Identification and quantitative detection of Legionella spp. in various aquatic environments by real-time PCR assay

In this study, a SYBR green quantitative real-time PCR was developed to quantify and detect the Legionella spp. in various environmental water samples. The water samples were taken from watershed, water treatment plant, and thermal spring area in Taiwan. Legionella was detected in 13.6 % (24/176), and the detection rate for river water, raw drinking water, and thermal spring water was 10, 21.4, and 16.6 %, respectively. Using real-time PCR, concentration of Legionella spp. in detected samples ranged between 9.75?×?10⁴ and 3.47?×?10⁵?cells/L in river water, 6.92?×?10⁴ and 4.29?×?10⁵?cells/L in raw drinking water, and 5.71?×?10⁴ and 2.12?×?10⁶?cells/L for thermal spring water samples. The identified species included Legionella pneumophila (20.8 %), Legionella jordanis (4.2 %), Legionella nautarum (4.2 %), Legionella sp. (4.2 %), and uncultured Legionella sp. (66.6 %). The presence of L. pneumophila in aquatic environments suggested a potential public health threat that must be further examined.     

Occurrence of diarrheagenic Escherichia coli genes in raw water of water treatment plants

Purpose

The high incidences of waterborne diseases are frequently associated with diarrheagenic Escherichia coli (DEC). DEC may pose a health risk to people who contact surface water for recreation or domestic use. However, there is no published report on the monitoring of DEC in drinking water sources in Taiwan. In this study, the occurrence of DEC genes in raw water for water treatment plants in Taiwan was investigated.

Method

Raw water samples were taken from water treatment plants adjacent to the Kaoping River in southern Taiwan. Each water sample was treated with membrane filtration followed by DNA extraction from the concentrate and concentrate enrichment, respectively. The target genes for various DEC strains of genes were identified, including enteroaggregative E. coli (EAEC), enterohemorrhagic E. coli (EHEC), enteroinvasive E. coli (EIEC), enteropathogenic E. coli (EPEC), and enterotoxigenic E. coli (ETEC).

Results

Among 55 water samples analyzed, DEC genes were detected in 16 (29.1%) samples. Strain-specific genes for EAEC, EHEC, EIEC, and EPEC were found in the percentages of 3.6%, 10.9%, 9.1%, and 9.1%, respectively. The specific gene for ETEC is not detected in the study. By looking at the presence/absence of specific genes and water sample characteristics, water temperature was found to differ significantly between samples with and without EHEC gene. In addition, pH levels differed significantly for EHEC and EPEC presence/absence genes, and turbidity was significantly different for water with and without EPEC genes.

Conclusion

DEC genes were detected in 29.1% of the raw water samples in the study location. The potential health threat may be increased if the treatment efficiencies are not properly maintained. Routine monitoring of DEC in drinking water sources should be considered.     

Using FeGAC/H2O2 process for landfill leachate treatment

Due to the growing concern of highly contaminated landfill leachate problems in Taiwan, an innovative advanced catalytic oxidation (FeGAC/H(2)O(2)) process was developed and employed in this research to treat the landfill leachate from central Taiwan. Experimental results indicated that the FeGAC/H(2)O(2) process could effectively remove organic compounds from landfill leachate. The presence of iron oxide coated granular activated carbon (FeGAC) greatly improved the oxidative ability of H(2)O(2) for the removal of humic acids, fulvic acids and non-humic substance from leachate. For instance, at pH 6, the removal efficiencies of FeGAC/H(2)O(2) and H(2)O(2) processes were 70% and 8%, respectively. FeGAC/H(2)O(2) combined both advantages of FeGAC and H(2)O(2) where FeGAC had good organics adsorption ability and could effectively catalyse the hydrogen peroxide oxidation reaction for organics removal.     

