MONTANA STATE LIBRARY 3 0864 0015 4554 3 BIOLOGICAL INTEGRITY OF SQUIRREL CREEK, BIG HORN COUNTY BASED ON THE COMPOSITION AND STRUCTURE OF THE BENTHIC ALGAE COMMUNITY Prepared for: State of Montana Department of Environmental Quality P.O. Box 2 00901 Helena, Montana 59620-0901 Project Officer: Patrick Newby STATE DOCUMENTS COLLECTION Prepared by: Loren L. Bahls, Ph.D. Hannaea 1032 Twelfth Avenue Helena, Montana 59601 August 15, 2000 'AY 2 8 2002 MONTANA STATE LIBRARY 1515 E. 6th AVE. HELENA, MONTANA 59520 Printed on 100\ Reayaled Pott-ConMwnar Pmptr SUMMARY On May 4, 2000, composite periphyton samples were collected from natural substrates at 3 sites on lower Squirrel Creek in southeastern Montana for the purpose of evaluating the biological effects of groundwater being discharged from wells used to test the feasibility of coal-bed methane development. Samples were collected, processed and analyzed following standard methods. Upper Squirrel Creek (si«-.e #4) had excellent algal diversity and species richness, indicating a stable and healthy aquatic community. Diatom association metrics at site #4 indicated good to excellent biological integrity and full support of aquatic life uses. Site #4 proved to be a suitable reference site. Squirrel Creek at the culvert (site #3) was moderately impaired by siltation and had significantly depressed diatom diversity. Besides siltation, elevated concentrations of organic nutrients and salts were the likely causes of impairment at this site. , The culvert site was significantly different, in terms of diatom species composition and diatom metrics, from the upstream reference site. Site #1 near the mouth of Squirrel Creek was also moderately impaired but it had better biological integrity than did site #3. The composition of the diatom associations at sites #1 and #3 indicated that site #1 had cooler water temperatures, higher dissolved oxygen concentrations and lower nutrient concentrations than site #3 . Discharges of pumped groundwater between sites #3 and #1 did not cause a further decline in biological integrity. The two duplicate samples collected from site #1 had essentially identical diatom floras. A comparison of metrics generated from samples collected this year with metrics generated from samples collected in 1978-80 indicated little or no change in biological integrity at this site in the intervening 20 years. INTRODUCTION This report evaluates the biological integrity, support of aquatic life uses, and probable causes of impairment to those uses, in lower Squirrel Creek near Decker, Montana. This evaluation is part of a study being conducted by staff of the Montana Department of Environmental Quality (MDEQ) to determine the chemical and biological effects of groundwater discharges to Squirrel Creek. The discharged groimdwater originates from test wells that are being used to dewater underlying coal beds for the purpose of determining the capacity and economic feasibility of methane gas production. Evaluation of use support in this report is based on the species composition and structure of periphyton (phytobenthos, benthic algae) communities at 3 stream sites that were sampled on May 4, 2000. 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 of streams: • 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. 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 irrigation intakes, create tastes and odors in drinking water, and cause other problems. ^ 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) . t PROJECT AREA AND SAMPLING SITES The project area is located in southeastern Big Horn County- near the town of Decker, Montana. Squirrel Creek heads on the Crow Indian Reservation east of Wyola and flows southeasterly for about 20 miles to where it enters the Tongue River less than a mile from the Wyoming state line. The project area encompasses the lower reach of Squirrel Creek below the Reservation boundary. The project area is within the Northwestern Great Plains Ecoregion (Woods et al . 