Priscilla; Sarkarati, Nassim

    2018-02-01

    Most dental hygiene educational programs include cultural competence education, but may not evaluate student outcomes. The aim of this study was to design and implement a mixed-methods evaluation to measure dental hygiene students' progression toward cultural competence. Two cohorts consisting of consecutive classes in one U.S. dental hygiene program participated in the study. A total of 47 dental hygiene students (100% response rate) completed self-assessments to measure their attitudes and knowledge at three time points between 2014 and 2016. Mean scores were calculated for three domains: Physical Environment, Communication, and Values. Qualitative analysis of the students' cultural diversity papers was also conducted to further evaluate students' knowledge and skills. Bennett's five-level conceptual framework was used to code phrases or sentences to place students in the general categories of ethnocentric or ethno-relative. The quantitative and qualitative results yielded different outcomes for Cohort 1, but not for Cohort 2. The Cohort 1 students assessed themselves statistically significantly lower over time in one of the three measured domains. However, the Cohort 2 students assessed themselves as statistically significantly more culturally competent in all three domains. Qualitative results placed 72% of Cohort 1 students and 83% of Cohort 2 students in the more desirable ethno-relative category. Since quantitative methods consisting of student self-assessments may not adequately measure students' cultural competence, adding qualitative methods to measure skills specific to patient care in this study added a robust dimension to evaluating this complex dental hygiene student competence.

  15. A Simple Method to Reduce both Lactic Acid and Ammonium Production in Industrial Animal Cell Culture

    Science.gov (United States)

    Freund, Nathaniel W.; Croughan, Matthew S.

    2018-01-01

    Fed-batch animal cell culture is the most common method for commercial production of recombinant proteins. However, higher cell densities in these platforms are still limited due to factors such as excessive ammonium production, lactic acid production, nutrient limitation, and/or hyperosmotic stress related to nutrient feeds and base additions to control pH. To partly overcome these factors, we investigated a simple method to reduce both ammonium and lactic acid production—termed Lactate Supplementation and Adaptation (LSA) technology—through the use of CHO cells adapted to a lactate-supplemented medium. Using this simple method, we achieved a reduction of nearly 100% in lactic acid production with a simultaneous 50% reduction in ammonium production in batch shaker flasks cultures. In subsequent fed-batch bioreactor cultures, lactic acid production and base addition were both reduced eight-fold. Viable cell densities of 35 million cells per mL and integral viable cell days of 273 million cell-days per mL were achieved, both among the highest currently reported for a fed-batch animal cell culture. Investigating the benefits of LSA technology in animal cell culture is worthy of further consideration and may lead to process conditions more favorable for advanced industrial applications. PMID:29382079

  16. A hybrid microfluidic-vacuum device for direct interfacing with conventional cell culture methods

    Directory of Open Access Journals (Sweden)

    Monuki Edwin S

    2007-09-01

    Full Text Available Abstract Background Microfluidics is an enabling technology with a number of advantages over traditional tissue culture methods when precise control of cellular microenvironment is required. However, there are a number of practical and technical limitations that impede wider implementation in routine biomedical research. Specialized equipment and protocols required for fabrication and setting up microfluidic experiments present hurdles for routine use by most biology laboratories. Results We have developed and validated a novel microfluidic device that can directly interface with conventional tissue culture methods to generate and maintain controlled soluble environments in a Petri dish. It incorporates separate sets of fluidic channels and vacuum networks on a single device that allows reversible application of microfluidic gradients onto wet cell culture surfaces. Stable, precise concentration gradients of soluble factors were generated using simple microfluidic channels that were attached to a perfusion system. We successfully demonstrated real-time optical live/dead cell imaging of neural stem cells exposed to a hydrogen peroxide gradient and chemotaxis of metastatic breast cancer cells in a growth factor gradient. Conclusion This paper describes the design and application of a versatile microfluidic device that can directly interface with conventional cell culture methods. This platform provides a simple yet versatile tool for incorporating the advantages of a microfluidic approach to biological assays without changing established tissue culture protocols.

  17. Optimization of an Efficient Non-Tissue Culture Transformation Method for Brassica Juncea

    International Nuclear Information System (INIS)

    Naeem, I.; Munir, I.; Iqbal, A.; Ullah, F.

    2016-01-01

    The major hurdles in successful in vitro transformation of Brassica juncea through standard tissue culture (STC) method are: culture contamination, somaclonal variations, and lack of expertise. Moreover, the current STC method is time consuming and needs continuous electricity. In the present study, the in planta transformation method through floral dip with or without vacuum infiltration was optimized for successful transformation of B. juncea. The B. juncea CV RAYA Anmol was used for transformation through Agrobacterium tumefaciens strain GV3101 harboring the binary vector plasmid pBinGlyBar4-EADcT. Based on the resistance reaction to the herbicide Basta, 20 and 40 resistant seedlings were obtained from 2000 seed germinated from the plants transformed through floral dip and vacuum infiltration methods, respectively. The PCR analyses further confirmed the presence of transgene in 3 floral dipped plants without vacuum infiltration and 17 floral dipped plants with vacuum infiltration, giving the transformation frequencies of 1.5*10/sup -3/ and 8.5*10/sup -3/, respectively. This method, which avoids tissue culture, will reduce the somaclonal variation accompanying prolonged culture of cells in a dedifferentiated state, will facilitate functional genomics and improvement of Brassica juncea with novel desirable traits while reducing time and expense. (author)

  18. cultural

    Directory of Open Access Journals (Sweden)

    Irene Kreutz

    2006-01-01

    Full Text Available Es un estudio cualitativo que adoptó como referencial teorico-motodológico la antropología y la etnografía. Presenta las experiencias vivenciadas por mujeres de una comunidad en el proceso salud-enfermedad, con el objetivo de comprender los determinantes sócio-culturales e históricos de las prácticas de prevención y tratamiento adoptados por el grupo cultural por medio de la entrevista semi-estructurada. Los temas que emergieron fueron: la relación entre la alimentación y lo proceso salud-enfermedad, las relaciones con el sistema de salud oficial y el proceso salud-enfermedad y lo sobrenatural. Los dados revelaron que los moradores de la comunidad investigada tienen un modo particular de explicar sus procedimientos terapéuticos. Consideramos que es papel de los profesionales de la salud en sus prácticas, la adopción de abordajes o enfoques que consideren al individuo en su dimensión sócio-cultural e histórica, considerando la enorme diversidad cultural en nuestro país.

  19. Transferring methods to teach business administration from one cultural context to another

    Directory of Open Access Journals (Sweden)

    Marie Catalo

    2015-12-01

    Full Text Available What happens when a teaching method is transferred from one cultural context to another? In this article we investigate this question by looking at how Computer Based Simulations (CBS were transposed from a French context to an Egyptian one. In this article we demonstrate, through the case of Egypt, how culture and the characteristics of the school system impact learning abilities. We describe what happens when Egyptian students are confronted with learning modes they have not encountered prior to University, in the context of an Egyptian-French dual-degree programme in business administration and business informatics. We show that the transfer of CBS as a teaching method revealed cultural differences between French and Egyptian students. As a consequence the teaching objectives of CBS were redefined in order to take the Egyptian context into account.

  20. A safety culture assessment by mixed methods at a public maternity and infant hospital in China

    Science.gov (United States)

    Listyowardojo, Tita Alissa; Yan, Xiaoling; Leyshon, Stephen; Ray-Sannerud, Bobbie; Yu, Xin Yan; Zheng, Kai; Duan, Tao

    2017-01-01

    Objective To assess safety culture at a public maternity hospital in Shanghai, China, using a sequential mixed methods approach. The study was part of a bigger study looking at the application of the mixed methods approach to assess safety culture in health care in different organizations and countries. Methodology A mixed methods approach was utilized by first distributing the Safety Attitudes Questionnaire measuring six safety culture dimensions and five independent items to all hospital staff (n=1482) working in 18 departments at a single hospital. Afterward, semistructured interviews were conducted using convenience sampling, where 48 hospital staff from nine departments at the same hospital were individually interviewed. Results The survey received a response rate of 96%. The survey findings show significant differences between the hospital departments in almost all safety culture dimensions and independent items. Similarly, the interview findings revealed that there were different, competing priorities between departments perceived to result in a reduced quality of collaboration and bottlenecks in care delivery. Another major finding was that staff who worked more hours per week would perceive working conditions significantly more negatively. Issues related to working conditions were also the most common concerns discussed in the interviews, especially the issue on high workload. High workload was also reflected in the fact that 91.45% of survey respondents reported that they worked 40 hours or longer per week. Finally, interview findings complemented survey findings, thus providing a more complete and accurate picture of safety culture. Conclusion Hospital leaders need to prioritize interventions focused on improving the quality of cross-department collaboration and reducing workload. A mixed methods assessment of safety culture provides more meaningful, targeted results, enabling leaders to prioritize and tailor improvement efforts to increase the impact of

  1. Teaching cross-cultural communication skills online: a multi-method evaluation.

    Science.gov (United States)

    Lee, Amy L; Mader, Emily M; Morley, Christopher P

    2015-04-01

    Cultural competency education is an important and required part of undergraduate medical education. The objective of this study was to evaluate whether an online cross-cultural communication module could increase student use of cross-cultural communication questions that assess the patient's definition of the problem, the way the problem affects their life, their concerns about the problem, and what the treatment should be (PACT). We used multi-method assessment of students assigned to family medicine clerkship blocks that were randomized to receive online cultural competency and PACT training added to their standard curriculum or to a control group receiving the standard curriculum only. Outcomes included comparison, via analysis of variance, of number of PACT questions used during an observed Standardized Patient Exercise, end-of-year OSCE scores, and qualitative analysis of student narratives. Students (n=119) who participated in the online module (n=60) demonstrated increased use of cross-cultural communication PACT questions compared to the control group (n=59) and generally had positive themes emerge from their reflective writing. The module had the biggest impact on students who later went on to match in high communication specialties. Online teaching of cross-cultural communication skills can be effective at changing medical student behavior.

  2. Establishment and Characterization of Primary Cultures from Iranian Oral Squamous Cell Carcinoma Patients by Enzymatic Method and Explant Culture

    Directory of Open Access Journals (Sweden)

    Meysam Ganjibakhsh

    2017-10-01

    Full Text Available Objectives: Oral Squamous Cell Carcinoma (OSCC is the most frequent oral cancer worldwide. It is known as the eighth most common cancer in men and as the fifth most common cancer in women. Cytogenetic and biochemical studies in recent decades have emphasized the necessity of providing an appropriate tool for such researches. Cancer cell culture is a useful tool for investigations on biochemical, genetic, molecular and immunological characteristics of different cancers, including oral cancer. Here, we explain the establishment process of five primary oral cancer cells derived from an Iranian population.Materials and Methods: The specimens were obtained from five oral cancer patients. Enzymatic, explant culture and magnetic-activated cell sorting (MACS methods were used for cell isolation. After quality control tests, characterization and authentication of primary oral cancer cells were performed by short tandem repeats (STR profiling, chromosome analysis, species identification, and monitoring the growth, morphology and the expression of CD326 and CD133 markers.Results: Five primary oral cancer cells were established from an Iranian population. The flow cytometry results showed that the isolated cells were positive for CD326 and CD133 markers. Furthermore, the cells were free from mycoplasma, bacterial and fungal contamination. No misidentified or cross-contaminated cells were detected by STR analysis.Conclusions: Human primary oral cancer cells provide an extremely useful platform for studying carcinogenesis pathways of oral cancer in Iranian population. They may be helpful in explaining the ethnic differences in cancer biology and the individuality in anticancer drug response in future studies.

  3. Anthropological Methods of Formation of University Students' Spiritual and Moral Culture

    Science.gov (United States)

    Kurbanov, Rashad A.; Nikonova, Elina I.; Gurbanov, Ramin A.; Svechnikova, Natalia V.; Tumarov, Konstantin B.; Marin, Evgeniy M.

    2016-01-01

    The relevance of the study is reasoned by the increasing complexity of life activity in modern society, which results in distortion of the moral and value criteria and norms. The purpose of the article is to reveal anthropological methods of formation of university students' spiritual and moral culture. The leading approach to the study is the…

  4. Digital Storytelling: A Method for Engaging Students and Increasing Cultural Competency

    Science.gov (United States)

    Grant, Natalie S.; Bolin, Brien L.

    2016-01-01

    Digital storytelling is explored as a method of engaging students in the development of media literacy and cultural competency. This paper describes the perceptions and experiences of 96 undergraduate students at a large Midwestern university, after completing a digital storytelling project in a semester-long diversity course. Digital storytelling…

  5. Rapid radiorespirometric method for the estimation of bacterial counts of cellulomonas cultures

    International Nuclear Information System (INIS)

    Jakubick-Neradova, V.

    1975-01-01

    The determination of bacterial counts in cultures of Cellulomonas species C-72-4 by a radiorespirometric technique was twenty times faster than the conventional method. A linear correlation was found between the bacterial counts and the 14 CO 2 metabolized from labelled glucose added to the cellulose medium. (orig.) [de

  6. X-ray and synchrotron methods in studies of cultural heritage sites

    Science.gov (United States)

    Koval'chuk, M. V.; Yatsishina, E. B.; Blagov, A. E.; Tereshchenko, E. Yu.; Prosekov, P. A.; Dyakova, Yu. A.

    2016-09-01

    X-ray and synchrotron methods that are most widely used in studies of cultural heritage objects (including archaeological sites)—X-ray diffraction analysis, X-ray spectroscopy, and visualization techniques— have been considered. The reported examples show high efficiency and informativeness of natural science studies when solving most diverse problems of archaeology, history, the study of art, museology, etc.

  7. Learning as Researchers and Teachers: The Development of a Pedagogical Culture for Social Science Research Methods?

    Science.gov (United States)

    Kilburn, Daniel; Nind, Melanie; Wiles, Rose

    2014-01-01

    In light of calls to improve the capacity for social science research within UK higher education, this article explores the possibilities for an emerging pedagogy for research methods. A lack of pedagogical culture in this field has been identified by previous studies. In response, we examine pedagogical literature surrounding approaches for…

  8. X-ray and synchrotron methods in studies of cultural heritage sites

    International Nuclear Information System (INIS)

    Koval’chuk, M. V.; Yatsishina, E. B.; Blagov, A. E.; Tereshchenko, E. Yu.; Prosekov, P. A.; Dyakova, Yu. A.

    2016-01-01

    X-ray and synchrotron methods that are most widely used in studies of cultural heritage objects (including archaeological sites)—X-ray diffraction analysis, X-ray spectroscopy, and visualization techniques— have been considered. The reported examples show high efficiency and informativeness of natural science studies when solving most diverse problems of archaeology, history, the study of art, museology, etc.

  9. X-ray and synchrotron methods in studies of cultural heritage sites

    Energy Technology Data Exchange (ETDEWEB)

    Koval’chuk, M. V.; Yatsishina, E. B.; Blagov, A. E.; Tereshchenko, E. Yu., E-mail: elenatereschenko@yandex.ru; Prosekov, P. A.; Dyakova, Yu. A. [National Research Centre “Kurchatov Institute” (Russian Federation)

    2016-09-15

    X-ray and synchrotron methods that are most widely used in studies of cultural heritage objects (including archaeological sites)—X-ray diffraction analysis, X-ray spectroscopy, and visualization techniques— have been considered. The reported examples show high efficiency and informativeness of natural science studies when solving most diverse problems of archaeology, history, the study of art, museology, etc.

  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. Critical assessment of extracellular polymeric substances extraction methods from mixed culture biomass

    DEFF Research Database (Denmark)

    Pellicer i Nàcher, Carles; Domingo Felez, Carlos; Mutlu, Ayten Gizem

    2013-01-01

    Extracellular polymeric substances (EPS) have a presumed determinant role in the structure, architecture, strength, filterability, and settling behaviour of microbial solids in biological wastewater treatment processes. Consequently, numerous EPS extraction protocols have recently been published....... This study presents a rigorous and critical assessment of existing physical and chemical EPS extraction methods applied to mixed-culture biomass samples (nitrifying, nitritation-anammox, and activated sludge biomass). A novel fluorescence-based method was developed and calibrated to quantify the lysis...

