Repetitive extragenic palindromic PCR (REP-PCR) as a method used for bulking process detection in activated sludge

Bulking of activated sludge is a world-widely prevalent problem and can lead to loss of bio-oxidation, further deterioration of effluent quality, and even to a complete breakdown of the entire treatment process. Most common reasons of bulking are bacterial community changes, especially excessive growth of filamentous bacteria or excess of biopolymers on surface of non-filamentous microbes. Because of complex nature of the bulking phenomenon, the successful bulking control strategy finding is still a very important need awaiting new options and advices. The repetitive extragenic palindromic PCR (REP-PCR) fingerprinting method has been applied to distinguish bacterial community in non-bulking and bulking activated sludge. The characteristic REP-PCR fingerprinting patterns, using the Ward??s clustering method, have been analyzed to determine homology/similarity relation between particular non-bulking and bulking sludge sampling. The received clustering results were in high concordance with activated sludge typing done based on physicochemical sludge analysis. The choice and application of molecular typing method in sludge analysis will depend upon the needs, skill level, and resources of the laboratory. The proposed REP-PCR method and statistical analysis of fingerprinting patterns seems to be simple, rapid, and effective methods to show differences between population in non-bulking and bulking activated sludge. It is easy to implement, and it may be useful for routinely activated sludge monitoring as well as may be helpful in early detection of bulking process.     

FIA-coupled spectrophotometric method for determination of Cr (VI) traces in natural waters: application of in-line dissolution of 1,5-diphenylcarbazide after heat treatment and activated alumina as adsorbent for preconcentration

This work proposes the quantification of Cr (VI) ions in natural waters in trace level, using activated alumina (Al₂O₃) as preconcentration support, controlled in-line dissolution of the solidified chromophore diphenylcarbazide after heat treatment and spectrophotometric detection. The manifold ensures high sensitivity of analytical response, good repeatability, and stability. In this work, optimization of experimental conditions of a flow injection system was chosen as the parameters for greater sensitivity and better selectivity. The selected optimized conditions were 0.30 mol L^?1 for H₂SO₄ concentration, system flow rate as 0.40 mL min^?1, sample injection volume of 192.50 μL, 2 min for preconcentration time, and 0.10 mol L^?1 for eluent concentration. The analytical curves obtained for real sample analysis show linear range from 0.192 to 0.961 μM, linear correlation coefficient R?=?0.9997 and LOD?=?0.024 μM. The preconcentration factor of about four times was obtained through the passage of 800 μL of a standard solution containing 0.961 μM of Cr (VI) through mini-column of preconcentration followed by elution at 192.5 μL of NH₄OH 0.1 mol L^?1 solution. The solid chromogenic reagent presented high durability (weeks in daily use with mass of 0.0993 g) and good reproducibility in analytical signal. The reactivation of the mini-column of alumina should be executed after ten injections of eluent, using 800 μL of HCl 0.02 mol L^?1 solution in flow through the same. Each cycle of injection and elution of the sample takes about 5 min on the proposed terms. Despite the length of each cycle still be high, low concentrations can be detected using a technique of relatively low cost. This is due in part, the association dissolution of the chromogenic reagent directly in the line and the preconcentration step. Another important factor is the economy of reagent chromogenic, low generation of reject contributing to better quality of the environment, and the high potential for applications to work in field.     

Ultracentrifugation as a direct method to concentrate viruses in environmental waters: virus-like particle enumeration as a new approach to determine the efficiency of recovery

Some health important enteric viruses are considered to be emerging waterborne pathogens and so the improvement of detection of these viruses in the aquatic environment is one of the most important steps in dealing with these pathogens. Since these viruses may be present in low numbers in water, it is necessary to concentrate water samples before viral detection. Although there are several methods to concentrate viruses in environmental waters, all present some drawbacks and consequently the method should be chosen that, despite its limitations, is adequate to achieve the aim of each study. As the effectiveness of the concentration methods is evaluated by determining the efficiency of viral recovery after concentration, it is important to use a simple and effective approach to evaluate their recovery efficiency. In this work ultracentrifugation, usually used as a secondary step for virus concentration, was evaluated as the main method to concentrate directly viruses in environmental water samples, using the microscopic enumeration of virus-like particles (VLP) as a new approach to estimate the efficiency of recovery. As the flocculation method is currently employed to concentrate viruses in environmental waters, it was also used in this study to assess the efficiency of the ultracentrifugation as the main viral concentration method in environmental waters. The results of this study indicate that ultracentrifugation is an adequate approach to concentrate viruses directly from environmental waters (recovery percentages between 66 and 72% in wastewaters and between 66 and 76% in recreational waters) and that the determination of VLP by epifluorescence microscopy is a simple, fast and cheap alternative approach to determine the recovery efficiency of the viral concentration methods.     

