MONTANA STATE UBRARV 3 0864 0015 4567 5 SUPPORT OF AQUATIC LIFE USES IN BIG SPRING CREEK, FERGUS COUNTY, MONTANA BASED ON THE COMPOSITION AND STRUCTURE OF THE BENTHIC ALGAE COMMUNITY Prepared for: State of Montana Department of Environmental Quality P.O. Box 200901 Helena, Montana 59620-0901 Project Officer: Rebecca Ridenour DEQ Contract No. 200012-3 TATE DOCUMENTS COLLECTION MAY 2 8 2002 MONTANA STATE LIBRARY 1515 E. 6th AVE. HELENA, MONTANA 59620 Prepared by: Loren L. Bahls, Ph.D. Hannaea 1032 Twelfth Avenue Helena, Montana 59601 December 14, 2001 Thio report ie printed on paper made from 100% recycled poet-consumer fiber SUMMARY In May and August 2001, periphyton samples were collected at six stations on Big Spring Creek near Lewistown, Montana for the purpose of assessing whether the creek is water-quality limited and in need of TMDLs . The samples were collected following DEQ standard operating procedures, processed and analyzed using standard methods for periphyton, and evaluated following modified USEPA rapid bioassessment protocols for wadeable streams. Results from the samples collected in May could not be compared to biological criteria Montana streams because these samples were collected outside of the summer sampling "window" . The sample collected below Lewistown in May contained an unusually large number of motile, mesotrophic to eutrophic diatoms that may indicate siltation and nutrient problems. In August, the site just below Big Springs supported a cold- water flora that is typical of a spring creek. The moderate impairment noted here is caused by natural thermal stress. Diatom metrics for site 02 above Lewistown indicated minor impairment from disturbance and possible low levels of toxic chemicals. Aquatic life uses were fully supported. Site 03 below Lewistown also provided full support of aquatic life uses, with minor impairment indicated from siltation and toxics. Diatom metrics for site 05 near the mouth of Big Spring Creek indicated moderate impairment and partial support of aquatic life uses . The probable cause of this impairment is nutrient enrichment . The bioassessment ratings for sites 03 and 05 would be the same if diatom metrics for these sites were compared to criteria for prairie streams instead of mountain streams . INTRODUCTION This report evaluates the biological integrity, support of aquatic life uses, and probable causes of impairment to those uses, in Big Spring Creek near Lewistown, Montana. The purpose of this report is to provide information that will help the State of Montana determine whether Big Spring Creek is water-quality limited and in need of TMDLs . The federal Clean Water Act directs states to develop water pollution control plans (Total Maximum Daily Loads or TMDLs) that set limits on pollution loading to water-quality limited waters. Water-quality limited waters are lakes and stream segments that do not meet water-quality standards, that is, that do not fully support their beneficial uses. The Clean Water Act and USEPA regulations require each state to (1) identify waters that are water-quality limited, (2) prioritize and target waters for TMDLs, and (3) develop TMDL plans to attain and maintain water- quality standards for all water-quality limited waters. Evaluation of use support in this report is based on the species composition and structure of the periphyton (benthic algae, phytobenthos) community at six stream sites that were sampled in May and August, 2001. The periphyton community is a basic biological component of all aquatic ecosystems. Periphyton accounts for much of the primary production and biological diversity in Montana streams (Bahls et al . 1992). Plafkin et al . (1989) and Stevenson and Bahls (1999) list several advantages of using periphyton in biological assessments: • Algae are universally present in large numbers in all streams and unimpaired periphyton assemblages typically support a large number (>30) of species,- • Algae have rapid reproduction rates and short life cycles, making them useful indicators of short-term impacts; As primary producers, algae are most directly affected by physical and chemical factors, such as temperature, nutrients, dissolved salts, and toxins; Sampling is quick, easy and inexpensive, and causes minimal damage to resident biota and their habitat; Standard methods and criteria exist for evaluating the composition, structure, and biomass of algal associations; Identification to species is straightforward for the diatoms, for which there is a large body of taxonomic and ecological literature; Excessive algae