1999). The surface geology of the watershed consists of Paleocene coal -bearing deposits of the Fort Union Formation and Eocene deposits of the Wasatch Formation (Renfro and Feray 1972) . Vegetation is mixed grassland (USDA 1976) . The main land use is livestock grazing. Periphyton samples were collected at 3 sites on Squirrel Creek on May 4, 2000 (Table 1, maps). The uppermost site (#4) and the lowest site (#1) bracket a 10-mile section of lower Squirrel Creek that extends from just below the Crow Reservation boundary to the Tongue River. The middle site (#3) is about 2 miles above the point where pumped groundwater from coal-bed methane development was being discharged into Squirrel Creek. Elevations at the sampling sites range from about 3,900 feet at the upper site to about 3,400 feet at the mouth of Squirrel Creek near Decker. Squirrel Creek is classified C-3 in the Montana Surface Water Quality Standards. METHODS Periphyton samples were collected by Patrick Newby of the MDEQ Monitoring and Data Management Bureau following standard I operating procedures of the MDEQ Planning, Prevention, and Assistance Division. A duplicate sample was collected at station #1 (mouth) for quality assurance purposes. 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. 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 in 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 (1977) . After the identification of soft algae, raw periphyton samples were cleaned of organic matter using sulfuric acid, 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) . For each slide, between 402 and 443 diatom cells (804 to 886 valves) were counted at random and identified to species. The following were used as the main taxonomic and autecological references for the diatoms: Krammer and Lange-Bertalot 1986, 1988, 1991a, 1991b; Patrick and Reimer 1966, 1975., Lowe (1974) was also used as an ecological reference for the diatoms. Bahls et al . (1984) provide autecological information on important diatom species that live in the Southern Fort Union Coal Region of Montana, including many of the diatom species in Squirrel Creek. t 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 from Squirrel Creek were compared to numeric biocriteria developed for streams in the Great Plains Ecoregions 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 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. Besides the ecoregional biocriteria listed in Table 3, Squirrel Creek metrics were also compared to metrics generated from an upstream control site (#4) using Protocol II in Bahls (1993) . Protocol II may be used on relatively short segments of stream where an upstream control site fully supports its aquatic life uses, that is, if it has a rating of "good" or "excellent" biological integrity using Protocol I. Protocol II is based on the percentage of change in metric values at study sites from values measured at the upstream control site. Criteria for evaluating biological integrity using Protocol II are given in Table 12 in Bahls (1993) . For Protocol I, only periphyton samples collected in summer (June 21-September 21) can be compared with confidence to reference stream samples because metric values change seasonally and summer is the season in which reference streams and impaired » streams were sampled for the purpose of biocriteria development. Protocol II can be used at any time of the year. Quality Assurance. Several steps were taken to assure that the study results are accurate and reproducible. First, a duplicate sample was collected at one of the sites (#1) and analyzed independently. A comparison of the metrics generated from these duplicate samples is a test of the reproducibility of the field sampling procedure. Upon receipt of the samples, station and sample information were recorded in a laboratory notebook and samples were assigned a unique number compatible with the Montana Diatom Database, e.g., 0418-32. The first part of this