Profiling of environmental Legionella pneumophila strains by randomly amplified polymorphic DNA method isolated from geographically nearby buildings

Legionella pneumophila (L. pneumophila) which is also known as etiologic agent Legionnaires Disease lives in natural water and man made water systems. These bacteria belonging to Legionellaceae family are divided 15 serogroups. Phenotypical methods used for the identification of Legionella isolates are not very discriminatory. In this study we investigated genotypic features of eight L. pneumophila serogroup 1 and 18 L. pneumophila serogroup 2-14 strains isolated from different buildings in Istanbul by randomly amplified polymorphic DNA (RAPD) method. Eight L. pneumophila serogroup 1 strains (37.5%) were similar RAPD profile and they were isolated from buildings located in a short distance (about 500 m). Four L. pneumophila serogroup 2-14 strains (22%) were identical genotypically. Three of these strains were isolated from buildings located in a short distance.     

Comparison of the microbial load of incoming and distal outlet waters from dental unit water systems in Istanbul

This is a cross-sectional study of the incoming and distal outlet water quality from 41 dental units in Istanbul, carried out to compare the total microbial loads using traditional culture method versus epifluorescence microscopy. The possible presence of Legionella pneumophila using traditional culture method was also analyzed. One hundred and twenty three samples were taken from the high-speed handpiece lines, air-water syringe lines and source (incoming) water supplies from 41 dental units. The samples were assayed for live/dead bacteria, heterotrophic bacterial counts and presence of L. pneumophila bacteria. Thirty nine out of 41 dental units (91%) were not able to meet the standard limit of 200 CFU/ml in dental unit waters. The live bacterial counts were 1-1.5 orders of magnitude higher than aerobic mesophilic heterotrophic bacteria. L. pneumophila (serogroup 2-14) was isolated from five out of 41 units. Some dental units were using commercially bottled (19 l) drinking water as a source. The source water of eight dental unit was heavily contaminated which were fed up by commercially bottled drinking water.     

Deposition of Inorganic Particulate Aerosols to Vegetation - A New Method of Estimating

A new, direct method was developed for quantifying inorganic particulate aerosols trapped by the forest canopy, and for determining the resulting input of elements to a forest ecosystem. The method is based on direct measurements of only six parameters. Using this method, it is possible to determine the load of aerosols trapped by the forest canopy and deposited to leaves, as well as the load of aerosols falling to the forest floor by impaction on plants. It is also possible to estimate the aerosol input of soluble and insoluble elements to an ecosystem. With this new method it has been found that the load of aerosols trapped by the canopy of a mixed forest located in the Rybnik Coal Basin averaged 189.0 kg x ha-1 x growing season-1, or 39.3% of the total inorganic particles reaching the ecosystem. The trapped aerosols provided 13.4 kg x ha-1 of soluble nitrogen and 0.91 kg x ha-1 of insoluble nitrogen over the growing season. At the same time, the input of soluble nitrogen from the atmosphere with rainfall to an open area averaged 13.9 kg x ha-1, and the input of insoluble nitrogen with inorganic dusts averaged 1.4 kg x ha-1.     

Evaluating seasonal dynamics of bacterial communities in marine fish aquaculture: a preliminary study before applying phage therapy

The increasing problem of antibiotic resistance in common pathogenic bacteria and the concern about the spreading of antibiotics in the environment bring the need to find new methods to control fish pathogens. Phage therapy represents a potential alternative to antibiotics, but its use in aquaculture requires a detailed understanding of bacterial communities, namely of fish pathogenic bacteria. Therefore, in this study the seasonal dynamics of the overall bacterial communities, microbiological water quality and disease-causing bacteria were followed in a marine aquaculture system of Ria de Aveiro (Portugal). Analysis of the bacterial diversity of the water samples by denaturing gradient gel electrophoresis (DGGE) of 16S rRNA gene fragments indicates that the bacterial community structure varied seasonally, showing a higher complexity during the warm season. The diversity of the main fish pathogenic bacteria, assessed by DGGE targeting the Vibrio genus, showed lower seasonal variation, with new dominating populations appearing mainly in the spring. Bacterial indicators, faecal coliforms and enterococci, enumerated by the filter-membrane method, also varied seasonally. The fluorescent in situ hybridization (FISH) results showed that the specific groups of bacteria varied during the study period and that the non-indigenous Enterobactereaceae family was the most abundant group followed by Vibrio and Aeromonas. The seasonal variation detected in terms of density and structure of total and pathogenic bacterial communities demonstrates the need for a careful monitoring of water through the year in order to select the suitable phages to inactivate fish pathogenic bacteria. The spring season seems to be the critical time period when phage therapy should be applied.     