growth in streams is often correctly perceived as a problem by the public. Periphyton and other biological communities reflect the biological integrity^ of waterbodies; restoring and maintaining the biological integrity of waterbodies is a goal of the federal Clean Water Act; Periphyton and other biological communities integrate the effects of different stressors and provide a measure of their aggregate impact; and Periphyton and other biological communities may be the only practical means of evaluating impacts from non-point sources of pollution where specific ambient criteria do not exist (e.g., impacts that degrade habitat or increase nutrients). Periphyton is a diverse assortment of simple photosynthetic organisms, called algae, and other microorganisms that live attached to or in close proximity of the stream bottom. Most algae, such as the diatoms, are microscopic. Diatoms are distinguished by having a cell wall composed of opaline glass- - hydrated amorphous silica. Diatoms often carpet a stream bottom with a slippery brown film. ^ Biological integrity is defined as "the ability of an aquatic ecosystem to support and maintain a balanced, integrated, adaptive community of organisms having a species composition, diversity, and functional organization comparable to that of natural habitats within a region" (Karr and Dudley 1981) . Some algae, such as the filamentous greens, are conspicuous and their excessive growth may be aesthetically displeasing, deplete dissolved oxygen, interfere with fishing and fish spawning, clog water filters and irrigation intakes, create tastes and odors in drinking water, and cause other problems. PROJECT AREA AND SAMPLING SITES The project area is located in Fergus County near the city of Lewistown (pop. 6,368) in central Montana. The Big Springs, seven miles southeast of Lewistown, generate most of the streamflow in Big Spring Creek. From Big Springs, Big Spring Creek flows northwesterly for about 30 miles through Lewistown and on to its confluence with the Judith River. Major tributaries of Big Spring Creek, including aquifers feeding the Big Springs, head in the Big Snowy Mountains (maximum elevation 8,730 feet), an outlier of the Northern Rockies Ecoregion (Woods et al . 1999) . Although the segment of Big Spring Creek addressed here is located in the Montana Valley and Foothill Prairies Ecoregion, water quality is dominated by the Big Springs. In the Montana Surface Water Quality Standards, the stream is classified B-l above Lewistown and B-2 below Lewistown. The surface geology of the Big Spring Creek watershed consists of Big Snowy dolomite and limestone in the headwaters, metamorphic rocks of the Kootenai Formation in the middle reach, and shales of the Colorado Group in the lower reach (Renfro and Feray 1972) . Vegetation is alpine tundra and spruce-fir forest in the headwaters, mixed forest and grassland in the middle reach, and mixed grassland at lower elevations (USDA 1976) . Periphyton samples were collected at 2 sites on May 25 and at another 4 sites on August 20 and 22, 2001. The 2 sites sampled on May 25 bracket the City of Lewistown (Map 1, Table 1) . The four sites sampled in August span the reach from just below Big Springs to near the mouth of the stream (Maps 2 and 3, Table 1) . Elevations at the sampling sites range from 4250 feet below Big Springs to 3475 feet near the mouth of the creek. Recreation, fish and aquatic life uses in upper Big Spring Creek are threatened by land development and discharges from a fish hatchery (MDEQ 1998) . Below Lewistown, aquatic life, fish and recreational uses are partially impaired by agriculture, channelization, on-site domestic wastewater, the outfall from the Lewistown wastewater treatment plant, stormwater runoff, animal confinement facilities, and silviculture (MDEQ 1998). Three previous reports on Big Spring Creek and tributaries have been prepared for MDEQ by this consultant (Bahls 1999a, 1999b, and 2001) . METHODS Periphyton samples were collected in May by Tom Pick, NRCS/DNRC, and in August by Rebecca Ridenour, MDEQ Monitoring and Data Management Bureau, following standard operating procedures of the MDEQ Planning, Prevention, and Assistance Division. Using appropriate tools, microalgae were scraped, brushed, or sucked from natural substrates in proportion to the rank of those substrates at the study site. Macroalgae were picked by hand in proportion to their abundance at the site. All collections of microalgae and macroalgae were pooled into a common container and preserved with Lugol ' s solution. The samples