number (0418) designates the sampling site (Squirrel Creek near mouth) ; the second part of the number (32) 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. On completion of the project, station information, sample information, and diatom proportional count data will be entered into the Montana Diatom Database. One set of diatom slides will be deposited in the University of Montana Herbarium in Missoula. The other set of slides will be retained by Hannaea in Helena. RESULTS AND DISCUSSION Results are presented in Figure 1 and Tables 4, 5 and 6, located near the end of this report following the Literature Cited section. Spreadsheets containing completed diatom proportional counts, with species' pollution tolerance classes (PTC) and percent abundances, are attached as Appendix A. SAMPLE NOTES Squirrel Creek, upper (#4) . The Vaucheria in this sample was senescent. » Squirrel Creek at culvert (#3) . This sample consisted mostly of macrophytes and was beginning to decompose. Squirrel Creek near mouth (#1) . This sample contained macrophytes . Diatoms occurred mostly as epiphytes on other algae . Squirrel Creek near mouth (#1, duplicate) . This sample contained mostly macrophytes. Diatoms occurred mostly as epiphytes on other algae. NON- DIATOM ALGAE The siphonaceous chrysophyte Vaucheria was prominent in all oZ the samples (Table 4) . This attached filamentous alga forms dark-green velvety mats on damp soil and in springs, seeps, and slow-moving streams (Prescott 1978) . It needs a constant supply of moisture and thrives only where flows are perennial. It is generally considered to be an indicator of good quality water, although the genus also has marine representatives (Palmer 1977) . Vaucheria and four genera of filamentous green algae were the only non-diatom algae present at the upper site (#4) . The common non- diatom genera here (Hormidiim and Oedogonium) , along with the absence of cyanobacteria, may indicate moderate nitrogen enrichment in upper Squirrel Creek. Vaucheria and diatoms dominated the algal flora at the middle site at the culvert (Table 4) . This site (#3) had the fewest genera of non-diatom algae (3) of all the Squirrel Creek sites, and was probably subject to the greatest amount of stress. The filamentous green alga Cladophora and the filamentous red alga Audouinella appeared only at the mouth of Squirrel Creek, where they were very common to abundant (Table 4) . Although Cladophora often becomes a nuisance, an abundance of Cladophora and Audouinella typically indicates cool, flowing, and > fresh waters (Palmer 1977). Vaucheria was also abundant here. The two samples from the mouth of Squirrel Creek contained very similar non-diatom floras. Only one common genus {Oedogonium) did not appear in both samples . DIATOMS Upper Squirrel Creek (#4) supported a large number of diatom species (68) and individuals were equitably distributed among the species: the dominant diatom species accounted for less than a quarter of the cells and the diversity index was a healthy 4.34 (Table 5) . Except for a slightly depressed pollution index, all of the diatom metrics at site #4 indicated excellent biological integrity and no impairment when compared to other prairie streams. Upper Squirrel Creek site #4 proved to be a suitable local reference site for use in Protocol II (see below) . The number of diatom species in Squirrel Creek declined by more than half from site #4 to site #3 at the culvert, to a minimum of 28 species (Table 5) . Together with an elevated siltation index, this low species richness indicated moderate stress and only fair biological integrity. Increases in the percent abundances of Achnanthes lanceolata, Navicula gregaria, and Navicula veneta (all pollution-tolerant diatoms) probably indicate a significant increase in the concentration of nutrients and dissolved ions between sites #4 and #3. These two sites were very different f loristically : they shared less than a quarter of their diatom floras. Diatom