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Ecosystem functions within a mine drainage passive treatment system

Clint M. Porter , Robert W. Nairn
University of Oklahoma, School of Civil Engineering and Environmental Science, 202 West Boyd Street, Room 334, Norman, OK 73019-1024, United States

a r t i c l e
Article history:

i n f o

a b s t r a c t
The impacts of acid mine drainage (AMD) on receiving water bodies have been extensively studied. Water quality improvement provided by constructed wetlands is well known, but little is known about the ecological function of these treatment systems. A passive treatment system was constructed in eastern Oklahoma to treat AMD from an abandoned coal mine. Multiple water quality parameters were monitored. Chlorophyll a concentrations were determined throughout the system to estimate productivity. Multi-plate articial substrate samplers were deployed to quantify and evaluate the macroinvertebrate assemblage

Received 27 February 2007 Received in revised form 11 December 2007 Accepted 22 December 2007

Keywords: Coal mine drainage Ecosystem functions Macroinvertebrates Chlorophyll a

within each cell. Productivity between treatment cells varied signicantly. The cells receiving efuent from anoxic organic substrate had elevated nutrient levels, which led to elevated productivity. Chlorophyll a levels reached hypereutrophic conditions into nal treatment cells. Macroinvertebrate community structure was signicantly different among treatment cells. Primary cells produced more diverse and evenly distributed taxa, including the presence of moderately intolerant taxa. The treatment system exhibited early successional stage ecosystem functions and trends toward destructive oscillations. 2008 Elsevier B.V. All rights reserved.



Acid mine drainage (AMD) is the product of oxidation reactions between water and sulde-laden geologic strata. However, net-alkaline waters also are common, having circum-neutral pH and greater alkalinity than acidity (e.g., Hedin et al., 1994). Aluminum, iron, manganese, zinc, lead, and copper are metals that have been measured in AMD (Soucek et al., 2000a,b; Cherry et al., 2001; Mays and Edwards, 2001; DeNicola and Stapleton, 2002; Schmidt et al., 2002; Younger et al., 2002). Passive and active systems are both currently used in the treatment of mine drainage. Active treatment processes are labor intensive and costly and usually include the addition of limestone, sodium hydroxide, ammonia, sodium bicarbonate,

or other alkaline substances (Wilmoth et al., 1979; Hedin et al., 1994; Brown et al., 2002; Younger et al., 2002). These processes can require multiple additions and mechanical homogenization over an extended treatment period. In recent years, the injection of coal combustion products (CCPs) into underground mine pools has been demonstrated to increase pH and precipitate metals via decreases in the oxidation of pyrite (Canty and Everett, 2006). CCPs are formed primarily during the burning of coal for electricity and include uidized bed ash (FBA), y ash, and ue gas desulfurization ash. FBA is formed by combining highly alkaline substances, e.g., limestone, to pulverized coal before combustion. During combustion, calcium oxides react with sulfur released from the coal, forming calcium sulfates. This process prevents harmful emissions

Corresponding author at: Enercon Services, Inc., 6525 North Meridian Avenue, Suite 400, Oklahoma City, OK 73116, United States. Tel.: +1 405 640 3011. E-mail address: cporter@enercon.com (C.M. Porter). 0925-8574/$ see front matter 2008 Elsevier B.V. All rights reserved. doi:10.1016/j.ecoleng.2007.12.013