were examined to estimate the relative abundance and rank by biovolume of diatoms and genera of soft (non-diatom) algae according to the method described by Bahls (1993) . Soft algae were identified using Dillard (1999) , Prescott (1978) , Smith (1950), and Whitford and Schumacher (1984). These books also served as references on the ecology of the soft algae, along with Palmer (1969, 1977) . After the identification of soft algae, the raw periphyton samples were cleaned of organic matter using sulfuric acid and postassium dichromate, and permanent diatom slides were prepared using Naphrax, a high refractive index mounting medium, following Standard Methods for the Examination of Water and Wastewater (APHA 1998) . Between 410 and 458 diatom cells (820 to 916 valves) were counted at random and identified to species. The four volume series by Krammer and Lange-Bertalot (1986, 1988, 1991a, 1991b) was used as the main taxonomic and autecological reference for the diatoms. Lowe (1974), Bahls et al . (1984), Van Dam et al . (1994), and Lange-Bertalot (1996) were also used as ecological references for the diatoms. The diatom proportional counts were used to generate an array of diatom association metrics (Table 2) . A metric is a characteristic of the biota that changes in some predictable way with increased human influence (Barbour et al . 1999) . Metric values generated from the August samples were compared to numeric biocriteria or threshold values developed for streams in the Rocky Mountain Ecoregion of Montana (Table 3) . These criteria are based on metric values measured in least- impaired reference streams (Bahls et al . 1992) and on metric values measured in streams that are known to be impaired by various sources and causes of pollution (Bahls 1993) . The biocriteria in Table 3 do not apply to the May samples, which were collected outside of the summer sampling "window" . The criteria in Table 3 distinguish among four levels of impairment and three levels of aquatic life use support: no impairment or only minor impairment (full support); moderate impairment (partial support) ; and severe impairment (nonsupport) . These impairment levels correspond to excellent, good, fair, and poor biological integrity, respectively. Quality Assurance. Several steps were taken to assure that the study results are accurate and reproducible. Replicate periphyton samples were collected from site 03. Both samples from this site were analyzed by Hannaea and the results were compared side-by-side (Tables 4 and 5) . Diatom metrics generated from the replicate samples resulted in identical bioassessment ratings for this site: minor impairment and full support of aquatic life uses. Upon receipt of the samples, station and sample information were recorded in a laboratory notebook and the samples were assigned a unique number compatible with the Montana Diatom Database, e.g., 2250-01. The first part of this number (2250) designates the sampling site (Big Spring Creek at County Farm below Lewistown) ; the second part of this number (01) designates the number of periphyton samples that have been collected at this site to date for which data have been entered into the Montana Diatom Database. Sample observations and analyses of soft (non-diatom) algae were recorded in a lab notebook along with station and sample information provided by MDEQ . A portion of the raw sample was used to make duplicate diatom slides. After completing the diatom proportional count, the slide used for the count will be deposited in the University of Montana Herbarium in Missoula. The be retainedSby Hannaea in Helena Station information, sample information, and diatom proportional count data have been entered into the Montana Diatom Database, maintained on a PC by Hannaea in Microsoft Access. RESULTS AND DISCUSSION Results are presented in Tables 4 and 5, which are located near the end of this report following the References section. Spreadsheets containing completed diatom proportional counts, with species' pollution tolerance classes (PTC) and percent abundances, are attached as Appendix A. SAMPLE NOTES BS-1. The Cladophora in this sample was senescent. Oscillatoria v/as present both as an epiphyte and as free-living trichomes . BS-2. Sample is silty,- contains Elodea and other plant material . M22BSPRC01. Some moss is present. M22BSPRC02. Visible mats of Phormidium are present. M22BSPRC05. Sample is silty. NGN- DIATOM ALGAE Diatoms dominated periphyton samples collected from the 8 Burleigh and County Farm sites in May (Table 4) . Green algae and cyanobacteria were