Development of a new method for the determination of residues of the neonicotinoid insecticide imidacloprid in juvenile chinook (Oncorhynchus tshawytscha) using ELISA detection

The neonicotinoid insecticide imidacloprid (IMI) has been proposed as an alternative to carbaryl for controlling indigenous burrowing shrimp on commercial oyster beds in Willapa Bay and Grays Harbor, Washington. A focus of concern over the use of this insecticide in an aquatic environment is the potential for adverse effects from exposure to non-target species residing in the Bay, such as juvenile Chinook (Oncorhynchus tshawytscha) and cutthroat trout (O. clarki). Federal registration and State permiting approval for the use of IMI will require confirmation that the compound does not adversely impact these salmonids following field applications. This will necessitate an environmental monitoring program for evaluating exposure in salmonids following the treatment of beds. Quantification of IMI residues in tissue can be used for determining salmonid exposure to the insecticide. Refinement of an existing protocol using liquid-chromatography mass spectrometry (LC-MS) detection would provide the low limits of quantification, given the relatively small tissue sample sizes, necessary for determining exposure in individual fish. Such an approach would not be viable for the environmental monitoring effort in Willapa Bay and Grays Harbor due to the high costs associated with running multiple analyses, however. A new sample preparation protocol was developed for use with a commercially available enzyme-linked immunosorbent assay (ELISA) for the quantification of IMI, thereby providing a low-cost alternative to LC-MS for environmental monitoring in Willapa Bay and Grays Harbor. Extraction of the analyte from the salmonid brain tissue was achieved by Dounce homogenization in 4.0 mL of 20.0 mM Triton X-100, followed by a 6 h incubation at 50-55 °C. Centrifugal ultrafiltration and reversed phase solid phase extraction were used for sample cleanup. The limit of quantification for an average 77.0 mg whole brain sample was calculated at 18.2 μg kg(-1) (ppb) with an average recovery of 79%. This relatively low limit of quantification allows for the analysis of individual fish. Using controlled laboratory studies, a curvelinear relationship was found between the measured IMI residue concentrations in brain tissue and exposure concentrations in seawater. Additonally, a range of IMI brain residue concentrations was associated with an overt effect; illustrating the utility of the IMI tissue residue quantification approach for linking exposure with defined effects.     

Development of a PCR protocol for the detection of Escherichia coli O157:H7 and Salmonella spp. in surface water

Escherichia coli O157:H7 and Salmonella are pathogenic microorganisms that can cause severe gastrointestinal illness in humans. These pathogens may be transmitted in a variety of ways, including food and water. The presence of Salmonella and E. coli O157:H7 in surface waters constitutes a potential threat to human health when used for either drinking or recreation. As with most waterborne pathogens, Salmonella and E. coli O157:H7 are difficult to detect and enumerate with accuracy in surface waters due to methodological limitations. The aim of this study was to develop a protocol for the detection of Salmonella spp., E. coli O157:H7 and E. coli virulence genes (stx ₁, stx ₂ and eae) in water using a single enrichment step and PCR. In spiked water samples, PCR results showed high sensitivity (<3 CFU/L) for both microorganisms. The protocol developed in this study has been applied in different surface waters in association with microbiological and physical analysis. The frequency of PCR positive samples was 33% for Salmonella and 2% for E. coli O157:H7 producing intimin (eae) and Shiga-like toxin I (stx ₁). Moreover, the finding of amplicons corresponding to eae and stx ₁ genes in the absence of E. coli O157:H7 suggested the possible presence of other pathogenic bacteria that carry these genes (e.g. EHEC, Shigella strains). The results obtained showed that the developed protocol could be applied as a routine analysis of surface water for the evaluation of microbiological risks.     

