RIVER MONITOR MANUAL

Transcription

1 JAMES RIVER WATCH RIVER MONITOR MANUAL With all issues and questions, please contact: Pat Calvert, Upper James RIVERKEEPER Text or Voice: (434) Jamie Brunkow, Lower James RIVERKEEPER Text or Voice: (757) RiverRats: James River s first line of citizen protection. James River Watch Citizen Monitoring Program

2 Standard Operating Procedures 1.0 Introduction The Standard Operating Procedures (SOP) outlined in this manual are designed to help guide and sustain a high quality water monitoring program. The procedures are utilized by the James River Association (JRA) and volunteers to assure that each individual or sampling crew follow the same protocols. Quality assurance in a monitoring program is essential for providing representative and accurate data for a water body, which is vital for the continuation of a monitoring program. The SOP s are listed by section, with added instructions for safety and tips on keeping up proper maintenance of testing equipment. All volunteers are to follow these guidelines and procedures to ensure reliable, high quality data. JRA uses the SOP as a tool for training new volunteers at introductory training classes, and during coordinated periodic training sessions. The SOP also serves as a tool for staff and volunteers to routinely reference during sampling activities. 2.0 Safety Safety to staff and volunteers is of the upmost importance to the James River Association. There are hazards in both the field and the lab, so please use caution. For instance, during inclement weather conditions, the sampling site may become a hazard due to wet and/or slippery conditions, high winds, or torrential water that can be dangerous to the individuals sampling. It is also recommended that more than one person be on site in case of an emergency. If at any time a volunteer feels that unsafe conditions exist, they are advised to terminate that activity immediately. The tests that are performed on water samples require the use of materials that must be handled appropriately. Follow basic handling procedures such as washing hands before and after use, using appropriate cleaning materials for spills, disposing of wastes properly and supervising others that may come in close contact with petri dishes or test kits. While sampling, JRA recommends using gloves or having alcohol sanitizer available to keep hands clean. There is always an inherent risk of harmful microorganisms being present in the river. 3.0 Testing Time Frame The time frame for collecting water samples and performing tests is important for maintaining a quality monitoring program. When samples are collected from all sites during the same time period, it allows for the comparability of data across the watershed. Clearly defined schedules are also crucial in maintaining coordination between JRA and volunteers. Volunteers will collect samples between 10:00 AM and 2:00 PM, on each Thursday, from May 30 to August weekends. Collection of samples on Thursdays will allow ample time for plates to be incubated with results ready by Friday afternoon allowing results to be posted before each weekend when 3

3 recreational river use is highest. Each sampling site is assigned a team of volunteers responsible for monitoring activities. Team members should coordinate schedules and communication to ensure samples are collected on time each week. Volunteers must receive prior approval from JRA to sample outside of the sampling window, or should contact JRA for help finding an alternate volunteer for any particular week. 4.0 Sampling Locations Prior thought has already been used in locating each monitoring site, and volunteers must sample only at JRA s predefined sites, listed below in Table 1. Site Number Location Latitude Longitude J05: Jamestown Beach James City County A01: City Point City of Hopewell A02: Pocahontas Canoe Launch City of Petersburg J10: 14 th Street Public Landing City of Richmond J15: Pipeline Rapids City of Richmond J20: Rope Swing at Tredegar City of Richmond J25: Tucker Park in Maidens Goochland County J30: Scottsville at DGIF Landing Albemarle County J35:Riveredge Park public boat landing J40: Buchanan at DGIF public boat landing Amherst County Botetourt County Table 1. JRA Sampling Locations. 5.0 Sampling Procedures JRA sampling procedures are divided into two types of activities below: field collection and lab testing. This section will cover procedures that are necessary to provide accurate, high quality data that represents the body of water being sampled. Training will be provided by JRA prior to volunteers taking on a monitoring site, but volunteers may request further training at any time. The water samples collected will be used to quantify the level of contamination or loading of pollutants (e.g., E. coli), to the James River and its tributaries. Therefore, it is imperative that samples are collected following protocols to accurately represent the body of water being sampled. 4