also common at both sites. The Burleigh site above Lewistown supported more than twice the number of algal genera than did the County Farm site below Lewistown. In August, diatoms (Diatoma hyemalis) dominated the site above Castle Creek just below the Big Springs. Two other chrysophycean algae- -Tribonema and l/aucheria- -ranked 2nd and 3rd in biomass here. All of these algae are typical of cold, spring- fed streams. Green algae were absent from this site. The mat -forming cyanobacterium Phormidium dominated the algal flora below Pike Creek in August (Table 4) . Diatoms were common here and ranked second. Green algae were also absent at this site, which supported only 2 genera of non-diatom algae. The filamentous and potentially nuisance- forming green alga Cladophora dominated the algal flora of Big Spring Creek at the two sites below Lewistown that were sampled in August (Table 4) . Diatoms ranked 2nd in biovolume at both sites and cyanobacteria were also present, especially as epiphytes on Cladophora at BSPRC05. Replicate samples from BSPRC03 generated very similar results (Table 4) . DIATOMS May Samples. Since they were collected outside of the summer sampling "window", diatom metrics generated from the May samples cannot be compared with the biocriteria in Table 3 . This is because samples collected during the cool seasons in Montana tend to be dominated by a single species. Nitzschia fonticola dominated the diatom assemblages at both sites sampled in May (Table 5) . This is a cool-season, but clean water diatom that indicates meso-eutrophic conditions (Van Dam et al . 1994). N. fonticola is also motile and is adapted to living on unstable substrates. The two periphyton samples that were collected in May from above and below Lewistown shared 64% of their diatom floras (Table 5) . This indicates that the two sites were very similar, floristically, and that little or no environmental change occurred between them. However, the two sites both supported a handful of teratological diatom valves, which may indicate the presence of toxic chemicals in the water. August Samples. The sample collected just below Big Springs in August was dominated by Diatoms hyemalis (Table 5) . This oligotrophic diatom indicates cold waters that do not vary in temperature by more than a few degrees seasonally. Although the Shannon species diversity and percent dominant species metrics for this site both indicated moderate impairment and partial support of aquatic life uses, the thermal stresses that cause this "impairment" may be considered natural and result from the discharge of the Big Springs. Diatom metrics at sites 02 and 03 indicated minor impairment but full use support in August (Table 5) . At site 02, a slightly elevated disturbance index and percent dominant species were due to an abundance -of Achnanthidiuw minutissimum. Site 02 only shared about 12% of its flora with site 01, indicating that the flora of Big Spring Creek had changed from a spring flora to more of a stream flora at this point . A somewhat elevated number of motile diatoms at site 03 indicated minor impairment from siltation here (Table 5) . Sites 02 and 03 shared only about a quarter of their floras, indicating that a moderate amount of environmental change had occurred between them. These sites bracket the City of Lewistown and the 10 outfall from the Lewistown wastewater treatment plant. The two replicate samples from site 03 generated a percent community similarity index of 84% (Table 5) . In addition, diatom metrics from both sites yielded the same bioassessment rating: minor impairment but with full support of aquatic life uses. These results are acceptable from a quality assurance standpoint. A large percent dominant species indicated moderate impairment and partial support of aquatic life uses at 3SPRC05 (Table 5) . The dominant species here was the eutrophic diatom Cocconeis pediculus . This diatom has a concave valve surface, by which it is adapted to living as an epiphyte on Cladophora and other filamentous algae. Its abundance at site 05 is directly related to dominance at this site by Cladophora (Table 4) . Cladophora and Cocconeis indicate that the probable cause of impairment at this site is nutrient enrichment. Sites 03 and 05 shared only about a third of their diatom floras, indicating that a moderate amount of environmental change (degradation) occurred between them. Both sites are downstream from the outfall of the Lewistown wastewater treatment plant. All of the sites sampled in August, except the one just below Big Springs, had a few