STANDARD METHODS For tl1e Exatnination of Water and Wastewater

SIXTEENTH EDITION

Prepared and published jointly by:

AMERICAN PUBLIC HEALTH ASSOCIATION

AMERICAN WATER WORKS ASSOCIATION

W A TER POLLUTION CONTRO L FEDERA TION

Joint Editoria! Board

ARNOLD E. GREENBERG, APRA, Cbairman

R. RHODES TRUSSELL, A WW A

L ENORE S. CLESCERI, WPCF

MARY ANN H. FRANSON

Managing Editor

Publication Office: American Public Health Association 1015 Fifteenth Street NW Washington, DC 20005

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TABLE OF CONTENTS

100 GENERAL INTRODUCIION PAGE

3 101 APPLICATIONS ... . .... .

102 LABORATORY APPARATUS, R.EAGENTS, ANO TECHNIQUES. 3

103 EXPRESSION OF REsUL TS . . . . . . . . . . . . . • . . . 17

104 PRECISION, ACCURACY, ANO CORRECTNESS OF ANALYSES 20

105

106

107

108

109

200

201

202

203

204

A. Precision and Accuracy . . . . . . . . 20 B. Quality Contro) in Chemical Analysis . . . 25 C. Checking Correctness of Analyses . . . . . 32

CoLLECTION ANO PRESERVATION OF SAMPLES 37

ION-EXCHANGE RESINS . . . . . . 44

REAGENT W ATER (TENTA TIVE) 44

SAFETY. . . . . . . . . . . . 46

BIBLIOGRAPHY . . . . . . . . 58

PHYSICAL EXAMINATION

INTROOUCTION . . . . . . . .

APPEARANCE. . . . . . . . . . . . . . . . . . . .

CALCIUM CARBONATE SATURATION (fENTATIVE).

COLOR .. . ....... . . .

A . Visual Comparison Method . B. Spectrophotometric Method . C. Tristimulus Filter Method .. D. ADMI Tristimulus Filter Method (fENTATIVE) .

E. Bibliography

205 CoNDUCTIVITY . . . . . . . . . . . . . .

206 Fl.OA T ABLES . . . . . . . . . . . . . . .

63

63

63

67

67 69 73 74 76

76

80

80 A . Particulate Floatab1es (TENT ATIVE) . B. Trichlorotrifluo:roethane-Soluble Floatable Oil and

Grease (TENTA TIVE) .

C. Bibliography . .

207 000R . ... . . .

208 0XYGEN TRANSFER

209 Souos ...... .

A. Total So1ids Dried at 103-lOs·c B. Total Dissolved Solids Dried at 180"C . C. Total Suspended Solids Dried at 103-to5•c .

xxi

83 85 85 92

92 93 95 96

Xxii

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TABLE OF CONTENTS

PAGE

D. Fixed and Volatile Solids Ignited at sso•c . . . . . . 97 E. Settleable Solids . . . . . . . . . . . . . . . . 98 F. Total, Fixed, and Volatile Solids in Solid and Semisolid

Samples . 99 G. Reference. . 100 H. Bibliography 100

210 SALINITY . . . . 101

102

102

109 122 122

A. Electrica1 Condu.ctivity Method . B. Hydrometric Method . C. Argentometric Method D. Reference. . E. Bibliography . . . . .

211 TASTE . . . . . . . . . . 122

A. Taste Threshold Test . 124 B. Taste Rating Test. 124 C. References . 125 D. Bib1iography . . 126

212 TEMPERATURE . . . 126

213 TESTS ON SLUDGES . 127

A. Oxygen-Consumption Rate 127 B. Settled Sludge Volume 129 C. S1udge Volume Index. 130 D. Zone Settling Rate 130 E. Specific Gravity. 132 F. References . 133 G. Bibliography . . 133

214 T URBIDITY. . . . . 133

A. Nephelometric Method-Nephelometric Turbidity Units . . . . . . . . . . . . . . . . . 134

B. Visual Methods-Jackson Turbidity Units. 137 C. Bibliography . . . . . . . . . . . . . . . 139

300 DETERMINATION OF METALS

301 lNTRODUCTION . 143

302 PRELIMINARY TREA TMENT OF SAMPLES 145

A. Preliminary Filtration . 145 B. Preliminary Treatment for Acid-Extractab1e Metals 146

c. Preliminary Digestion for Meta1s 146

D. Nitric Acid Digestion . 148 E. Nitric Acid-Sulfuric Acid Digestion . 148

F . Nitric Acid-Perchloric Acid Digestion . 149

T ABLE OF CONTENTS

G. Nitric Acid-Perchloric Acid-Hydrofluoric Acid Digestion ... ......... .... . .

H. Dry Ashing. . . . . . . . . . . . . . . . . .

303 METALS BY ATOMIC ABSORPTION SPECTROMETRY

A. Determination of Antimony, Bismuth, Cadmium, Calcium, Cesium, Chromium, Cobalt, Copper, Gold, lridium, Iron, Lead, Lithium, Magnesium, Manganese, Nickel, Palladium, Platinum, Potassium, Rhodium, Ruthenium, Silver, Sodium, Strontium,