  12. Simple Sample Preparation Method for Direct Microbial Identification and Susceptibility Testing From Positive Blood Cultures

    Directory of Open Access Journals (Sweden)

    Hong-wei Pan

    2018-03-01

    Full Text Available Rapid identification and determination of the antibiotic susceptibility profiles of the infectious agents in patients with bloodstream infections are critical steps in choosing an effective targeted antibiotic for treatment. However, there has been minimal effort focused on developing combined methods for the simultaneous direct identification and antibiotic susceptibility determination of bacteria in positive blood cultures. In this study, we constructed a lysis-centrifugation-wash procedure to prepare a bacterial pellet from positive blood cultures, which can be used directly for identification by matrix-assisted laser desorption/ionization-time-of-flight mass spectrometry (MALDI-TOF MS and antibiotic susceptibility testing by the Vitek 2 system. The method was evaluated using a total of 129 clinical bacteria-positive blood cultures. The whole sample preparation process could be completed in <15 min. The correct rate of direct MALDI-TOF MS identification was 96.49% for gram-negative bacteria and 97.22% for gram-positive bacteria. Vitek 2 antimicrobial susceptibility testing of gram-negative bacteria showed an agreement rate of antimicrobial categories of 96.89% with a minor error, major error, and very major error rate of 2.63, 0.24, and 0.24%, respectively. Category agreement of antimicrobials against gram-positive bacteria was 92.81%, with a minor error, major error, and very major error rate of 4.51, 1.22, and 1.46%, respectively. These results indicated that our direct antibiotic susceptibility analysis method worked well compared to the conventional culture-dependent laboratory method. Overall, this fast, easy, and accurate method can facilitate the direct identification and antibiotic susceptibility testing of bacteria in positive blood cultures.

  13. Comparison of culture and PCR methods in the diagnosis of bacterial meningitis.

    Science.gov (United States)

    Başpınar, Emel Ödemiş; Dayan, Saim; Bekçibaşı, Muhammed; Tekin, Recep; Ayaz, Celal; Deveci, Özcan; Hoşoğlu, Salih

    Our aim in this study is to compare the standard culture method with the multiplex PCR and the Speed-Oligo ® Bacterial Meningitis Test (SO-BMT) - a hybridization-based molecular test method - during the CSF examination of the patients with the pre-diagnosis of acute bacterial meningitis. For the purposes of this study, patients with acute bacterial meningitis treated at the Dicle University Medical Faculty Hospital, Infectious Diseases and Clinical Microbiology Clinic between December 2009 and April 2012 were retrospectively evaluated. The diagnosis of bacterial meningitis was made based on the clinical findings, laboratory test anomalies, CSF analysis results, and the radiological images. Growth was observed in the CSF cultures of 10 out of the 57 patients included in the study (17.5%) and Streptococcus pneumoniae was isolated in all of them. The CSF samples of 34 patients (59.6%) were positive according to the SO-BMT and S. pneumoniae was detected in 33 of the samples (97.05%), while Neisseria meningitidis was found in 1 sample (2.95%). In a total of 10 patients, S. pneumoniae was both isolated in the CSF culture and detected in the SO-BMT. The culture and the SO-BMT were negative in 23 of the CSF samples. There was no sample in which the CSF culture was positive although the SO-BMT was negative. While SO-BMT seems to be a more efficient method than bacterial culturing to determine the pathogens that most commonly cause bacterial meningitis in adults, further studies conducted on larger populations are needed in order to assess its efficiency and uses. Copyright © 2016 Sociedade Brasileira de Microbiologia. Published by Elsevier Editora Ltda. All rights reserved.

  14. 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.

  15. 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.

  16. Comparison of Real-time PCR method and blood culture in diagnosis of septicemia

    Directory of Open Access Journals (Sweden)

    Ali Gholami

    2016-02-01

    Full Text Available Background: Bloodstream infections (BSI have a high incidence and high mortality in the worldwide. The mortality rate is variable between 20-70%. Therefore, early and timely detection of BSI agent in clinical laboratories is necessary. The aim of this study was to determine an efficient diagnostic tool to septicemia in accompany of blood culture method by Real-time PCR (using panbacterial 23S rRNA gene. Methods: This cross-sectional study was conducted in two analytical and clinical stages in Hamadan University of Medical Sciences, Iran, from October 2014 to June 2015. In analytical stage, sensitivity (by serial dilution from 104 to 1 CFU/ml and specificity of the primer were evaluated with the Staphylococcus aureus (as Gram positive indicator bacteria and Escherichia coli (as Gram-negative indicator bacteria, human genome (from Hella cell culture, Candida albicans yeast and Aspergillus fumigatus fungus. In clinical stage, 121 blood samples were collected from patients suspected to sepsis in intensive care unit (ICU from Hamadan University Hospitals. Finally, the results of Real-time PCR and blood culture methods were compared. Results: The Real-time PCR showed a sensitivity ranging from 2 to 10 target copies per reaction to the whole blood for Escherichia coli and Staphylococcus aureus respectively. The specificity of this method was evaluated and no false positive amplification was identified. 57.85% (70 cases of the samples were positive by Real-time PCR and 13.22% (16 cases of the samples were positive by blood culture. However, none of the cases that were positive by blood culture were negative in Real-time PCR. As well as, 44.62% (54 cases of cases were positive by Real-time PCR but blood culture showed no bacteria in the samples, and 42.15% (51 cases were negative by both methods. Correlation or agreement of Kappa was 0.20, that indicating poor agreement between the two methods. Conclusion: Real-time PCR is more sensitive than blood

  17. Comparison of the lysis centrifugation method with the conventional blood culture method in cases of sepsis in a tertiary care hospital.

    Science.gov (United States)

    Parikh, Harshal R; De, Anuradha S; Baveja, Sujata M

    2012-07-01

    Physicians and microbiologists have long recognized that the presence of living microorganisms in the blood of a patient carries with it considerable morbidity and mortality. Hence, blood cultures have become critically important and frequently performed test in clinical microbiology laboratories for diagnosis of sepsis. To compare the conventional blood culture method with the lysis centrifugation method in cases of sepsis. Two hundred nonduplicate blood cultures from cases of sepsis were analyzed using two blood culture methods concurrently for recovery of bacteria from patients diagnosed clinically with sepsis - the conventional blood culture method using trypticase soy broth and the lysis centrifugation method using saponin by centrifuging at 3000 g for 30 minutes. Overall bacteria recovered from 200 blood cultures were 17.5%. The conventional blood culture method had a higher yield of organisms, especially Gram positive cocci. The lysis centrifugation method was comparable with the former method with respect to Gram negative bacilli. The sensitivity of lysis centrifugation method in comparison to conventional blood culture method was 49.75% in this study, specificity was 98.21% and diagnostic accuracy was 89.5%. In almost every instance, the time required for detection of the growth was earlier by lysis centrifugation method, which was statistically significant. Contamination by lysis centrifugation was minimal, while that by conventional method was high. Time to growth by the lysis centrifugation method was highly significant (P value 0.000) as compared to time to growth by the conventional blood culture method. For the diagnosis of sepsis, combination of the lysis centrifugation method and the conventional blood culture method with trypticase soy broth or biphasic media is advocable, in order to achieve faster recovery and a better yield of microorganisms.

  18. New method for exopolysaccharide determination in culture broth using stirred ultrafiltration cells.

    Science.gov (United States)

    Bergmaier, D; Lacroix, C; Guadalupe Macedo, M; Champagne, C P

    2001-10-01

    A new method to remove simple carbohydrates from culture broth prior to the quantification of exopolysaccharides (EPS) was developed and validated for the EPS-producing strain, Lactobacillus rhamnosus RW-9595M. This method uses ultrafiltration (UF) in stirred cells followed by polysaccharide detection in the retentate by the phenol-sulfuric acid method. The UF method was compared with a conventional method based on ethanol extraction, dialysis, protein removal by trichloroacetic acid (TCA) and freeze-drying. EPS production during pH-controlled batch fermentations in basal minimum medium, whey permeate (WP). and whey permeate supplemented with yeast extract, minerals and Tween-80 (SWP) was determined by the new UF and conventional methods. EPS recovery by the new method ranged from 83% to 104% for EPS added in the concentration range 40-1,500 mg/l in 0.1 M NaCl solution or culture medium. The UF method was rapid (8 h), accurate and simple, and required only a small sample volume (1-5 ml). A very high maximum EPS production was measured in SWP by both the UF and conventional methods (1,718 and 1,755 mg/l).

  19. [Detection of norovirus RNA in bivalve molluscs by using bacteria-culture-employed method (A3T method)].

    Science.gov (United States)

    Akiba, Tetsuya; Tanaka, Tatsuya; Nagano, Miyuki; Mori, Kohji; Hayashi, Yukinao; Obata, Hiromi; Chiba, Takashi; Ikuta, Yasuhisa; Kamiya, Yoriko; Nakama, Akiko; Hosaka, Mitsugu; Kai, Akemi

    2010-01-01

    Norovirus (NV) RNA has rarely been detected in foods despite the use of highly sensitive methods such as RT-PCR and real-time RT-PCR. In the modified method (A3T method) reported previously, a bacterial culture process was introduced into the standard protocol for NV detection to remove some inhibitor(s) present in food ingredients. To confirm the efficiency of the A3T method and to examine NV contamination in bivalve molluscs, we tried to detect NV RNA in bivalve molluscs on the market and in oyster samples associated with foodborne outbreaks by using the standard method and the A3T method. NV RNAs were detected in 20 samples (18.0%) of 111 bivalve molluscs, including oysters, on the market by use of the A3T method, while only one sample (0.9%) was positive according to the standard method. NV RNA was also detected in 10 of 35 oyster samples related to foodborne outbreaks by the A3T method. Those results show that the A3T method is suitable for the detection of NV in bivalve molluscs in general laboratories.

  20. Monitoring the ecology of Bacillus during Daqu incubation, a fermentation starter, using culture-dependent and culture-independent methods.

    Science.gov (United States)

    Yan, Zheng; Zheng, Xiao-Wei; Han, Bei-Zhong; Han, Jian-Shu; Nout, M J Robert; Chen, Jing-Yu

    2013-05-01

    Daqu, a traditional fermentation starter, has been used to produce attractively flavored foods such as vinegar and Chinese liquor for thousands of years. Although Bacillus spp. are one of the dominant microorganisms in Daqu, more precise information is needed to reveal why and how Bacillus became dominant in Daqu, and next, to assess the impact of Bacillus sp. on Daqu and its derived products. We combined culture-dependent and culture-independent methods to study the ecology of Bacillus during Daqu incubation. Throughout the incubation, 67 presumptive Bacillus spp. isolates were obtained, 52 of which were confirmed by 16S rDNA sequencing. The identified organisms belonged to 8 Bacillus species: B. licheniformis, B. subtilis, B. amyloliquefaciens, B. cereus, B. circulans, B. megaterium, B. pumilus, and B. anthracis. A primer set specific for Bacillus and related genera was used in a selective PCR study, followed by a nested DGGE PCR targeting the V9 region of the 16S rDNA. Species identified from the PCR-DGGE fingerprints were related to B. licheniformis, B. subtilis, B. amyloliquefaciens, B. pumilus, B. benzoevorans, and B. foraminis. The predominant species was found to be B. licheniformis. Certain B. licheniformis strains exhibited potent antimicrobial activities. The greatest species diversity occurred at the Liangmei stage of Daqu incubation. To date, we lack sufficient knowledge of Bacillus distribution in Daqu. Elucidating the ecology of Bacillus during Daqu incubation would enable the impact of Bacillus on Daqu to be accessed, and the quality and stabilization of Daqu-derived products to be optimized.

  1. Reflexive photography: an alternative method for documenting the learning process of cultural competence.

    Science.gov (United States)

    Amerson, Roxanne; Livingston, Wade G

    2014-04-01

    This qualitative descriptive study used reflexive photography to evaluate the learning process of cultural competence during an international service-learning project in Guatemala. Reflexive photography is an innovative qualitative research technique that examines participants' interactions with their environment through their personal reflections on images that they captured during their experience. A purposive sample of 10 baccalaureate nursing students traveled to Guatemala, where they conducted family and community assessments, engaged in home visits, and provided health education. Data collection involved over 100 photographs and a personal interview with each student. The themes developed from the photographs and interviews provided insight into the activities of an international experience that influence the cognitive, practical, and affective learning of cultural competence. Making home visits and teaching others from a different culture increased students' transcultural self-efficacy. Reflexive photography is a more robust method of self-reflection, especially for visual learners.

  2. Methods for Culture-Independent Identification of Lactic Acid Bacteria in Dairy Products

    Directory of Open Access Journals (Sweden)

    Tomislav Pogačić

    2010-01-01

    Full Text Available Culture-independent molecular tools have been introduced into food microbiology during the last ten years. Most of them are based on the amplification of a bulk bacterial DNA extracted directly from a sample, the targeting of a selected gene, or a variable region of the selected gene. Many studies have explored indigenous lactic acid bacteria in dairy products by culture-independent molecular approaches. It is well known that indigenous microbiota significantly contribute to the uniqueness of artisanal cheeses. However, there is no molecular method that can provide complete qualitative and quantitative insight into the microbiota associated with a certain ecosystem. Therefore, a combination of molecular approaches should be applied to get a more objective picture of the microbiota. This paper aims to present the most widely used culture-independent molecular tools for identifying lactic acid bacteria in dairy products.

  3. Express Method for Redox Potential and pH Measuring in Microbial Cultures

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    Oleksandr Tashyrev

    2014-09-01

    Full Text Available The new modification of potentiometric pH and Eh measuring was developed. The essence of the method is to measure pH and Eh in compact hermetic redox cell. This modification provides reliable and fast pH and Eh measuring with minimum expense of culture liquid (1 cm3 for each measuring. The application of the developed modification in microbiology was shown on the example of bacterial culture Kocuria carniphila isolated from Negev desert soil (Israel. Dynamics of the Eh changing in the growing culture allows establishing the influence of the carbon and energy sources concentration on the microbial growth, the rate of oxygen consumption and character of microbial interaction with toxic compounds. Copiotriphic conditions (800 mg/l C were shown to be optimal for K. carniphila. It was shown that K. carniphila was resistant to 200 ppm of Cu2+, nevertheless it did not extract Cu2+ out of liquid medium.

  4. The efficient method for simultaneous monitoring of the culturable as well as nonculturable airborne microorganisms.

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    Barbara Hubad

    Full Text Available Cultivation-based microbiological methods are a gold standard for monitoring of airborne micro-organisms to determine the occupational exposure levels or transmission paths of a particular infectious agent. Some highly contagious microorganisms are not easily culturable but it is becoming evident that cultivation and molecular methods are complementary and in these cases highly relevant. We report a simple and efficient method for sampling and analyzing airborne bacteria with an impactor-type high-flow-rate portable air sampler, currently used for monitoring culturable bacteria or fungi. A method is reported for extraction of nucleic acids from impacted cells without prior cultivation and using agarose as a sampling matrix. The DNA extraction efficiency was determined in spiked samples and, samples taken from a wastewater treatment plant and an alpine area. The abundance, diversity and quantity of total bacteria were analysed by a quantitative polymerase chain reaction, and by construction and analysis of clone libraries. The method does not interfere with downstream PCR analysis and can cover the gap between traditional culture and molecular techniques of bioaerosol monitoring.