teratological diatom, valves. This may indicate small concentrations of toxic chemicals, including heavy metals. None of the sites sampled in May and August supported a large number of diatoms in the family Epithemiaceae . This indicates that phosphorus, not nitrogen, was likely the limiting nutrient in Big Spring Creek. The overall impairment ratings for Big Spring Creek would not change if metrics for sites 03 and 05 in the B-2 segment below Lewistown were compared to criteria for plains streams instead of mountain streams. Site 03 would still have good 11 biological integrity (minor impairment due to teratological cells) and site 05 would have fair biological integrity due to elevated percent dominant species and depressed diatom diversity REFERENCES APHA. 1998. Standard Methods for the Examination of Water and Wastewater. 20th Edition. American Public Health Association, Washington, D.C. Bahls, L.L. 1979. Benthic diatom diversity as a measure of water quality. Proc . Mont. Acad. Sci . 38:1-6. Bahls, L.L. 1993. Periphyton Bioassessment Methods for Montana Streams (Revised) . Montana Department of Health and Environm.ental Sciences, Helena. Bahls, L.L. 1999a. Use Support in Big Spring Creek Based on Periphyton Composition and Community Structure. Prepared for the Montana Department of Environmental Quality, Helena. Bahls, L.L. 1999b. Effects Of An Oil Spill on The Composition and Structure of the Periphyton Community in Casino Creek and Big Spring Creek near Lewistown, Montana. Prepared for the Montana Department of Environmental Quality, Helena, Contract No. 200012. Bahls, L.L. 2001. Biological Integrity of Cottonwood Creek, Fergus County, Montana, Based on the Composition and Structure of the Benthic Algae Community. Prepared for the Montana Department of Environmental Quality, Helena, Contract No. 200012-3. Bahls, L.L., E.E. Weber, and J. 0. Jarvie. 1984. Ecology and Distribution of Major Diatom Ecotypes in the Southern Fort Union Coal Region of Montana. U.S. Geological Survey Professional Paper 1289, U.S. Government Printing Office, Washington . Bahls, L.L., Bob Bukantis, and Steve Tralles. 1992. Benchmark Biology of Montana Reference Streams. Montana Department of Health and Environmental Sciences, Helena. Barbour, M.T., J. Gerritsen, B.D. Snyder, and J.B. Stribling. 1999. Rapid Bioassessment Protocols for Use in Streams and Wadeable Rivers: Periphyton, Benthic Macroinvertebrates and Fish. Second Edition. EPA/841-B-99-002 . U.S. EPA, Office of Water, Washington, D.C. 12 Dillard, G.E. 1999. Common Freshwater Algae of the United States. J. Cramer, Berlin. Johansen, J.R. 1999. Diatoms of Aerial Habitats. Chapter 12 in Stoermer, E.F., and J. P. Smol (eds.), The Diatoms, Cambridge University Press, New York. Karr, J.R., and D.R. Dudley. 1981. Ecological perspectives on water quality goals. Environmental Management 5:55-69. Krammer, K., and H. Lange-Bertalot . 1986. Bacillariophyceae, Part 2, Volume 1: Naviculaceae . In Ettl, H., J. Gerloff, H. Heynig, and D. Mollenhauer (eds.), Freshwater Flora of Middle Europe. Gustav Fischer Publisher, New York. Krammer, K., and H. Lange-Bertalot. 1988. Bacillariophyceae, Part 2, Volume 2: Bacillariaceae, Epithemiaceae , Surirellaceae . In Ettl, H., J. Gerloff, H. Heynig, and D. Mollenhauer (eds.). Freshwater Flora of Middle Europe. Gustav Fischer Publisher, New York. Krammer, K., and H. Lange-Bertalot. 1991a. Bacillariophyceae, Part 2, Volume 3: Centrales, Fragilariaceae, Eunotiaceae. In Ettl, H., J. Gerloff, H. Heynig, and D. Mollenhauer (eds.), Freshwater Flora of Middle Europe. Gustav Fischer Publisher, Stuttgart. Krammer, K., and H. Lange-Bertalot. 1991b. Bacillariophyceae, Part 2, Volume 4: Achnanthaceae, Critical Supplement to Navicula (Lineolatae) and Gomphonema, Complete List of Literature for Volumes 1-4. In Ettl, H., G. Gartner, J. Gerloff, H. Heynig, and D. Mollenhauer (eds.). Freshwater Flora of Middle Europe. Gustav Fischer Publisher, Stuttgart. Lange-Bertalot, Horst. 1979. Pollution tolerance of diatoms as a criterion for water quality estimation. Nova Hedwigia 64 :285-304 . Lange-Bertalot, Horst. 1996. Red list of limnic diatoms from Germany. Schr.-R. f. Vegetationskde . , H. 28, pp. 633-677. BfN, Bonn-Bad Godesberg. Lowe, R.L. 1974. Environmental Requirements and Pollution Tolerance of Freshwater Diatoms. EPA-670/4- 74-005 . McFarland, B.H., B.H. Hill, and W.T. Willingham. 1997. Abnormal Fragilaria spp . (Bacillariophyceae) in streams impacted by mine drainage. Jour, of Freshwater Ecology 12 (1) : 141- 149 . MDEQ. 1998. Waterbodies in Need of TMDL Development. Montana Department of Environmental Quality, Helena. 