xxiii

PAGE

150 150

151

Thallium, Tin, and Zinc by Direct Aspiration into an Air-Acetylene Flame . . . . . . . . . . . . . . . . 157

B. Determination of Low Concentrations of Cadmium, Chromium, Cobalt, Copper, Iron, Lead, Manganese, Nickel, Silver, and Zinc by Chelation with Ammonium Pyrrolidine Dithiocarbamate (APDC) and Extraction into Methyl Isobutyl Ketone (MIBK). 160

C. Determination of Aluminum, Barium, Beryllium, Molybdenum, Osmium, Rhenium, Silioon, Thorium, Titanium, and Vanadium, by Direct Aspiration into a Nitrous Oxide-Acetylene Flame . . . . . . . . . . 162

D. Determination of Low Concentrations of Aluminum and Beryllium by Chelation with 8-Hydroxyquinoline and Extraction into Methyl lsobutyl Ketone. . . . . 164

E. Determination of Arsenic and Selenium by Conversion to Their Hydrides by Sodium Borohydride Reagent and Aspiration into an Atomic Absorption Atomizer. 165

F. Determinationof Mercuryby theCold VaporTechnique. 171

304 D ETERMINATION OF MICRO QUANTITIES OF ALUMINUM,

ANTIMONY, ARSENIC, BARIUM, B ERYLLIUM, CADMIUM,

CHROMJUM, CoBALT, COPPER, lRON, LEAD, MANGANESE, MOLYBDENUM, NICKEL, SELENIUM,

SiLVER ANO TIN BY ELECTROTHERMAL A TOMIC

ABSORPTION SPECTROMETRY

305 METALS BY EMISSION SPECTROSCOPY USING AN

INDUCTIVEL Y COUPLED PLASMA SOURCE

(TENTATIVE).

306 ALUMINUM. . . .

A. Atomic Absorption Spectrometric Method . B. Eriochrome Cyanine R Method . . . . . .

307 ARSENIC . . . . . . . . . . . . . . . . . . .

A. Atomic Absorption Spectrometric Method . B. Silver Diethyldithiocarbamate Method . . .

173

180

182

183 183

187

187 187

xxiv T ABLE OF CONTENTS

C. Mercuric Bromide Stain Method D. Bibliography

308 BARIUM ..

309 B ERYLLIUM

A. Atomic Absorption Spectrometric Method . B. Aluminon Method

310 CADMIUM . . ... . ..... . .. .. . .

A. Atomic Absorption Spectrometric Method . B. Dithizone Method

311 CALCIUM . . . . . . . . . . . . . . . . . . .

P AGE

189 191

191

191

192 192

193

194 194

196

A. Atomic Absorption Spectrometric Method . 196 B. Permanganate Titrimetric Method . 197 C. EDT A Titrimetric Method 199 D. Bibliography . . . . . . . . . . . 200

312 C HROMIUM . . • . . . . . . . . . . . 201

A. Atomic Absorption Method for Total Chromium. 201 B. Colorimetric Method . 20 l

313 CoPPER • . . • . . . . • . . . . • . . • • • 204

A. Atomic Absorption Spectrometric Method . 205 B. Neocuproine Method . 205 C. Bathocuproine Method 207 D. Bibliography . . . . . 208

314 HARDNESS . . . . . . . . 209

A. Hardness by Calculation 209 B. EDTA Titrimetric Method 210 C. Bibliography . . . . . . . 214

315 IRON. . . . . . . . . . . . . 214

A. Atomic Absorption Spectrometric Method . 21 S B. Phenanthroline Method. 215 C. References . 220 D. Bibliography . . . . . . 220

316 LEAD . . . . . . . . . . . 221

A. Atomic Absorption Spectrometric Methòd. 221 B. Dithizone Method 221 C. References 223

317 LITHIUM . . . . . . . 224

A. Atomic Absorption Spectrometric Method . 224 B. Flame Emission Photometric Method . . . 224

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318 MAGNESIUM . . . . . . . . . . . . . . . . .