Documentation of bioaerosol concentrations in an indoor composting facility in France

Bioaerosol concentrations were investigated in a totally indoor composting facility processing fermentable household and green wastes to assess their variability. Stationary samples were collected by filtration close to specific composting operations and then were analysed for cultivable mesophilic bacteria, thermophilic bacteria, mesophilic fungi, thermophilic fungi, endotoxins and total airborne bacteria (DAPI-staining). Indoor concentrations exceeded the background levels, between 500 and 5400 EU m(-3) for endotoxins, 10(4) and 10(6) CFU m(-3) for cultivable bacteria and generally below 10(5) CFU m(-3) for airborne cultivable fungi. No significant (p > 0.05) differences were observed between the indoor composting operations. Successive 30 minute bioaerosol samples were collected to investigate the variation of cultivable mesophilic microorganisms over the work shift. Concentrations of mesophilic bacteria and fungi varied up to 1 log unit depending on the time at which they were collected in the day. Total airborne particles, counted using an optical particle counter, were present at up to 10(8) particles m(-3) and several concentration peaks were noted. Values for total airborne bacteria were roughly 70-fold higher than cultivable bacteria. These results raise the question of the sampling strategy (duration of sampling; number of samples to be collected) used in similar studies. They provide new bioaerosol concentration data in a composting facility and suggest that the filtration sampling method might be a useful tool for exposure measurements in that occupational environment.     

Intervention study of airborne fungal spora in homes with portable HEPA filtration units

The concentrations and composition of airborne fungal spores in homes fitted with portable HEPA filtration units were examined to provide information to evaluate the importance of varying levels of fungal spores in residential environments in Perth, Australia. A novel method for simulating activity/impaction on carpeted environments was also investigated. Reductions in fungal (35%) and particulate (38%) levels were achieved in the air filter homes. Penicillium, Cladosporium and yeasts were the most common and widespread fungi recovered indoors and outdoors. Fungal range decreased over the study period but this could be due to an overall reduced dissemination of spores (less spores in the air).     

Activity monitoring for nitrifying bacteria by fluorescence in situ hybridization and respirometry

The activities of two families of nitrifying bacteria weremeasured by applied respirometer and fluorescence insitu hybridization (FISH) for the purpose of monitoring nitrogenremoval. Oxygen uptake rates (OUR) for Nitrosomonas andNitrobacter were separately measured with the dosing ofselective inhibitors during respirometric monitoring. The FISHmethod was applied to analyze microorganism communities. In 5mg NH₄ ⁺-N/L solution, the total nitrogen oxygendemand (NOD) was found to be 6,672 mg DO/L-day with ammoniaand nitrite oxidation rate of 4,512 and 2,160 mg DO/L-day,respectively. The oxygen consumption by endogenous oxidationwas 288 mg DO/L-day and the biological oxygen demand 936 mgDO/L-day. Respirometric measurements were well matched by theFISH result, which described the presence and quantity of eachnitrifying bacteria as nitrification proceeded. It was foundthat when coupled with OUR data, FISH could provide valuableinformation about the nitrogen removal process in manypractical wastewater treatment systems.     

