Matching Items (48)

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A Strategy to Establish a Quality Assurance/Quality Control Plan for the Application of Biosensors for the Detection of E. Coli in Water

Description

Rapid bacterial detection using biosensors is a novel approach for microbiological testing applications. Validation of such methods is an obstacle in the adoption of new bio-sensing technologies for water testing. Therefore, establishing a quality assurance and quality control (QA/QC) plan

Rapid bacterial detection using biosensors is a novel approach for microbiological testing applications. Validation of such methods is an obstacle in the adoption of new bio-sensing technologies for water testing. Therefore, establishing a quality assurance and quality control (QA/QC) plan is essential to demonstrate accuracy and reliability of the biosensor method for the detection of E. coli in drinking water samples. In this study, different reagents and assay conditions including temperatures, holding time, E. coli strains and concentrations, dissolving agents, salinity and pH effects, quality of substrates of various suppliers of 4-methylumbelliferyl glucuronide (MUG), and environmental water samples were included in the QA/QC plan and used in the assay optimization and documentation. Furthermore, the procedural QA/QC for the monitoring of drinking water samples was established to validate the performance of the biosensor platform for the detection of E. coli using a culture-based standard technique. Implementing the developed QA/QC plan, the same level of precision and accuracy was achieved using both the standard and the biosensor methods. The established procedural QA/QC for the biosensor will provide a reliable tool for a near real-time monitoring of E. coli in drinking water samples to both industry and regulatory authorities.

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Date Created
2017-01-03

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Impact of Environmental Factors on Legionella Populations in Drinking Water

Description

To examine the impact of environmental factors on Legionella in drinking water distribution systems, the growth and survival of Legionella under various conditions was studied. When incubated in tap water at 4 °C, 25 °C, and 32 °C, L. pneumophila

To examine the impact of environmental factors on Legionella in drinking water distribution systems, the growth and survival of Legionella under various conditions was studied. When incubated in tap water at 4 °C, 25 °C, and 32 °C, L. pneumophila survival trends varied amongst the temperatures, with the stable populations maintained for months at 25 °C and 32 °C demonstrating that survival is possible at these temperatures for extended periods in oligotrophic conditions. After inoculating coupons of PVC, copper, brass, and cast iron, L. pneumophila colonized biofilms formed on each within days to a similar extent, with the exception of cast iron, which contained 1-log less Legionella after 90 days. L. pneumophila spiked in a model drinking water distribution system colonized the system within days. Chlorination of the system had a greater effect on biofilm-associated Legionella concentrations, with populations returning to pre-chlorination levels within six weeks. Biofilms sampled from drinking water meters collected from two areas within central Arizona were analyzed via PCR for the presence of Legionella. Occurrence in only one area indicates that environmental differences in water distribution systems may have an impact on the survival of Legionella. These results document the impact of different environmental conditions on the survival of Legionella in water.

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Date Created
2015-05-19

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The Impact of Capsid Proteins on Virus Removal and Inactivation During Water Treatment Processes

Description

This study examined the effect of the amino acid composition of protein capsids on virus inactivation using ultraviolet (UV) irradiation and titanium dioxide photocatalysis, and physical removal via enhanced coagulation using ferric chloride. Although genomic damage is likely more extensive

This study examined the effect of the amino acid composition of protein capsids on virus inactivation using ultraviolet (UV) irradiation and titanium dioxide photocatalysis, and physical removal via enhanced coagulation using ferric chloride. Although genomic damage is likely more extensive than protein damage for viruses treated using UV, proteins are still substantially degraded. All amino acids demonstrated significant correlations with UV susceptibility. The hydroxyl radicals produced during photocatalysis are considered nonspecific, but they likely cause greater overall damage to virus capsid proteins relative to the genome. Oxidizing chemicals, including hydroxyl radicals, preferentially degrade amino acids over nucleotides, and the amino acid tyrosine appears to strongly influence virus inactivation. Capsid composition did not correlate strongly to virus removal during physicochemical treatment, nor did virus size. Isoelectric point may play a role in virus removal, but additional factors are likely to contribute.

