Appendix A Standard Operating Procedures - University of Rhode ...

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Standard Operating Procedure 008 (Prior number URIWW-SOP-4B) ISDS Microbiological Procedure University of Rhode Island Watershed Watch 1.0 PURPOSE AND DESCRIPTION Date: Revision: Author: 08/08 3 Elizabeth Herron The purpose of this method is to determine the concentration of fecal coliform bacteria within an Individual Sewage Disposal System (ISDS) sample. Samples are collected in sterile bottles, an aliquot of sample is filtered and the resulting filter placed into a media filled Petri dish and incubated. After incubation, the number of bacteria colonies are then counted, and the concentration calculated. This procedure is utilized for ISDS samples only. This method is applicable to undiluted samples in the range of
handling these chemicals. Material Safety Data Sheets (MSDS) are located in each laboratory for the materials stored in the specific laboratory. MSDS are contained in plastic file folders in rooms 019 and 018 and a notebook in room 002. 2.2 Technician Training/Qualifications General training in laboratory technique, use of an autoclave and sterile technique as well as specific training regarding procedures specific to this method must be completed prior to analyzing samples using this method. Technician training will be provided either by Elizabeth Herron (Laboratory Project Manager – Microbiology) or Linda Green (Laboratory Project Manager – Nutrients). 3.0 REQUIRED MATERIALS Required Material Notes Re-order information Coastal Institute in Kingston, room 081 Autoclave and maximum temperature thermometer University of Rhode Island (URI), Kingston Coastal Institute Watershed Watch (WW) Laboratory, room 002 Autoclave safe white plastic sample bottles 125 – 500 mL Autoclave safe flasks Autoclave gloves (Insulated) Metal autoclave tray Combination hot plate and magnetic stirrer Magnetic stirring bar 2 L Sodium Hydroxide (NaOH) URI – WW Computer Caustic. Causes eye, skin, digestive and respiratory tract burns. Hygroscopic (absorbs moisture from the air). Microbiology worksheets Project data sheets are found on the URIWW computer. See Section 8.0 Documentation Coastal Institute in Kingston, room 018 Balance Calibration weights in drawer beneath the balance Coastal Institute in Kingston, room 019 35 C Incubator Thermolyne Type 142300 Incubator Thomas catalog #6118L50 44.5 C Water bath Precision Fecal coliform bath 66855 Baxter catalog #W3182-2 Petri dish rack for water bath UV Sterilization box Vacuum pump and manifold Baxter catalog #W3182-11 2 of 22 ISDS Microbiological Analysis SOP 008 S:\WW\awwword\LABPROC\all QAPPs\LABQAPPs\QAPP Rev5 -0609\SOPs\SOP 008 ISDSMicrobes.doc
Page 1 and 2: Appendix A Standard Operating Proce
Page 3 and 4: 4. Report ANY accidents IMMEDIATELY
Page 5 and 6: SUBJECT: Hazardous Materials and Ch
Page 7 and 8: 2.0 METHOD USED TO OBTAIN ULTRAPURE
Page 9 and 10: Standard Operating Procedure 003 Ge
Page 11 and 12: 3.2.2 Full Description of Labware C
Page 13 and 14: Standard Operating Procedure 004 Ge
Page 15 and 16: 4. Open the door and remove the loa
Page 17 and 18: Standard Operating Procedure 005 Bo
Page 19 and 20: 3 of 3 Autoclave Use / Maintenance
Page 21 and 22: 7. Close the door, and lock it by t
Page 23 and 24: Standard Operating Procedure 007 (P
Page 25 and 26: Several chemicals are utilized in t
Page 27 and 28: Required Material Notes Re-order in
Page 29 and 30: 5.2 Quality Assurance/Quality Contr
Page 31 and 32: mixed it will be filtered and treat
Page 33 and 34: Procedure All equipment to come int
Page 35 and 36: 5. After completion of the autoclav
Page 37 and 38: 5. Label at least one plate from ea
Page 39 and 40: 18. After 2 hours in the incubator
Page 41 and 42: 7.0 REFERENCES APHA, AWWA & WEF. St
Page 43 and 44: Dil. (mLs): The volume of sample fi
Page 45 and 46: Bacterial Sample Log & Worksheet: B
Page 47: Standard Operating Procedure 008 (P
Page 51 and 52: Required Material 4 of 22 Notes Re-
Page 53 and 54: 5.2 Quality Assurance/Quality Contr
Page 55 and 56: 5.2.8 QA Check on New Plates E. col
Page 57 and 58: 7. While the mTEC media is autoclav
Page 59 and 60: 12 of 22 5.3.2.3 Preparation of 1N
Page 61 and 62: 4. Label the bottom (half holding t
Page 63 and 64: 20. Transfer the plates to the 44.5
Page 65 and 66: 8.0 DOCUMENTATION Example Project D
Page 67 and 68: URI Watershed Watch Laboratory Quar
Page 69 and 70: Date 22 of 22 URI Watershed Watch L
Page 73 and 74: Disposal Samples are archived for a
Page 75 and 76: 3. Aluminum weigh dishes, pans, and
Page 77 and 78: a. If a sample clogs a filter, remo
Page 79 and 80: 8.0 DOCUMENTATION Example Data Shee
Page 83 and 84: 4.2 pH and Alkalinity Matrix Sample
Page 85 and 86: are stored in bulk in the cabinet a
Page 87 and 88: 5.3.3 Procedure - Analysis of pH on
Page 89 and 90: 5. Use a non-acid washed graduated
Page 91 and 92: 8.0 DOCUMENTATION Data sheet for an
Page 95 and 96: 3.0 REQUIRED MATERIALS Required Mat
Page 97 and 98: field blank water and 100 mL of the
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procedure. The DO probe should not
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5.3.2 Procedure - Day of Sample Col