4 5.1 Sampling Materials Field Kit (gray tool box) Sterile Plastic Sample Bottles (at least one per site) Thermometer Turbidity sample bottle Nitrile Gloves Sharpie marker Pencil Hand sanitizer Roll of labeling tape Other Field Materials Cooler Frozen ice pack Sampling pole Field instructions (laminated cards) Copy of field data sheet Lab Materials Incubator with thermometer Sterile Petri dishes Sterile Pipettes Coliscan media Turbidity meter o Calibration blank samples (contained in turbidity case) o 5mL glass sample vials (contained in turbidity case) Kimwipes (for cleaning glass turbidity vials) SOP manual and/or laminated instructions cards Tape (to seal plates during incubation) 5.2 Prior to Sampling 1. Ensure field kit is complete and contains properly functioning materials. Contact JRA to make arrangements for any replacement materials in advance of sampling. 2. Prepare a cooler with a frozen ice pack, in order to keep water samples cool. 3. Label bacteria sample bottle (one per site) with site name. 4. Dress appropriately for sampling activities, monitor weather and river levels to make certain conditions are safe for sampling. 5. Allow approximately 30 minutes for Coliscan media to thaw at room temperature. Media should be in liquid form before water samples are added and plated. Do not shake media bottles in order to speed thawing, as this may cause air bubbles to appear in the plated sample. 6. Allow time for incubator to warm to 35 o -40 o C. This typically requires 30 min. 5

5 5.3 Field Sampling Method At each sampling site, volunteers will measure air temperature, water temperature, and will collect two water samples (one sample for turbidity and one for E. coli bacteria). General site observations will also be recorded. Sampling poles are supplied with each kit, which allow volunteers to collect samples from the shore. In order to maintain high quality data, volunteers must follow the best practices outlined below during sampling activities. Collecting a representative water sample. If possible, look for a part of the river that is flowing. Avoid sampling behind an obstruction in the water column, or in a non-flowing or stagnant area. Always face upstream when collecting samples. If wading into the river to collect your sample, avoid disturbance of the stream bed, which could lead to inaccurate readings. Fill sample bottles to the shoulder, leaving an air gap for mixing later in the lab. The Rule of Three. The rule of three is simply a process that one should use in the field, in the lab, during clean-up, and when preparing a sample for testing. In the field, always rinse out the turbidity water sample bottles with water from the site. Do this three times before finally filling the bottle a fourth time, capping, and storing for transport. This process will ensure that carry over from the previous sample will have no effect on the new sample. The rule of three is not required for collecting bacteria samples, since bacteria sample bottles have been sterilized. Step 1. Temperature Measurement. Air Temperature Measure air temperature in an area that is out of direct sunlight. If possible you may suspend the thermometer in the shade from a nearby tree or structure. Allow at least 2 minutes to pass before reading thermometer, and record air temperature to the nearest one-tenth degree Celsius on the data log sheet. Water Temperature Measure water temperature using the same thermometer used for air temperature. You may measure temperature in the water source itself by setting the thermometer in the water s edge. Alternatively, you may measure the temperature of your turbidity water sample immediately following sample collection. Do not measure water temperature of the bacteria sample, as this may contaminate the sample. Again, allow at least 2 minutes for the thermometer to adjust, and record water temperature to the nearest one-tenth degree Celsius on the data log sheet. Step 2. Turbidity Sample Collection. Locate the plastic bottle labeled turbidity sample bottle in field kit (gray toolbox). This bottle is reused each week of sampling. Securely attach bottle to sampling pole, fill bottle to shoulder using the Rule of Three. Cap the bottle, remove from sampling pole, and store for transport. It is not necessary to keep the turbidity sample on ice. 6