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Created

Date Created
2015-11-08

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Aerosolization Methods for Dispersion of Bacterial Cells in Air

Description

Legionella is a gram-negative bacterium with the ability for human infection by inhalation or aspiration of water containing the bacteria. Legionella live in aquatic environments and have been identified in cooling towers, humidifiers and respiratory therapy treatments, among others. Infection

Legionella is a gram-negative bacterium with the ability for human infection by inhalation or aspiration of water containing the bacteria. Legionella live in aquatic environments and have been identified in cooling towers, humidifiers and respiratory therapy treatments, among others. Infection with Legionella bacteria leads to Legionnaire’s Disease or Pontiac Fever (Edelstein, 1993). Information regarding the means of aerosolization of Legionella bacteria has not yet been reported, therefore the relevance of experimentation was defined. The objective of this study is to determine the modes by which bacteria may be aerosolized under laboratory conditions. Specifically, to measure the amount of bacteria transported over a specific distance in a given amount of time and determine the most effective mode of bacterial aerosolization. Three methods of bacterial aerosolization were tested, these included an electric paint sprayer, an air paint sprayer and a hand-held spray bottle. E. coli was used as a surrogate for Legionella in experimentation due to its similar bacterial properties. Both bacteria are gram-negative, aerobic bacilli while Legionella is approximately 2 μm in length (Botzenhart, 1998), and E. coli is between 1 and 3 μm in length (Reshes, 2007). The accessibility and non-pathogenicity of E. coli also served as factors for the substitution.
In order to measure the aerosolization efficiency of each spray method, an air sampler was placed opposite to the position of the sprayer, on either side of a sealed box. Each sprayer was filled with E. coli concentrated at 104 CFU/ml in a PBS solution and sprayed for a time span of 1 and 5 seconds. For each of these time intervals an air sample was collected immediately following the spray as well as 5 minutes after the spray. Compared to the other two methods, the air spray method consistently showed the highest number of bacterial cells aerosolized. While all three methods resulted in the aerosolization of bacteria, the results determined the Air Spray method as the most efficient means of bacterial aerosolization. In this study, we provide a practical and efficient method of bacterial aerosolization for microbial dispersion in air. The suggested method can be used in future research for microbial dispersion and transmission studies.
In addition, a humidifier was filled with a spiked solution of E. coli and operated for a period of 1 and 5 seconds at its maximum output. Air samples were collected after 0 and 5 minutes. Immediately after the humidifier operation was stopped a small number of colonies were detected in the air sample and no colonies were detected in the air sample collected after a 5-minute elapsed time. This experiment served as a proof of concept for airborne pathogen’s transmission by a humidifier.

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Created

Date Created
2015-12

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Physical Water Treatment Effects on Evaporation Rates, Hardness, and Microbial Life

Description

The rising need for water reuse in the Southwest United States has increased awareness of the quality of wastewater. This need is caused by an increased population having basic water needs; inefficient water use, such as overwatering lawns and leaking