A. Atomic Absorption Spectrometric Method . B. Gravimetrie Method . . . C. Magnesium by Calculation D. Bibliography . . . . . . .

319 MANGANESE (TOTAL) ... .

A. Atomic Absorption Spectrometric Method . B. Persulfate Method C. Bibliography . . . . . . . . . . . . . .

320 MERCURY . . . . . . . . . . . . . . . . .

A. Cold Vapor Atomic Absorption Method . B. Dithizone Method . . . . . . . . . . .

321 NICKEL ......... ... ... . . .

A. Atomic Absorption Spectrometric Method . B. Heptoxime Method (TENT ATIVE) . C. D imethylglyoxime Method D. Bibliography . . . . . . . . . . . .

322 POTASSIUM. . . . . . . . . . . . . . .

A. Atomic Absorption Spectrometric Method . B. Flame Photometric Method . C. Bibliography . . . . . . . . . . . . . . .

323 SELENIUM . . . . . . . . . . . . . . . . . .

A. Atomic Absorption Spectrometric Method . B. Diaminobenzidine Method . . . . . . . . C. Distillation and D iaminobenzidine Method. D. Bibliography . . . . . . . . . . . . . . .

324 SILVER .... . . ............. .

A. Atomic Absorption Spectrometric Method . B. Dithizone Method C. Bibliography . . . . . . . . . . . . . . .

325 SODIUM ...... . . .. ...... . . .

A. Atomic Absorption Spectrometric Method . B. Flame Emission Photometric Method . . .

326 STRONTIUM . . . . . . . . . . . . . . . . .

A. Atomic Absorption Spectrometric Method . B. Flame Emission Photometric Method C. Bibliography . . . . . . . . . . . . . . .

327 V ANADI UM . . . . . . . . . . . . . . . . . .

A. Atomic Absorption Spectrometric Method . B. Gallic Acid Method . .. ........ .

P AGE

226

226 226 228 228 228

229 229 231

232 232 232

234

234 234 236 236

236 236 237 237

238

238 238 240 241

242

242 242 245

245

246 246

249

249 250 251

252

252 252

xxvi T ABLE OF CONTENTS

PAGE

328 ZINC . 254

A. Atomic Absorption Spectrometric Method . 254 B. Dithizone Method I. 255 c. Dithizone Method II 257 D. Zincon Method . 259 E. Bibliography . 260

P art 400 DETERMINATION OF INORGANIC NONMETALLIC

CONSTITUENTS

401 INORGANIC NONMETALS-INTRODUCTION 265

402 ACIDITY. 265

403 ALKALINITY . 269 404 BORON. 274

A. Curcumin Metbod 274 B. Carmine Method . 276 c. Bibliography . 277

405 BROMIDE. 278

406 CARBON DIOXIDE 279 A. Nomographic Determination of Free Carbon Dioxide

and the Three Forms of Alkalinity 280 B. Titrimetric Method for F ree Carbon Dioxide. 285 c. Carbon Dioxide and Fonns of Alkalinity by

Calculation . 285 D. Bibliography . 286

407 CHLORIDE . 286 A. Argentometric Method 287 B. Mercuric Nitrate Method . 288 c. Potentiometric Method . 290 D. Automated Ferricyanide Method 292 E. Bibliography . 294

408 CHLORINE (REsiDUAL) . 294

A. lodometric Method I . 298 B. lodometric Method Il. 300 c. Amperometric Titration Method. 303 D. DPD Ferrous Titrimetric Method . 306 E. DPD Colorimetric Method . 309 F. Leuco Crystal Violet Method . 310 G. Syringaldazine (F ACTS) Method (TENTA TIVE) 313 H. References . 314 I. Bibliography . 31 5

T ABLE OF CONTENTS xxvii

409 CHLORINE 0 EMAND . .

PAGE

316

316 318 319

A. Laboratory Method . B. Field Method . . C. Bibliography . . . .

410 CHLORINE 0IOXIOE ..

A. lodometric Method . B. Amperometric Method C. DPD Method (TENTATIVE) . D. Bibliography . . . . . . . . .

319

320 322 323 324

411 CHLORINE R EQUIREMENT . . . . 325

A. Method for Contro! of Disinfection 325 B. Methods for Pu.rposes Other Than Disinfection Contro!. 327

412 CYANIDE. . . . . . . . . . . . . . . 327

A. Preliminary Treatment of Samples . 331 B. Total Cyanide a.fter Distillation 334 C. Titrimetric Method . . . . . . . . 336 D. Colorimetric Method . . . . . . . 337 E. Cyanide-Selective Electrode Method . 338 F. Cyanides Amenable to Chlorination after Distillation. 340 G. Cyanides Amenable to Chlorination without

