SRB Project – Sulfite Reducing Bacteria Primary Chilled Water Loop Product: WBS (Water Bacteria Solution) Location: Major Southwest Texas University Cisne Enterprises, Inc. Purely Elemental 1
Mr. Assistant Director Assistant Director Plant Operations/HVAC Nov 2.2009 Reference: SRB Contract ________Campus Mr. The treatment program was begun on October the 16th 2009. Injection points were selected on the University campus: Kelly Hall, Miners Village and Satellite Plant. The method of monitoring has been by ATP. ATP testing provides a rapid method for the detection of all living cells in the water as well as a method for determination of the biocide and bio-dispersant efficacy in a very short time period. A short time period, being the area of focus at this point. Biological plate counts are very good. They however do require on the average of two weeks for determination, due to the fact that the Bacterial Colony Forming Unit counts need to reach eye level determination before the result can be known. The method of determination by ATP is fifteen seconds and results are as real time biological monitoring. ATP analysis offers several advantages as an assay for active biomass: It can be performed in a short period of time, and it has a sensitive limit of detection. ATP is an energy cell molecule. ATP (adenosine triphosphate) A nucleotide that is of fundamental importance as a carrier of chemical energy in all living organisms. It consists of adenine linked to D-ribose (i.e. adenosine); the D-ribose component bears three phosphate groups, linearly linked together by covalent bonds. These bonds can undergo hydrolysis to yield either a molecule of ADP (adenosine diphosphate) and inorganic phosphate or a molecule of AMP. Both these reactions yield a large amount of energy (about 30.6 kJ mol –1 ) that is used to power the living cell with resultant ATP determination. The standard PLATE TECHNIQUE FOR BACTERIAL ENUMERATION will be used as the program progresses. Short time SRB auger methods are being used as a rapid detection method to give an assessment of total SRB colonies after the first chemical charge has been accomplished. An initial injection is in progress to establish our treatment objectivities. Five initial monitoring locations have been selected as control points, Kelly Hall-Miners Village- Central Plant-Satellite Plant-Main Thermal Water Storage Tank are the locations established. Initial ATP in energy source units Fmol before program start 10/15/09 Date 10/13/09 2
Location Central Plant Satellite Plant Miners Village Kelly Hall Main Storage Tank ATP fmol 688 287 285 206 186 SRB > 10-3 < 10-2 > 10-3 > 10-4 > 10-3 Location: Central Plant Satellite Plant Miners Village Kelly Hall Main Storage Tank Program Start Date 10/16/09 Chemical Injection start up. Reading on 10/28/09 ATP 12 10 8 7 16 SRB < 10-3 < 10-1 10-3 < 10-3 > 10-3 Guidelines SRB 10-3 CFU = 1000 colonies ATP Less than 20 is considered very good for aerobic and non aerobic bacteria bio mass this may not include all SRB or NRB types of bacteria, rapid test cultures are used for quick determination. The next phase of monitoring will be a more definitive quantification for SRB CFU, conjunctive with ATP fmol energy units. Injection is scheduled to begin directly into the thermal storage tank. This will be followed by a system drain down and recharge of biocide into the systems. The units will be tested for biomass and direct injection will start on a localized basis in units found to be contaminated. Additional recirculation of some units may be required, so as to remove larger biomass concentrations should they be found. Definitive reports will be forthcoming, it would be somewhat misleading to list reduction percentages and or pass/fail locations at this time, based upon a two week start up time. As more information becomes collected, it will offer an objective evaluation of the progress being made. Thank You 3
Summary Primary Chilled Water Loop The Problem : Infected water with SRB Reason : Contaminated water in the chilled water loop. Time Line: Step #1: Injection Time 14 days 2 week Treatment Two Step #2: Application Time 60 days Months Application Time Step # 3: Maintenance Flush 30 days Monthly System Cleaning Step # 4: Shock Time 365 days One Year Projected efficacy time to disinfect the water in Chilled Water Conclusion : Diminish the SRB problem in the system, reducing the overall repair and replacement problems to HVAC department. 4
Primary Chilled Water Loop Is Something in Your Water ? THE PROBLEM is that the chilled water loop has developed Sulfite Reducing Bacteria (SRB) problem. This problem was determined with field testing and an in-depth analysis by an independent metallurgy laboratory, specializing in these types of problems, made it possible for us to concentrate in a more refined area of the problem. This project outline comes as a result of those findings. Chemical selection for this type of problem becomes complex. Primarily in regard to the environmental issues involved with the treatment of four million gallons of water at a major university campus. The purpose of the prolonged treatment exposure time is to avoid the protein excretion from the bacteria. When a quick rupture of the cell membrane wall occurs the cell attempts to avoid intrusion via this excretion method, further compounding the problem. By applying a differential charge across the cell wall it will slowly lose its permeability without the protective filming, which might otherwise occur from the bacteria. This is accomplished by the highly cationic charge, imparted by the treatment process, allowing the unit to lose the nutrients necessary to sustain life. The bacteria have been present a long time and have the ability to live and thrive under many and varied conditions. Our job is to “out smart” the cell during the treatment process in order to have successful elimination. The surfactant properties of the material being used will assist us in maintaining the fluid of the dead cells. This maintains the cells in the water phase, allowing removal through the system bleed. Outlined below is the method that will be used for biological growth, Adenosine Triphosphate (ATP). The use of firefly luciferase to assay adenosine triphosphate (ATP) extracted from microorganisms provides an easy means to enumerate microbes within minutes. The small amount of light produced is proportional to ATP and thus microbial number. The average bacterium contains around 10(-15) g ATP per cell. Present reagents permit detection of 10(3) cells per tube. Luminometers currently on the market detect about 10(-12) g ATP. Proper extraction of ATP from the microbes is an essential part of any protocol, as is the removal of non-microbial ATP from, for example, somatic cells also present in samples. The technique may be applied to a wide range of samples, for example food and beverages and clinical samples such as urine. The ATP assay gives a global measure of microbial numbers. This test is a cell biomarker and is considered the Gold standard in Bio testing. By applying the treatment as outlined and using ATP, the problem can be controlled, eliminating future issues and concerns. 5
Primary Chilled Water Loop Example of SRB Determination of Potential SRB Population - observe daily for reaction. Date of Test: 01-12-09 thru 02-12-09 / Sample taken from: Kelly Hall / Chemical: WBS SRB Test showing (+) Positive. Treated sample The treatment being used will rupture the cell wall of the bacteria and accelerate the loss of cell nutrients resulting in the extinction of the cell. For this to occur, some resident contact time will be required as the chemical imparts a cationic charge to the cell wall, resulting in the parting of the protein structure and allowing for the loss of the required food sourcing occurring. Based on laboratory observations, this treatment will be effective for the systems considered for treatment. Page 4 of 9 6
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