species richness and diversity rebounded somewhat at the mouth of Squirrel Creek (#1) , although siltation index values were still on the threshold of moderate impairment (Table 5) . The diatom assemblage here was dominated by Navicula gregaria. This species tolerates a wide range of conductivities, from fresh 8 to brackish, and prefers cooler water temperatures (Krammer and Lange-Bertalot 1986) . Navicula gregaria is one of the most common diatoms in the southern Fort Union Region. In this area, it prefers cool, mid- size streams (1-100 cfs) , very hard and slightly brackish waters, and pH values between 8.10 and 8.35. It tolerates some suspended sediment and turbidity and reaches peak numbers in May and June (Bahls et al. 1984) . The increase in Achnanthes minutissima at site #1 also indicates cooler, better oxygenated water compared to site #3. The decline in Navicula veneta between sites #3 and #1 indicates lower concentrations of nutrients at site #1. Sites #3 and #1 shared a bit more than half of their diatom floras. The two replicate samples from site #1 contained very similar diatom assemblages that generated very similar metrics (Table 5) . The similarity index for the two replicate samples was 85.72. This compares favorably with a mean similarity index of 81 for counts conducted on replicate slides made from the same sample (Bahls 1993) . One sample indicated moderate impairment from siltation and the replicate indicated only minor impairment, but both values were very close to the threshold. The station at the mouth of Squirrel Creek was one of nine stream sites that were intensively monitored over a span of three years as part of the Southern Fort Union biological baseline study (Bahls 1980, Klarich and Regele 1980) . The site was also included in an aquatic inventory of the CX Ranch conducted by Olson-Elliott and Associates for Consolidation Coal Company. Under these studies, a total of 31 periphyton samples were collected at site #1 from June 1978 through October 1980. Diatom proportional counts were completed on 30 of these samples and Shannon species diversity and other diatom association metrics were calculated from these counts (Bahls 1980, Montana Diatom Database) . A comparison of metrics generated from this year's samples to those generated from the 1978-80 samples will help determine whether and by how much biological integrity has changed at site #1 over the intervening 20 years. Such comparison will also help to ascertain whether pumped groundwater discharges were affecting biological integrity when the samples were collected this year. The mean diatom species diversity index for 30 samples collected at site #1 between 1978 and 1980 was 3.69, with a standard deviation of 0.70. Although slightly lower, the diversity index values calculated for the two samples collected at site #1 in 2000 were both within one standard deviation of the baseline mean (Figure 1) . One may conclude from this analysis that there has been no measurable change in diatom species diversity at site #1 from 1978-80 to this year. Other metrics generated from the 1978-80 Squirrel Creek samples may also be compared with metrics generated from this year's samples. For example, the nine samples collected at site #1 during the Southern Fort Union study contained a maximum of 3 non-diatom genera, a minimum of 0 genera, and an average of 1.7 genera (Bahls 1980) . In 2000, the two samples collected here had 5 and 4 genera of non-diatom algae (Table 4) . Moreover, in 1978-80 site #1 produced a maximum of 40 diatom species, a minimum of 17 species, and an average of 32 species. In 2000, the replicate samples collected at site #1 contained 44 and 55 species (Table 5) . By all of the above measures- -diatom species diversity, non-diatom genus richness, and diatom species richness- -there has been no decline in genetic diversity at the lower Squirrel Creek station in the last 20 years. 