  5. In vitro culture of functionally active buffalo hepatocytes isolated by using a simplified manual perfusion method.

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    Santanu Panda

    Full Text Available In farm animals, there is no suitable cell line available to understand liver-specific functions. This has limited our understanding of liver function and metabolism in farm animals. Culturing and maintenance of functionally active hepatocytes is difficult, since they survive no more than few days. Establishing primary culture of hepatocytes can help in studying cellular metabolism, drug toxicity, hepatocyte specific gene function and regulation. Here we provide a simple in vitro method for isolation and short-term culture of functionally active buffalo hepatocytes.Buffalo hepatocytes were isolated from caudate lobes by using manual enzymatic perfusion and mechanical disruption of liver tissue. Hepatocyte yield was (5.3 ± 0.66×107 cells per gram of liver tissue with a viability of 82.3 ± 3.5%. Freshly isolated hepatocytes were spherical with well contrasted border. After 24 hours of seeding onto fibroblast feeder layer and different extracellular matrices like dry collagen, matrigel and sandwich collagen coated plates, hepatocytes formed confluent monolayer with frequent clusters. Cultured hepatocytes exhibited typical cuboidal and polygonal shape with restored cellular polarity. Cells expressed hepatocyte-specific marker genes or proteins like albumin, hepatocyte nuclear factor 4α, glucose-6-phosphatase, tyrosine aminotransferase, cytochromes, cytokeratin and α1-antitrypsin. Hepatocytes could be immunostained with anti-cytokeratins, anti-albumin and anti α1-antitrypsin antibodies. Abundant lipid droplets were detected in the cytosol of hepatocytes using oil red stain. In vitro cultured hepatocytes could be grown for five days and maintained for up to nine days on buffalo skin fibroblast feeder layer. Cultured hepatocytes were viable for functional studies.We developed a convenient and cost effective technique for hepatocytes isolation for short-term culture that exhibited morphological and functional characteristics of active hepatocytes

  6. 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

  7. 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.

  8. Water Quality Improvement of Media Culture for Tilapia (Oreochromis niloticus) with Cleaner Production Method

    Science.gov (United States)

    Haeruddin; Supriharyono; Febrianto, S.

    2018-02-01

    The tilapia (Oreochromis niloticus), is known as a high adaptability and brackish water tolerance fish. This fish is also has a meat with high protein content, that ranges about 65 -75%. Generally the tilapia is cultured using a conventional system with high density. It is caused degradation of water quality of media culture, and finally increase mortality rate of fish cultured. The application of tilapia cultivation with cleaner production method by giving enzyme into the feed to upgrade the efficiency of feed utilization, presumed that could improve the water quality of cultivation media. It is due to the lower of feed and feces residues. Therefore the concentration of toxic compounds, such as ammonia, nitrite and sulfide, will be lower. The experiments were conducted for 35 days with a completely factorial randomized design. The first factor was the dosage of enzyme in the feed, consisting of 4 dosages, and the second factor was the duration of the test fish maintenance (5 weeks). Water quality variables examined included ammonia, nitrite and sulfide. The results showed that enzyme dosage had no significantly impact on ammonia, nitrite and sulfide concentrations in the test media culture. However, the feeding with enzyme in low dosage, resulted less concentration of ammonia, nitrite and sulfide than it was without enzyme). The duration of fish cultured has significantly effect on the concentration of ammonia, nitrite and sulfide in the test media. While it is no significantly correlation between dosage and duration of maintenance.

  9. Molecular biology-based methods for quantification of bacteria in mixed culture: perspectives and limitations.

    Science.gov (United States)

    Nagarajan, Karthiga; Loh, Kai-Chee

    2014-08-01

    Species-specific enumeration of mixed community is invaluable as it facilitates a better understanding of the significance of the individual strains, their interactions, and the underlying mechanisms of community dynamics. Mixed microbial community has been characterized by microbiological, biochemical, or molecular biology-based methods. While microbiological and biochemical techniques do not provide adequate quantitative information of the members of the consortia and require additional techniques for a more comprehensive analysis, molecular biology-based methods analyze the microbial consortium based on specific DNA sequences and do not require isolation and culturing of bacteria for quantitative analysis. These methods outshine conventional culture-based techniques in terms of better sensitivity, reproducibility, and reliability. Quantitative molecular biology methods have been classified as PCR-based and probe hybridization methods. The PCR-based methods includes quantitative real-time PCR and terminal restriction fragment length polymorphism, while fluorescent in situ hybridization and DNA microarrays fall under probe hybridization methods. The workflow, the quantification methods, and their potential applications are discussed in this review by highlighting their advantages and possible limitations.

  10. Safe operation of nuclear power plants - Is safety culture an adequate management method?

    International Nuclear Information System (INIS)

    Piirto, A.

    2012-01-01

    One of the characteristics of a good safety culture is a definable commitment to the improvement of safety behaviours and attitudes at all organisational levels. A second characteristic of an organisation with excellent safety culture is free and open communication. The general understanding has been that safety culture is a part of organisation culture. In addition to safety culture thinking, proactive programmes and displays of proactive work to improve safety are required. This work needs to include, qt a minimum, actions aiming at reducing human errors, the development of human error prevention tools, improvements in training, and the development of working methods and the organisation's activities. Safety depends not only on the technical systems, but also on the organisation. There is a need for better methods and tools for organisational assessment and development. Today there is universal acceptance of the significant impact that management and organisational factors have over the safety significance of complex industrial installations such as nuclear power plants. Many events with significant economic and public impact had causes that have been traced to management deficiencies. The objective of this study is development of new methods to increase safety of nuclear power plant operation. The research has been limited to commercial nuclear power plants that are intended for electrical power generation in Finland. Their production activities, especially operation and maintenance, are primarily reviewed from a safety point of view, as well as human performance and organisational factors perspective. This defines the scope and focus of the study. The research includes studies related to knowledge management and tacit knowledge in the project management context and specific studies related to transfer of tacit knowledge in the maintenance organization and transfer of tacit knowledge between workers of old generation and young generation. The empirical results

  11. Safe operation of nuclear power plants - Is safety culture an adequate management method?

    Energy Technology Data Exchange (ETDEWEB)

    Piirto, A.

    2012-07-01

    One of the characteristics of a good safety culture is a definable commitment to the improvement of safety behaviours and attitudes at all organisational levels. A second characteristic of an organisation with excellent safety culture is free and open communication. The general understanding has been that safety culture is a part of organisation culture. In addition to safety culture thinking, proactive programmes and displays of proactive work to improve safety are required. This work needs to include, qt a minimum, actions aiming at reducing human errors, the development of human error prevention tools, improvements in training, and the development of working methods and the organisation's activities. Safety depends not only on the technical systems, but also on the organisation. There is a need for better methods and tools for organisational assessment and development. Today there is universal acceptance of the significant impact that management and organisational factors have over the safety significance of complex industrial installations such as nuclear power plants. Many events with significant economic and public impact had causes that have been traced to management deficiencies. The objective of this study is development of new methods to increase safety of nuclear power plant operation. The research has been limited to commercial nuclear power plants that are intended for electrical power generation in Finland. Their production activities, especially operation and maintenance, are primarily reviewed from a safety point of view, as well as human performance and organisational factors perspective. This defines the scope and focus of the study. The research includes studies related to knowledge management and tacit knowledge in the project management context and specific studies related to transfer of tacit knowledge in the maintenance organization and transfer of tacit knowledge between workers of old generation and young generation. The empirical

  12. A rapid plaque method using vertical tube cultures for titration of viruses and neutralizing antibodies.

    Science.gov (United States)

    Wallis, C; Parks, W; Sakurada, N; Melnick, J L

    1965-01-01

    Standard methods for titrating and typing enteroviruses and other viruses, or for assaying antibodies against them, are based on observation of metabolic inhibition of infected cells or on direct microscopic reading of cytopathogenic effects. Incubation of cell cultures for at least a week, with two or three readings during this period, is usually required before assessment is completed. This report describes a vertical-tube method in which cell monolayers are confined to the bottom end of a serological tube; an agar overlay is used after virus or virus-serum inoculation. The reduced monolayer area allows seeding with only about 30% of the cells required for standard tube cultures, and 5% of those required for plaque assay in bottle cultures. The new method requires only a single macroscopic reading one to three days after the test is set up. This method has proved economical, simple and rapid in epidemiological studies on rapidly growing viruses of the entero-, reo-, herpes-, myxo- and poxvirus groups, and for tests of the genetic markers of live poliovirus vaccine.

  13. Studies of Ancient Russian Cultural Objects Using the Neutron Tomography Method

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    Sergey Kichanov

    2018-01-01

    Full Text Available Neutron radiography and tomography is a non-destructive method that provides detailed information about the internal structure of cultural heritage objects. The differences in the neutron attenuation coefficients of constituent elements of the studied objects, as well as the application of modern mathematical algorithms to carry out three-dimensional imaging data analysis, allow one to obtain unique information about the spatial distribution of different phases, the presence of internal defects, or the degree of structural degradation inside valuable cultural objects. The results of the neutron studies of several archaeological objects related to different epochs of the Russian history are reported in order to demonstrate the opportunities provided by the neutron tomography method. The obtained 3D structural volume data, as well as the results of the corresponding data analysis, are presented.

  14. Prevalence of periodontopathogenic bacteria in patients suffering from periodontitis using culture and PCR methods

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    Amir Aliramezani

    2012-01-01

    Full Text Available Background and Aims: Periodontitis is one of the most common oral diseases with the various incidence rates in different populations. A number of bacteria are considered as the major etiologic agents of periodontitis. The aim of the present study was to determine the prevalence of periodontopathogen bacteria in patients using both PCR and culture techniques.Materials and Methods: In this study, one-hundred patients (including 62 females and 38 males with an average age of 49±11.5 years with adult periodontitis referred to periodontics department of School of Dentistry/Tehran University of Medical Sciences were investigated. The samples were taken and sent immediately to the laboratory for culture and molecular evaluation. The PCR was performed using specific primers and the statistical analysis of data was performed using SPSS statistic software and McNemar test.Results: The results demonstrated that the total detection rate in culture method was 64%. The rate of Aggregatibacter actinomycetemcomitans (Aa was 28% which was significantly higher than that of Porphyromonas gingivalis (Pg (6% and Prevotella intermedia (Pi (3%. 27% of cases showed mixed bacterial growth. 65% of patients were positive using molecular method. The rate of Aa (30% was significantly higher than that of Pg (7% and Pi (5%. The mixed PCR positive rate containing of Aa, Pg and Pi was (23%.Conclusion: In this study, it was found that most of the bacteria isolated using culture and molecular methods were Aa, Pg and Pi, respectively. Although the detection frequencies of both techniques were similar, the specificity, sensitivity and bacterial detection speed of the PCR technique is obviously higher. Therefore, the use of molecular techniques is strongly recommended. However, both techniques seem to be suitable for microbiological diagnostics.

  15. Growth and accumulation of flavan-3-ol in Camellia sinensis through callus culture and suspension culture method

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    Sutini Sutini

    2017-02-01

    Full Text Available This study was aimed to assess flavan-3-ol biomass in C. sinensis through callus cultures and suspension cultures derived from leaf explants. Callus initiation of both cultures were using Murashige and Skoog medium were enriched with plant growth regulators Naphtha-lene Acetic Acid 3.0 mg/L and kinetin 2.0 mg/L. The procedures in this study were: (1 callus initiation by cutting the leaves of C. sinen-sis shoots then planted on Murashige and Skoog medium that were enriched with plant growth regulators, (2 sub callus culture on fresh medium that enriched with the same growth regulators, (3 suspension culture initiation of liquid callus, (4 growth examination of callus and suspension cultures in week 12, (5 examination of qualitative-quantitative content of flavan-3-olin suspension cultures at week 4. The results show that suspension cultures contain biomass flavan-3-ol that increase in the same manner of the increase of callus age and weight

  16. 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.

  17. Bacterial ecology of artisanal Minas cheeses assessed by culture-dependent and -independent methods.

    Science.gov (United States)

    Perin, Luana Martins; Savo Sardaro, Maria Luisa; Nero, Luís Augusto; Neviani, Erasmo; Gatti, Monica

    2017-08-01

    Artisanal Minas cheese is produced in Minas Gerais state, Brazil and its varieties are named according to their geographical origin (Serro, Canastra, Serra do Salitre, Araxá and Campo das Vertentes). The cheese is produced with raw cow's milk and the whey from the previous cheese production ("pingo"). The high economic and cultural importance of artisanal cheese in Brazil justifies the efforts to ensure its safety, quality and provenance. This study aimed to characterize the microbial diversity composition, and geographical distribution of artisanal Minas cheese, focusing on the characterization of its autochthonous lactic acid bacteria (LAB) microbiota. Artisanal Minas cheese varieties from Serro, Canastra, Serra do Salitre, Araxá and Campo das Vertentes were analyzed by culture-dependent (culturing and LAB sequencing) and -independent (repetitive extragenic palindromic-PCR (rep-PCR) and length heterogeneity-PCR, LH-PCR) methods to characterize the microbiota. The microbial counts were variable between cheese samples, and some samples presented high number of coagulase positive bacteria and coliforms that may be associated with hygienic issues. In all samples was observed a prevalence of LAB. 16S rRNA sequencing and rep-PCR of the LAB strains identified four genus (Lactobacillus, Lactococcus, Enterococcus and Weissella), ten species and more than one strain per species. Lactobacillus was the most prevalent genera in all the cheeses. LH-PCR revealed a further six genera and ten species that were not identified by culturing, highlighting the importance of combining both culture-dependent and -independent methods to fully characterize microbiota diversity. Principal component analysis of the LH-PCR data and cluster analysis of rep-PCR data revealed that the artisanal Minas cheese microbiota was influenced not only by their geographical origin but also by the cheese farm. The lack of standardization in the milking and cheese manufacturing procedures between

  18. How guiding coalitions promote positive culture change in hospitals: a longitudinal mixed methods interventional study.

    Science.gov (United States)

    Bradley, Elizabeth H; Brewster, Amanda L; McNatt, Zahirah; Linnander, Erika L; Cherlin, Emily; Fosburgh, Heather; Ting, Henry H; Curry, Leslie A

    2018-03-01

    Quality collaboratives are widely endorsed as a potentially effective method for translating and spreading best practices for acute myocardial infarction (AMI) care. Nevertheless, hospital success in improving performance through participation in collaboratives varies markedly. We sought to understand what distinguished hospitals that succeeded in shifting culture and reducing 30-day risk-standardised mortality rate (RSMR) after AMI through their participation in the Leadership Saves Lives (LSL) collaborative. We conducted a longitudinal, mixed methods intervention study of 10 hospitals over a 2-year period; data included surveys of 223 individuals (response rates 83%-94% depending on wave) and 393 in-depth interviews with clinical and management staff most engaged with the LSL intervention in the 10 hospitals. We measured change in culture and RSMR, and key aspects of working related to team membership, turnover, level of participation and approaches to conflict management. The six hospitals that experienced substantial culture change and greater reductions in RSMR demonstrated distinctions in: (1) effective inclusion of staff from different disciplines and levels in the organisational hierarchy in the team guiding improvement efforts (referred to as the 'guiding coalition' in each hospital); (2) authentic participation in the work of the guiding coalition; and (3) distinct patterns of managing conflict. Guiding coalition size and turnover were not associated with success (p values>0.05). In the six hospitals that experienced substantial positive culture change, staff indicated that the LSL learnings were already being applied to other improvement efforts. Hospitals that were most successful in a national quality collaborative to shift hospital culture and reduce RSMR showed distinct patterns in membership diversity, authentic participation and capacity for conflict management. © Article author(s) (or their employer(s) unless otherwise stated in the text of the

  19. A safety culture assessment by mixed methods at a public maternity and infant hospital in China

    Directory of Open Access Journals (Sweden)

    Listyowardojo TA

    2017-07-01

    Full Text Available Tita Alissa Listyowardojo,1 Xiaoling Yan,2,3 Stephen Leyshon,1 Bobbie Ray-Sannerud,1 Xin Yan Yu,4 Kai Zheng,4 Tao Duan2,3 1Life Sciences Program, Group Technology and Research, DNV GL, Hovik, Norway; 2Quality and Safety Department, Shanghai First Maternity and Infant Hospital, 3Tongji University School of Medicine, Shanghai, 4Healthcare Department, Business Assurance, DNV GL, Beijing, China Objective: To assess safety culture at a public maternity hospital in Shanghai, China, using a sequential mixed methods approach. The study was part of a bigger study looking at the application of the mixed methods approach to assess safety culture in health care in different organizations and countries.Methodology: A mixed methods approach was utilized by first distributing the Safety Attitudes Questionnaire measuring six safety culture dimensions and five independent items to all hospital staff (n=1482 working in 18 departments at a single hospital. Afterward, semistructured interviews were conducted using convenience sampling, where 48 hospital staff from nine departments at the same hospital were individually interviewed.Results: The survey received a response rate of 96%. The survey findings show significant differences between the hospital departments in almost all safety culture dimensions and independent items. Similarly, the interview findings revealed that there were different, competing priorities between departments perceived to result in a reduced quality of collaboration and bottlenecks in care delivery. Another major finding was that staff who worked more hours per week would perceive working conditions significantly more negatively. Issues related to working conditions were also the most common concerns discussed in the interviews, especially the issue on high workload. High workload was also reflected in the fact that 91.45% of survey respondents reported that they worked 40 hours or longer per week. Finally, interview findings complemented

  20. Ethnonursing: A Qualitative Research Method for Studying Culturally Competent Care across Disciplines

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    Marilyn R. McFarland PhD, RN, FNP-BC, CTN

    2012-07-01

    Full Text Available Nurse anthropologist, Madeleine Leininger, developed the culture care theory and ethnonursing research method to help researchers study transcultural human care phenomena and discover the knowledge nurses need to provide care in an increasingly multicultural world. The authors propose that the ethnonursing method can be useful for research that addresses providing care in other disciplines, including education, administration, physical, occupational, and speech therapy, social work, pharmacy, medicine, and other disciplines in which research findings have implications for human care and health. The authors discuss the culture care theory and describe the ethnonursing research method's enablers, data analysis phases, and qualitative evaluation criteria. The theory is presented as a guide for using research findings to design culturally competent and congruent care to promote well-being among diverse people, groups, communities, and institutions. Resources include a reference list of key source publications, a discussion of exemplar studies, and samples of a theory-based, open-ended interview guide and data coding system.