Distillation (Short-Cut Method). 341 H. Weak and Dissociable Cyanide . 344 l. Cyanogen Chloride . . . . . . . 345 J. Spot Test for Sample Screening . 345 K. Cyanates . . 34 7 L. Thiocyanate 348 M. References 351

413 FLUORIDE . . 352

A. Preliminary Distillation Step 355 B. Electrode Method. . . . 357 C. SPADNS Metbod. . . . 359 D. Alizarin Visual Method. 361 E. Complexone Method 362 F. Bibliography . . . . . . 364

414 IODIDE. . . . . . . . . . . 364

A. Leuco Crystal Violet Method . 365 B. Catalytic Reduction Method. 367 C. References . 368 D. Bibliography . . . . . . . . 368

-xxviii T ABLE OF CONTENTS

PAGE

415 lODINE . 369

A. Leuco Crystal Vio1et Method (TENTA TIVE) 369 B. Amperometric Titration Method. 371

c. Bib1iography . 372

416 NITROOEN . 373

417 NITROGEN (AMMONIA) . 374

A. Preliminary Distillation Step 377 B. Nesslerization Method (Direct and Following

Distillation) 379 c. Phenate Method 382 D. Titrimetric Method . 383 E. Ammonia-Se1ective Electrode Method . 384 F . Ammonia-Se1ective E1ectrode Method Using Known

Addition. 386 G. Automated Phenate Method. 388 H. Bibliography . 390

418 NITROOEN (NITRATE) 391

A. Ultraviolet Spectrophotometric Screening Method 392 B. Nitrate E1ectrode Screening Method . 393 c. Cadmium Reduction Method . 394 D. Chromotropic Acid Method . 397 E. Devarda's AJloy Reduction Method . 398 F. Automated Cadmium Reduction Method

(TENTA TIVE). 400 G. Bibliography . 403

419 NITROOEN (NITRITE). 404

420 NITROOEN (ORGANIC) 406

A. Macro-Kjeldahl Method. 408 B. Semi-Micro-Kje1dahl Method (TENTATIVE) 411

421 OXYGEN (DISSOLVED) 413

A. lodometric Methods 416 B. Azide Modification . 418 c. Permanganate Modification . 420 D. Alum Floccu1ation Modification. 421 E. Copper Sulfate-Sulfamic Acid Floccu1ation

Modification . 421 F . Membrane Electrode Method . 422 G. References . 425

422 0ZONE (RESIDUAL). 426

423 PH VALUE. 429

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PAGE

424 PHOSPHORUS . . . . . . . . . . . . . . . . . 437

A. Preliminary Filtration Step . . . . . . . . 441 B. Preliminary Acid Hydrolysis Step for Acid-

Hydrolyzable Phosphorus . . . . . . . . . . . . . . 442 C. Preliminary Digestion Steps for Total Phosphorus . . . 442 D. Vanadomolybdophosphoric Acid Colorimetric Method . 445 E. Stannous Chloride Method . . . . . . . . . 446 F. Ascorbic Acid Method . . . . . . . . . . . 448 G. Automated Ascorbic Acid Reduction Method 450 H. Bibliography . . . . . . . . . . . . . . . 452

425 SILICA . . . . . . . . . . . . . • . . . . . . 453

A. Atornic Absorption Speetrometric Method . 455 B. Gravimetrie Method . . 455 C. Molybdosilicate Method. . . . . . . . . . 457 D. Heteropoly Blue Metbod . . . . . . . . . 460 E. Automated Method for Molybdate-Reactive Silica 461 F. References . 463 G. Bibliography . . . . . . . . . . . . . . . . 463

426 SULFATE . . . . . . . . . . . . • . . . . . . . 464

A. Gravimetrie Method with Ignition of Residue 464 B. Gravimetrie Method with Drying of Residue. 466 C. Turbidimetric Method . . . . . . . . . 467 D. Automated Methylthymol Blue Method . 468 E. Bibliography . . . . . . . . . . . . . . 470