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13. The water seal of the BOD bottl
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7.0 REFERENCES 100 - 350 350 - 500
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14 of 14 BI - 12 Fla STE -S 5 BI -
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Standard Operating Procedure 012 (P
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Required Material Notes Re-order in
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Filter Blank Filter blanks consist
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8. When the fluorometer calls for t
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5.2.5 Calibration Check/Laboratory
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6. Check the calibration of the rep
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5.3.2.2 Preparation of 100 mL of 1N
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5.3.3.1 Procedure for Diluting Off-
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7.0 REFERENCES APHA, AWWA, WEF. Sta
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Chlorophyll-a Calibration Record Ca
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Chlorophyll-a Analysis Data Sheet N
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samples are analyzed until an accep
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5.3.1.1 Preparation of 1000 ppm NaC
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2. Prepare a filter housing by plac
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2. Pipette 100 µl of the chloride
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parts and eyes. Sulfuric Acid (H2SO
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The available volume of each reagen
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Corrective Action If the %D is grea
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. Values returned by the balance sh
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5. Shake the unfiltered sample bott
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5.3.2.3 Sample Analysis Detailed in
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2.0 HEALTH AND SAFETY CONSIDERATION
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6 of 19 5.2.1 Method Detection Limi
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and the samples analyzed between th
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10 of 19 serviced and re-calibrated
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X 20 400 X 25 500 X 50 1000 (1.0 mL
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they must be placed in the freezer
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Th ese instruction allow for the cr
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6. After the computer results are p
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USample Log Sheet SR1 SYNC (High st
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1 of 25 Standard Operating Procedur
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Corrective Action If any method (di
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Corrective Action If the %D is grea
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The percent recovery is calculated
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All phosphorus glassware should be
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adjustable pipette to add intermedi
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Preparation of Nitrate/Nitrite Work
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c. Enter SPK2 for the spike number
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Digesting Reagent for Total Phospho
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. The standard curve is rejected if
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Sample Log Sheet SR1 SYNC (High std
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Standard Operating Procedure 017 Sa
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5.2.3 Sample Replication Sample rep
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Sample Data Sheet Analysis Tech's R
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Salinity Analysis SOP 017 S:\WW\aww
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Standard Operating Procedure 018 En
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4.0 SAMPLE STORAGE, PRESERVATION AN
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eanalyzed. The field samples will b
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c. Dilute to the 100 mL mark on the
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7.0 CALCULATIONS Record the number
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11 of 21 Enterolert SOP 018 filenam
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Sample Bottle Sterility Check Bacte
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URI Watershed Watch Laboratory Quar
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Figure 1: Balance Calibration Data
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6.0 DOCUMENTATION 4 of 5 Bacterial
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