10 Figure 1 and the data in Table 5 also demonstrate that the most significant changes in the diatom flora and in associated diatom metrics occurred between the upstream reference site (#4) and the culvert site (#3) . By comparison, changes between sites #3 and #1 were quite small. These two sites had much more in common with one another, in terms of floristics and diatom community structure, than they did with the upstream reference site (Figure 1 and Table 5) . Using Protocol II and the upstream site (#4) on Squirrel Creek as the local reference site, biological integrity at the two downstream sites was rated as "fair" with moderat? impairment and partial support of aquatic life uses (Table 6) . These low ratings were due to the low index of community similarity between the reference site and the study sites. This assessment also indicates that the most significant impairment of biological integrity in Squirrel Creek occurs between site #4 and site #3, and that little or no additional impairment occurs downstream from site #3 . 11 % LITERATURE CITED 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. 1980. Salinity and The Structure of Benthic Algae (Periphyton) Communities in Streams of the Southern Fort Union Region, Montana. Environmental Sciences Division, Montana Department of Health and Environmental Sciences, Helena . Bahls, L.L. 1993. Periphyton Bioassessment Methods for Montana Streams (Revised) . Montana Department of Health and Environmental Sciences, Helena. Bahls, L.L., Bob Bukantis, and Steve Tralles. 1992. Benchmark Biology of Montana Reference Streams . Montana Department of Health and Environmental Sciences, Helena. 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. 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. 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. Klarich, D.A., and S.M. Regele. 1980. Structure, General Characteristics, and Salinity Relationships of Benthic Macroinvertebrate Associations in Streams Draining the Southern Fort Union Coalfield Region of Southeastern Montana. Environmental Sciences Division, Montana Department of Health and Environmental Sciences, Billings. 12 • • 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 . 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 . 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 . Patrick, Ruth, and C.W. Reimer. 1966. The Diatoms of The United States Exclusive of Alaska and Hawaii. Volume 1: Fragilariaceae, Eunotiaceae, Achnanthaceae, Naviculaceae. Monograph Number 13, The Academy of Natural Sciences, Philadelphia . Patrick, Ruth, and C.W. Reimer. 1975. The Diatoms of The United States Exclusive of Alaska and Hawaii. Volume 2, Part 1: Entomoneidaceae, Cymbellaceae, Gomphonemaceae, Epithemiaceae. Nonograph Number 13, The Academy of Natural Sciences, Philadelphia. 13 fl 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. Stevenson, R.J., and Y. Pan. 1999. Assessing Environmental Conditions in Rivers and Streams with Diatoms. Chapter 2 in Stoermer, E.F., and J. P. Smol (eds.). The Diatoms: Applications for the Environmental and Earth Sciences. Cambridge University Press, New York. USDA. 1976. Climax Vegetation of Montana (map). U. S. Department of Agriculture, Soil Conservation Service, Cartographic Unit, Portland. 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. 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J)418 3 3Mtzschiajvittea_ 041833|Pinnularia microstauron v. brebissonii rQsigaia_deJicatuium_ 041833Rhoicosphenia cun/ata MliB33lSlaurQneis tackeL 041833iSurirella brebissonii _041833!SurirelJa minula 041833;SyDedfa iamelica. jDA1833JSynedraJasciculata_ Q41833!Synedra ulna PoHutJon Tolerance Cia$$ Count Percent 71 A&. An. A&. _ia_ 405 _2l a_ J. J2_ _25. 