427 SULFIDE . . . . . . . . . . . . . . . . . . 470

A. Separation of Soluble and lnsoluble Sulfides . 473 B. Sample Pretreatment to Remove lnterfering Substances

or to Concentrate the Sulfide . 473 C. Methylene Blue Method. . . . . . . . . . . 475 D. Iodometrie Method . . . . . . . . . . . . . 476 E. Caleulation of Un-ionized Hydrogen Sulfide . 477 F. Bibliography . . . . 478

428 SULFITE . . . . . . . . . . . . . . • • . . 479

A. Iodometric Method . . . . . . . . . . . 479 B. Phenanthroline Method (TENTA TIVE) . 480 C. References . . . . . . . . . . . . . . . 483

429 DETERMINATION OF ANIONS BY ION CHROMATOORAPHY

WITH CoNDUCTIVITY MEASUREMENT 483

Part 500 DETERMINATION OF ORGANIC

CONSTITUTENTS

501 INTRODUCTION . . . . . . . . . . . . . . . . . . . . • . 491

xxx T ABLE OF CONTENTS

PAOE 502 METHANE . . . . . . . . . . . . . . 492

A . Combustible-Gas Indicator Method 492 B. Volumetric Method. 495 C. Bibliography . . . . . . . . . 496

503 0IL AND GREASE. . . . 496

A. Partition-Gravimetric Method . 497 B. Partition-Infrared Method . . . 498 C. Soxhlet Extracùon Method . . 499 D . Ext raction Metbod for Sludge Samples. 501 E. Hydrocarbons . . . . . . . 501 F. Bibliography . . . . . . . . . . . . . 503

504 0RGANIC AND VOLATILE ACIDS . . . . . 503

A . Chromatographic Separation Method for Organic Acids 503 B. Distilla tion Method . . . 505 C. Bibliography . . . . . . . . 506

505 0ROANIC CARBON (TOTAL). 507 A. Combustion-Infrared Method 508 B. Persulfate-Ultraviolet Oxidation Method . 511 c. Wet-Oxidation Method . 513 D. Bibliography . 515

506 0ROANIC HALOGEN (TOTAL) Adsorption-Pyrolysis-Titrimetic Method (TENTA TIVE) 516

507 0XYGEN D EMAND (BIOCHEMICAL) 525

508 OXYOEN DEMAND (CHEMICAL). 532 A. Open Reflux Method . 533 B. Closed Reflux, T itrimetric Method 535 c. Closed Reflux, Colorimetric Method. 537 D . References . 538 E. Bibliography . 538

509 PESTICIDES (0RGANIC) . 538 A. Organochlorine Pesticides . 538

Appendix-Standardization of Magnesia-Silica Gel Column by Weight Adjustment Based on Adsorption of Lauric Acid . 548

B. Chlorinated Phenoxy Acid Herbicides . 550 c. Bibliography . 555

510 PHENOLS . 556

A. Cleanup Procedure . 557 B. Chloroform Extraction Method 558 c. Direct Photometric Method . 560

T ABLE OF CONTENTS xxxi

PAOE

D. Gas-Liquid Chromatographic Method . 561 E. Bibliography 570

511 SLUDOE DIOESTER GAS 571 A. Volumetric Method. 572 B. Gas Chromatographic Method 574 c. References . 577 D. Bibliography . 577

512 SURFACTANTS 577 A. Surfactant Separation by Sublation. 578 B. Anionic Surfactants as MBAS . 581 c. Nonionic Surfactants as CT AS 585 D . References . 588 E. Bibliography . 589

513 T ANNIN ANO L IONIN. 590

514 H ALOOENATED M ETHANES AND ETHANES BY PUROE

AND TRAP. 591

515 0ROANIC CoMPOUNDS CAUSINO T ASTE AND ÙDOR . 602

516 OROANIC CoNTAMINM'TS, Gas Chromatographic/ M ass

Spectrometric M ethod 612

P art 600 AUTOMATE D LABORATORY ANALYSES (deleted,

see specific constituents)

P art 700 EXAMINATION OF WATER AND WASTE WATER

FOR RADIOACfiVITY

701 INTRODUCTION . 625

A. Collection of Samples . 626 B. Counting Room 627 c. Counting Instruments. 627 D . Laboratory Reagents and Apparatus. 633 E. Expression of R esults . 634 F. Statistics . 634 G . Quality Assur ance 636 H . References . 637 l. Bibliography 637

702 RADIOACTIVITY IN WASTEWATER 638

703 GROSS ALPHA AND GROSS BETA RADIOACTIVITY (fOTAL,

SUSPENDED, AND DISSOLVED) 640

704 T OTAL RADIOACTIVE STRONTIUM AND STRONTIUM 90 IN

W ATER 646

705 RADIUM IN WATER BY PRECJPITATION. 652

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706 RADIUM 226 BY R.ADON IN WATER (SOLUBLE, SUSPENOED, ANO TOTAL) 657