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and signicantly increases the sulfate in the product (Kilgour, 1992). In terms of passive treatment, constructed or treatment wetland systems have been demonstrated to be effective for water quality improvement (Hedin et al., 1994; Watzlaf et al., 2004). In recent years, vertical-ow systems, relying on bacterial sulfate reduction under anaerobic conditions and limestone dissolution, have become common (Mercer, 2000; Watzlaf et al., 2004). Aerobic surface ow systems are typically used along with vertical-ow cells to successfully remove metals from AMD and signicantly increase pH (Mercer, 2000; Rose and Dietz, 2002). This combination of unit processes is commonly dened as a reducing and alkalinity producing system (RAPS). These systems are also known as successive alkalinity producing systems (SAPSs). The water treatment properties of RAPS are well known, but little is understood about the presence or absence of ecosystem functions within these treatment systems. Odum (1969) outlined ecosystem attributes and trends of developing ecosystems in an article on successional stages. Ecosystem attributes dened by Odum (1969) include primary productivity, community diversity, community composition, nutrient cycling, and complexity of food chains. Extensive work has been completed over the years dening primary productivity within diverse surface water bodies (e.g., Odum, 1956, 1968; Mitsch, 1988; Mitsch and Reeder, 1991; Mitsch and Gosselink, 2000). The ratio between total photosynthesis and community respiration has been a valued metric in dening production within ecosystems. Chlorophyll a concentrations within water column samples have also been extracted to assess indirect measures of productivity (Carpenter et al., 1986; Carpenter, 1988; Cronk and Mitsch, 1994; Spieles and Mitsch, 2000). Aquatic community structure encompasses a multiple number of aquatic organisms, from zooplankton to macroinvertebrates and also shes. Like productivity, community structure and function within aquatic ecosystems have been extensively researched (e.g., Forbes, 1907; Hutchinson, 1969; Cummins, 1978; Karr, 1981; Matthews, 1982; Hilsenhoff, 1987). From a number of these studies, metrics and indices have been created to quantify the quality of these different communities. Karr (1981) developed an index to assess sh community structure within lotic systems, and an index was developed by Hilsenhoff (1987) to assess impacts of organic pollutants to macroinvertebrate communities within lentic systems. These studies have been successful in collecting empirical data that characterize ecosystem attributes as dened by Odum (1969). Wetland ecosystem functions are important for human and ecological health. Models have been developed that predict macroinvertebrate trophic structure and oxygen demand in freshwater wetlands (Spieles and Mitsch, 2003), but few studies have tried to evaluate ecosystem functions in constructed wetlands receiving mine drainage. The objective of our study was to identify different ecosystem functions within a RAPS created to receive mine drainage. This research will help to determine if these systems provide auxiliary benets to the improvement of metal-contaminated water.


Materials and methods

Site selection

Coal has been mined in eastern Oklahoma for over 100 years. The HoweWilburton Coal District was an economically important area of coal extraction in the early 1900s. The Bache and Denman Coal Company operated an underground coal mine from 1907 until around 1927 near the town of Red Oak, OK. This site is located in the Arkansas Valley Ecoregion and is situated between the Ozark Highlands to the north and the Ouachita Mountains to the south. Grassland and forest communities are scattered throughout the area (Woods et al., 2005). As part of a separate research project, a highly alkaline CCP was injected into the Bache and Denman mine void space to limit pyrite oxidation in situ (Canty, 1999; Canty and Everett, 2006). FBA was homogenized with mine water into an injectable slurry and pumped into the mine void through several wells. An initial FBA injection of 417 tons was performed in July 1997; a second injection of 2400 tons of FBA was completed in December 2001. In autumn 2001, a multi-celled passive treatment system was constructed to receive the AMD from the abandoned mine seep. The original design of the system was based on pre-injection pH values and metal loading. The RAPS was composed of ve cells, including three aerobic or oxidation ponds, and two vertical-ow cells. AMD ows by gravity rst through an oxidation pond then into a sequence of verticalow cells and oxidation ponds. In the vertical-ow cells, AMD ows through a mixture of horse manure (the organic substrate) and limestone into a limestone sub-drain containing perforated pipes which transmit water to the next cell.


In situ water quality analyses

Physiochemical water quality data were collected approximately every 2 weeks for 1 year at the inow and outow of each treatment cell. The acronym ROW (Red Oak Wetland) was used to label the sampling locations and a numeric value was provided for each ROW site based on location within the system. Mine waters entering the rst treatment cell were analyzed at ROW 1, waters entering the second treatment cell were analyzed at ROW 2, and this labeling system continued through ROW 6 where waters leaving the fth treatment cell were analyzed (Fig. 1). Dissolved oxygen, pH, electrical conductivity, and temperature measurements were collected with Thermo Orion Model 1230 handheld eld meters with all probes manufactured by Thermo Orion (Thermo Orion, Beverly, MA). Alkalinity was determined via titration with a Hach digital titrator following modied methods from APHA (1998). Turbidity measurements were determined with a Hach 2100P Turbidimeter (Hach, Loveland, CO) following APHA (1998). Flow measurements were calculated at each site using a calibrated bucket and stopwatch. Two 250-mL grab samples (polypropylene bottles) were collected for metal, anion, and oxygen demand analyses. Samples for metals analysis were acidied with trace metal grade nitric acid to pH 2 (APHA, 1998). One 500-mL grab sample was collected at each site for chlorophyll a extraction (APHA, 1998).