  1. Tissue culture methods for the clonal propagation and genetic improvement of Spanish red cedar (Cedrela odorata).

    Science.gov (United States)

    Peña-Ramírez, Yuri; Juárez-Gómez, Juan; González-Rodríguez, José Antonio; Robert, Manuel L

    2012-01-01

    The choice of a method to culture red cedar tissues depends on the final objectives pursued. If homogeneous clonal material is required for experimental purposes, the easiest way is to generate the lines through adventitious shoot induction from seedlings germinated from seeds. If the objective is to generate high yielding material for plantation purposes, the choice will be the same method but starting from mature vegetative tissues from selected elite plants. Most of the process are the same, but the initial steps are less efficient and much more elaborate. If the purpose is to generate lines with new genetic characteristics through somaclonal variation, mutagenesis, or genetic transformation, somatic embryogenesis will be required. No single method in its present form is suitable for all purposes. Eventually, the efficient production of somatic embryos from rejuvenated shoots collected from mature selected plants is the ideal way to culture this species, but for the time being we have to choose one or the other. In this chapter, we present a grafting procedure to rejuvenate and maintain mother plants in the greenhouse and the in vitro culture systems we have developed for the production of Cedrela odorata propagules using explants from both young seedlings and mature tissues from selected old trees. Using a modified TY17 medium and the BioMINT(®) temporary immersion system, we obtained high multiplication and ex vitro transplantation rates for efficient large-scale propagation of this species.

  2. Enlargement of induced variations by combined method of chronic irradiations with callus culture in sugarcane

    International Nuclear Information System (INIS)

    Nagatomi, Shigeki

    1993-01-01

    The present study was conducted to elucidate the effects of gamma ray irradiation and callus culture upon induced variation of the regeneratives. The populations regenerated from young leaf tissue of chronic irradiated plnats grown under a gamma field receiving a total dose of 300 and 100 Gy, showed rather wider variation on quantitative characters than plants from populations of the non-irradiated. This variation extended in both negative and positive directions. Analysis of variance also revealed that variation and heritability in broad sense of most agronomic characters increased significantly among the subclones as the irradiation done rose. Principal component analysis also indicated that the subclones from the irradiated population were more variable than the non-irradiated. Such variation with higher heritability could be transmitted to the following generations by clonal propagation and utilized as genetic sources in mutation breeding. The combined method with chronic irradiation followed by tissue culture is evaluated as an effective method of widening mutation spectrum and increasing mutation frequency in regenerated plants. In addition, this method is valid to improve any crop species which can regenerate plants through callus culture. (author)

  3. Cultures of psychiatry and the professional socialization process: the case of containment methods for disturbed patients.

    Science.gov (United States)

    Bowers, Len; Alexander, Jane; Simpson, Alan; Ryan, Carl; Carr-Walker, Paola

    2004-08-01

    Acute mental disorder necessitating admission to hospital is often accompanied by disturbed behaviour that threatens the health of the person concerned or that of those around them. A range of containment methods are used by psychiatric professionals to keep patients and staff safe. These strategies are strongly emotive and attract strong moral valuations, yet differ sharply between countries. This paper reports a study to investigate the relationship between attitudes to these containment methods, and exposure to psychiatric education and practice. It was hypothesized that the culture of psychiatry in the study country would socialise students' views towards the locally dominant pattern of relative evaluations. Nine cohorts of student psychiatric nurses at different stages of their training at one UK University were asked to complete ratings on 11 containment methods. Containment methods fell into five groups, with mechanical restraint and net beds attracting the most severe disapproval. Neither the relative evaluation of methods, nor the intensity of those evaluations, changed systematically with duration of training. The findings support the interpretation that the relative evaluations of psychiatric containment methods are a property of wider national cultures, rather than an isolated tradition of professional psychiatric practice.

  4. 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.

  5. 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

  6. 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.

  7. 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.

  8. 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 ...

  9. 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 ...

  10. 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...

  11. 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.

  12. 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...

  13. 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...

  14. The State of the Art of Teaching Research Methods in the Social Sciences: Towards a Pedagogical Culture

    Science.gov (United States)

    Wagner, Claire; Garner, Mark; Kawulich, Barbara

    2011-01-01

    No formal pedagogical culture for research methods in the social sciences seems to exist and, as part of the authors' endeavour to establish such a culture, this article reviews current literature about teaching research methods and identifies the gaps in the research. Articles in academic journals spanning a 10-year period were collected by…

  15. Simple and high yielding method for preparing tissue specific extracellular matrix coatings for cell culture.

    Science.gov (United States)

    DeQuach, Jessica A; Mezzano, Valeria; Miglani, Amar; Lange, Stephan; Keller, Gordon M; Sheikh, Farah; Christman, Karen L

    2010-09-27

    The native extracellular matrix (ECM) consists of a highly complex, tissue-specific network of proteins and polysaccharides, which help regulate many cellular functions. Despite the complex nature of the ECM, in vitro cell-based studies traditionally assess cell behavior on single ECM component substrates, which do not adequately mimic the in vivo extracellular milieu. We present a simple approach for developing naturally derived ECM coatings for cell culture that provide important tissue-specific cues unlike traditional cell culture coatings, thereby enabling the maturation of committed C2C12 skeletal myoblast progenitors and human embryonic stem cells differentiated into cardiomyocytes. Here we show that natural muscle-specific coatings can (i) be derived from decellularized, solubilized adult porcine muscle, (ii) contain a complex mixture of ECM components including polysaccharides, (iii) adsorb onto tissue culture plastic and (iv) promote cell maturation of committed muscle progenitor and stem cells. This versatile method can create tissue-specific ECM coatings, which offer a promising platform for cell culture to more closely mimic the mature in vivo ECM microenvironment.

  16. Irradiation as a Method of Salvation of Cultural Heritage Objects Under Massive Jeopardy

    International Nuclear Information System (INIS)

    Katusin-Razem, B.; Braun, M.; Jagic, R.

    2013-01-01

    Cultural heritage artefacts made of organic materials are susceptible to deterioration caused by the action of insects, moulds, fungi and bacteria. The problem of biodeterioration becomes especially acute after natural catastrophes and weather inclemencies, or after human activities conducive to an abrupt development of pests. Usually a large number of objects are imperilled at the same time and their sheer number aggravates any organized preservation effort. Irradiation has proven an effective method of preservation under the circumstances, e.g. for the prevention of massive proliferation of fungal infestation of books wetted by flood. War destructions in Croatia 1991 - 1995 seriously jeopardized many cultural objects. Their treatment by 60Co gamma rays in the Radiation Chemistry and Dosimetry Laboratory of the Ruðer Boškoviæ Institute played a significant role in the prevention of massive biodeterioration. In co-operation with the Croatian Conservation Institute, one third of 5000 evacuated objects, mostly polychromic wooden sculptures, were irradiated for desinsection and disinfection, enabling their joint accommodation in depots until restoration. This contribution to the preservation of jeopardized cultural heritage objects has been recognized internationally as a specially significant and successful case of the application of irradiation to cultural heritage. The presentation describes in more detail the preservation and restoration of the altar of the Holy Cross from the church of the Blessed Virgin Mary of the Snow in Kamensko.(author)

  17. 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.

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

    Directory of Open Access Journals (Sweden)

    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.

  19. 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.

  20. 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.

  1. 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

    Directory of Open Access Journals (Sweden)

    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

  2. Methods to induce primary and secondary traumatic damage in organotypic hippocampal slice cultures.

    Science.gov (United States)

    Adamchik, Y; Frantseva, M V; Weisspapir, M; Carlen, P L; Perez Velazquez, J L

    2000-04-01

    Organotypic brain slice cultures have been used in a variety of studies on neurodegenerative processes [K.M. Abdel-Hamid, M. Tymianski, Mechanisms and effects of intracellular calcium buffering on neuronal survival in organotypic hippocampal cultures exposed to anoxia/aglycemia or to excitotoxins, J. Neurosci. 17, 1997, pp. 3538-3553; D.W. Newell, A. Barth, V. Papermaster, A.T. Malouf, Glutamate and non-glutamate receptor mediated toxicity caused by oxygen and glucose deprivation in organotypic hippocampal cultures, J. Neurosci. 15, 1995, pp. 7702-7711; J.L. Perez Velazquez, M.V. Frantseva, P.L. Carlen, In vitro ischemia promotes glutamate mediated free radical generation and intracellular calcium accumulation in pyramidal neurons of cultured hippocampal slices, J. Neurosci. 23, 1997, pp. 9085-9094; L. Stoppini, L.A. Buchs, D. Muller, A simple method for organotypic cultures of nervous tissue, J. Neurosci. Methods 37, 1991, pp. 173-182; R.C. Tasker, J.T. Coyle, J.J. Vornov, The regional vulnerability to hypoglycemia induced neurotoxicity in organotypic hippocampal culture: protection by early tetrodotoxin or delayed MK 801, J. Neurosci. 12, 1992, pp. 4298-4308.]. We describe two methods to induce traumatic cell damage in hippocampal organotypic cultures. Primary trauma injury was achieved by rolling a stainless steel cylinder (0.9 g) on the organotypic slices. Secondary injury was followed after dropping a weight (0.137 g) on a localised area of the organotypic slice, from a height of 2 mm. The time course and extent of cell death were determined by measuring the fluorescence of the viability indicator propidium iodide (PI) at several time points after the injury. The initial localised impact damage spread 24 and 67 h after injury, cell death being 25% and 54%, respectively, when slices were kept at 37 degrees C. To validate these methods as models to assess neuroprotective strategies, similar insults were applied to slices at relatively low temperatures (30

  3. Co-interviewing across gender and culture: expanding qualitative research methods in Melanesia.

    Science.gov (United States)

    Redman-MacLaren, Michelle L; Api, Unia K; Darius, Matupit; Tommbe, Rachael; Mafile'o, Tracie A; MacLaren, David J

    2014-09-06

    The social and cultural positions of both researchers and research participants influence qualitative methods and study findings. In Papua New Guinea (PNG), as in other contexts, gender is a key organising characteristic and needs to be central to the design and conduct of research. The colonial history between researcher and participant is also critical to understanding potential power differences. This is particularly relevant to public health research, much of which has emerged from a positivist paradigm. This paper describes our critical reflection of flexible researcher responses enacted during qualitative research in PNG. Led by a senior male HIV researcher from PNG, a male from a PNG university and a female from an Australian university conducted qualitative interviews about faith-based responses to HIV in PNG. The two researchers planned to conduct one-on-one interviews matching gender of participants and interviewer. However, while conducting the study, four participants explicitly requested to be interviewed by both researchers. This experience led us to critically consider socially and culturally situated ways of understanding semi-structured interviewing for public health research in Melanesia. New understandings about public health research include: (i) a challenge to the convention that the researcher holds more power than the research participant, (ii) the importance of audience in Melanesia, (iii) cultural safety can be provided when two people co-interview and (iv) the effect an esteemed leader heading the research may have on people's willingness to participate. Researchers who occupy insider-outsider roles in PNG may provide participants new possibilities to communicate key ideas. Our recent experience has taught us public health research methods that are gender sensitive and culturally situated are pivotal to successful research in Melanesia. Qualitative research requires adaptability and reflexivity. Public health research methods must continue

  4. Different Donor Cell Culture Methods Can Influence the Developmental Ability of Cloned Sheep Embryos.

    Directory of Open Access Journals (Sweden)

    LiBing Ma

    Full Text Available It was proposed that arresting nuclear donor cells in G0/G1 phase facilitates the development of embryos that are derived from somatic cell nuclear transfer (SCNT. Full confluency or serum starvation is commonly used to arrest in vitro cultured somatic cells in G0/G1 phase. However, it is controversial as to whether these two methods have the same efficiency in arresting somatic cells in G0/G1 phase. Moreover, it is unclear whether the cloned embryos have comparable developmental ability after somatic cells are subjected to one of these methods and then used as nuclear donors in SCNT. In the present study, in vitro cultured sheep skin fibroblasts were divided into four groups: (1 cultured to 70-80% confluency (control group, (2 cultured to full confluency, (3 starved in low serum medium for 4 d, or (4 cultured to full confluency and then further starved for 4 d. Flow cytometry was used to assay the percentage of fibroblasts in G0/G1 phase, and cell counting was used to assay the viability of the fibroblasts. Then, real-time reverse transcription PCR was used to determine the levels of expression of several cell cycle-related genes. Subsequently, the four groups of fibroblasts were separately used as nuclear donors in SCNT, and the developmental ability and the quality of the cloned embryos were compared. The results showed that the percentage of fibroblasts in G0/G1 phase, the viability of fibroblasts, and the expression levels of cell cycle-related genes was different among the four groups of fibroblasts. Moreover, the quality of the cloned embryos was comparable after these four groups of fibroblasts were separately used as nuclear donors in SCNT. However, cloned embryos derived from fibroblasts that were cultured to full confluency combined with serum starvation had the highest developmental ability. The results of the present study indicate that there are synergistic effects of full confluency and serum starvation on arresting fibroblasts in

  5. Different Donor Cell Culture Methods Can Influence the Developmental Ability of Cloned Sheep Embryos.

    Science.gov (United States)

    Ma, LiBing; Liu, XiYu; Wang, FengMei; He, XiaoYing; Chen, Shan; Li, WenDa

    2015-01-01

    It was proposed that arresting nuclear donor cells in G0/G1 phase facilitates the development of embryos that are derived from somatic cell nuclear transfer (SCNT). Full confluency or serum starvation is commonly used to arrest in vitro cultured somatic cells in G0/G1 phase. However, it is controversial as to whether these two methods have the same efficiency in arresting somatic cells in G0/G1 phase. Moreover, it is unclear whether the cloned embryos have comparable developmental ability after somatic cells are subjected to one of these methods and then used as nuclear donors in SCNT. In the present study, in vitro cultured sheep skin fibroblasts were divided into four groups: (1) cultured to 70-80% confluency (control group), (2) cultured to full confluency, (3) starved in low serum medium for 4 d, or (4) cultured to full confluency and then further starved for 4 d. Flow cytometry was used to assay the percentage of fibroblasts in G0/G1 phase, and cell counting was used to assay the viability of the fibroblasts. Then, real-time reverse transcription PCR was used to determine the levels of expression of several cell cycle-related genes. Subsequently, the four groups of fibroblasts were separately used as nuclear donors in SCNT, and the developmental ability and the quality of the cloned embryos were compared. The results showed that the percentage of fibroblasts in G0/G1 phase, the viability of fibroblasts, and the expression levels of cell cycle-related genes was different among the four groups of fibroblasts. Moreover, the quality of the cloned embryos was comparable after these four groups of fibroblasts were separately used as nuclear donors in SCNT. However, cloned embryos derived from fibroblasts that were cultured to full confluency combined with serum starvation had the highest developmental ability. The results of the present study indicate that there are synergistic effects of full confluency and serum starvation on arresting fibroblasts in G0/G1 phase

  6. Physicochemical characteristics and biological activities of polysaccharide fractions from Phellinus baumii cultured with different methods.