13 Palmer, CM. 1969. A composite rating of algae tolerating organic pollution. Journal of Phycology 5:78-82. Palmer, CM. 1977. Algae and Water Pollution: An Illustrated Manual on the Identification, Significance, and Control of Algae in Water Supplies and in Polluted Water. EPA-600/9-77-036. Plafkin, J.L., M.T. Barbour, K.D. Porter, S.K. Gross, and R.M. Hughes. 1989. Rapid Bioassessment Protocols for Use in Rivers and Streams: Benthic Macroinvertebrates and Fish. EPA 440-4-89-001. Prescott, G.W. 1978. How to Know the Freshwater Algae. Third Edition. Wm. C Brown Company Publishers, Dubuque, Iowa. Renfro, H.B., and D.E. Feray. 1972. Geological Highway Map of the Northern Rocky Mountain Region. American Association of Petroleum Geologists, Tulsa, Oklahoma. Smith, G.M. 1950. the Fresh-Water Algae of The United States. McGraw-Hill Book Company, New York. Stevenson, R.J., and L.L. Bahls. 1999. Periphyton Protocols. Chapter 6 in Barbour, M.T., J. Gerritsen, B.D. Snyder, and J.B. Stribling. Rapid Bioassessment Protocols for Use in Streams and Wadeable Rivers: Periphyton, Benthic Macroinvertebrates and Fish. Second Edition. EPA/841-B-99- 002. U.S. EPA, Office of Water, Washington, D.C USDA. 1976. Climax Vegetation of Montana (map). U. S. Department of Agriculture, Soil Conservation Service, Cartographic Unit, Portland. Van Dam, H., A. Mertens, and J. Sinkeldam. 1994. A coded checklist and ecological indicator values of freshwater diatoms from the Netherlands. Netherlands Journal of Aquatic Ecology, 28 (1) : 117-133 . Whitford, L.A., and G.J. Schumacher. 1984. A Manual of Fresh- Water Algae (Revised) . Sparks Press, Raleigh, North Carolina . Whittaker, R.H. 1952. A study of summer foliage insect communities in the Great Smoky Mountains. Ecological Monographs 22:1-44. Woods, A.J., Omernik, J.M., Nesser, J. A., Shelden, J., and Azevedo, S.H. 1999. Ecoregions of Montana (color poster with map), U.S. Geological Survey, Reston, Virginia. 14 i'jf^ o Q. 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W 4J •H 4-) iH C fC 0 r-\ V -H IH E 0 -rH Oj CO o u ■H 4J c 0 x: (0 iH o ft3 ""^ in o E o l*H 0 4J fO 4J tn u ■H 4-1 c ^^ ro iH 4J o tn 0 o iH 4-1 0 c fO iH 0 n •H fO l*-l o u H 4J Is 4-1 :3 0 o 4J II APPENDIX A: DIATOM PROPORTIONAL COUNTS Big Spring Creek at Burleigh (BS-1) 11/23/01 Sample Genus/SpeciesA/arJety PoUution Tolerance Ctass CjHtnt^ Percent 225101 Achnanthes lanceolata 2 3 0.36 225101 Achnanthidium biasolettianum 3 17 2.02 225101 Achnanthidium minutissimum 3 143 17.02 225101 Amphipleura pellucida 2 1 0.12 225101 Amphora libyca 3 1 0.12 2251 01 Amphora pediculus 3 19 2.26 225101 Aulacoseira distans 3 1 0.12 225101 Cocconeis pediculus 3 0 0.00 225101 Cocconeis placentula 3 16 1.90 225101 Cymbella affinis 3 23 2.74 225101jCymbeila cistula 3 1 0.12 0.00 225101 Diatoma hyemalis 3 0 225101 Dialoma moniliformis 2 7 0.83 225101 Diatoma vulgaris 3 9 1.07 225101 Encyonema silesiacum 2 11 1.31 225101 Encyonopsis microcephala 2 10 1.19 22510l'Frustulia vulgaris 2 2 0.24 225101 Gomphonema angustatum 2 1 0.12 225101iGomphonema minutum 3 2 0.24 225101 Gomphonema olivaceum 3 9 ' 1.07 225101 Gomphonema parvulum 1 2 0.24 225101 Gyrosigma acuminatum 3 1 0.12 225101 Melosira varians 2 8 0.95 225101iNavicula acceptata 2 0 0.00 225101 Navicula capitata 2 2 0.24 225101 Navicula capitatoradiata 2 11 1.31 225101 Navicula cryptotenella 2 60 7.14 225101 Navicula pupula 2 2 0.24 225101 Navicula reichardtiana 2 10 1.19 225101 Navicula tripunctata 3 36 4.29 225101 Nitzschia acicularis 2 5 0.60 225101 Nitzschia apiculata 2 2 0.24 225101 Nitzschia dissipata 3 34 4.05 225101 Nitzschia fonticola 3 300 35.71 225101 Nitzschia frustulum 2 2 0.24 225101iNitzschia heufleriana 3 8 0.95 225101 Nitzschia linearis 2 1 0.12 225101 Nitzschia palea 1 6 0.71 225101 Nitzschia paleacea 2 2 0.24 225101 Nitzschia sociabilis 2 14 1.67 225101 Nitzschia vermicularis 2 1 0.12 225101 Pseudostaurosira brevistriata 3 8 0.95 225101 Reimeria sinuata 3 2 0.24 225101 Staurosira construens 3 2 0.24 225101 Staurosirella ieptostauron 3 28 3.33 225101 Staurosirella pinnata 3 6 0.71 225101 Surirella minuta 2 3 0.36 225101 Synedra nana 3 4 0.48 225101 Synedra ulna 2 4 0.48 Page 1 Big Spring Creek at County Farm (BS-2) 11/23/01 Sample Genus/Species/Variety PoJlutJon Tolerance Class Count Pereeitt'"!! 