The rising need for water reuse in the Southwest United States has increased awareness of the quality of wastewater. This need is caused by an increased population having basic water needs; inefficient water use, such as overwatering lawns and leaking pipes; and recent drought conditions all over the southwestern US. Reclaimed water is a possible solution. It's used for a variety of non-potable, or non-drinkable, reasons. These uses include: cooling power plants, concrete mixing, artificial lakes, and irrigation for public parks and golf courts. Cooling power plants utilizes roughly 41% of the total water consumed by the United States, which makes it the highest user of water in the US. The attention is turned to optimizing mechanical processes and reducing the amount of water consumed. Wet-recirculating systems reuse cooling water in a second cycle rather than discharging it immediately. Cooling towers are commonly used to expose water to ambient air. As the water evaporates, more water is withdrawn while the rest continues to circulate. These systems have much lower water withdrawals than once-through systems, but have higher water consumption. The cooling towers in wet-recirculating plants and other warm machinery have two major limitations: evaporation of pumped water and scale formation in the components. Cooling towers circulate water, and only draw as it evaporates, which conserves water. The scale formation in the components is due to the hardness of the water. Scale occurs when hard water evaporates and forms solid calcium carbonate. This formation can lead to reduced flow or even clogging in pipes, fouling of components or pipes, and reduced cooling efficiency. Another concern from the public over the use of reclaimed water is the possibility of there being fecal contamination. This fear stems from the stigma associated with drinking water that essentially came from the toilet. An emerging technology, in order to address these three issues, is the use of an electromagnetic device. The wires have a current flowing through which induces a magnetic field perpendicular to the direction of the flow, while the electrical field is proportional to the flow velocity. In other words, the magnetic and electrical fields will create an effect that will concentrate cations at the center of the pipe and anions at the wall of the pipe or the other way depending on the direction of the flow. Reversing the field will then cause the cations and anions to move toward one another and increase the collision frequency and energy. The purpose of these experiments is to test the effects of the electromagnetic device on the aforementioned topics. There are three tests that were performed, a surface tension test, a hardness test, and a microbial test. The surface tension test focused on the angle of a water droplet until it burst. The angle would theoretically decrease as the bond between water molecules increased due to the device. The results of this test shows a lower angle for the treated water but a higher angle for the untreated one. This means the device had an effect on the surface tension of the water. Hard water is caused by calcium and magnesium ions in the water. These ions are dissolved in the water as it travels past soil and rocks. The purpose of this test is to measure the free calcium ion amount in the water. If the free calcium number lowers, then it can be assumed it collided with the carbonate and formed calcium carbonate. This calcium carbonate causes a reduction in hardness in the water. The result of the test showed no correlation between ion concentrations in the treated/untreated system. The e. coli test focused on testing the effects of an electromagnetic device on inhibiting fecal contamination in water/wastewater at a treatment facility. In order to detect fecal contamination, we test for bacteria known as fecal coliforms, more specifically e. coli. The test involved spiking the system with bacteria and testing its concentrations after time had passed.The e. coli results showed no trend in the inactivation of the bacteria. In conclusion, the device had varying results, but multiple steps can be taken in the future in order to continue research.

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Created

Date Created
2014-12

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Impact of copper nanoparticles on inactivation and toxicity pathway on model bacteria

Description

Nanotechnology is a scientific field that has recently expanded due to its applications in pharmaceutical and personal care products, industry and agriculture. As result of this unprecedented growth, nanoparticles (NPs) have become a significant environmental contaminant, with potential to impact

Nanotechnology is a scientific field that has recently expanded due to its applications in pharmaceutical and personal care products, industry and agriculture. As result of this unprecedented growth, nanoparticles (NPs) have become a significant environmental contaminant, with potential to impact various forms of life in environment. Metal nanoparticles (mNPs) exhibit unique properties such as increased chemical reactivity due to high specific surface area to volume ratios. Bacteria play a major role in many natural and engineered biogeochemical reactions in wastewater treatment plants and other environmental compartments. I have evaluated the laboratory isolates of E. coli, Bacillus, Alcaligenes, Pseudomonas; wastewater isolates of E. coli and Bacillus; and pathogenic isolate of E. coli for their response to 50 & 100 nm sized Cu nanoparticles (CuNPs). Bactericidal tests, scanning electron microscopy (SEM) analyses, and probable toxicity pathways assays were performed. The results indicate that under continuous mixing conditions, CuNPs are effective in inactivation of the selected bacterial isolates. In general, exposure to CuNPs resulted in 4 to >6 log reduction in bacterial population within 2 hours. Based on the GR, LDH and MTT assays, bacterial cells showed different toxicity elicitation pathways after exposure to CuNPs. Therefore, it can be concluded that the laboratory isolates are good candidates for predicting the behavior of environmental isolates exposed to CuNPs. Also, high inactivation values recorded in this study suggest that the presence of CuNPs in different environmental compartments may have an impact on pollutants attenuation and wastewater biological treatment processes. These results point towards the need for an in depth investigation of the impact of NPs on the biological processes; and long-term effect of high load of NPs on the stability of aquatic and terrestrial ecologies.