UPLC-MS/MS法同时测定地表水中多种有机污染物

采用超高效液相色谱-串联质谱法直接进样测定地表水中26种有机污染物,通过优化试验条件,使各目标化合物在1.00μg/L^100μg/L范围内线性良好,相关系数(r 2)>0.995,方法检出限为0.003μg/L^1.0μg/L。将该方法用于地表水样的测定,加标回收率为79.0%~120%,6次测定结果的RSD均<15%。     

Monitoring of Biofilm-associated Legionella pneumophila on Different Substrata in Model Cooling Tower System

Cooling towers have the potential to develop infectious concentrations of Legionella pneumophila. Legionella counts increases where biofilm and warm water temperatures are present. In this study, biofilm associated L. pneumophila and heterotrophic bacteria were compared in terms of material dependence. Model cooling tower system was experimentally infected by L. pneumophila standard strain and monthly monitored. Different materials were tested for a period of 180 days. The lowest L. pneumophila and heterotrophic plate counts were measured on plastic polymers, whereas L. pneumophila and heterotrophic bacteria were accumulated rapidly on galvanized steel surfaces. It can be concluded that selection of plastic polymers, as a manufacturing material, are suitable for recirculating water systems.     

Investigation of the bacterial load and antibiotic susceptibility of dental units

The aim of this study was to evaluate the bacterial contamination level and to determine the antibiotic susceptibility of the isolated bacteria from dental unit waterlines (DUWLs) in Istanbul. Bacterial quality of DUWLs is very important, as patients and dental staff are regularly exposed to water and aerosols generated by the unit. If opportunistic pathogens such as Staphylococcus aureus, Pseudomonas aeruginosa, and Legionella pneumophila are present in DUWLs, patient and dental staff can be infected. One hundred water samples were collected from high-speed drills and input waters from 50 dental units. Aerobic heterotrophic bacteria counts and the presence of Legionella, Pseudomonas, oral streptococci, and Staphylococcus were investigated in dental unit waters and aerosol samples. In addition, the antibiotic susceptibility of the isolated and identified bacteria from DUWLs was examined. This research found that 37 out of 50 dental unit water samples exceeded the American Dental Association’s limit of 200 colony-forming units (CFU)/mL^?1. Legionella, oral streptococci, and S. aureus were not detected in any water or aerosol samples, but P. aeruginosa was isolated in three DUWLs. Also, Pseudomonas and Staphylococcus were found in water and aerosol samples. Cefoperazone, ofloxacin, gentamicin, ciprofloxacin, and piperacillin were the most effective antibiotics against the isolated bacteria from DUWLs.     

Bacterial communities in urban aerosols collected with wetted-wall cyclonic samplers and seasonal fluctuations of live and culturable airborne bacteria

Airborne transmission of bacterial pathogens from point sources (e.g., ranches, dairy waste treatment facilities) to areas of food production (farms) has been suspected. Determining the incidence, transport and viability of extremely low levels of pathogens require collection of high volumes of air and characterization of live bacteria from aerosols. We monitored the numbers of culturable bacteria in urban aerosols on 21 separate days during a 9 month period using high volume cyclonic samplers at an elevation of 6 m above ground level. Culturable bacteria in aerosols fluctuated from 3 CFU to 6 million CFU/L of air per hour and correlated significantly with changes in seasonal temperatures, but not with humidity or wind speed. Concentrations of viable bacteria determined by fluorescence staining and flow cytometry correlated significantly with culturable bacteria. Members of the phylum Proteobacteria constituted 98% of the bacterial community, which was characterized using 16S rRNA gene sequencing using DNA from aerosols. Aquabacterium sp., previously characterized from aquatic environments, represented 63% of all clones and the second most common were Burkholderia sp; these are ubiquitous in nature and some are potential human pathogens. Whole genome amplification prior to sequencing resulted in a substantial decrease in species diversity compared to characterizing culturable bacteria sorted by flow cytometry based on scatter signals. Although 27 isolated colonies were characterized, we were able to culture 38% of bacteria characterized by sequencing. The whole genome amplification method amplified DNA preferentially from Phyllobacterium myrsinacearum, a minor member of the bacterial communities, whereas Variovorax paradoxus dominated the cultured organisms.     