Modular evaluation method for subsurface activities (MEMSA). A novel approach for integrating social acceptance in a permit decision-making process for subsurface activities

We investigate how the decision support system ‘Modular Evaluation Method Subsurface Activities’ (MEMSA) can help facilitate an informed decision-making process for permit applications of subsurface activities. To this end, we analyze the extent the MEMSA approach allows for a dialogue between stakeholders in a transparent manner. We use the exploration permit for the underground gas storage facility at the Pieterburen salt dome (Netherlands) as a case study. The results suggest that the MEMSA approach is flexible enough to adjust to changing conditions. Furthermore, MEMSA provides a novel way for identifying structural problems and possible solutions in permit decision-making processes for subsurface activities, on the basis of the sensitivity analysis of intermediate rankings. We suggest that the planned size of an activity should already be specified in the exploration phase, because this would allow for a more efficient use of the subsurface as a whole. We conclude that the host community should be involved to a greater extent and in an early phase of the permit decision-making process, for example, already during the initial analysis of the project area of a subsurface activity. We suggest that strategic national policy goals are to be re-evaluated on a regular basis, in the form of a strategic vision for the subsurface, to account for timing discrepancies between the realization of activities and policy deadlines, because this discrepancy can have a large impact on the necessity and therefore acceptance of a subsurface activity.     

Bioluminescent yeast estrogen assay (BLYES) as a sensitive tool to monitor surface and drinking water for estrogenicity

Estrogenic Endocrine Disrupting Chemicals (EDCs) are a concern due to their ubiquity and recognized adverse effects to humans and wildlife. Methods to assess exposure to and associated risks of their presence in aquatic environment are still under development. The aim of this work is to assess estrogenicity of raw and treated waters with different degrees of pollution. Chemical analyses of selected EDCs were performed by liquid chromatography-tandem mass spectrometry, and estrogenic activity was evaluated using in vitro bioluminescent yeast estrogen assay (BLYES). Most raw water samples (18/20) presented at least one EDC and 16 rendered positive in BLYES. When EDCs were detected, the bioassay usually provided a positive response, except when only bisphenol A was detected at low concentrations. The highest values of estrogenic activity were detected in the most polluted sites. The maximum estrogenic activity observed was 8.7 ng equiv. of E2 L(-1). We compared potencies observed in the bioassay to the relative potency of target compounds and their concentrations failed to fully explain the biological response. This indicates that bioassay is more sensitive than the chemical approach either detecting estrogenic target compounds at lower concentrations, other non-target compounds or even synergistic effects, which should be considered on further investigations. We have not detected either estrogenic activity or estrogenic compounds in drinking water. BLYES showed good sensitivity with a detection limit of 0.1 ng equiv. E2 L(-1) and it seems to be a suitable tool for water monitoring.     

Freshwater leeches (Hirudinea) as a screening tool for detecting organic contaminants in the environment

In earlier work, we found that leeches from an industrially polluted creek bioaccumulated chlorophenols to much higher concentrations than other resident benthic invertebrates and fish. We suggested that leeches may have significant potential as biomonitors for these and other organic contaminants in the environment. In this study, we compared the bioaccumulation and depuration of 16 organic compounds, including eight chlorophenols (CPs), lindane, DDT and four derivatives, benzothiazole (BT) and 2-(Methylthio)benzothiazole (MMBT) for three species of leeches. Dina dubia had the highest bioaccumulation capacity for most contaminants, but residues persisted longest in Erpobdella punctata. Helobdella stagnalis appeared capable of degrading some compounds. Half lives of CPs, DDT and DDT derivatives were generally longer than one month. In contrast, half lives were only 1 day for lindane, 1–2.5 days for MMBT and 7 days for BT despite very high initial tissue concentrations of the latter two compounds. Bioconcentration factors for contaminants in leeches were higher than those reported for other aquatic organisms. Half lives for lindane, DDT and DDT derivatives were consistent with the literature for other organisms, but half lives for CPs were much longer. The results suggest that leeches would be excellent biomonitors of both continuous and intermittent contamination of a waterway with CPs and DDT, as they retain these compounds for long periods after exposure. Their usefulness as a screening tool for lindane and benzothiazoles would be limited to chronically contaminated environments.     

亚铵法制浆造纸厂中段废水处理技术分析研究

通过对采用两级气浮做预处理 ,生物硫化床串联接触氧化做深度处理的治理技术的分析研究 ,结合废水处理工程实例 ,说明了用两级串联气浮 +硫化床 +接触氧化工艺处理造纸中段水技术切实可靠 ,该治理工艺对处理亚铵法麦草制浆造纸中段废水有较强的针对性和可行性。     

Determination of landscape beauties through visual quality assessment method: a case study for Kemaliye (Erzincan/Turkey)