6 Step 3. Bacteria Sample Collection. Locate sterile plastic bacteria sample bottle, and ensure bottle is labeled with site name. Securely attach bottle to sampling pole, remove cap, and fill bottle to shoulder. Cap the bottle, remove from sampling pole, and store in cooler for transport. Avoid touching the inside of the bottle cap and the mouth of the bottle. Step 4. Other Site Data and Observations. Record basic site information on the data log sheet (e.g., volunteer names, site, time, date, etc.). Use the data log sheet as a reference in the field to ensure all information is collected. Volunteers that have access to a digital camera or smartphone may take a picture of the site to submit later with the monitoring report. Observations Section: Make any general observations note things such as water level, recent rain activity, weather conditions. Look for life and signs of environmental changes: e.g. disturbances, erosion, ducks, geese, sea nettles, crabs, minnows, etc. Give numbers or estimates when possible. 6.0 Lab Testing Procedures 6.1 General Lab Testing Practices 1. Indicate on the Data Log (Appendix A.) the time & date of sample, sampler namess and any activity (e.g. sample collection, sample analysis). Make sure log sheets are completely filled out, and kept with the sampling kit. 2. Make sure all lab equipment and instrumentation is clean, unstained and dry from prior testing cycles (e.g. turbidity vials cleaned and dried from prior use). 3. Make sure work surface is clean and free of clutter. 4. If using JRA s office space in Richmond, Williamsburg or Lynchburg for lab testing, contact staff ahead of time to make arrangements for office access. 6.2 Turbidity Lab Method Calibration (to be performed each week that the turbidity meter is used) 1. Turn on turbidity meter; follow menu prompts and select Measure, then press ENTER, select Turbidity With Blank, then press ENTER. 2. Turbidity is measured by light penetration through the sample, and results can be skewed by smudges or stray fingerprints left on a vial. Before inserting any vials into the turbidity meter, wipe clean with Kimwipe to remove any moisture or fingerprints. 3. Insert vial labeled blank into the meter, with white vertical line on vial positioned to face user. Close turbidity meter cap, and select Scan Blank. Push ENTER. 4. After a few seconds, once unit has completed blank calibration, remove blank vial. 5. Insert the vial containing 10 NTU standard, again with white vertical line on the vial positioned to face user and after again wiping with Kimwipe. Close turbidity meter cap, select Scan Sample and press ENTER. 7

7 6. If the reported turbidity value equals the standard value of 10 NTU, calibration is complete and you may move on to the steps below titled Turbidity Measurement. 7. If the reported value does not equal the standard value of 10 NTU, press the down arrow key to highlight Calibrate, then press ENTER. Use the arrow keys to adjust the value to NTU, then press ENTER, then Set Calibration will be highlighted and you must press ENTER one more time to complete calibration. Turbidity Measurement 1. Gently shake turbidity sample to mix thoroughly. 2. Locate an empty vial in the turbidity meter case, and fill to white horizontal line. Use the Rule of Three in filling the vial. 3. Turn on turbidity meter; follow menu prompts and select Measure then Turbidity With Blank. 4. Turbidity is measured by light penetration through the sample, and results can be skewed by smudges or stray fingerprints left on a vial. Before inserting any vials into the turbidity meter, wipe clean with Kimwipes to remove any moisture, or fingerprints. 5. Insert vial labeled blank into the meter, with white vertical line on vial positioned to face user. Close turbidity meter cap, and select Scan Blank Press ENTER. 6. After a few seconds, once unit has completed blank calibration, remove blank vial. 7. Insert the vial containing river water, again with white vertical line on vial positioned to face user. Close turbidity meter cap, select Scan Sample and press ENTER. 8. Record reported turbidity value on data log sheet. 9. Rinse used vial(s) with distilled water, again using the Rule of Three, dry using Kimwipe, and store in turbidity meter case. 6.3 Bacteria Lab Method Step 1. Sample Plating. 1. Label the bottom (smaller, taller piece) of the Petri dish using a permanent marker. It is best to label the dishes using small lettering on the outer rim of the dish. The minimum information needed should be the site ID number, sample volume, and replicate number (if needed). 2. Mix the water sample in the sterile bottle and then transfer exactly 3.0 ml of the river water sample to a bottle of Coliscan medium using a sterile pipette. 3. Gently swirl the bottle of Coliscan media so that it mixes with the sample water. Do not shake the bottle as this will cause the medium to foam and make reading the colonies difficult. 4. Pour the entire contents of the bottle into a Petri dish. It is important to perform this step on a level surface so the solution forms an even layer across the plate. 5. Allow the solution to solidify (approximately 30 minutes) prior to incubation. For safety purposes, it is a good idea to loosely tape each Petri dish shut after the media solidifies. Step 2. Incubation. Incubate the Petri dishes upside down for approx. 24 hours at 35 o - 40 o Celsius. This is approximately 95 o o F. JRA has supplied incubators with each sampling kit, but if no incubator is available for some reason, place the Petri dishes in the safest warmest spot you can find. Depending 8