21 _aa_ 3&. Al. Al SL2A 8.49 JL24 _£L12 JiAZ _£L96 JL12 JL9J _(L84 J1.12 A^Q _Q.48 J).84 J1.12 _1.Z9 J}.48 JL12 JL96 JZ2J J1.96 036 Q.12 0 24 JL12 JL12 JL96 48 44 024 JLQQ JL24 JLSe _QJ12 JLQQ JU2 AAA 0.36 JL36 _2^ _2^ JL4a 024 JL24 JLQQ JL24 JL36 JL24 JU2 JL4a 0.24 JLQQ JL12 036 _3^ _QJL2 JLM 3M JL56 1 32 _Qd2l Page 1 Squirrel Creek at culvert (Site #3) 8/1 5/00 • Sample \ Genus/8 peclesA/arfety PolUttion Tolefance Class CcMunt Percem 19510liAchnanthes lanceolata 2 81 9.93 1951oVcaloneis amphisbaena 2 0 0.00 195101 Cocconeis placentula 3 3 0.37 195101 Cyclotella meneghiniana 2 23 2.82 195101 Entomoneis paludosa 2 2 0.25 195101 Fragilaria vaucheriae 2 0 0.00 195101 Gomphonema angustatum 2 54 6.62 195101 Gomphonema mexicanum 2 0 0.00 195101 Gomphonema parvulum 1 12 1.47 195101 Gomphonema sp. 3 13 1.59 195101 Navicula atomus 1 2 0.25 195101 Navicula cincta 1 23 2.82 195101 Navicula erifuga 2 16 1.96 195101 Navicula gregaria 2 311 38.11 195101 Navicula incertata 2 2 0.25 195101 Navicula lanceolata (Ag.) E. 2 4 0.49 195101 Navicula minima 1 2 0.25 195101 Navicula omissa 1 2 0.25 195101 Navicula peregrina 2 2 0.25 195101 Navicula phyllepta 2 6 0.74 195101 Navicula recens 2 1 0.12 195101 Navicula salinarum 1 1 0.12 195101 Navicula veneta 1 136 16.67 195101 Nitzschia communis 1 1 0.12 195101 Nitzschia debiiis 2 2 0.25 195101 Nitzschia frustulum 2 15 1.84 195101 Nitzschia hungarica 2 1 0.12 195101 Nitzschia palea 1 1 0.12 195101 Nitzschia paleacea 2 0 0.00 195101 Rhoicosphenia curvata 3 1 0.12 195101 Surirella minuta 2 71 8.70 195101 Synedra famelica 2 28 3.43 Page 1 Squirrel Creek, upper (Site #4) 8/15/00 Sample Qenus/8p«cJesA/ariety Polkrtion Tolcfance Class Count Percertt 195201 Achnanthes lanceolata 2 13 1.47 1 95201 jAchnanthes minutissima 3 38 4.29 195201 Amphora inariensis 3 4 0.45 195201 Amphora libyca 3 1 0.11 195201 Amphora pediculus 3 2 0.23 195201 Amphora veneta 1 2 0.23 195201 Caloneis bacillum 2 7 0.79 195201 Cocconeis pediculus 3 3 0.34 195201 195201 Cocconeis placentula 3 55 6.21 Cyclotella meneghiniana 2 207 23.36 195201 Cymbella affinis 3 27 3.05 195201 Cymbelia cistula 3 3 0.34 195201 Cymbtila cymbiformis 3 1 0.11 195201 Cymbella mexicana 3 1 0.11 195201 Cymbella mueiiori 2 6 0.68 195201 Cymbella silesiaca 2 108 12.19 195201 Cymbella sinuata 3 1 0.11 195201 Diatoma tenuis 2 6 0.68 195201 Diploneis pseudovalis 2 0 0.00 195201 Diploneis puella 2 6 0.68 195201 Epithemia sorex 3 2 0.23 195201 Eunotia sp. 3 1 0.11 195201 Fragilaria capucina 2 2 0.23 195201 Fragilaria vaucheriae 2 11 1.24 195201 Frustulia vulgaris 2 1 0.11 195201 Gomphonema angustatum 2 1 0.11 195201 Gomphonema mexicanum 2 2 0.23 195201 Gomphonema olivaceum 3 13 1.47 195201 Gomphonema parvulum 1 13 1.47 195201 Gyrosigma spencerii 2 4 0.45 195201 Navicula capitata 2 1 0.11 195201 Navicula capitatoradiata 2 12 1.35 195201 Navicula cincta 1 3 0.34 195201 Navicula circumtexta 1 0 0.00 195201 Navicula cryptotenella 2 6 0.68 195201 Navicula gregaria 2 20 2.26 195201 Navicula lanceolata (Ag.) E. 2 16 1.81 195201 Navicula libonensis 2 2 0.23 195201 Navicula phyllepta 2 2 0.23 195201 Navicula radiosa 3 2 0.23 195201 Navicula recens 2 1 0.11 195201 Navicula reichardtiana 2 65 7.34 195201 Navicula schroeterii 2 2 0.23 195201 Navicula tripunctata 3 18 2.03 195201 Navicula veneta 1 23 2.60 195201 Nitzschia amphibia 2 0 0.00 195201 Nitzschia angustatula 2 1 0.11 Page 1 Squirrel Creek, upper (Site #4) 8/15/00 San^le tSfmmJP&fmUiiiNmiety Polfettion Tolerance Class Count P^rcerrt 195201 Nitzschia apiculata 2 10 1.13 195201 Nitzschia calida 2 0 0.00 195201 Nitzschia communis 1 1 0.11 195201 Nitzschia debilis 2 1 0.11 195201 Nitzschia dissipata 3 33 3.72 195201 Nitzschia frustulum 2 7 0.79 195201 Nitzschia gracilis 2 2 0.23 195201 Nitzschia hungarica 2 2 0.23 195201 Nitzschia linearis 2 3 0.34 195201 Nitzschia recta 3 4 0.45 195201 Nitzschia sigma 2 2 0.2'3 195201 Nitzschia solita 1 3 0.34 195201 Nitzschia vitrea 1 1 0.11 195201 Pinnularia borealis 2 1 0.11 195201 Pleurosigma delicatulum 2 0 0.00 195201 Rhoicosphenia curvata 3 2 0.23 195201 Rhopaiodia gibba 2 1 0.11 195201 Stauroneis kriegeri 3 1 0.11 195201 Surirella minuta 2 74 8.35 195201 Synedra famelica 2 9 1.02 195201 Synedra ulna 2 14 1.58 Page 2 • RECEIVED AUG 1 6 lOOQ OEQ / PPA W-'nlfnrlr^ n rtrt^ M.«.,,, "'^^MUufinii