Assessing the effects of severe rainstorm-induced mixing on a subtropical,subalpine lake

Severe rainstorms cause vertical mixing that modifies the internal dynamics (e.g., internal seiche, thermal structure, and velocity filed) in warm polymictic lakes. Yuan Yang Lake (YYL), a subtropical, subalpine, and seasonally stratified small lake in the north-central region of Taiwan, is normally affected by typhoons accompanied with strong wind and heavy rainfall during the summer and fall. In this study, we used the field data, statistical analysis, spectral analysis, and numerical modeling to investigate severe rainstorm-induced mixing in the lake. Statistical determination of the key meteorological and environmental conditions underlying the observed vertical mixing suggests that the vertical mixing, caused by heat loss during severe rainstorms, was likely larger than wind-induced mixing and that high inflow discharge strongly increased heat loss through advection heat. Spectral analysis revealed that internal seiches at the basin scale occurred under non-rainstorm meteorological conditions and that the internal seiches under the rainstorm were modified on the increase of the internal seiche frequencies. Based upon observed frequencies of the internal seiches, a two-dimensional model was simulated and then appropriate velocity patterns of the internal seiches were determined under non-rainstorm conditions. Moreover, the model implemented with inflow boundary condition was conducted for rainstorm events. The model results showed that the severe rainstorms promoted thermal destratification and changed vertical circulation of the basin-scale, internal seiche motion into riverine flow.     

Molecular detection and comparison of Acanthamoeba genotypes in different functions of watersheds in Taiwan

The protistan genus Acanthamoeba is a free-living amoeba existing in various environments. Within this protistan genus, there are some species recognized as potential human pathogens, which may cause granulomatous amoebic encephalitis, Acanthamoeba keratitis and chronic granulomatous lesions of the skin. In this study, 211 water samples were collected from two watersheds in southern Taiwan. We detected Acanthamoeba based on the PCR amplification with a genus-specific primer pair and investigation of Acanthamoeba in Puzih River and Kaoping River in southern Taiwan. Acanthamoeba species were detected in 34 (16.1%) samples. The presence or absence of Acanthamoeba within the water samples showed significant difference with the levels of water temperature and total coliforms. The most frequently identified Acanthamoeba genotype was T4 (n = 19), followed by T5 (n = 8), and then T15 (n = 3). Genotype T6, T7/T8, T11 and T12 were all detected once. Genotype T4, T5, T6, T11 and T15 of Acanthamoeba are responsible for Acanthamoeba keratitis and should be considered a potential health threat associated with human activities in environmental surface water watersheds.     

Quantifying carbon footprint for ecological river restoration

Environment, Development and Sustainability - RIVER restoration is a popular technique to rehabilitate degraded river habitat. Given the nature of these types of engineering projects, using...     

Changes in the grassland-forest boundary at Ta-Ta-Chia long term ecological research (LTER) site detected by stable isotope ratios of soil organic matter

The carbon isotope analysis [delta13C values] of organic samples can be a useful research in ecological studies because delta13C values are indicative of the plant source. This study investigated the changes in plant communities along the grassland-forest boundary in the alpine forest at Ta-Ta-Chia long term ecological research (LTER) site in central Taiwan using carbon isotope data. The aim of this study was focused on the forest fire affected the change of vegetation community. Four pedons from grassland dominated by Miscanthus transmorrisonensis (pedons 1 and 2), transition zone by Tsuga and Yushania nittakeyamensis (pedon 3), and forest zone by Tsuga and nittakeyamensis (pedon 4) were examined. Soil organic matter (SOM) delta13C values in the upper soil horizon were similar to delta13C values of the overlaying vegetation types. This indicates that the boundary between these plant communities remained the same in the past decades. The delta13C values of the grassland SOM ranged from -19.4 per thousand to -24.1 per thousand, showing decrease with soil depth. This suggests that C4 plants (transmorrisonensis) have replaced C3 plants of Tsuga and nittakeyamensis. The delta13C values of the Tsuga forest area (pedon 4) range from -27.0 per thousand to -23.5 per thousand and showed only slight change with soil depth, implying that C3 plants have remained the major species in the forest.     

