Biological field and laboratory methods for measuring the quality of ...

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BIOLOGICAL METHODS Diatom Species Proportional Count Before preparing the diatom slides, use an oxidizing agent to digest the gelatinous stalks and other extracellular organic materials causing cell clumping. Before the oxidant is added, however, centrifuge or settle the sample to remove the formalin. If centrifugation is preferred, transfer the sample to a conical tube and centrifuge 10 minutes at 1000 X G. Decant the formalin, resuspend the sample in 10 ml of distilled water, and recentrifuge. Decant, take up the sample in 8 ml of 5 percent potassium (or ammonium) persulfate, and transfer back to the (rinsed) sample vial. If the settling method is preferred, follow the instructions given in the Plankton Section for removing salt from the diatom concentrate, but add persulfate or hydrogen peroxide instead of distilled water. After the formalin is replaced by the oxidant, heat the sample to 95°C for 30 minutes (do not boil). Cool, remove the oxidant by centrifugation or settling, and take up the diatoms in 2 to 3 ml of distilled water. Proceed with the preparation of the permanent diatom mount as described in the Plankton Section. Label the slide with the station location and inclusive sample dates. Carry out the diatom strip count as described in the Plankton Section, except that separated, individual valves (half cell walls) are tallied as such, and the tally is divided by two to obtain cell numbers. 3.2.3 Biomass Cell Volume See the Plankton Section. Dry and Ash-free Weight See the Plankton Section. Centrifugation, Sedimentation and Displacement Centrifugation. Place sample in graduated centrifuge tube and centrifuge for 20 minutes at 1000 X G. Relate the volume in milliliters to the area sampled. Sedimentation. Place sample in graduated cylinder and allow sample to settle at least 24 hours. Relate the volume in milliliters to the area sampled. 4 Displacement. Use displacement for large growths of periphyton when excess water can be readily removed. Once the excess water is removed, proceed as per Plankton Section; however, do not pour sample through a No. 20 mesh, nylon bolting cloth. Chlorophy11 The chlorophyll content of the periphyton is used to estimate the algal biomass and as an indicator of the nutrient content (or trophic status) or toxicity of the water and the taxonomic composition of the community. Periphyton growing in surface water relatively free of organic pollution consists largely of algae, which contain approximately 1 to 2 percent chlorophyll a by dry weight. If dissolved or particulate organic matter is present in high concentrations, large populations of filamentous bacteria, stalked protozoa, and other nonchlorophyll bearing microorganisms develop and the percentage of chlorophyll a is then reduced. If the biomass-chlorophyll a relationship is expressed as a ratio (the autotrophic index), values greater than 100 may result from organic pollution (Weber and McFarland, 1969; Weber, 1973). . Ash-free Wgt (mg/m 2 ) AutotrophiC Index =Chlorophyll a (mg/m2) To obtain information on the physiological condition (or health) of the algal periphyton, measure the amount of pheophytin a, a physiologically inactive degradation product of chlorophyll a. This degradation product has an absorption peak at nearly the same wavelength as chlorophyll a and, under severe environmental conditions, may be responsible for most if not all of the OD 66 3 in the acetone extract. The presence of relatively large amounts of pheophytin a is an abnormal condition indicating water quality degradation. (See the Plankton Section.) To extract chlorophyll, grind and steep the periphyton in 90 percent aqueous acetone (see Plankton Section). Because of the normal seasonal succession of the algae, the taxonomic composition and the efficiency of extraction by steeping change continually during the year. Although mechanical or other cell disruption may not increase the recovery of pigment from