225001 Aclinanthidium biasolettianum 3 2 0.23 225001, Achnanthidium minutissimum 3 46 5.25 225001 Amphipleura pellucida 2 1 0.11 225001 Amphora pediculus 3 16 1.82 225001 Cocconeis placentula 3 2 0.23 225001 Cymbella affinis 3 12 1.37 225001 Denticula tenuis 3 2 0.23 225001 biatoma mesodon 3 1 0.11 225001 Diatoma moniliformis 2 8 0.91 225001 biatoma vulgaris 3 18 2.05 225001 Encyonopsis microcephala 2 4 0.46 225001 Gomphonema minutum 3 22 2.51 225001 Gomphonema olivaceum 3 25 2.85 225001 Melosira varians 2 2 0.23 225001 'Navicuia acceptata 2 2 0.23 225001 Navicuia capitatoradiata 2 9 1.03 225001 Navicuia cryptotenella 2 18 2.05 225001 Navicuia gregaria 2 2 0.23 225001 Navicuia halophila 2 2 0.23 225001 Navicuia menisculus 2 3 0.34 225001 Navicuia pelliculosa 1 2 0.23 225001 Navicuia reichardtiana 2 5 0.57 225001 Navicuia tripunctata 3 60 6.84 225001jNavicula trivialis 2 2 0.23 225001 Nitzschia acicularis 2 9 1.03 225001 Nitzschia archibaldii 2 4 0.46 225001 Nitzschia dissipata 3 24 2.74 225001 Nitzschia fonticola 3 460 52.45 225001 Nitzschia heufleriana 3 3 0.34 225001 Nitzschia linearis 2 4 0.46 225001 Nitzschia palea 1 64 7.30 225001 Nitzschia pusilla 1 10 1.14 225001 Nitzschia recta 3 1 0.11 225001 Nitzschia sociabilis 2 6 0.68 225001 Nitzschia vermicularis 2 2 0.23 225001 Reimeria sinuata 3 3 0.34 225001 Rhoicosphenia curvata 3 7 0.80 225001 Staurosirella leptostauron 3 7 0.80 225001 Staurosirella pinnata 3 2 0.23 225001 Synedra nana 3 1 0.11 225001 Synedra ulna 2 4 0.46 Page 1 Big Spring Creek above Castle Creek (BSPRC 01) 10/21/01 Sample Genus/SpeciesA/ariety Pollution Tolerance Class Count Percent li 1 03803 Achnanthes lanceolate 2 11 1.32 1 03803 Achnanthidium biasolettianum 3 5 0.60 103803Achnanthidium minutissinnum 3 19 2.28 103803 Amphora pediculus 3 10 1.20 1 03803 Cocconeis pediculus 3 0 0.00 1 03803 Cocconeis placentula 3 7 0.84 1 03803 Cyclotella distinguenda 2 3 0.36 1 03803 Cymbella cistula 3 2 0.24 1 03803 Cymbella falaisensis 3 36 4.32 103803 Denticula tenuis 3 0 0.00 103803 Diatoma hyemalis 3 592 70.98 103803piatoma mesodon 3 5 0.60 103803 Diatoma vulgaris 3 4 0.48 1 03803 Encyonema silesiacum 2 2 0.24 1 03803 Encyonopsis microcephala 2 0 0.00 1 03803 Fragilaria capucina 2 8 0.96 1 03803 Fragilaria vaucheriae 2 0 0.00 1 03803 Gomphonema angustatum 2 37 4.44 103803Gomphonema minutum 3 3 0.36 103803|Gomphonema parvulum 1 13 1.56 103803'Karayevia clevei 3 2 0.24 1 03803 Meridion circulare 3 6 1 0.72 1 03803 Navicula capitatoradiata 2 0 0.00 103803 Navicula decussis 3 0 0.00 103803 Navicula lundii 2 0 0.00 103803 Navicula minima 1 0 0.00 103803 Navicula tripunctata 3 4 0.48 1 03803 Nitzschia fonticola 3 53 6.35 1 03803 Rhoicosphenia curvata 3 0 0.00 1 03803 Staurosirella leptostauron 3 0 0.00 1 03803 Staurosirella pinnata 3 5 0.60 1 03803 Synedra ulna 2 7 0.84 Page 1 Big Spring Creek below Pike Creek (BSPRC 02) 10/21/01 Sample Genus/SpeciesA/ariety Pottutiott Totefance Class Count Percent 1 75602 Achnanthidium biasolettianum 3 68 8.10 175602Achnanthidium miriLrtissimum 3 389 46.31 175602Amphipleura pellucida 2 0 0.00 175602 Amphora inariensis 3 4 0.48 175602Amphora pediculus 3 20 2.38 1 75602 Aulacoseira distans 3 3 0.36 1 75602 Cocconeis placentula 3 23 2.74 1 75602 Cymbella affinis 3 89 10.60 1 75602 Cymbella cistula 3 2 0.24 1 75602 Denticula kuetzingii 3 2 0.24 1 75602 Denticula tenuis 3 2 0.24 1 75602 Diatoma moniliformis 2 31 3.69 1 75602 Diatoma vulgaris 3 2 0.24 1 75602 Encyonema brehmii 2 6 0.71 1 75602 Encyonema silesiacum 2 22 2.62 1 75602 Encyonopsis microcephala 2 41 4.88 1 75602 Fragilaria vaucheriae 2 4 0.48 1 75602 Gomphonema angustatum 2 4 0.48 1 75602 Gomphonema olivaceum 3 2 0.24 175602Gomphonema pan/ulum 1 3 0.36 175602 Gomphonema pumilum 3 9 1.07 1 75602 Navicula acceptata 2 0 0.00 1 75602 Navicula capitatoradiata 2 4 0.48 175602 Navicula cryptotenella 2 13 1.55 175602 Navicula pupula 2 0 0.00 175602 Navicula reichardtiana 2 2 0.24 175602 Navicula tripunctata 3 8 0.95 1 75602 Nitzschia acicularis 2 2 0.24 1 75602 Nitzschia dissipata 3 2 0.24 175602 Nitzschia fonticola 3 36 4.29 175602 Nitzschia palea 1 4 0.48 175602 Nitzschia sociabilis 2 2 0.24 1 75602 Reimeria sinuata 3 11 1.31 1 75602 Staurosirelia leptostauron 3 7 0.83 1 75602 Synedra ulna 2 23 2.74 Page 1 Big Spring Creek below Lewistown (BSPRC 03) 10/21/01 SampJe Genus/SpeciesA/anety Pollution Tolerance Class CounI Percent 182202Achnanthes lanceolata 2 2 0.22 1 82202 Achnanthidium biasolettianum 3 6 0.66 182202Achnanthidium minutissimum 3 39 4.29 182202 Amphora inariensis 3 2 0.22 182202 Amphora pediculus 3 46 5.05 1 82202 Aulacoseira islandica 3 1 0.11 1 82202 Cocconeis pediculus 3 55 6.04 1 82202 Cocconeis placentula 3 31 3.41 1 82202 Cymbella affinis 3 9 0.99 1 82202 Denticula kuetzingii 3 2 0.22 1 82202 Diatoma moniliformis 2 18 1.98 1 82202 