707 RADIUM 228 (SOLUBLE) (TENTA TIVE) 667

708 TRITIUM . 670

709 RADIOACTIVE CESIUM 673

710 RADIOACTIVE lODINE 674

A. Precipitation Method . 675

B. Ion-Exchange Method. 676

c. Distillation Method . 678

D. Bibliography . 679

711 URANIUM 679

A. Radiochemical M ethod (TENTA TIVE) 680

B. Fluorometric Method (TENTA TIVE) . 682

c. Bibliography . 684

P art 800 TOXICITY TEST METHODS FOR AQUATIC

ORGANISMS

801 INTRODUCTION . 689 A. Terminology . 689 B. Basic Requirements for Toxicity Tests . 691

c. Conducting Toxicity Tests. 691

D. Preparing Organisms for Toxicity Tests 695

E. Toxicity Test Systems, Materials, and Procedures 707 F. Calculating, Analyzing, and Reporting Results of

Toxicity Tests 715

G. Interpreting and App1ying Results of T oxicity Tests 719

H. References . 720 l. Bibliography . 725

802 BIOSTJMULATION (ALGAL PRODUCTIVITY) 726 A. Generai Principles 726 B. Planning and Evaluating Algal Assays. 727 c. Apparatus 727 D . Sample Handling . 728 E. Synthetic Alga! Culture Medium 729 F. Inoculum. 729 G. Test Conditions and Procedures . 731 H. Effect of Additions . 733 I. Data Analysis and Interpretation 734 ] . References . 735 K. Bibliography . 735

803 TOXJCJTY TESTING WITH PHYTOPLANKTON (TENTA TIVE). 735

804 TOXICITY TEST PROCEDURES USING ZOOPLANKTON. 737

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A. Toxicity Test Procedures for Ciliated Protozoa (TENTATIVE) . . . . . . . . . . . . . . . 737

B. Toxicity Test Procedures for Daphnia (TENTA TIVE) . 739 C. Toxicity Test Procedures for the Calanoid Copepod,

Acartia tonsa (Dana) (TENTA TIVE) . . . . . 742

805 TOXICITY T EST PROCEDURES USJNG SCLERACTINIAN

CORAL (TENTATIVE). . . . . . . . . . 748

A. Selecting and Preparing Test Organisms . 749 B. Toxicity Test Procedures . . . . . 751 C. Evaluating and Reporting Results . . . . 753 D. References . . . . . . . . . . . . . . . 754

806 TOXICITY TEST PROCEDURES FOR ANNELIDS

(TENTATIVE). . . . . . . . . . . . . . 756

A. Selecting and Preparing Test Organisms . 757 B. Toxicity Test Procedures 760 C. Data Evaluation . . . . . . . . . . . . 763 D. Bibliography . . . . . . . . . . . . . . 763

807 TOXICITY 'fEsnNG PROCEDURES FOR CRUSTACEANS

(TENTATIVE). . . . . . . . . . . . 764

A. Selecting and Preparing Test Species. 765 B. Conducting the Toxicity Tests. 774 C. Reporting Results. 782 D. References . . . . . . . . . . 782 E. Bibliography . . . . . . . . . 785

808 TOXICITY TEST PROCEDURES FOR AQUA TIC I NSECTS

(TENTATIVE). . . . . . . . . . . . . . 786

A. Selecting and Preparing Test Organisms . 786 B. Toxicity Test Procedures 789 C. Data Evaluation 791 D. References . . . . . . . 791 E. Bibliography . . . . . . 791

809 TOXlCITY TEST PROCEDURES USING M OLLUSKS

(TENTA TI VE). . . . . . . . . . . . . . 792

A. Selecting and Preparing Test Organisms . 793 B. Conducting the Toxicity Tests . . 795 C. Reporting and Analyzing Results 798 D. References . . . . . . . . . . . 798 E. Bibliography . . . . . . . . . . 799

810 TOXICITY TEST PROCEDURES FOR FISH . 800

A. Fish Selection and Preparation . . . 800

xxxiv T ABLE OF CONTENTS

P AGE

B. Test Procedures. . . . . . . . . . . . . . . . . . 810 C. References . . . . . . . . . . . . . . . . . 819

Part 900 MICROBIOLOGICAL EXAMINATION OF WATER

901 I NTRODUCTION. . . . . . . . • . . • . . . . . . . . 827

A. Generai Discussion . . . . . . . . . . . . . . . . 827 B. U.S. EPA Regulations for Drinking Water Quality . 829

902 LABORATORY QUALITY AsSURANCE .....

A. Intralaboratory Quality Contro) Guidelines B. Interlaboratory Quality Contro! . C. References . . . . . . D. Bibliography . . . . . . .