Enhanced uptake of As, Zn, and Cu by Vetiveria zizanioides and Zea mays using chelating agents

Vetiveria zizaniodes (vetiver) is commonly known for its effectiveness in soil and sediment erosion control. It can tolerate to extreme soil conditions and produce a high biomass even growing in contaminated areas. Zea mays (maize) can also produce a very high biomass with a fast growth rate and possesses some degree of metal tolerance. A greenhouse study was conducted to investigate the feasibility of using vetiver and maize for remediation of arsenic (As)-, zinc (Zn-), and copper (Cu)-amended soils and evaluate the effects of chelating agents on metal uptake by these plants. Vetiver had a better growth (dry weight yield of root and shoot) than maize under different treatment conditions. The effects of different chelating agents on As, Zn, and Cu extraction from soil to soil solution were studied. Among the nine chelating agents used, it was noted that 20 mmol NTA could maximize As and Zn bioavailability, while 20 mmol HEIDA could maximize Cu bioavailability in the soil solution. The surge time in maximizing metal uptake ranged from 16 to 20 days which indicated that timing on plant harvest was an important factor in enhanced metal accumulation. In general, vetiver was a more suitable plant species than maize in terms of phytoextraction of metals from metal-contaminated soil. Application of NTA in As-amended soil and HEIDA in Cu-amended soil at the rate of 20 mmol kg(-1) increased 3-4-fold of As and Cu in shoot of both plants, whereas application of NTA (20 mmol kg(-1)) increased 37- and 1.5-fold of Zn accumulation in shoot of vetiver and maize, respectively. The potential environmental risk of metal mobility caused by chelating agents used for phytoextraction should not be overlooked.     

Effect of copper chloride on the emissions of PCDD/Fs and PAHs from PVC combustion

The influences of temperature, air flow and the amount of copper chloride upon the types and amount of the toxic emissions such as polychlorinated dibenzo-p-dioxins and polychlorinated dibenzofurans (PCDD/Fs) and polycyclic aromatic hydrocarbons (PAHs) during combustion of polyvinyl chloride (PVC) were investigated. The mechanism concerning the effect of temperature and copper chloride on the PCDD/Fs and PAHs formation was discussed. The results shown that without copper chloride, trace amounts of PCDD/Fs and large amounts of PAHs were found in the emissions from the pure PVC combustion under various combustion conditions. The addition of copper chloride enhanced PCDD/Fs formation, but it seems that the formation of PAHs decreased with increasing amount of copper chloride, and greater total amount of PAHs were produced at the higher temperature under our experimental conditions.     

Studies on the carbohydrases in the digestive tract of the milkfish chanos chanos

Crude extracts from various regions of the digestive tract of pond grown milkfish were tested forttheir ability to catalyze the hydrolysis of various carbohydrates. The most active carbohydrases were those involved in the hydrolysis of -glucosidic bonds. Maltose, trehalose, dextrin, starch and glycogen were rapidly hydrolyzed in the presence of crude extracts from the intestines and the pyloric caeca. High amylase activity was observed in extracts from the intestines, pancreas, pyloric caeca and liver. The intestinal amylase had optimum activity at pH 6.2 and at a temperature of about 50°C. It was active at a chloride concentration of 10 to 40 ppt. The amylase activity in the intestines consistently peaked daily at about noon when the milkfish gut was full. In contrast, enzyme activity was significantly lower at 0030 hrs when the gut was empty. These results are consistent with earlier observations that the milkfish is a daytime feeder and suggest further that intestinal amylase secretion is in phase with the feeding activity of the milkfish. Although the fishes used in this study fed mostly on the naturally occurring algae in the ponds, no cellulase activity was detected in any region of the digestive tract. Less active carbohydrases that were detected include a -glucosidase and -galactosidase, both of which were of limited substrate specificity.SEAFDEC Contribution No. 71