    Science.gov (United States)

    Li, Tingting; Yang, Yan; Liu, Yanfang; Zhou, Shuai; Yan, Meng Qiu; Wu, Di; Zhang, Jingsong; Tang, Chuanhong

    2015-11-01

    Nine polysaccharide fractions were obtained from the fruiting bodies, submerged mycelia, and solid state fermented products of Phellinus baumii using different concentrations of ethanol precipitation. The chemical characteristics and in vitro immunological activities of the nine polysaccharide fractions were compared and studied. Results indicated that the fractions precipitated with 50% ethanol had higher yields of polysaccharides and submerged mycelia contributed to high extraction yields of polysaccharides and possessed higher polysaccharide contents. HPSEC-MALLS-RI analysis showed that the molecular weight (Mw) of polysaccharide fractions from these three materials decreased with the increasing of precipitated ethanol concentration. The Mw of fruiting body polysaccharide fractions ranged from 1.98×10(4)Da to 1.89×10(6)Da. Large-molecular-weight polysaccharides (from 2.11×10(6)Da to 2.01×10(7)Da) were found in submerged mycelia. Some lower-molecular-weight polysaccharide components were found in solid fermented products. Different culture methods contributed to significant differences in monosaccharide components and molar ratios. The 50% ethanol precipitated fractions exhibited more complexity on monosaccharide compositions comparing with fractions precipitated with 30% and 70% ethanol. Polysaccharide fractions derived from submerged mycelia exhibited higher macrophages stimulation activities. Submerged culture was found to be a suitable method to prepare active polysaccharides because of its short culture span and reasonable cost. Copyright © 2015 Elsevier B.V. All rights reserved.

  7. Robust Microplate-Based Methods for Culturing and in Vivo Phenotypic Screening of Chlamydomonas reinhardtii

    Directory of Open Access Journals (Sweden)

    Timothy C. Haire

    2018-03-01

    Full Text Available Chlamydomonas reinhardtii (Cr, a unicellular alga, is routinely utilized to study photosynthetic biochemistry, ciliary motility, and cellular reproduction. Its minimal culture requirements, unicellular morphology, and ease of transformation have made it a popular model system. Despite its relatively slow doubling time, compared with many bacteria, it is an ideal eukaryotic system for microplate-based studies utilizing either, or both, absorbance as well as fluorescence assays. Such microplate assays are powerful tools for researchers in the areas of toxicology, pharmacology, chemical genetics, biotechnology, and more. However, while microplate-based assays are valuable tools for screening biological systems, these methodologies can significantly alter the conditions in which the organisms are cultured and their subsequent physiology or morphology. Herein we describe a novel method for the microplate culture and in vivo phenotypic analysis of growth, viability, and photosynthetic pigments of C. reinhardtii. We evaluated the utility of our assay by screening silver nanoparticles for their effects on growth and viability. These methods are amenable to a wide assortment of studies and present a significant advancement in the methodologies available for research involving this model organism.

  8. Gulliver's Eggs: Why Methods are not an Issue of Qualitative Research in Cultural Psychology.

    Science.gov (United States)

    Tateo, Luca

    2015-06-01

    The future of qualitative methods regards the kind of object cultural psychology is interested and the kind of questions it can ask. I propose that the object should be experiencing, understood as a complex whole, consisting of lived-by action and counter-action, that is contextual inter-action with the world in the form of an experiencing subject and otherness. The kind of questions cultural psychology can ask is instead related to the epistemological status attributed to both researcher and participant. Probably few scholars such as Vygotsky, Piaget and Lewin understood to what extent experiencing is always changing, because the relationship between mind, alterity and culture is co-generative. This also implies a relativization and a decentralization of the psychology's perspective. Finally, I provide some examples from the history of psychology and some suggestions to work at the level of such complexity by using methods that can work with complex objects such as products of human activity (e.g., art, literature, architecture, etc.).

  9. Development of a Novel Nuclear Safety Culture Evaluation Method for an Operating Team Using Probabilistic Safety Analysis

    Energy Technology Data Exchange (ETDEWEB)

    Han, Sangmin; Lee, Seung Min; Seong, Poong Hyun [KAIST, Daejeon (Korea, Republic of)

    2015-05-15

    IAEA defined safety culture as follows: 'Safety Culture is that assembly of characteristics and attitudes in organizations and individuals which establishes that, as an overriding priority, nuclear plant safety issues receive the attention warranted by their significance'. Also, celebrated behavioral scientist, Cooper, defined safety culture as,'safety culture is that observable degree of effort by which all organizational members direct their attention and actions toward improving safety on a daily basis' with his internal psychological, situational, and behavioral context model. With these various definitions and criteria of safety culture, several safety culture assessment methods have been developed to improve and manage safety culture. To develop a new quantitative safety culture evaluation method for an operating team, we unified and redefined safety culture assessment items. Then we modeled a new safety culture evaluation by adopting level 1 PSA concept. Finally, we suggested the criteria to obtain nominal success probabilities of assessment items by using 'operational definition'. To validate the suggested evaluation method, we analyzed the collected audio-visual recording data collected from a full scope main control room simulator of a NPP in Korea.

  10. Development of a Novel Nuclear Safety Culture Evaluation Method for an Operating Team Using Probabilistic Safety Analysis

    International Nuclear Information System (INIS)

    Han, Sangmin; Lee, Seung Min; Seong, Poong Hyun

    2015-01-01

    IAEA defined safety culture as follows: 'Safety Culture is that assembly of characteristics and attitudes in organizations and individuals which establishes that, as an overriding priority, nuclear plant safety issues receive the attention warranted by their significance'. Also, celebrated behavioral scientist, Cooper, defined safety culture as,'safety culture is that observable degree of effort by which all organizational members direct their attention and actions toward improving safety on a daily basis' with his internal psychological, situational, and behavioral context model. With these various definitions and criteria of safety culture, several safety culture assessment methods have been developed to improve and manage safety culture. To develop a new quantitative safety culture evaluation method for an operating team, we unified and redefined safety culture assessment items. Then we modeled a new safety culture evaluation by adopting level 1 PSA concept. Finally, we suggested the criteria to obtain nominal success probabilities of assessment items by using 'operational definition'. To validate the suggested evaluation method, we analyzed the collected audio-visual recording data collected from a full scope main control room simulator of a NPP in Korea

  11. Evaluation of methods to prepare samples of leafy green vegetables for preenrichment with the Bacteriological Analytical Manual Salmonella culture method.

    Science.gov (United States)

    Jacobson, Andrew Patrick; Gill, Vikas Singh; Irvin, Kari Anne; Wang, Hua; Hammack, Thomas Seyliard

    2012-02-01

    Three sample preparation procedures, soak, stomach, and blend, were evaluated using the Bacteriological Analytical Manual Salmonella culture method with eight types of leafy green produce. In the soak method, test portions were added to lactose broth without homogenization; in the stomach method, test portions were stomached with lactose broth; and in the blend method, test portions were blended with lactose broth. Twenty artificially contaminated test portions were analyzed with each procedure in individual experimental trials. The number of test portions identified as positive were compared among the procedures. Statistically significant differences were identified with Fisher's exact two-tailed F test (P < 0.05). Where differences did occur (P < 0.05), the soak procedure was the most effective or was at least as effective as homogenization by either blending or stomaching. Statistically significant differences most frequently occurred with romaine lettuce and cabbage; for these items, blending was significantly less effective than the soak procedure. Overall, for all of the produce types examined, results showed that the soak procedure was more effective than either of the homogenization procedures in recovering Salmonella from leafy green produce. Of the 540 test portions examined by each sample preparation method, 344 were positive for the presence of Salmonella by soaking, 293 by stomaching, and 232 by blending. We recommend that the soak procedure replace homogenization for the analysis of leafy green produce because the soak procedure is more productive than homogenization by either blending or stomaching of the leafy green produce types as reported herein.

  12. Features of method of medical physical culture at insufficiency of aortic valve

    Directory of Open Access Journals (Sweden)

    S.A. Kalmykov

    2013-01-01

    Full Text Available The basic approaches are considered to application of facilities of medical physical education at aortic insufficiency on the stages of physical rehabilitation. An analysis is conducted more than 20 literary sources. The mechanisms of medical action of physical exercises are specified - restorative influence, forming of temporal indemnifications, trophic action, normalization of the broken functions. It is set that task, forms, facilities, the methods of medical physical culture depend on the degree of weight of disease, degree of cardio-vascular insufficiency and stage of physical rehabilitation. Engaged in a medical physical culture conducted in form morning hygienical gymnastics, medical gymnastics, independent employments, dosed walks, walking on steps, mobile and sporting games. It is marked that sparing training and training the motive modes are instrumental in the gradual training of the cardio-vascular system. Recommended the dosed walking to lead to a to 5-8 km on the sparing training and to 8-12 km on training modes.

  13. [Application of a modified culturing method for lymphocytes in cytogenetic research].

    Science.gov (United States)

    Gao, Chaoxian; Hui, Changye; Zhang, Wen; Guo, Yan; Li, Limei; Chen, Yuting; Zhang, Liuzhuo; Yang, Xinyue; Huang, Xianqing

    2016-08-01

    To establish a modified method for microculturing whole human blood for cytogenetic analysis. A novel tube rack was designed to overcome the drawbacks of directly culturing the cells within centrifuge tubes. The fractions of human plasma, human serum and two commercial fetal bovine sera were analyzed with 15% sodium dodecyl sulfate-polyacrylamide gel electrophoresis (SDS-PAGE). The influence of adding 0%, 5%, 10%, 15%, 20%, 25% and 30% autologous plasma to the culture on lymphocyte transformation rate and mitotic index (MI) was examined. The SDS-PAGE analysis showed a significant difference between commercial fetal bovine sera, and that the components of human plasma were similar to those of fetal bovine serum. The value of MI in lymphocyte was evidently increased along with addition of autologous plasma. However, this has exerted no significant effect on the transformation rate. With the addition of 10% autologous plasma, the MI value has become much higher than the conventional method. A modified method was established by application of a novel tube inclined rack and optimization of whole blood inoculation. This method is easier and cheaper, and is suitable for application in clinical practice.

  14. Comparison of the Lysis Centrifugation Method with the Conventional Blood Culture Method in Cases of Sepsis in a Tertiary Care Hospital

    OpenAIRE

    Parikh, Harshal R; De, Anuradha S; Baveja, Sujata M

    2012-01-01

    Introduction : Physicians and microbiologists have long recognized that the presence of living microorganisms in the blood of a patient carries with it considerable morbidity and mortality. Hence, blood cultures have become critically important and frequently performed test in clinical microbiology laboratories for diagnosis of sepsis. Objectives: To compare the conventional blood culture method with the lysis centrifugation method in cases of sepsis. Materials and Methods: Two hundred ...

  15. Oral rinse as a potential method to culture Candida isolate from AIDS patients

    Directory of Open Access Journals (Sweden)

    Desiana Radithia

    2011-12-01

    Full Text Available Background: Candida isolate is easily sampled from oral cavity by swabbing directly on the candidiasis lesion, to be smeared onto slides for direct examination or cultured in a growth medium. This method is by far the gold standard for defining candidiasis diagnosis. However it is difficult to apply on sensitive patients and almost impossible on patients showing no clinical appearance of oral candidiasis. AIDS patients are very prone to candida infection and have a tendency of repetitive infection involving mixed species. As many candida species show different susceptibility to anti-fungal agents, it is necessary to identify the species causing the infection in the management of oral candidiasis. Oral rinse is a suggested method to obtain candida isolate to be cultured for further analysis such as species identification. This method is simple and less risky on infection transmission as less tools are required in the procedure. Purpose: This study aimed to assess the application of oral rinse as an alternative method to culture Candida isolate from AIDS patients. Methods: A cross-sectional observative study was conducted in HIV/AIDS in-Patient Facility of Intermediate Care Unit for Infection Disease, Dr. Soetomo Hospital Surabaya. Fourteen stadium 4 AIDS patients matching criteria were swabbed on 1/3-posterior of the tongue, and then given 10 ml phosphate buffer saline to rinse vigorously for 15 seconds. Both specimens were cultured on Sabouraud’s dextrose agar and colony growth was observed. Results: Candida colonies were able to grow from all 14 isolates (100% by both methods. Qualitatively, cultures from oral rinse specimens were more populated than cultures from swab specimens. Conclusion: Oral rinse is an applicable technique to obtain Candida species isolate. This technique is safe, easy, non-invasive, and needs less tools therefore less risky for HIV transmission.Latar belakang: Isolat Candida mudah diambil dengan cara mengusap lesi

  16. ANALYSIS OF GROWTH OF LALAWAK Barbonymus balleroides (VALENCIENNES, 1842 IN THREE CULTURE METHODS

    Directory of Open Access Journals (Sweden)

    Deni Radona

    2017-09-01

    Full Text Available In fish culture, optimal growth could be influenced by various culture methods. Aim of the study was to evaluate the productivity of Barbonymus balleroides, lalawak in floating net cages, concrete ponds, and earthen ponds. Cultivation was designed with the circulation water system. Experiment was conducted using completely randomized design with three treatments and three replications for each treatment. The experimental fish, sized of 4.20 ± 0.64 cm SL and weight of 2.14 ± 0.99 g, were obtained from induced breeding. The stocking density used was 20 individuals/m3. Fish were fed 3% of total weight two times every day using commercial pellet with 35% protein content for 90 day. The result showed that lalawak reared in earthen pond was no significant difference on length, weight, and biomass compared with that one in concrete pond (P>0.05, but significantly different (P0.05 among the three different culture systems for survival rate and FCR. Lalawak reared on earthen pond system supported with optimal water quality could increase productivity value.

  17. The double pedigree: a method for studying culturally and genetically inherited behavior in tandem.

    Directory of Open Access Journals (Sweden)

    Etienne Danchin

    Full Text Available Transgenerational sources of biological variation have been at the center of evolutionary studies ever since Darwin and Wallace identified natural selection. This is because evolution can only operate on traits whose variation is transmitted, i.e. traits that are heritable. The discovery of genetic inheritance has led to a semantic shift, resulting in the tendency to consider that only genes are inherited across generations. Today, however, concepts of heredity are being broadened again to integrate the accruing evidence of non-genetic inheritance, and many evolutionary biologists are calling for the inclusion of non-genetic inheritance into an inclusive evolutionary synthesis. Here, we focus on social heredity and its role in the inheritance of behavioral traits. We discuss quantitative genetics methods that might allow us to disentangle genetic and non-genetic transmission in natural populations with known pedigrees. We then propose an experimental design based on cross-fostering among animal cultures, environments and families that has the potential to partition inherited phenotypic variation into socially (i.e. culturally and genetically inherited components. This approach builds towards a new conceptual framework based on the use of an extended version of the animal model of quantitative genetics to integrate genetic and cultural components of behavioral inheritance.