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Created

Date Created
2012

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Multivariable Analysis for Irrigation with Gray Water, Impact of Turbidity and Organic Content in Gray Water on Bacterial Inactivation

Description

The impact of physical/chemical properties of gray water on microbial inactivation in gray water using chlorine was investigated through creating artificial gray water in lab, varying specific components, and then measuring microbial inactivation. Gray water was made through taking autoclaved

The impact of physical/chemical properties of gray water on microbial inactivation in gray water using chlorine was investigated through creating artificial gray water in lab, varying specific components, and then measuring microbial inactivation. Gray water was made through taking autoclaved nanopure water, and increasing the concentration of surfacants, the turbidity, the concentration of organic content, and spiking E. coli grown in tryptic soy broth (TSB); chlorine was introduced using Clorox Disinfecting Bleach2. Bacteria was detected using tryptic soy agar (TSA), and E. coli was specifically detected using the selective media, brilliance. The log inactivation of bacteria detected using TSA was shown to be inversely related to the turbidity of the solution. Complete inactivation of E. coli concentrations between 104-105 CFU/100 ml in gray water with turbidities between 10-100 NTU, 0.1-0.5 mg/L of humic acid, and 0.1 ml of Dawn Ultra, was shown to occur, as detected by brilliance, at chlorine concentrations of 1-2 mg/L within 30 seconds. These result in concentration time (CT) values between 0.5-1 mg/L·min. Under the same gray water conditions, and an E. coli concentration of 104 CFU/100 ml and a chlorine concentration of 0.01 mg/L, complete inactivation was shown to occur in all trials within two minutes. These result in CT values ranging from 0.005 to 0.02. The turbidity and humic acid concentration were shown to be inversely related to the log inactivation and directly related to the CT value. This study shows that chlorination is a valid method of treatment of gray water for certain irrigation reuses.

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Date Created
2020-05

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Impact of Turbidity on the UV Inactivation of Escherichia coli

Description

Turbidity is a known problem for UV water treatment systems as suspended particles can shield contaminants from the UV radiation. UV systems that utilize a reflective radiation chamber may be able to decrease the impact of turbidity on the efficacy

Turbidity is a known problem for UV water treatment systems as suspended particles can shield contaminants from the UV radiation. UV systems that utilize a reflective radiation chamber may be able to decrease the impact of turbidity on the efficacy of the system. The purpose of this study was to determine how kaolin clay and gram flour turbidity affects inactivation of Escherichia coli (E. coli) when using a UV system with a reflective chamber. Both sources of turbidity were shown to reduce the inactivation of E. coli with increasing concentrations. Overall, it was shown that increasing kaolin clay turbidity had a consistent effect on reducing UV inactivation across UV doses. Log inactivation was reduced by 1.48 log for the low UV dose and it was reduced by at least 1.31 log for the low UV dose. Gram flour had a similar effect to the clay at the lower UV dose, reducing log inactivation by 1.58 log. At the high UV dose, there was no change in UV inactivation with an increase in turbidity. In conclusion, turbidity has a significant impact on the efficacy of UV disinfection. Therefore, removing turbidity from water is an essential process to enhance UV efficiency for the disinfection of microbial pathogens.