903 LABORATORY APPARATUS ..

904 WASHING ANO STERILIZATION

905 PREPARATJON OF CULTURE MEDIA .

A. Generai Procedures . B. Water ... . .. . C. Media Specifications D. Bibliography .

906 SAMPLES . . . . . . . •

A. Collection . . . . . B. Preservation and Storage C. Bibliography . . . . . .

907 HETEROTROPHIC PLATE COUNT

A. Pour Plate Method . . . . B. Spread Plate Method . . . . C. Membrane Filter Method . .

908 M ULTIPLE-TUBE fERMENTATION TECHNIQUE FOR

830

831 848 848 849

849

853

853

853 854 855 856

856

856 858 859

860

864 866 869

MEMBERS OF THE COLIFORM GROUP . . . . • . 870

A. Standard Total Coliform Multiple-Tube MPN Tests 872 B. Application of Tests to Routine Examinations . 876 C. Fecal Coliform MPN Procedure. . . . . . . . . . 876 D. Estimation of Bacterial Density . . . . . . . . . . 880 E. Presence-Absence (PA) Coliform Test (TENTATIVE) . 882 F. Bibliography . . . . . . . . . . . . . . . . . . . . 885

909 MEMBRANE fiL TER TECHNIQUE FOR MEMBERS OF THE

COLIFORM GROUP . . . . . . . . . . . . . . . . . . 886

A. Standard Total Coliform Membrane Filter Procedure. 887 B. Delayed-Incubation Total Coliform Procedure . 894 C. Fecal Coliform Membrane Filter Procedure . . 896 D. Delayed-lncubation Fecal Coliform Procedure . 898

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PAGE

E. Klebsiella Membrane Filter Procedure (TENTATIVE) . 899 F. Bibliography . 901

910 TESTS FOR THE FECAL STREPTOCOCCUS GROUP. 902

A . Multiple-Tube Technique . 904 B. Membrane Filter Technique. 905

c. Fecal Streptococcal Plate Count (TENTA TIVE) . 908 D. Bibliography . 909

9 11 D fFFERENTIATION OF THE COLIFORM G ROUP . 910

A. Culture Purifica tion. 910

B. Differentiation 910 c. Significance of Coliform Types 913

D. Media, Reagent-s, and Procedures 914

E. References . 916 F. Bibliography . 917

912 DETECTION OF PATHOGENIC MICROORGANISMS. 917

A. Generai Qualitative lsolation and Identification Procedures for Salmone/la 919

B. Immunofluorescence Identification Procedure for Sa/monella . 923

c. Quantitative Sa/monella Procedures . 926

D . Shigella 927

E. Enteropathogenic Escherichia coli . 928

F. Campylobacter jejuni . 928 G . Vibrio cholerae and NAG Vibrios . 930

H . Pathogenic Leptospires 932

I. Legionellaceae 934

J. Yersinia enterocolitica . 936 K . Pathogenic Protozoa 937

L. References . 941

M . Bibliography . 944

913 DETECTION OF ENTERIC VIRUSES . 946

A. Virus Concentration by Adsorption to and Elution from Microporous Filters (TENT ATIVE) . 948

B. Virus Concentration by Aluminum Hydroxide Adsorption-Precipitation (TENTA TIVE) 959

c. Hydroextraction-Dia1ysis with Polyethylene Glyco1 (TENTA TIVE). 961

D . Recovery of Viruses from Suspended Solids in Water and Wastewater (TENTA TIVE) 963