  18. An in vitro monocyte culture method and establishment of a human monocytic cell line (K63).

    Science.gov (United States)

    Kadoi, Katsuyuki

    2011-01-01

    A novel method of monocyte culture in vitro was developed. The fraction of monocytes was obtained by density centrifugation of heparinised human venous blood samples. Monocytes were suspended in a modified Rosewell Park Memorial Institute medium (RPMI)-1640 (mRPMI) supplemented with 10% non-inactivated autologous serum added to the feeder cells. An avian cell line was used for feeder cells. Only those monocytes that settled on feeder cells grew rapidly at 37°C-38°C into a formation of clumped masses within two to three days. The cell mass was harvested and subcultures were made without feeder cells. A stable cell line (K63) was established from subcultures using a limited dilution method and cell cloning in microplates. K63 cells were adapted for later growth in the mRPMI medium supplemented with 10% foetal calf serum. The cells were well maintained at over 50th passage levels. This method proved to be applicable for monocyte cultures of animals as well.

  19. A new method for the diagnosis of Trichomonas gallinae infection by culture.

    Science.gov (United States)

    Cover, A J; Harmon, W M; Thomas, M W

    1994-07-01

    Diagnosis of Trichomonas gallinae infection can reliably be made by the demonstration of organisms in material taken from a bird's mouth and crop. This can be examined directly or incubated in a growth medium until organisms are numerous enough to be easily found in aliquots examined with the microscope. We compared two methods of culture diagnosis of Trichomonas gallinae infection in pigeons. A commercially available kit, In-Pouch TF, proved as sensitive for this purpose as in vitro Diamond's medium, but had practical advantages over the usual in vitro system.

  20. Comparison of immunochromatography, PCR and culture methods for the detection of Campylobacter bacteria.

    Science.gov (United States)

    Dey, Shuvra Kanti; Nishimura, Shuichi; Okitsu, Shoko; Hayakawa, Satoshi; Mizuguchi, Masashi; Ushijima, Hiroshi

    2012-12-01

    A total of 463 fecal specimens from human patients were tested for the presence of Camphylobacter by a commercial immunochromatography kit (ImmunoCard STAT! CAMPY), polymerase chain reaction (PCR) and conventional bacteriological (CB) methods. Using culture as the standard of reference, the ImmunoCard STAT! CAMPY assay had a sensitivity of 86% and a specificity of 100%. On the other hand, using PCR as the standard of reference, ImmunoCard STAT! CAMPY assay had a sensitivity of 90.5% and a specificity of 100%. Copyright © 2012. Published by Elsevier B.V.

  1. Cultural border crossing in three urban classrooms: A mixed methods study

    Science.gov (United States)

    Roopnarine, Rupnarain

    This study examined the effects of the instruction of four youth cultural border crossing behaviors: flexibility, being at ease, playfulness, and citizenship as an intervention aimed at helping students to transition across three borders, student to student, student to science, and student to teacher. The research involved 12 ninth- and 10th-grade students in a large urban school district in three diverse classrooms, A, B, and C. Four students in each classroom volunteered for the study. The students in Groups A and B were in 9th grade Living Environment and students in Group B were in 10th grade chemistry. These students participated in this instructional intervention for three months. The study was conducted using both quantitative and qualitative methods based on participant observations, interviews, and questionnaire. The result indicated that there was no significant effect of the cultural border crossing instructions on the students' interactions across the three borders examined. However, the instructions helped Group A and Group B to be more flexible but not group C. Also, the instructions helped Group A to be more playful and at ease but not Group B and C. The instructions also helped Group A to show more citizenship but not Group B and C. In addition, there was no difference between the pretest and posttest cultural bother crossing behavior. Moreover, qualitative data analysis showed that the participants were more flexible, at ease, and playful among peers than across student to teacher and student to science borders. Also, the use of citizenship in the three groups showed no effect on the participants' interaction with peers. Although, the findings showed no effect of cultural border crossing instructions on students' interactions, it is suggested that we continue to find ways to help students feel more comfortable in science.

  2. An improved culturing method for opiine fruit fly parasitoids and its application to parasitoid monitoring in the field.

    Science.gov (United States)

    Masry, Ayad; Furlong, Michael J; Clarke, Anthony R; Cunningham, John Paul

    2018-02-01

    Good culturing methods play an important role in the study of insect behavior and its application to pest management. Here, we describe and validate a new method for rearing the parasitoid wasp, Diachasmimorpha kraussii, which attacks some of the world's worst fruit fly pests and is an internationally used biological control agent. Our method differs from standard culturing approaches by presenting adult wasps with host-infested artificial media within a "culturing bag," which mimics a natural (fruit) oviposition substrate. In laboratory trials using wild collected D. kraussii, the culturing bag method was compared to the use of host-infested nectarines, and a commonly used laboratory method of presenting host-infested artificial media within Petri dishes. The culturing bag method proved to be a significant improvement on both methods, combining the advantages of high host survival in artificial media with parasitism levels that were the equivalent to those recorded using host-infested fruits. In our field study, culturing bags infested with the Queensland fruit fly, Bactrocera tryoni, and hung in a mixed peach and nectarine orchard proved to be effective "artificial fruits" attracting wild D. kraussii for oviposition. Significantly more adult wasps were reared from the culturing bags compared to field collected fruits. This was shown to be due to higher fruit fly larval density in the bags, as similar percentage parasitism rates were found between the culturing bags and ripe fruits. We discuss how this cheap, time-efficient method could be applied to collecting and monitoring wild D. kraussii populations in orchards, and assist in maintaining genetic variability in parasitoid laboratory cultures. © 2016 Institute of Zoology, Chinese Academy of Sciences.

  3. Using the House-Tree-Person (H-T-P) as a Method to Understanding Individual Cultural Differences among Children.

    Science.gov (United States)

    Lambeth, Pauline J.

    Finding innovative and effective methods to infer the ability of culturally diverse students continues to present a problem for educators. This paper proposes the House-Tree-Person (HTP) projective technique as a way for school educators to gather important data about student functioning without the use of culturally biased instruments. This…

  4. 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.

  5. Comparative analysis of effectiveness of some students’ physical culture training methodic

    Directory of Open Access Journals (Sweden)

    A.Yu. Osipov

    2017-08-01

    Full Text Available Introduction: scientific researches witness about presence of serious tendency to weakening students’ everyday motor activity and absence of motivation to physical culture and sports practicing. Some specialists offer to solve this problem by making physical culture classes more oriented on sports. At the same time there are studies, offering to use methodic of cardio-power training. Purpose: comparative analysis of effectiveness of different programs on Physical culture trainings with the help of sport oriented approach and functional-power training. The main criterion of trainings’ effectiveness was the level of physical and functional fitness of students’ organism to physical loads. Material: in the research students (n=600: boys - n=300, girls - n=300, age - 19-20 years participated. Selection of students was fulfilled, considering indicators of body mass. We used the methods of physical condition control and assessment (control tests and functional fitness (functional tests. Besides, we considered indicators of busy mass by students’ regular weighing. Results: we found some increase of body mass of students, who attended different sport specialization trainings. Boys and girls (athletic gymnastic and ping pong demonstrated confident body mass increase in average by 3-4 kg during academic year Students, practicing cardio-power training - HOT IRON- practically did not increase body mass. Difference in body mass indicators of such students between beginning of trainings and at the end of the researches was insignificant (±0.3 kg. Tests’ results witness about general reduction of indicators of students’ main physical qualities during academic year. Significant increase of physical strength was found only in students, specializing in athletic gymnastic (boys and girls. Students of cardio-power training - HOT IRON-demonstrated confident increase of indicators of physical strength, general endurance and flexibility. Conclusions: our

  6. The relationship between glass ceiling and power distance as a cultural variable by a new method

    Directory of Open Access Journals (Sweden)

    Naide Jahangirov

    2015-12-01

    Full Text Available Glass ceiling symbolizes a variety of barriers and obstacles that arise from gender inequality at business life. With this mind, culture influences gender dynamics. The purpose of this research was to examine the relationship between the glass ceiling and the power distance as a cultural variable within organizations. Gender variable is taken as a moderator variable in relationship between the concepts. In addition to conventional correlation analysis, we employed a new method to investigate this relationship in detail. The survey data were obtained from 109 people working at a research center which operated as a part of the non-profit private university in Ankara, Turkey. The relationship between the variables was revealed by a new method which was developed as an addition to the correlation in survey. The analysis revealed that the female staff perceived the glass ceiling and the power distance more intensely than the male staff. In addition, the medium level relation was determined between the power distance and the glass ceiling perception among female staff.

  7. Proteomic profiling of an undefined microbial consortium cultured in fermented dairy manure: Methods development.

    Science.gov (United States)

    Hanson, Andrea J; Paszczynski, Andrzej J; Coats, Erik R

    2016-03-01

    The production of polyhydroxyalkanoates (PHA; bioplastics) from waste or surplus feedstocks using mixed microbial consortia (MMC) and aerobic dynamic feeding (ADF) is a growing field within mixed culture biotechnology. This study aimed to optimize a 2DE workflow to investigate the proteome dynamics of an MMC synthesizing PHA from fermented dairy manure. To mitigate the challenges posed to effective 2DE by this complex sample matrix, the bacterial biomass was purified using Accudenz gradient centrifugation (AGC) before protein extraction. The optimized 2DE method yielded high-quality gels suitable for quantitative comparative analysis and subsequent protein identification by LC-MS/MS. The optimized 2DE method could be adapted to other proteomic investigations involving MMC in complex organic or environmental matrices. © 2016 WILEY-VCH Verlag GmbH & Co. KGaA, Weinheim.

  8. Comparing culture and molecular methods for the identification of microorganisms involved in necrotizing soft tissue infections

    DEFF Research Database (Denmark)

    Rudkjøbing, Vibeke Børsholt; Thomsen, Trine Rolighed; Xu, Yijuan

    2016-01-01

    libraries with subsequent Sanger sequencing for most samples, 3) the Ibis T5000 biosensor and 4) 454-based pyrosequencing. Furthermore, quantitative PCR (qPCR) was used to verify and determine the relative abundance of Streptococcus pyogenes in samples. RESULTS: For 70 % of the surgical samples...... to culture. Although the molecular methods generally gave concordant results, our results indicate that Microseq may misidentify or overlook microorganisms that can be detected by other molecular methods. Half of the patients were found to be infected with S. pyogenes, but several atypical findings were also...... made including infection by a) Acinetobacter baumannii, b) Streptococcus pneumoniae, and c) fungi, mycoplasma and Fusobacterium necrophorum. CONCLUSION: The study emphasizes that many pathogens can be involved in NSTIs, and that no specific "NSTI causing" combination of species exists. This means...

  9. Optimization of Ex Vivo Murine Bone Marrow Derived Immature Dendritic Cells: A Comparative Analysis of Flask Culture Method and Mouse CD11c Positive Selection Kit Method

    Directory of Open Access Journals (Sweden)

    Rahul Ashok Gosavi

    2018-01-01

    Full Text Available 12–14 days of culturing of bone marrow (BM cells containing various growth factors is widely used method for generating dendritic cells (DCs from suspended cell population. Here we compared flask culture method and commercially available CD11c Positive Selection kit method. Immature BMDCs’ purity of adherent as well as suspended cell population was generated in the decreasing concentration of recombinant-murine granulocyte-macrophage colony-stimulating factor (rmGM-CSF in nontreated tissue culture flasks. The expression of CD11c, MHCII, CD40, and CD86 was measured by flow cytometry. We found significant difference (P<0.05 between the two methods in the adherent cells population but no significant difference was observed between the suspended cell populations with respect to CD11c+ count. However, CD11c+ was significantly higher in both adhered and suspended cell population by culture method but kit method gave more CD11c+ from suspended cells population only. On the other hand, using both methods, immature DC expressed moderate level of MHC class II molecules as well as low levels of CD40 and CD86. Our findings suggest that widely used culture method gives the best results in terms of yield, viability, and purity of BMDCs from both adherent and suspended cell population whereas kit method works well for suspended cell population.

  10. 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...

  11. Scientific methods and cultural heritage an introduction to the application of materials science to archaeometry and conservation science

    CERN Document Server

    Artioli, Gilberto

    2010-01-01

    Artioli provides an introduction to the methods and rationales of the scientific investigation of cultural heritage materials, with an emphasis placed on the analytical strategies, modes of operation and resulting information rather than on technicalities.

  12. A large-scale automated method for hepatocyte isolation: effects on proliferation in culture.

    Science.gov (United States)

    Nieuwoudt, M J; Kreft, E; Olivier, B; Malfeld, S; Vosloo, J; Stegman, F; Kunneke, R; Van Wyk, A J; Van der Merwe, S W

    2005-01-01

    Large-scale sterile methods for isolating hepatocytes are desirable for the development of bioartificial liver support systems. In this study the traditional centrifuge method was compared with the use of a Baylor Rapid Autologous Transfusion (BRAT) machine for isolating large quantities of porcine hepatocytes. After isolating hepatocytes, the methods were evaluated in terms of cell viability and yield per liver, proliferation over 7 days, and the effects on the cell cycle using the trypan blue exclusion test, conventional phase-contrast light microscopy, the lactate to pyruvate ratio, the leakage of lactate dehydrogenase (LD) and aspartate aminotransferase (AST), lidocaine clearance, albumin production, and flow cytometry. With the centrifuge method the mean cell viability was 92.5%, while with the BRAT method the viability was 95.9%. The minimal cell yields with the BRAT procedure were 7.3 x 10(9) for 250-ml centrifuge bowls and 2.8 x 10(9) for 165-ml bowls, which compares well with that found by other authors. Because the same initial procedures were employed in both methods the total hepatocyte yield per liver was comparable. Flow cytometry confirmed that the proliferation of hepatocytes was facilitated by oxygenation during the isolation procedure. The recovery of hepatocytes in culture following isolation was similar after either method. Daily microscopic investigation indicated that cytoplasmic vacuolization and granularities were present after either procedure and these disappeared following 3-4 days of culturing. Flow cytometry indicated that the hepatocyte cell cycle was similar after either method; at 7 days the profile indicated that the cells were still proliferating. Trends in the lactate to pyruvate ratio and the leakage of LD and AST indicated that the functional polarity of hepatocytes was regained after approximately 3 days. Lidocaine clearance at 4 days indicated that the cytochrome P450 system was active, while significant albumin production was

  13. Estudio sobre la efectividad para la prevención de la legionelosis del sistema de calentamiento instantáneo, instalado en la red de agua sanitaria de un hospital Effectiveness study of a pasteurization system in controlling contamination with Legionella installed in a hospital's hot water system

    Directory of Open Access Journals (Sweden)

    Laura Gavaldà Mestre

    2006-12-01

    Full Text Available Se ha estudiado la efectividad de un sistema de pasteurización en el control de la contaminación por Legionella en la red de agua caliente de un hospital.El hospital había optado por este sistema debido a que los acumuladores convencionales originales presentaban importantes problemas en la capacidad de producción y en el mantenimiento de las temperaturas de distribución.El estudio fue iniciado después de haberse instalado los pasteurizadores y ha consistido en la realización de controles mensuales de Legionella y temperatura durante un período de 11 meses. De los resultados, se puede valorar que el sistema de pasteurización se considera efectivo, siempre y cuando las condiciones de la red (circulación, material, diseño, etc sean las adecuadas. El sistema no se considera efectivo en redes antiguas y con ramales sin circulación como se ha podido constatar en este estudio en el edificio de servicios, donde las conducciones presentaban un estado de conservación deficiente. En esta zona de servicios los controles de Legionella han mostrado una mejora únicamente después de que se iniciara un programa de purgas junto con la paulatina sustitución de los tramos en mal estado.The effectiveness of a pasteurization system in controlling contamination with Legionella has been evaluated in a hospital’s hot water system.The hospital acquired these equipments because the original system -conventional hot water tanks– had problems in producing and maintaining the distribution temperatures in the hot water circuit.The study started 11 months after the pasteurizers had been installed. The study consisted on Legionella and temperature controls which were conducted monthly during an 11-month period. Results have proved that a pasteurization system method to be an effective system of instantaneous warming provided that there are adequate conditions (circulation, materials, design, etc.. This system has no effectiveness in old nets or nets with