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Date Created
2020-05

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Biosensor platform for rapid detection of E. coli in drinking water

Description

The need for rapid, specific and sensitive assays that provide a detection of bacterial indicators are important for monitoring water quality. Rapid detection using biosensor is a novel approach for microbiological testing applications. Besides, validation of rapid methods

The need for rapid, specific and sensitive assays that provide a detection of bacterial indicators are important for monitoring water quality. Rapid detection using biosensor is a novel approach for microbiological testing applications. Besides, validation of rapid methods is an obstacle in adoption of such new bio-sensing technologies. In this study, the strategy developed is based on using the compound 4-methylumbelliferyl glucuronide (MUG), which is hydrolyzed rapidly by the action of E. coli β-D-glucuronidase (GUD) enzyme to yield a fluorogenic product that can be quantified and directly related to the number of E. coli cells present in water samples. The detection time required for the biosensor response ranged from 30 to 120 minutes, depending on the number of bacteria. The specificity of the MUG based biosensor platform assay for the detection of E. coli was examined by pure cultures of non-target bacterial genera and also non-target substrates. GUD activity was found to be specific for E. coli and no such enzymatic activity was detected in other species. Moreover, the sensitivity of rapid enzymatic assays was investigated and repeatedly determined to be less than 10 E. coli cells per reaction vial concentrated from 100 mL of water samples. The applicability of the method was tested by performing fluorescence assays under pure and mixed bacterial flora in environmental samples. In addition, the procedural QA/QC for routine monitoring of drinking water samples have been validated by comparing the performance of the biosensor platform for the detection of E. coli and culture-based standard techniques such as Membrane Filtration (MF). The results of this study indicated that the fluorescence signals generated in samples using specific substrate molecules can be utilized to develop a bio-sensing platform for the detection of E. coli in drinking water. The procedural QA/QC of the biosensor will provide both industry and regulatory authorities a useful tool for near real-time monitoring of E. coli in drinking water samples. Furthermore, this system can be applied independently or in conjunction with other methods as a part of an array of biochemical assays in order to reliably detect E. coli in water.

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Created

Date Created
2015

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Antibiotics as environmental pollutants: associated public health threats and residues in animal protein and biosolids

Description

This dissertation studies the larger issue of antibiotic resistance with respect to how antibiotics are being introduced into the environment, focusing on two major anthropogenic pathways: animal husbandry for human consumption, and the recycling of wastewater and municipal sludge generated

This dissertation studies the larger issue of antibiotic resistance with respect to how antibiotics are being introduced into the environment, focusing on two major anthropogenic pathways: animal husbandry for human consumption, and the recycling of wastewater and municipal sludge generated during conventional biological sewage treatment.

For animal production on land (agriculture) antibiotics are often used for growth enhancement and increased feed efficiency. For animal production in water (aquaculture) antibiotics are often used as a prophylactic. I found that the same antibiotics are being used in both industries and that the same strains of human pathogens have also been isolated from both sources, expressing identical resistance mechanisms. In U.S. seafood, five out of 47 antibiotics screened for were detected at levels of 0.3 to 7.7 ng/g fresh weight. Although compliant with FDA regulations, the risk for resistance still exists, as even low antibiotic concentrations have been shown to exert selective pressure on bacteria.

Similarly low concentrations of antibiotics were found in U.S. biosolids at levels of 0.6 to 19.1 ng/g dry weight. Of the five antibiotics detected, two have never been reported before in biosolids. Three have never been reported before in U.S. biosolids. Using the raw numbers obtained from antibiotic screenings in biosolids, I assessed the impact of employing four different LC-MS/MS methods, concluding that analysts should experimentally determine the most appropriate quantitation method based on the analyte targeted, matrix investigated, and research goals pursued. Preferred quantitation approaches included the isotope dilution method with use of an analogous standard and, although time and resource demanding, the method of standard addition.

In conclusion, antibiotics introduced into the environment via agriculture, aquaculture, and wastewater recycling pose a combination of chemical and biological threats. Aside from exerting outright chemical toxicity to non-target organisms, antibiotic residues can promote the development of multi-drug resistance in human pathogens. Public health protection approaches to stem the risks posed by animal husbandry may include reserving drugs for exclusive, human use, decreasing their usage altogether, improving reporting efforts, reevaluating existing regulations on agricultural and aquacultural antibiotic usage, and improved risk assessment for biosolids application on land.

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Created

Date Created
2015