8 on the exact temperature, the plates may need to be incubated hours prior to analysis. Step 3. Data Analysis. It is recommended to use white or graph paper as a background to make identifications easier. If there is a large number of colonies, drawing quadrants on this paper can help in counting the number of colonies. 1. Count the number of dark blue to royal purple colonies on each plate and record this number on the data log sheet. Do not count teal colored or pink to dark red colonies. If you count more than 60 colonies, you can stop and report the value as greater than > Calculate the number of E. coli cells per 100 milliliters and record on the data form. Use the following formula: (total # E. coli colonies/ml sample size) x 100 Figure 1. Petri dish after incubation. The green circled colonies are E. coli and the yellow circled ones are very likely to be E. coli based on the photo. The red circled are not E. coli. 9

9 Step 4. Safe Disposal of Wastes 1. Used pipettes and sample bottles may be disposed of in household trash. These items are also recyclable, and may be returned to JRA for recycling. 2. Rinse empty bottles of Coliscan medium one to three times with tap water and dispose in your household trash. (This is to wash out all of the media to prevent pathogens from growing.) 3. Wipe down the area where you poured the media into the Petri dishes with cleaning agent to kill any bacteria from the sample bottles. It is recommended to not perform this test where food is present or prepared. 4. After the results have been recorded, add just enough bleach or rubbing alcohol to each Petri dish to completely cover the solid media. Allow to stand for at least 10 minutes to ensure all bacteria have been killed. Plates can then be disposed in household trash. 6.4 Bacteria Quality Control (QC) Samples Quality Control protocols are an integral part of all analytical procedures to ensure that results are reliable. Volunteers will be asked to test additional split samples on alternating weeks throughout the sampling season from May-September. Comparison of these split samples helps JRA to rate the precision of our sampling methods, and to evaluate the technique of all staff and volunteers. See Appendix B of the SOP for the schedule of QC samples by sampling team/site. 7.0 Data Recording 1. All volunteers are required to record data on the provided data log sheets (Appendix A.). Data sheets from each week must be kept within JRA supplied notebooks that stay with sampling kits at all times. 2. Data and photographs must also be uploaded to JRA s James River Watch (JRW) website, on each Friday and immediately following analysis of bacteria results. A smartphone may be utilized to upload data & photos. 3. Each volunteer is provided a username (i.e. your registered address) and password to access JRW. The web link to upload data is: 4. Questions or errors experienced with the JRW website should be forwarded to JRA staff promptly, to ensure issues are resolved and data is added each week in a timely manner. Thank You Your volunteerism for James River Association is tremendously appreciated. We at JRA hope you enjoy your active role in monitoring the James and in helping to provide a detailed record and reference of water quality. You are helping a great deal by providing us with the tools to identify and track sources of pollutants in the watershed. Your James RIVERKEEPERS 10