  14. The comparison of two methods to obtain human oral keratinocytes in primary culture

    International Nuclear Information System (INIS)

    Klingbeil, Maria Fatima Guarizo

    2006-01-01

    The therapeutic procedures frequently used in oral treatments for the pathological diseases are surgical, resulting in failures of the mucosal continuity.The possibility to obtain transplantable oral epithelia from an in vitro cell culture opens new utilization perspectives not only to where it comes from, but also as a reconstructive material for other parts of the human body, such as: urethra, epithelia corneo-limbal, cornea, ocular surface. Many researchers still use controversial methods for obtaining cells. It was therefore evaluated and compared the efficiency in both methods: enzymatic and direct explant to obtain oral keratinocytes from human oral mucosa. Fragments of intra oral epithelial tissues from healthy human subjects, undergoing dental surgeries, were donated to the research project. The keratinocytes were cultivated over a feeder-layer from a previously irradiated 3T3 Swiss albino fibroblasts. In this study it was compared the time needed in the cell obtention, the best cell amount between both methods, the life-span, the cell capacity to form an in vitro epithelia and its morphologic structure. The results in the assessment of both methods have shown the possibility to obtain keratinocytes from a small oral fragment, but at the same time we may verify the advantages and peculiar restrictions for each one of both analyzed methods. (author)

  15. Counting Legionella cells within single amoeba host cells

    Science.gov (United States)

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

  16. A rapid and sensitive method for measuring N-acetylglucosaminidase activity in cultured cells.

    Directory of Open Access Journals (Sweden)

    Victor Mauri

    Full Text Available A rapid and sensitive method to quantitatively assess N-acetylglucosaminidase (NAG activity in cultured cells is highly desirable for both basic research and clinical studies. NAG activity is deficient in cells from patients with Mucopolysaccharidosis type IIIB (MPS IIIB due to mutations in NAGLU, the gene that encodes NAG. Currently available techniques for measuring NAG activity in patient-derived cell lines include chromogenic and fluorogenic assays and provide a biochemical method for the diagnosis of MPS IIIB. However, standard protocols require large amounts of cells, cell disruption by sonication or freeze-thawing, and normalization to the cellular protein content, resulting in an error-prone procedure that is material- and time-consuming and that produces highly variable results. Here we report a new procedure for measuring NAG activity in cultured cells. This procedure is based on the use of the fluorogenic NAG substrate, 4-Methylumbelliferyl-2-acetamido-2-deoxy-alpha-D-glucopyranoside (MUG, in a one-step cell assay that does not require cell disruption or post-assay normalization and that employs a low number of cells in 96-well plate format. We show that the NAG one-step cell assay greatly discriminates between wild-type and MPS IIIB patient-derived fibroblasts, thus providing a rapid method for the detection of deficiencies in NAG activity. We also show that the assay is sensitive to changes in NAG activity due to increases in NAGLU expression achieved by either overexpressing the transcription factor EB (TFEB, a master regulator of lysosomal function, or by inducing TFEB activation chemically. Because of its small format, rapidity, sensitivity and reproducibility, the NAG one-step cell assay is suitable for multiple procedures, including the high-throughput screening of chemical libraries to identify modulators of NAG expression, folding and activity, and the investigation of candidate molecules and constructs for applications in

  17. 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.

  18. 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

  19. 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

  20. 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.

  1. 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.

  2. 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.

  3. 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

  4. 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.

  5. 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...

  6. Urine samples from disposable diapers: an accurate method for urine cultures.

    Science.gov (United States)

    Cohen, H A; Woloch, B; Linder, N; Vardi, A; Barzilai, A

    1997-03-01

    The method of collection of the urine sample is of paramount importance in making a diagnosis of urinary tract infection in infants and children. Squeezing urine out of disposable diapers can provide a urine sample that can be used to detect chemical abnormalities as well as a specimen suitable for microscopic examination. To date there have been no reported studies on the use of this technique for urine culture as compared with samples collected by suprapubic aspiration and catheterization. Urine was obtained from 38 infants aged under 2 years who presented with fever with no obvious cause. All infants had urine collected either by catheterization or suprapubic aspiration and by extraction from a disposable diaper. The urine samples were cultured using standard bacteriologic techniques. Five infants had a urinary tract infection, as shown by a pure growth of more than 10(5) colonies/mL of a single species of bacterium. In all the cases the same result was obtained from both the diaper urine sample and the sample obtained by suprapubic aspiration or catheter. In 31 infants the urine samples collected by both techniques (diaper and catheter or suprapubic aspiration) were negative, and in only 2 infants did the diaper specimen yield a positive result, while the urine obtained by suprapubic aspiration or catheter was sterile. Urine obtained from a disposable diaper can provide a valid sample for diagnosing urinary tract infection. The technique is simple, and can be carried out readily in ambulatory settings with minimal equipment and expense.

  7. Improving the Measurement of Shared Cultural Schemas with Correlational Class Analysis: Theory and Method

    Directory of Open Access Journals (Sweden)

    Andrei Boutyline

    2017-05-01

    Full Text Available Measurement of shared cultural schemas is a central methodological challenge for the sociology of culture. Relational Class Analysis (RCA is a recently developed technique for identifying such schemas in survey data. However, existing work lacks a clear definition of such schemas, which leaves RCA’s accuracy largely unknown. Here, I build on the theoretical intuitions behind RCA to arrive at this definition. I demonstrate that shared schemas should result in linear dependencies between survey rows—the relationship usually measured with Pearson’s correlation. I thus modify RCA into a “Correlational Class Analysis” (CCA. When I compare the methods using a broad set of simulations, results show that CCA is reliably more accurate at detecting shared schemas than RCA, even in scenarios that substantially violate CCA’s assumptions. I find no evidence of theoretical settings where RCA is more accurate. I then revisit a previous RCA analysis of the 1993 General Social Survey musical tastes module. Whereas RCA partitioned these data into three schematic classes, CCA partitions them into four. I compare these results with a multiple-groups analysis in structural equation modeling and find that CCA’s partition yields greatly improved model fit over RCA. I conclude with a parsimonious framework for future work.

  8. Mimicking Muscle Stem Cell Quiescence in Culture: Methods for Synchronization in Reversible Arrest.

    Science.gov (United States)

    Arora, Reety; Rumman, Mohammed; Venugopal, Nisha; Gala, Hardik; Dhawan, Jyotsna

    2017-01-01

    Growing evidence supports the view that in adult stem cells, the defining stem cell features of potency and self-renewal are associated with the quiescent state. Thus, uncovering the molecular logic of this reversibly arrested state underlies not only a fundamental understanding of adult tissue dynamics but also hopes for therapeutic regeneration and rejuvenation of damaged or aging tissue. A key question concerns how adult stem cells use quiescence to establish or reinforce the property of self-renewal. Since self-renewal is largely studied by assays that measure proliferation, the concept of self-renewal programs imposed during non-proliferating conditions is counterintuitive. However, there is increasing evidence generated by deconstructing the quiescent state that highlights how programs characteristic of this particular cell cycle exit may enhance stem cell capabilities, through both cell-intrinsic and extrinsic programs.Toward this end, culture models that recapitulate key aspects of stem cell quiescence are useful for molecular analysis to explore attributes and regulation of the quiescent state. In this chapter, we review the different methods used to generate homogeneous populations of quiescent muscle cells, largely by manipulating culture conditions that feed into core signaling programs that regulate the cell cycle. We also provide detailed protocols developed or refined in our lab over the past two decades.

  9. Optimized FISH methods for visualizing RNA localization properties in Drosophila and human tissues and cultured cells.

    Science.gov (United States)

    Diot, Cédric; Chin, Ashley; Lécuyer, Eric

    2017-08-15

    Eukaryotic gene expression is orchestrated by a large number of regulatory steps to modulate the synthesis, maturation and fate of various families of protein-coding and non-coding RNA molecules. Defining the subcellular localization properties of an RNA molecule is thus of considerable importance for gleaning its function(s) and for elucidating post-transcriptional gene regulation pathways. For decades, fluorescent In Situ hybridization (FISH) has constituted the gold-standard technique for assessing RNA expression and distribution properties in cultured cells, tissue specimens, and whole mount organisms. Recently, several attempts aimed at advancing multiplex RNA-FISH experiments have been published. However, these procedures are both financially demanding and technically challenging, while their full potential remains unexploited. Here we describe an optimized RNA-FISH method employing the Tyramide Signal Amplification system that robustly enhances resolution and sensitivity needed for exploring RNA localization in Drosophila embryos, tissues and commonly cultured human and insect cell lines. Methodological details and key parameters are outlined for high-throughput analyses conducted in 96-well plate format. Copyright © 2017. Published by Elsevier Inc.

  10. An Improved Culture Method for Selective Isolation of Campylobacter jejuni from Wastewater

    Science.gov (United States)

    Kim, Jinyong; Oh, Euna; Banting, Graham S.; Braithwaite, Shannon; Chui, Linda; Ashbolt, Nicholas J.; Neumann, Norman F.; Jeon, Byeonghwa

    2016-01-01

    Campylobacter jejuni is one of the leading foodborne pathogens worldwide. C. jejuni is isolated from a wide range of foods, domestic animals, wildlife, and environmental sources. The currently available culture-based isolation methods are not highly effective for wastewater samples due to the low number of C. jejuni in the midst of competing bacteria. To detect and isolate C. jejuni from wastewater samples, in this study, we evaluated a few different enrichment conditions using five different antibiotics (i.e., cefoperazone, vancomycin, trimethoprim, polymyxin B, and rifampicin), to which C. jejuni is intrinsically resistant. The selectivity of each enrichment condition was measured with Ct value using quantitative real-time PCR, and multiplex PCR to determine Campylobacter species. In addition, the efficacy of Campylobacter isolation on different culture media after selective enrichment was examined by growing on Bolton and Preston agar plates. The addition of polymyxin B, rifampicin, or both to the Bolton selective supplements enhanced the selective isolation of C. jejuni. The results of 16S rDNA sequencing also revealed that Enterococcus spp. and Pseudomonas aeruginosa are major competing bacteria in the enrichment conditions. Although it is known to be difficult to isolate Campylobacter from samples with heavy contamination, this study well exhibited that the manipulation of antibiotic selective pressure improves the isolation efficiency of fastidious Campylobacter from wastewater. PMID:27617011

  11. METHODS FOR IMPROVING INFORMATION CULTURE IN THE DEVELOPMENT OF RESEARCH AND CREATIVE SKILLS STUDENTS BASED ON CREATIVE THINKING

    Directory of Open Access Journals (Sweden)

    Teplaya Naila Aligasanovna

    2013-05-01

    Full Text Available The article discusses methods to improve the information culture in the development of research and creative abilities of students on the basis of creative thinking. The purpose: consider ways to improve the information culture in the development of research and creative abilities of students on the basis of creative thinking. Method or methodology of carrying out of job: theoretical analysis of the psychological and educational literature. Results: The essence of the methods to improve the information culture, which leads to the development of abilities to work in a multi-factor systems in the face of uncertainty, to the construction of more complex hierarchical structures of their own activities, updating combinatorial abilities, that improve the development process of research and creativity, to generate the highest level IV information Culture - Professionally. Scope of results: preparation of the information for University students.

  12. A Non-Destructive Culturing and Cell Sorting Method for Cardiomyocytes and Neurons Using a Double Alginate Layer

    Science.gov (United States)

    Terazono, Hideyuki; Kim, Hyonchol; Hayashi, Masahito; Hattori, Akihiro; Nomura, Fumimasa; Kaneko, Tomoyuki; Yasuda, Kenji

    2012-01-01

    A non-destructive method of collecting cultured cells after identifying their in situ functional characteristics is proposed. In this method, cells are cultivated on an alginate layer in a culture dish and released by spot application of a calcium chelate buffer that locally melts the alginate layer and enables the collection of cultured cells at the single-cell level. Primary hippocampal neurons, beating human embryonic stem (hES) cell-derived cardiomyocytes, and beating hES cell-derived cardiomyocyte clusters cultivated on an alginate layer were successfully released and collected with a micropipette. The collected cells were recultured while maintaining their physiological function, including beating, and elongated neurites. These results suggest that the proposed method may eventually facilitate the transplantation of ES- or iPS-derived cardiomyocytes and neurons differentiated in culture. PMID:22870332

  13. A non-destructive culturing and cell sorting method for cardiomyocytes and neurons using a double alginate layer.

    Directory of Open Access Journals (Sweden)

    Hideyuki Terazono

    Full Text Available A non-destructive method of collecting cultured cells after identifying their in situ functional characteristics is proposed. In this method, cells are cultivated on an alginate layer in a culture dish and released by spot application of a calcium chelate buffer that locally melts the alginate layer and enables the collection of cultured cells at the single-cell level. Primary hippocampal neurons, beating human embryonic stem (hES cell-derived cardiomyocytes, and beating hES cell-derived cardiomyocyte clusters cultivated on an alginate layer were successfully released and collected with a micropipette. The collected cells were recultured while maintaining their physiological function, including beating, and elongated neurites. These results suggest that the proposed method may eventually facilitate the transplantation of ES- or iPS-derived cardiomyocytes and neurons differentiated in culture.

  14. Cultural Heritage Digitalization on Traditional Sundanese Music Instrument Using Augmented Reality Markerless Marker Method

    Directory of Open Access Journals (Sweden)

    Budi Arifitama

    2017-07-01

    Full Text Available Research into cultural heritage which implements augmented reality technology is limited. Most recent research on cultural heritage are limited on storing data and information in the form of databases, this creates a disadvantage for people who wants to see and feel at the same moment on actual cultural heritage objects. This paper, proposes a solution which could merge the existing cultural object with people using augmented reality technology. This technology would preserve traditional instrument in the form of 3D object which can be digitally protected. The result showed that the use of augmented reality on preserving cultural heritage would benefit people who try to protect their culture.

  15. [Fungal composition in massa medicata fermentata based on culture dependent method and independent PCR-SSCP technique].

    Science.gov (United States)

    Chen, Juan; Jiao, Xiao-lin; Yang, Chun-yong; Song, Mei-fang; Gao, Wei-wei

    2014-11-01

    To analyze the fungal composition in Massa Medicata Fermentata based on culture dependent method and independent PCR-SSCP technique. Fungi were directly isolated from Massa Medicata Fermentata samples. The obtained strains were identified according to morphology and DNA sequence. Meanwhile the total fungal DNA was extracted from Massa Medicata Fermentata samples, the cultural independent PCR-SSCP technique based on β-tubulin gene were used to identify the mycobiota. According to cultural method, Aspergillus flavus and Rhizopus oryzae were present in Massa Medicata Fermentata samples, while A. flavus and A. niger were present in fried Massa Medicata Fermentata samples. In contrast, 5 species were obtained by PCR-SSCP technique, A. flavus was overlapped with fungal taxa derived from culture dependent method; A. ambiguu and A. s ivoriensis were dominant with relative abundance of 57% and 35% respectively, while the relative abundance of A. flavus was as low as 4%. None species was obtained from fried Massa Medicata Fermentata samples. PCR-SSCP based on β-tubulin gene could distinguish fungi into species, culture dependent method combined with culture independent method could better understand the fungal composition associated with Massa Medicata Fermentata fermentation.