every sample, routine grinding will significantly increase (l0 percent or more) the average recovery of chlorophyll from samples collected over a period of several months. Where glass slides are used as substrates, place the individual slides bearing the periphyton directly in separate small bottles (containing 100 ml) of acetone when removed from the sampler. Similarly, place periphyton removed from other artificial or natural substrates in the field immediately in 90 percent aqueous acetone. (Samples should be macerated, however, when returned to the lab.) Acetone solutions of chlorophyll are ex 4.0 BIBLIOGRAPHY PERIPHYTON tremely sensitive to photodecomposition and lose more than 50 percent of their optical activity if exposed to direct sunlight for only 5 minutes. Therefore, samples placed in acetone in the field must be protected from more than momentary exposure to direct sunlight and should be placed immediately in the· dark. Samples not placed in acetone in the field should be iced until processed. If samples are not to be processed on the day collected, however, they should be frozen and held at - 20°C. For the chlorophyll analysis, see the Plankton Section. American Public Health Association. 1971. Standard methods for the exammation of water and wastewater, 13th ed., APHA, New York. Blum, 1. L. 1956. The ecology of river algae. Bot. Rev. 22(5):291. Butcher, R. W. 1932. Studies in the ecology of rivers. 11. The microflora of rivers with special reference to the algae on the nver bed. Ann. Bot. 46: 813-861. Butcher, R. W. 1940. Studies in the ecology of rivers. IV. Observations on the growth and distribution of sessile algae in the River Hull, Yorkshire. 1. Ecology, 28:210-223. Butcher, R. W. 1946. Studies in the ecology of nvers. VI1. The algae of organically enriched waters. J. Ecology, 35: 186-191. Butcher, R. W. 1959. Biological assessment of river pollution. Proceedings Linnean Society, 170:159-165; Abstract in: J. Sci. Food Agn.l0:(11):104. Cairns, J., Jr., D. W. Albough, F. Busey, and M. D. Chanay. 1968. The sequential companson mdex - A simplified method for nonbiologists to estimate relative differences in biological diversity in stream pollution studies. JWPCF,40(9): 1607-1613. Cooke, W. B. 1956. Colonization of artificial bare areas by microorganisms. Bot. Rev. 22(9):613-638. Cooke, W. B. 1959. Fungi in polluted water and sewage. IV. The occurrence of fungi in a tricklmg filter-type sewage treatment plant. In: Proceedmg, 13th Industrial Waste Conference, Purdue University, Series No. 96,43(3):26-45. Cummins, K. W., C. A. Tyron, Jr., and R. T. Hartman (Editors). 1966. Organism-substrate relationships in streams. Spec. Pub!. No.4, Pymatunmg Lab. of Eco!., Univ. Pittsburgh. 145 pp. Fjerdingstad, E. 1950. The microflora of the River Molleaa with specml reference to the relation of the benthal algae to pollution. Folia Limnol. Scand. No.5, Kobenhaven. 123 pp. Fjerdingstad, E. 1964. PollutIOn of stream estimated by benthal phytomicroorganisms. I. A saprobic system based on communities organisms and ecolOgical factors. Hydrobio!. 49(1):63-131. Fjerdmgstad, E. 1965. Taxonomy and saproblc valency of benthic phytomicroorganisms. Hydrobio!. 50(4):475-604. Hawkes, H. A. 1963. Effects of domestic and industrial discharge on the ecology of nffles in midland streams. Intern. J. Air Water Poll. 7(6/7):565-586. Holtje, R. H. 1943. The biology of sewage sprinkling filters. Sewage Works J. 15(1):14-29. Keup, L. E. 1966. Stream biology for assessing sewage treatment plant efficiency. Water and Sewage Works, 113: 11-411 . Kolkwitz, R., and M. Marsson. 1908. Oekologie der pflanzlichen Saprobien. Benchte Deutschen Botamschen Gesellschaft. 26a:505-519. Kolkwitz, R., and M. Marsson. 1909. Oekologie der Tierischen Saprobien. Intemationale Revue Gesamten HydrobiologlC Hydrographie, 2: 126-152. Lackey, 1. B. 1950. Aquatic biology and the water works engineer. Public Works. 81: 30-41 ,64. Mackenthun, K. M. 1969. The practice of water pollution biology. U. S, FWPCA, Washington, D.C. 281 pp. Mackenthun, K. M., and L. E. Keup. 1970. Biological problems encountered in water supplies. JAWWA, 62(8):520-526. O'Connell, J. M., and N. A. Thomas. 1965. Effect of benthic algae on stream dissolved oxygen. Proc. ASCE, J. Samt. Eng. Dlv. 91:1-16. Odum, H. T. 1957. Trophic structure and productivity of Silver Springs, Florida. Eco!. Monogr. 27:55-112. Parrish, L. P., and A. M. Lucas. 1970. The effects of waste waters on periphyton growths in the Missouri River. (Manuscript). U. S. FWPCA Nat'!. Field InvestigatIOns Center, Cincinnati. 5
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... EPA-610/4-13 -001 July 1973 BIO
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• FOREWORD Man and his environmen
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Name Anderson, Max Arthur, John W.