Diatoma vulgaris 3 86 9.45 1 82202 Encyonema silesiacum 2 27 2.97 1 82202 Encyonopsis microcephala 2 8 0.88 1 82202 Gomphoneis herculeana 3 1 0.11 1 82202 Gomphonema minutum 3 112 12.31 1 82202 Gomphonema olivaceum 3 2 0.22 182202 Gomphonema parvulum 1 4 0.44 1 82202 Gyrosigma attenuatum 3 2 0.22 1 82202 Karayevia clevei 3 2 0.22 1 82202 Melosira varians 2 1 0.11 1 82202 Navicula acceptata 2 2 0.22 1 82202 Navicula capitatoradiata 2 30 3.30 182202 Navicula cryptotenella 2 26 2.86 182202|Navicula gregaria 2 3 0.33 182202 Navicula menisculus 2 4 0.44 182202 Navicula pupula 2 2 0.22 182202 Navicula reichardtiana 2 31 3.41 182202 Navicula tripunctata 3 75 8.24 1 82202 Nitzschia capitellata 2 0 0.00 1 82202 Nitzschia dissipata 3 8 0.88 182202 Nitzschia fonticola 3 116 12.75 182202 Nitzschia inconspicua 2 2 0.22 182202 Nitzschia sociabilis 2 2 0.22 182202 Nitzschia supralitorea 2 4 0.44 1 82202 Rhoicosphenia curvata 3 139 15.27 1 82202!Staurosirella leptostauron 3 2 0.22 1 82202 Staurosirella pinnata 3 0 0.00 1 82202 Synedra ulna 2 8 0.88 Page 1 Big Spring Creek below Lewistown (BSPRC 03r) 10/21/01 Sample Genus/SpeciesA/ariety Pollution Totcfance Class Count Percent .ii 1 82203 Achnanthes lanceolata 2 2 0.22 1 82203 Achnanthidium biasolettianum 3 3 0.33 1 82203 Achnanthidium minutissimum 3 60 6.54 1 82203 Amphipleura pellucida 2 0 0.00 182203 Amphora inariensis 3 2 0.22 182203 Amphora pediculus 3 61 6.65 182203pocconeis pediculus 3 42 4.58 182203:Cocconeis placentula 3 45 4.91 1 82203 Cymbellaaffinis 3 8 0.87 1 82203 Diatoma moniliformis 2 14 1.53 1 82203 Diatoma vulgaris 3 96 10.47 182203iEncyonema auerswaldii 2 1 0.11 182203 Encyonema brehmii 2 5 0.55 1 82203 Encyonema silesiacum 2 16 1.74 1 82203 Encyonopsis microcephala 2 17 1.85 1 82203 Fragilaria capucina 2 2 0.22 1 82203 Fragilaria vaucheriae 2 2 0.22 1 82203 Gomphoneis herculeana 3 2 0.22 1 82203 Gomphonema minutum 3 117 12.76 1 82203 Gomphonema olivaceum 3 8 0.87 182203 Gomphonema parvulum 1 9 0.98 182203 Gomphonema pumilum 3 2 0.22 1 82203 Gomphonema truncatum 3 2 0.22 1 82203 Navicula acceptata 2 6 0.65 1 82203 Navicula capitatoradiata 2 29 L 3.16 182203 Navicula cryptotenella 2 15 1.64 182203 Navicula lundii 2 3 0.33 182203 Navicula menisculus 2 2 0.22 182203 Navicula recens 2 2 0.22 182203 Navicula reichardtiana 2 39 4.25 182203 Navicula stroemii 2 2 0.22 182203 Navicula tripunctata 3 36 3.93 1 82203 Nitzschia capitellata 2 2 0.22 1 82203 Nitzschia dissipata 3 0 0.00 1 82203 Nitzschia fonticola 3 123 13.41 182203 Nitzschia palea 1 6 0.65 182203 Nitzschia sociabilis 2 2 0.22 1 82203 Reimeria sinuata 3 2 0.22 1 82203 Rhoicosphenia curvata 3 109 11.89 1 82203 Stauroneis smithii 2 2 0.22 1 82203 Staurosira construens 3 2 0.22 1 82203 Staurosirella leptostauron 3 11 1.20 1 82203 Staurosirella pinnata 3 3 0.33 1 82203 Synedra acus 2 2 0.22 1 82203 Synedra ulna 2 3 0.33 Page 1 Big Spring Creek near mouth (BSPRC 05) 10/21/01 Sample Genus/SpeciesA/afiety PolJution Toterance Class Count Percent 1 75902 Achnanthidium minutissimum 3 79 9.63 175902 Amphora inariensis 3 2 0.24 175902 Amphora libyca 3 0 0.00 175902 Amphora pediculus 3 38 4.63 1 75902 Aulacoseira canadensis 3 0 0.00 1 75902 Aulacoseira distans 3 1 0.12 175902 Aulacoseira islandica 3 2 0.24 1 75902 Cocconeis pediculus 3 417 50.85 1 75902 Cocconeis placentula 3 27 3.29 1 75902 Cyclotella meneghiniana 2 1 0.12 175902 Cymbella affinis 3 2 0.24 1 75902 Encyonema silesiacum 2 3 0.37 1 75902 Encyonopsis microcephala 2 63 7.68 1 75902 Epithemia sorex 3 2 0.24 1 75902 Gomphonema minutum 3 1 0.12 1 75902 Gomphonema parvulum 1 10 1.22 175902 Gyrosigma attenuatum 3 1 0.12 1 75902 Navicula capitata 2 0 0.00 1 75902 Navicula capitatoradiata 2 26 3.17 175902 Navicula cryptoteneila 2 18 2.20 175902 Navicula menisculus 2 5 0.61 175902 Navicula peregrina 2 2 0.24 175902 Navicula perpusilla 2 2 0.24 175902 Navicula radiosa 3 1 0.12 175902 Navicula reichardtiana 2 14 1.71 175902 Navicula tripunctata 3 10 1.22 175902 Navicula veneta 1 0 0.00 175902 Nitzschia acicularis 2 2 0.24 175902 Nitzschia capitellata 2 0 0.00 1 75902 Nitzschia dissipata 3 39 4.76 1 75902 Nitzschia fonticola 3 2 0.24 175902 Nitzschia frustulum 2 6 0.73 175902 Nitzschia heufleriana 3 2 0.24 175902 Nitzschia palea 1 7 0.85 175902 Nitzschia paleacea 2 4 0.49 175902 Nitzschia penminuta 3 2 0.24 175902 Nitzschia sigmoidea 3 1 0,12 175902 Nitzschia sociabilis 2 2 0.24 1 75902 Reimeria sinuata 3 1 0.12 1 75902 Rhoicosphenia curvata 3 17 2.07 175902 Rhopalodia gibba 2 2 0.24 1 75902 Staurosirella leptostauron 3 2 0.24 1 75902 Staurosireila pinnata 3 4 0.49 Page 1