  16. 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

  17. Between Contemporary Art and Cultural Analysis: Alternative Methods for Knowledge Production

    Directory of Open Access Journals (Sweden)

    Billy Ehn

    2012-03-01

    Full Text Available Artistic research suggests alternative methods for producing various kinds of knowledge, whether within or without the confines of academe. These methods may involve either the production of investigative artworks or the writing by the artist of a doctoral dissertation about his or her own work. For cultural researchers, the methods employed by artists engaged in these processes are both familiar and challenging, as conventional ethnography is mixed with more unpredictable experiments. This article presents several contemporary artworks, including sculpture, film, dance, installation and performance that explore various aspects of reality. What can be learnt from these works? And what could be achieved by an open exchange between artists and academic researchers? Four methodological approaches are highlighted as being of particular interest. The first relates to artists’ tendency to live experimentally, using themselves both as actors and as research objects. The second arises from the very tangible ways in which contemporary artworks approach the theme of materiality. The third relates to the emotional nature of much of contemporary art – even when it is categorised as conceptual – in its creation, forms of presentation, and influence on the spectator. Finally, many artists are gifted with the ability to find and communicate surprising meanings in ordinary life. How do they do this? Part of the answer seems to be that although artists are open to the implementation of “wild whims”, they exploit their spontaneity in a highly professional manner.

  18. Isolation of adipose-derived stem cells by using a subfractionation culturing method.

    Science.gov (United States)

    Yi, TacGhee; Kim, Wang-Kyun; Choi, Joon-Seok; Song, Seung Yong; Han, Juhee; Kim, Ji Hye; Kim, Won-Serk; Park, Sang Gyu; Lee, Hyun-Joo; Cho, Yun Kyoung; Hwang, Sung-Joo; Song, Sun U; Sung, Jong-Hyuk

    2014-11-01

    Adipose-derived stem cells (ASCs) isolated from subcutaneous adipose tissue have been tested in clinical trials. However, ASCs isolated by enzyme digestion and centrifugation are heterogeneous and exhibit wide variation in regenerative potential and clinical outcomes. Therefore, we developed a new method for isolating clonal ASCs (cASCs) that does not use enzyme digestion or centrifugation steps. In addition to cell surface markers and differentiation potential, we compared the mitogenic, paracrine and hair growth-promoting effects of ASCs isolated by the gradient centrifugation method (GCM) or by the new subfractionation culturing method (SCM). We selected three cASCs isolated by SCM that showed high rates of proliferation. The cell surface markers expressed by ASCs isolated by GCM or SCM were very similar, and SCM-isolated ASCs could potentially differentiate into different cell lineages. However, cASC lines exhibited better mitogenic and paracrine effects than ASCs isolated by GCM. The expression of Diras3, Myb, Cdca7, Mki67, Rrm2, Cdk1 and Ccna2, which may play a key role in cASC proliferation, was upregulated in cASCs. In addition, cASCs exhibited enhanced hair growth-promoting effects in dermal papilla cells and animal experiments. SCM generates a highly homogeneous population of ASCs via a simple and effective procedure that can be used in therapeutic settings.

  19. Combination of culture-independent and culture-dependent molecular methods for the determination of bacterial community of iru, a fermented Parkia biglobosa seeds.

    Directory of Open Access Journals (Sweden)

    Gbenga Adedeji Adewumi

    2013-01-01

    Full Text Available In this study, bacterial composition of iru produced by natural, uncontrolled fermentation of Parkia biglobosa seeds was assessed using culture-independent method in combination with culture-based genotypic typing techniques. PCR-denaturing gradient gel electrophoresis (DGGE revealed similarity in DNA fragments with the two DNA extraction methods used and confirmed bacterial diversity in the sixteen iru samples from different production regions. DNA sequencing of the highly variable V3 region of the 16S rRNA genes obtained from PCR-DGGE identified species related to Bacillus subtilis as consistent bacterial species in the fermented samples, while other major bands were identified as close relatives of Staphylococcus vitulinus, Morganella morganii, B. thuringiensis, Staphylococcus saprophyticus, Tetragenococcus halophilus, Ureibacillus thermosphaericus, Brevibacillus parabrevis, Salinicoccus jeotgali, Brevibacterium sp. and Uncultured bacteria clones. Bacillus species were cultured as potential starter cultures and clonal relationship of different isolates determined using amplified ribosomal DNA restriction analysis (ARDRA combined with 16S-23S rRNA gene internal transcribed spacer (ITS PCR amplification, restriction analysis (ITS-PCR-RFLP and randomly amplified polymorphic DNA (RAPD-PCR. This further discriminated Bacillus subtilis and its variants from food-borne pathogens such as Bacillus cereus and suggested the need for development of controlled fermentation processes and good manufacturing practices (GMP for iru production to achieve product consistency, safety quality and improved shelf life.

  20. Combination of culture-independent and culture-dependent molecular methods for the determination of bacterial community of iru, a fermented Parkia biglobosa seeds.

    Science.gov (United States)

    Adewumi, Gbenga A; Oguntoyinbo, Folarin A; Keisam, Santosh; Romi, Wahengbam; Jeyaram, Kumaraswamy

    2012-01-01

    In this study, bacterial composition of iru produced by natural, uncontrolled fermentation of Parkia biglobosa seeds was assessed using culture-independent method in combination with culture-based genotypic typing techniques. PCR-denaturing gradient gel electrophoresis (DGGE) revealed similarity in DNA fragments with the two DNA extraction methods used and confirmed bacterial diversity in the 16 iru samples from different production regions. DNA sequencing of the highly variable V3 region of the 16S rRNA genes obtained from PCR-DGGE identified species related to Bacillus subtilis as consistent bacterial species in the fermented samples, while other major bands were identified as close relatives of Staphylococcus vitulinus, Morganella morganii, B. thuringiensis, S. saprophyticus, Tetragenococcus halophilus, Ureibacillus thermosphaericus, Brevibacillus parabrevis, Salinicoccus jeotgali, Brevibacterium sp. and uncultured bacteria clones. Bacillus species were cultured as potential starter cultures and clonal relationship of different isolates determined using amplified ribosomal DNA restriction analysis (ARDRA) combined with 16S-23S rRNA gene internal transcribed spacer (ITS) PCR amplification, restriction analysis (ITS-PCR-RFLP), and randomly amplified polymorphic DNA (RAPD-PCR). This further discriminated B. subtilis and its variants from food-borne pathogens such as B. cereus and suggested the need for development of controlled fermentation processes and good manufacturing practices (GMP) for iru production to achieve product consistency, safety quality, and improved shelf life.

  1. A Simple Alkaline Method for Decellularizing Human Amniotic Membrane for Cell Culture

    Science.gov (United States)

    Saghizadeh, Mehrnoosh; Winkler, Michael A.; Kramerov, Andrei A.; Hemmati, David M.; Ghiam, Chantelle A.; Dimitrijevich, Slobodan D.; Sareen, Dhruv; Ornelas, Loren; Ghiasi, Homayon; Brunken, William J.; Maguen, Ezra; Rabinowitz, Yaron S.; Svendsen, Clive N.; Jirsova, Katerina; Ljubimov, Alexander V.

    2013-01-01

    Human amniotic membrane is a standard substratum used to culture limbal epithelial stem cells for transplantation to patients with limbal stem cell deficiency. Various methods were developed to decellularize amniotic membrane, because denuded membrane is poorly immunogenic and better supports repopulation by dissociated limbal epithelial cells. Amniotic membrane denuding usually involves treatment with EDTA and/or proteolytic enzymes; in many cases additional mechanical scraping is required. Although ensuring limbal cell proliferation, these methods are not standardized, require relatively long treatment times and can result in membrane damage. We propose to use 0.5 M NaOH to reliably remove amniotic cells from the membrane. This method was used before to lyse cells for DNA isolation and radioactivity counting. Gently rubbing a cotton swab soaked in NaOH over the epithelial side of amniotic membrane leads to nearly complete and easy removal of adherent cells in less than a minute. The denuded membrane is subsequently washed in a neutral buffer. Cell removal was more thorough and uniform than with EDTA, or EDTA plus mechanical scraping with an electric toothbrush, or n-heptanol plus EDTA treatment. NaOH-denuded amniotic membrane did not show any perforations compared with mechanical or thermolysin denuding, and showed excellent preservation of immunoreactivity for major basement membrane components including laminin α2, γ1-γ3 chains, α1/α2 and α6 type IV collagen chains, fibronectin, nidogen-2, and perlecan. Sodium hydroxide treatment was efficient with fresh or cryopreserved (10% dimethyl sulfoxide or 50% glycerol) amniotic membrane. The latter method is a common way of membrane storage for subsequent grafting in the European Union. NaOH-denuded amniotic membrane supported growth of human limbal epithelial cells, immortalized corneal epithelial cells, and induced pluripotent stem cells. This simple, fast and reliable method can be used to standardize

  2. A simple alkaline method for decellularizing human amniotic membrane for cell culture.

    Directory of Open Access Journals (Sweden)

    Mehrnoosh Saghizadeh

    Full Text Available Human amniotic membrane is a standard substratum used to culture limbal epithelial stem cells for transplantation to patients with limbal stem cell deficiency. Various methods were developed to decellularize amniotic membrane, because denuded membrane is poorly immunogenic and better supports repopulation by dissociated limbal epithelial cells. Amniotic membrane denuding usually involves treatment with EDTA and/or proteolytic enzymes; in many cases additional mechanical scraping is required. Although ensuring limbal cell proliferation, these methods are not standardized, require relatively long treatment times and can result in membrane damage. We propose to use 0.5 M NaOH to reliably remove amniotic cells from the membrane. This method was used before to lyse cells for DNA isolation and radioactivity counting. Gently rubbing a cotton swab soaked in NaOH over the epithelial side of amniotic membrane leads to nearly complete and easy removal of adherent cells in less than a minute. The denuded membrane is subsequently washed in a neutral buffer. Cell removal was more thorough and uniform than with EDTA, or EDTA plus mechanical scraping with an electric toothbrush, or n-heptanol plus EDTA treatment. NaOH-denuded amniotic membrane did not show any perforations compared with mechanical or thermolysin denuding, and showed excellent preservation of immunoreactivity for major basement membrane components including laminin α2, γ1-γ3 chains, α1/α2 and α6 type IV collagen chains, fibronectin, nidogen-2, and perlecan. Sodium hydroxide treatment was efficient with fresh or cryopreserved (10% dimethyl sulfoxide or 50% glycerol amniotic membrane. The latter method is a common way of membrane storage for subsequent grafting in the European Union. NaOH-denuded amniotic membrane supported growth of human limbal epithelial cells, immortalized corneal epithelial cells, and induced pluripotent stem cells. This simple, fast and reliable method can be used to

  3. 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.

  4. 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.

  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. 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)

  7. Investigation of the in vitro culture process for skeletal-tissue-engineered constructs using computational fluid dynamics and experimental methods.

    Science.gov (United States)

    Hossain, Md Shakhawath; Chen, X B; Bergstrom, D J

    2012-12-01

    The in vitro culture process via bioreactors is critical to create tissue-engineered constructs (TECs) to repair or replace the damaged tissues/organs in various engineered applications. In the past, the TEC culture process was typically treated as a black box and performed on the basis of trial and error. Recently, computational fluid dynamics (CFD) has demonstrated its potential to analyze the fluid flow inside and around the TECs, therefore, being able to provide insight into the culture process, such as information on the velocity field and shear stress distribution that can significantly affect such cellular activities as cell viability and proliferation during the culture process. This paper briefly reviews the CFD and experimental methods used to investigate the in vitro culture process of skeletal-type TECs in bioreactors, where mechanical deformation of the TEC can be ignored. Specifically, this paper presents CFD modeling approaches for the analysis of the velocity and shear stress fields, mass transfer, and cell growth during the culture process and also describes various particle image velocimetry (PIV) based experimental methods to measure the velocity and shear stress in the in vitro culture process. Some key issues and challenges are also identified and discussed along with recommendations for future research.

  8. 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%).

  9. 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%.

  10. [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.

  11. 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.

  12. 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

  13. 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.

  14. 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.

  15. 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.

  16. Evaluation of culture methods for rapid screening of swine faecal samples for Yersinia enterocolitica O : 3 biotype 4

    DEFF Research Database (Denmark)

    Hoorfar, Jeffrey; Holmvig, C.B.F.

    1999-01-01

    In two studies, seven different culture protocols were compared to test naturally contaminated faecal samples from pigs for isolation of Y. enterocolitica serotype O; 3/biotype 4( n = 70 and n = 79). Four of the protocols were based on the Nordic Committee on Food Analysis (NMKL protocols), while...... indicate possibilities of shortening the culture methods by replacing most of the biochemical tests with an agglutination test based on a monoclonal antibody....

  17. Comparison of group B streptococci colonization in vaginal and rectal specimens by culture method and polymerase chain reaction technique

    Directory of Open Access Journals (Sweden)

    Shahrokh Bidgani

    2016-03-01

    Conclusion: The frequency of GBS culture from rectal samples was higher than vaginal samples. However, the detection percentage of GBS using PCR from vaginal samples was higher than rectal samples. By contrast, the culture is a time-consuming method requiring at least 48 hours for GBS fully identification but PCR is a sensitive and rapid technique in detection of GBS, with the result was acquired during 3 hours.

  18. Designing 3ERP to meet use and users - methods to handle cultural diversity and collaborative networking in SMEs

    DEFF Research Database (Denmark)

    Dirckinck-Holmfeld, Lone; Nielsen, Janni

    2007-01-01

    The purpose is to describe a project the aim of which is to develop and validate the scientific methodological foundation for design of user friendly and usable, flexible and configurable global Enterprise Resource Planning system for Small and Medium sized Enterprises (SMEs) which are adoptable ...... to cultural diversity and support the collaborative networking tasks embedded in 3gERP. Focus is on Human Centered Design methods and tools which must be developed to handle cultural diversity....

  19. Exploring the response process of culturally differing survey respondents with a response style: A sequential mixed-methods study

    NARCIS (Netherlands)

    Morren, M.H.; Gelissen, J.P.T.M.; Vermunt, J.K.

    2013-01-01

    This article presents a mixed methods approach that integrates quantitative and qualitative methods to analyze why the four largest minorities in the Netherlands-Turks, Moroccans, Antilleans, and Surinamese-respond differently to items treating cultural topics. First, we conducted latent class

  20. [Characterization of bacterial diversity in the Shengli-S12 oil reservoir by culture-dependent and culture-independent methods].

    Science.gov (United States)

    Yuan, Sangqing; Xue, Yanfen; Wang, Weidong; Li, Ximing; Ma, Yanhe

    2008-08-01

    Examining bacterial diversity in an oil reservoir of Shengli oil field by both culture-independent molecular technique and enrichment method. The heterotrophic bacteria, hydrocarbon-degrading bacteria and sulfate-reducing bacteria were enriched from S12-4 oil-well samples by the corresponding media. Then the genomic DNAs of the enrichments were extracted and the 16S rRNA gene clone libraries were constructed. The phylogenetic analysis revealed that the bacterial 16S rRNA gene clone libraries of the 3 enrichments were dominated by clones of Thermotoga, Thermaerobacter and Thermotoga, respectively. Sequences of the other co-dominant clones observed only in the enrichments of hydrocarbon-degrading bacteria and sulfate-reducing bacteria were, respectively, associated with Marinobacter and Moorella. The uncultured 16S rRNA gene library was also generated directly from total DNA of S12-4 oil-well samples by bacterial primer set. Sequence analysis of this bacterial library indicated that a large percentage of clones were highly related to the genus Pseudomonas and the dominant species emerging in the enrichment samples had a very low content in the tested oil reservoir. The significant difference of the bacterial composition between the samples obtained from independent-culture method and enrichment method implies that the specialized nutrient may lead to a distinctive selection of dominant organisms. Through culture-dependent and culture-independent methods, we acquired important information on the bacterial diversity of ShengLi oil reservoir. These results may expand our understanding of the microbial diversity of oil reservoir and provide useful information for MEOR(microbial enhancement of oil recovery).