Kemaliye (Erzincan/Turkey) is the member of European Association of Historic Towns and Regions. The aim of this study was to reveal the visual richness of the town; to identify the relationship between landscape spatial pattern and visual quality of the landscape and to offer some suggestions for the future planning in regarding to these visual beauties. The visual quality assessment method was used in this study. The results of the study revealed three landscape types that have the highest visual quality. Among those, the highest one is urban scenery 3 (US3; VQP = 5.9400), the second is geological structure scenery 5 (GSS 5; VQP = 5.9200) and the third natural scenery 3 (NS3; VQP = 5.9133). Visual quality assessment showed that urban pattern, geological structure and natural resources of the region also have visual value. The relationships between landscape spatial pattern and visual quality of landscape indicated that certain characteristics of landscape affected the quality. For instance, as the texture level decreased in natural landscapes and as the green areas increased in geological structure, visual preferences ratio increased. Some suggestions were also made regarding the visual resources use in the region.     

Chironomid mouthpart deformities as indicator of environmental quality: a case study in Lake Trasimeno (Italy)

The aim of the present study was to monitor mouthpart deformities of Chironomus plumosus (Insecta-Diptera) in Lake Trasimeno (2000-2010) in relation with sediment contamination by heavy metals, which is one of the main causes of deformity induction. In Lake Trasimeno, concentrations of heavy metals in the investigated littoral zone were low, in comparison with those of the central area. By contrast, the incidence of deformities was much higher in the littoral station (2006-2009) than in the central area. In the littoral zone, the deformities progressively decreased and by 2010 reached values similar to those observed in the central area (2000-2002, 2010). This decrease was mainly due to the reduction of severe deformities, thus indicating an environmental quality recovery of this littoral sector. Chemical analysis revealed that the higher incidence of deformities in the littoral zone was not caused by heavy metal contamination, unless they may have contributed to a joint action with other toxicants. The present study on Lake Trasimeno provided additional evidence that chironomid mouthpart deformities can be used in biomonitoring programs to evaluate sediment contamination by toxicants. Since mouthpart deformities may also be induced by compounds not included in routine chemical analyses, they may better reflect sediment quality than chemical analysis alone.     

Enteromorpha flexuosa (Wulfen) J. Agardh (Chlorophyta: Ulvales)--evaluation as an indicator of heavy metal contamination in a tropical estuary

The use of bioindicators for contaminant monitoring ispopular in all sectors of the environment but quite oftenbioindicators are utilised without rigorous evaluation oftheir viability as an indicator. We report field andlaboratory investigations into the value of a commonlyfound macroalga, Enteromorpha flexuosa (Wulfen) J.Agardh (Chlorophyta: Ulvales) as an indicator of copper,zinc and lead contamination in a tropical estuary inFiji. In the laboratory, metal content of E.flexuosa after 60 days of growth in seawater containingincreasing concentrations of the metals showed almostperfect correlation with metal concentrations in waterfor all three metals, and concentration factors obtainedwere generally higher than values reported for otherspecies of Enteromorpha. Performance in the fieldwas evaluated by a one-year monitoring of metals in E. flexuosa, water and sediments from a contaminatedestuary. Metal concentrations in water were always belowdetection limits and field concentration factors couldnot be determined but the concentration of lead in E. flexuosa showed a high correlation with the leadcontent of sediments. Lack of such correlation forcopper and zinc in the field suggests that otherenvironmental factors besides metal load in the physicalenvironment could be controlling the bioaccumulation ofthese two metals. Whilst demonstrating E. flexuosato be a viable bioindicator for lead, these results alsohighlight the need for proper assessment before anorganism is considered for environmental monitoring.     

Development of a solid phase microextraction (SPME) method for the sampling of VOC traces in indoor air

Solid-phase microextraction (SPME) was studied for the measurement of volatile organic compounds (VOCs) in indoor air. An adsorptive PDMS/Carboxen fibre was used and an analytical methodology was developed in order to overcome competitive adsorption. Kinetics and adsorption isotherms were investigated for different sample volumes and model compounds. In order to evaluate competitive adsorption on the fibre, these compounds were studied alone and in mixture. From the results obtained, the operating conditions allowing co-adsorption of the target compounds were determined: the air sample is enclosed in a 250 mL glass bulb where the SPME fibre is exposed until adsorption equilibrium. This procedure was combined with GC/MS analysis for the identification and quantification of VOCs in indoor air. The performances were determined by using a standard gas containing 10 VOCs representative of indoor environments (acetaldehyde, acetone, BTX, alpha-pinene, trichloroethylene, alkanes). The detection limits were determined in single ion monitoring mode and for a signal to noise ratio of 3. Except acetaldehyde (6 microg m(-3)), they are all below 0.5 microg m(-3). Calibration curves are linear up to 10 micromol m(-3) for all the compounds with good correlation coefficients (above 0.99). The reproducibility ranges from 6 to 12% according to the compound. The methodology was then applied to the comparison of the VOCs content in classrooms of two different schools.     