Simplification and validation of a large volume polyurethane foam sampler for the analysis of persistent hydrophobic compounds in drinking water

The use of a large volume polyurethane foam (PUF) sampler was validated for rapid extraction of persistent organic pollutants (POPs), such as polychlorinated dibenzo-p-dioxins and dibenzofurans (PCDD/Fs), in raw water and treated water from drinking water plants. To validate the recovery of target compounds in the sampling process, a (37)Cl-labeled standard was spiked into the 1st PUF plug prior to filtration. An accelerated solvent extraction method, as a pressurized liquid extractor (PLE), was optimized to extract the PUF plug. For sample preparation, tandem column chromatography (TCC) clean-up was used for rapid analysis. The recoveries of labeled compounds in the analytical method were 80-110% (n = 9). The optimized PUF-PLE-TCC method was applied in the analysis of raw water and treated potable water from seven drinking water plants in South Korea. The sample volume used was between 18 and 102 L for raw water at a flow rate of 0.4-2 L min(-1), 95 and 107 L for treated water at a flow rate of 1.5-2.2 L min(-1). Limit of quantitation (LOQ) was a function of sample volume and it decreased with increasing sample volume. The LOQ of PCDD/Fs in raw waters analyzed by this method was 3-11 times lower than that described using large-size disk-type solid phase extraction (SPE) method. The LOQ of PCDD/F congeners in raw water and treated water were 0.022-3.9 ng L(-1) and 0.018-0.74 ng L(-1), respectively. Octachlorinated dibenzo-p-dioxin (OCDD) was found in some raw water samples, while their concentrations were well below the tentative criterion set by the Japanese Environmental Ministry for drinking water. OCDD was below the LOQ in the treated drinking water.     

Measurements of dissolved methylmercury in natural waters using diffusive gradients in thin film (DGT)

A diffusive gradient in thin films (DGT) technique for measuring methylmercury (MeHg) concentrations in natural waters was developed using 3-mercaptopropyl-functionalized silica gel to preconcentrate the methylmercury. The new resin was characterized and calibrated. Methylmercury is efficiently accumulated at a pH range of 3-9. Basic performance tests of the new DGT device confirmed the applicability of Fick's first law for such DGT measurements. The diffusion coefficient of methylmercury in polyacrylamide gel was 5 x 10(-6) cm(2) s(-1). Methylmercury concentrations determined by DGT deployed for different time periods in the field are statistically not different from results obtained through direct measurements. The DGT technique represents therefore an alternative in situ sampling method for methylmercury. The detection limit of the overall method is 1 pg of MeHg, which correspond to approximately 30 pg L(-1) of MeHg in a water sample, when deploying a typical DGT device for 24 hours. Lower MeHg concentrations are measurable using longer deployment times or thinner diffusive gel layers.     

昆明市不同功能区夏季空气微生物污染监测

为了监测和评价昆明市空气微生物污染状况,通过平皿自然沉降法,于2010年夏季对其不同功能区空气真菌和细菌进行了初步监测。结果表明:昆明市夏季空气微生物以细菌为主;以空气真菌含量评价为清洁;8月空气真菌含量最高,7月空气细菌含量最低;空气微生物含量日变化不明显。     

Characteristics of aerosol types during large-scale transport of air pollution over the Yellow Sea region and at Cheongwon,Korea, in 2008