E. Assay and Identification of Viruses in Samp1e Concentrates (TENT ATIVE) . 964

E. References . 969

xxxvi T ABLE OF CONTENTS

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914 BACTERIOLOGICAL EXAMINATION OF RECREATIONAL WATERS. 974

A. Swimming Poo1s 974

B. Natural Bathing Beaches 976

c. Membrane Filter Technique for Pseudomonas

aeruginosa (TENT ATIVE) . 978

D. Multiple. Tube Technique for Pseudomonas aeruginosa

(TENTATIVE). 979

E. Bibliography . 980

915 DETECTION OF FUNGI 981

A. Pour Plate Technique. 982

B. Spread Plate Technique . 984

c. Membrane Filter Technique . 985

D. Technique for Y easts . 986

E. Zoosporic Fungi 987

F. Aquatic Hyphomycetes . 988

G. Pathogenic Fungi . 989

H. References . 990

l. Bibliography . 991

916 DETECTION OF ACTINOMYCETES 992

917 NEMATOLOGICAL EXAMINATION 995

A. Technique for Nematodes . 997 B. Illustrated Key to Fresh Water Nematodes 999 c. References . 1018

D . Bibliography . 1018

918 IDENTIFICATION OF IRON AND SULFUR BACTERIA. 1019

A. Iron Bacteria . 1020

B. Sulfur Bacteria . 1022

c. Enumeration, Enrichment, and Isolation of Iron and Sulfur Bacteria (TENTA TIVE) . 1025

D. References . 1029

919 RAPID DETECTJON METHODS. 1031

A. Seven-Hour Fecal Coliform Test (TENTATIVE) . 1031

B. Special Techniques (TENTA TIVE) 1032

c. Coliphage Detection (TENTA TIVE). 1033

D . References . 1035

920 STRESSED 0RGANISMS 1036

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PAGE

P art 1000 BIOLOGICAL EXAMINATION OF WATER

1001 INTRODUCTION , 1041

1002 PLANKTON. 1043

A. Introduction 1043 B. Sample Collection. 1044 c. Concentration Techniques . 1054 D. Mounting and Preparation for Examination 1055 E. Microscopes and Calibrations . 1057 F. Counting Techniques . 1060 G. Chlorophyll. 1067 H . Determination of Biomass (Standing Crop). 1072 l. Metabolic Rate Measurements. 1075 J. References . 1079

1003 PERIPHYTON . 1083

A. Introduction . 1083 B. Sample Collection. 1084 c. Sample AnaJysis 1085 D . Productivity 1088 E. Interpreting and Reporting Results 1099 F. References . 1100 G. Bib1iography . 1101

1004 MACROPHYTON. 1103

A. Introduction 1103 B. Preliminary Survey . 1104 c. Vegetation Mapping Methods . 1104 D . Population Estimates . 1106 E. Productivity 1110 F. References . 1111 G. Bibliography . 1111

1005 BENTHIC MACROINVERTEBRATES. 1113

A. Introduction . 1113 B. Samp1e Collection. 1114 c. Sarnple Processing and Analysis . 1125 D. Data Evaluation and Presentation . 1126 E. References . 1127 F. Bibliography . 1129

1006 F'IsH . 1131

A. Introduction . 1131

xxxvìiì

103:1 104:1 104:11

104:III 104:1V 105:1

1007

TABLE OF CONTENTS

PAGE

B. Sample Collection and Preservation . 1131

c. Quantitative Analysis of Fish Populations 1138

D. Sample Ana!ysis 1140

E. Fish ldentification 1141

F. lnvestigation of Fish Kills. 1145

G . References . 1146

H. Bibliography . 11 47

IDENTIFICATION OF AQUATIC 0RGANISMS. 1148

A. Procedure in Identification 1148

B. Key to Major Groups of Aquatic Organisms (Plates 1-38) . 1149

l. Key to Microscopic Organisms . 1149

2. Key t o Macroscopic Organisms. 1150

3. Key to M acroinvertebrates . 1150

4. Key to Crustaceans 1151 5. Key to Insect Pupae . 1152

6. Key to Insect Larvae, Nymphs, and Some Adults 1152

c. List of Common Types of Aquatic Organisms (Piates 1-38), by Trophic Leve! . 1153

ACKNOWLEDGMENTS . 1154

D. Key for ldentification of Freshwater Algae Common in Water Supplies and Polluted Waters (Color Plates A-F). . . . . . . 1194

l. Blue-Green Algae 1194 2. Diatoms . . . . . l I 96 3. Flagellate Algae . 1197 4. Green Algae and Associated Forms. l 198

E. Recent Changes in Names of Algae . 1201 F. lndex to Illustrations . . . . . 1203 G . Selected Taxonomic References 1207

INDEX . . . . . . . . . . . . . . 1217

TABLES

Conversion Factors . . . . . . . . . . . . . . . . . . . Factors for Computing Contro! Chart Lines . . . . . . . Data and Computations for Construction of Example x-R

18 27

Chart .................. . .... . Moving Average and Range Table . ......... . Conductivity Factors of lons Commonly Found in Water Summary of Special Sampling or Handling Requirements

29 30 34 42