Copper mine tailing disposal in northern Chile rocky shores: Enteromorpha compressa (Chlorophyta) as a sentinel species

The study assesses the ecological impact caused by the El Salvador untreated (1975–1990) and treated (1991–1994) copper mine tailings on rocky intertidal communities in and around the dumping site at Caleta Palito, northern Chile. Ecological changes are monitored for 16 years in polluted and unpolluted sites within a geographical area of 90 km. Copper concentration levels in water and the intertidal Chlorophyta E. compressa are presented. The results confirm a notorious reduction in the number of species and significant differences between polluted and unpolluted intertidal communities. At polluted sites, following the initiation of the disposal, all species of invertebrates and algae disappeared and primary space (rock) was partially or completely dominated by E. compressa along more than a decade. Its persistence in these sites supports the view that this taxon is a sentinel species resisting high levels of copper pollution. During the past four years, following the steps given to treat the tailings, at polluted sites there are preliminary indications showing increases in the number of species of algae and invertebrate. The need for future monitoring to elucidate ecosystem restoration processes is discussed.     

Accumulation of Endosulfan in Wild Rat, Rattus norvegious as a Result of Application to Soya Bean in Mazoe (Zimbabwe)

An assessment of levels of endosulfan and endosulfan sulphate in rats collected from a plot sprayed with endosulfan was carried out for a period of four months. The analysis was carried out in the livers, muscles and fatty tissues of the rats using Gas Chromatography. Samples were ground with sodium sulphate and aluminium oxide and the analyte soxhlet extracted with hexane. The extract was then cleaned by florisil column chromatography. Levels of endosulfan in rats were found to range in livers from 0.356 to 5.807 ppb, in muscles from 0.089 to 5.167 ppb and in fatty tissues from 0.083 to 4.785 ppb. Levels of endosulfan sulphate correlated well with those of endosulfan and ranged in livers from 0.00 to 3.96 ppb, in muscles from 0.00 to 3.37 ppb and in fatty tissue from 0.00 to 2.93 ppb. In the control rats samples, collected from an unsprayed area in the city of Harare (University of Zimbabwe), no endosulfan could be detected.     

Amplified ribosomal DNA restriction analysis as a routine tool to assess toxicant driven changes in hindgut bacterial populations of Porcellio dilatatus (Crustacea: Isopoda)

Changes in saprophytic organism's gut microbial communities may present a threat to organic matter breakdown which can ultimately lead to soil function impairment. In this study, Amplified Ribosomal DNA Restriction Analysis (ARDRA) was evaluated as a potential simple molecular tool to assess shifts in bacterial community structure in hindgut populations of Porcellio dilatatus exposed to contaminated food. This prospective tool can also be used for a variety of purposes and samples prior to the use of more specific and sophisticated methods.     

Application of cold-induced aggregation microextraction as a fast,simple, and organic solvent-free method for the separation and preconcentration of Se(IV) in rice and various water samples

The developed method is based on cold-induced aggregation microextraction of Se(IV) using the 1-butyl-3-methylimidazolium hexafluorophosphate ionic liquid as an extractant followed by spectrophotometry determination. The extraction of Se(IV) was performed in the presence of dithizone as the complexing agent. In this method, a very small amount of 1-butyl-3-methylimidazolium hexafluorophosphate was added to the sample solution containing Se-dithizone complex. Then, the solution was kept in a thermostated bath at 50 °C for 4 min. Subsequently, the solution was cooled in an ice bath and a cloudy solution was formed. After centrifuging, the extractant phase was analyzed using a spectrophotometric detection method. Some important parameters that might affect the extraction efficiency were optimized (HCl, 0.6 mol L^?1; dithizone, 4.0?×?10^?6 mol L^?1; ionic liquid, 100 μL). Under the optimum conditions, good linear relationship, sensitivity, and reproducibility were obtained. The limit of detection (LOD) (3S_b/m) was 1.5 μg L^?1, and the relative standard deviation (RSD) was 1.2 % for 30 μg L^?1 of Se(IV). The linear range was obtained in the range of 5–60 μg L^?1. It was satisfactory to analyze rice and various water samples.