Episodes of large-scale transport of airborne dust and anthropogenic pollutant particles from different sources in the East Asian continent in 2008 were identified by National Oceanic and Atmospheric Administration satellite RGB (red, green, and blue)-composite images and the mass concentrations of ground level particulate matter. These particles were divided into dust, sea salt, smoke plume, and sulfate by an aerosol classification algorithm. To analyze the aerosol size distribution during large-scale transport of atmospheric aerosols, aerosol optical depth (AOD) and fine aerosol weighting (FW) of moderate imaging spectroradiometer aerosol products were used over the East Asian region. Six episodes of massive airborne dust particles, originating from sandstorms in northern China, Mongolia, and the Loess Plateau of China, were observed at Cheongwon. Classified dust aerosol types were distributed on a large-scale over the Yellow Sea region. The average PM10 and PM2.5 ratio to the total mass concentration TSP were 70% and 15%, respectively. However, the mass concentration of PM2.5 among TSP increased to as high as 23% in an episode where dust traveled in by way of an industrial area in eastern China. In the other five episodes of anthropogenic pollutant particles that flowed into the Korean Peninsula from eastern China, the anthropogenic pollutant particles were largely detected in the form of smoke over the Yellow Sea region. The average PM10 and PM2.5 ratios to TSP were 82% and 65%, respectively. The ratio of PM2.5 mass concentrations among TSP varied significantly depending on the origin and pathway of the airborne dust particles. The average AOD for the large-scale transport of anthropogenic pollutant particles in the East Asian region was measured to be 0.42 ± 0.17, which is higher in terms of the rate against atmospheric aerosols as compared with the AOD (0.36 ± 0.13) for airborne dust particles with sandstorms. In particular, the region ranging from eastern China, the Yellow Sea, and the Korean Peninsula to the Korea East Sea was characterized by high AOD distributions. In the episode of anthropogenic polluted aerosols, FW averaged 0.63 ± 0.16, a value higher than that in the episode of airborne dust particles (0.52 ± 0.13) with sandstorms, showing that fine anthropogenic pollutant particles contribute greatly to atmospheric aerosols in East Asia.     

氨氮实验室空白偏高的影响因素研究

利用排除法对影响氨氮实验室空白测定的各种因素进行了考察。结果表明,固定剂硫酸的添加不会引起空白值偏高。滤纸因含有铵盐会引起空白偏高,但可采取用去离子水浸泡的方法消除。絮凝沉淀过程中应避免将水体pH调至9.0左右。在水体pH=9.0左右时,沉淀物颗粒尺寸最小,过滤时渗入滤后液中,影响吸光度;过滤后的滤液pH会降低,导致滤液中重新析出沉淀物,也影响吸光度。     

Development of an improved methodology to detect infectious airborne influenza virus using the NIOSH bioaerosol sampler

A unique two-stage cyclone bioaerosol sampler has been developed at NIOSH that can separate aerosols into three size fractions. The ability of this sampler to collect infectious airborne viruses from a calm-air chamber loaded with influenza A virus was tested. The sampler's efficiency at collecting aerosolized viral particles from a calm-air chamber is essentially the same as that from the high performance SKC BioSampler that collects un-fractionated particles directly into a liquid media (2.4 × 10(4) total viral particles per liter of sampled air (TVP/L) versus 2.6 × 10(4) TVP/L, respectively, after 15 min) and the efficiency is relatively constant over collection times of 15, 30 and 60 min. Approximately 34% of the aerosolized infectious virus collected after 15 min with the NIOSH bioaerosol sampler remained infectious, and infectious virus was found in all three size fractions. After 60 min of sampling, the infectious virus/liter air found in the NIOSH bioaerosol sampler was 15% of that found in the SKC BioSampler. This preservation of infectivity by the NIOSH bioaerosol sampler was maintained even when the initial infectivity prior to aerosolization was as low as 0.06%. The utility of the NIOSH bioaerosol sampler was further extended by incorporating an enhanced infectivity detection methodology developed in our laboratory, the viral replication assay, which amplified the infectious virus making it more readily detectable.     