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The role of aquatic biology in the
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FOREWORD 1 PREFACE CONTENTS BIOLOGI
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BIOMETRICS 1.0 INTRODUCTION 1.1 Ter
Page 13 and 14: BIOLOGICAL METHODS sample or a plan
Page 18: TABLE 1. RAW DATA ON PLANKTON COUNT
Page 33: BIOLOGICAL METHODS y POSITIVE INTER
Page 40: PLANKTON 1.0 INTRODUCTION . 2.0 SAM
Page 43 and 44: sampling is usually sufficient. In
Page 45 and 46: Several sampling methods can be use
Page 47 and 48: 100X. When combined with the ocular
Page 49 and 50: float, examine the underside of the
Page 51 and 52: of counts to numbers per ml is quit
Page 53 and 54: pipet with distilled water into a c
Page 56: BIOLOGICAL METHODS Calculate the ch
Page 59 and 60: PLANKTON REFERENCES Holmes, R. W. 1
Page 61 and 62: PERIPHYIOI
Page 68 and 69: BIOLOGICAL METHODS Patrick, R. 1957
Page 70: MACROPHYTON 1.0 INTRODUCTION . 2.0
Page 73 and 74: 3.0 REFERENCES MACROPHYTON Blackbur
Page 75 and 76: I '" MACROINVERTEBRATES 1.0 INTRODU
Page 77 and 78: 1.0 INTRODUCTION The aquatic macroi
Page 79 and 80: the limitations of available sampli
Page 81 and 82: Because of the extreme spatial and
Page 83: predetermined depth. Grabs with spr
Page 86 and 87: BIOLOGICAL METHODS Because of the s
Page 88 and 89: BIOLOGICAL METHODS fauna and hydrol
Page 90: BIOLOGICAL METHODS technique, or co
Page 94: BIOLOGICAL METHODS Equitability "e,
Page 102 and 103: BIOLOGICAL METHODS TABLE 7. CLASSIF
Page 104 and 105: BIOLOGICAL METHODS TABLE 7. (Contin
Page 107 and 108: TABLE 7. (Continued) MACROINVERTEBR
Page 109 and 110: MACROlNVERTEBRATE REFERENCES 33. Ll
Page 111 and 112: MACROINVERTEBRATE REFERENCES Hauber
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MACROINVERTEBRATE REFERENCES Robert
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FISH
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Deepwater seInIng usually requires
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BIOLOGICAL METHODS (Lonchocarpus ni
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BIOLOGICAL METHODS Figure 4. Gill n
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BIOLOGICAL METHODS the major univer
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BIOLOGICAL METHODS 7.0 BIBLIOGRAPHY
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Freshwater: Northeast FISH REFERENC
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FISH REFERENCES U.S. Department of
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BIOASSAY 1.0 GENERAL CONSIDERATIONS
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BIOLOGICAL METHODS When making wast
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BIOLOGICAL METHODS TABLE 1. STOCK C
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BIOLOGICAL METHODS the concentratio
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BIOLOGICAL METHODS Dimick, R. E., a
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Laboratory in Newtown, Ohio. Groups
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taining some yellow pigment, coarse
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Appendix A Test (Evansville, Indian
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2.3 Food Use a good frozen trout fo
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BIOLOGICAL METHODS 3.5 Methods When
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APPENDIX
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Item Source Cat. No. Unit Approx. C
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2.3 Fish Sources of information on
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Definitions In its original concept
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To Minims _ Liquid Ounces _ Gills _
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