Bioaerosol sampling by a personal rotating cup sampler CIP 10-M

High concentrations of bioaerosols containing bacterial, fungal and biotoxinic matter are encountered in many workplaces, e.g. solid waste treatment plants, waste water treatment plants and sewage networks. A personal bioaerosol sampler, the CIP 10-M (M-microbiologic), has been developed to measure worker exposure to airborne biological agents. This sampler is battery operated; it is light and easy to wear and offers full work shift autonomy. It can sample much higher concentrations than biological impactors and limits the mechanical stress on the microorganisms. Biological particles are collected in 2 ml of liquid medium inside a rotating cup fitted with radial vanes to maintain an air flow rate of 10 l min(-1) at a rotational speed of approximately 7,000 rpm. The rotating cup is made of sterilisable material. The sampled particles follow a helicoidal trajectory as they are pushed to the surface of the liquid by centrifugal force, which creates a thin vertical liquid layer. Sterile water or another collecting liquid can be used. Three particle size selectors allow health-related aerosol fractions to be sampled according to international conventions. The sampled microbiological particles can be easily recovered for counting, incubation or further biochemical analysis, e.g., for airborne endotoxins. Its physical sampling efficiency was laboratory tested and field trials were carried out in industrial waste management conditions. The results indicate satisfactory collection efficiency, whilst experimental application has demonstrated the usefulness of the CIP 10-M personal sampler for individual bioaerosol exposure monitoring.     

Determination of response of real-time SidePak AM510 monitor to secondhand smoke, other common indoor aerosols, and outdoor aerosol

The amount of light scattered by airborne particles inside an aerosol photometer will vary not only with the mass concentration, but also with particle properties such as size, shape, and composition. This study conducted controlled experiments to compare the measurements of a real-time photometer, the SidePak AM510 monitor (SidePak), with gravimetric mass. PM sources tested were outdoor aerosols, and four indoor combustion sources: cigarettes, incense, wood chips, and toasting bread. The calibration factor for rescaling the SidePak measurements to agree with gravimetric mass was similar for the cigarette and incense sources, but different for burning wood chips and toasting bread. The calibration factors for ambient urban aerosols differed substantially from day to day, due to variations in the sources and composition of outdoor PM. A field evaluation inside a casino with active smokers yielded calibration factors consistent with those obtained in the controlled experiments with cigarette smoke.     

Assessment of drinking water quality using ICP-MS and microbiological methods in the Bholakpur area,Hyderabad, India

A total of 16 people died and over 500 people were hospitalized due to diarrhoeal illness in the Bholakpur area of Hyderabad, India on 6th May 2009. A study was conducted with immediate effect to evaluate the quality of municipal tap water of the Bholakpur locality. The study consists of the determination of physico-chemical properties, trace metals, heavy metals, rare earth elements and microbiological quality of drinking water. The data showed the variation of the investigated parameters in samples as follows: pH 7.14 to 8.72, EC 455 to 769 μS/cm, TDS 303.51 to 515.23 ppm and DO 1.01 to 6.83 mg/L which are within WHO guidelines for drinking water quality. The water samples were analyzed for 27 elements (Li, Be, B, Na, Mg, Al, Si, K, Ca, V, Cr, Mn, Fe, Ni, Co, Cu, Zn, As, Se, Rb, Sr, Mo, Ag, Cd, Sb, Ba and Pb) using inductively coupled plasma-mass spectrometry (ICP-MS). The concentrations of Fe (0.12 to 1.13 mg/L), Pb (0.01 to 0.07 mg/L), Cu (0.01 to 0.19 mg/L), Ni (0.01 to 0.15 mg/L), Al (0.16 to 0.49 mg/L), and Na (38.36 to 68.69 mg/L) were obtained, which exceed the permissible limits of the World Health Organization (WHO) for drinking water quality guidelines. The remaining elements were within the permissible limits. The microbiological quality of water was tested using standard plate count, membrane filtration technique, thermotolerant coliform (TTC), and most probable number (MPN) methods. The total heterotrophic bacteria ranged from 1.0 × 10⁵ to 18 × 10⁷ cfu/ml. Total viable bacteria in all the water samples were found to be too numerable to count and total number of coliform bacteria in all water samples were found to be of order of 1,100 to >2,400 MPN index/100 ml. TTC tested positive for coliform bacteria at 44.2°C. All the water samples of the study area exceeded the permissible counts of WHO and that (zero and minimal counts) of the control site (National Geophysical Research Institute) water samples. Excessively high colony numbers indicate that the water is highly contaminated with microorganisms and is hazardous for drinking purposes. Bacteriological pollution of drinking water supplies caused diarrhoeal illness in Bholakpur, which is due to the infiltration of contaminated water (sewage) through cross connection, leakage points, and back siphoning.