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Water quality analyses

Acidied water samples were digested following procedures outlined by APHA (1998) for repeat nitric acid digestions. After digestion, all samples were analyzed for metal concentrations (aluminum, iron, manganese, magnesium, calcium, nickel, copper, and zinc) via a PerkinElmer 5100 atomic absorption spectrophotometer (APHA, 1998). Samples collected for anion analyses (uoride, chloride, nitrite, nitrate, bromide, phosphate, and sulfate) were ltered through 0.2-m lters and analyzed via ion chromatography with a Dionex 2010i IC (APHA, 1998). Samples collected for chlorophyll a extraction were ltered immediately upon return to the laboratory. Filters were placed into airtight bags and kept at 20 C until extraction following methods dened by APHA (1998). Five-day biochemical oxygen demand (BOD) tests were performed on non-preserved samples (APHA, 1998) upon return to the laboratory, using appropriate dilutions. Dissolved oxygen measurements were determined with a Thermo Orion Model 1230 handheld multimeter (Thermo Orion, Beverly, MA). Sulde concentrations were determined via colorimetric methods designed by Hach and modied from EPA method 376.2. A Hach DR2000 spectrophotometer was used to measure sulde concentrations (Hach, Loveland, CO; APHA, 1998).

tem, multi-plate articial substrate samplers were deployed for 30-day periods at each sampling location between June and October 2003 (Hester and Dendy, 1962). Multi-plate samplers consisted of 10 square pieces of tempered hardboard with a total surface area of 0.6 m2 . At the end of the 30-day period, samplers were retrieved and preserved in ethanol. All macroinvertebrates were collected and identied to the family level. Within the family Chironomidae, two distinct morphologies were observed and these organisms were identied to the genus level. All identication was based on Merritt and Cummins (1984) and Higgins et al. (1985). Several metrics were chosen from primarily two community indices to assess the functional and structural components of the macroinvertebrate assemblages. The metrics were identied by Plafkin et al. (1989) and Haysip (1992) for use in U.S. EPA Rapid Bioassessment Protocols. A total of eight metrics were included: taxa richness, contribution of the dominant taxa, ratio of EPT and Chironomidae abundance, EPT index, ratio of scraper and lter collector functional feeding groups, invertebrate density, Shannons diversity, and Hills evenness (Shannon, 1948; Washington, 1984).


Statistical analyses


Macroinvertebrate community structure

To evaluate the colonizing macroinvertebrate assemblages and dene community structure within the treatment sys-

Descriptive statistics were calculated for each parameter at each sampling location. One-way ANOVA analyses were run using Statistix 7 (Statistix 7 Software, 2000) for all eld collected and laboratory analyzed data. Tukeys multiple comparison tests were performed to determine signicant differences between sites, using an alpha value of 0.05.


Physiochemical eld data

Fig. 1 A diagram of the Red Oak reducing and alkalinity producing system. Diagram is not to scale. ROW designates the sampling locations.

Physiochemical eld data followed expected temporal trends for a small lentic system. Water temperatures were higher during the summer months and lower during the winter within the treatment system, but temperature values at the mine seep remained constant. Dissolved oxygen concentrations at each of the sampling locations uctuated with temperature. The sampling locations that received efuent from vertical-ow cells, ROW 3 and ROW 5, had dissolved oxygen concentrations consistently below 1.5 mg/L (Table 1). Turbidity values varied signicantly between locations. Efuent from the vertical-ow cells, and the mine seep, had relatively low turbidity. The mean values collected at these locations remained between 0.5 and 1.6 NTU for most of the year. The turbidity values collected in the efuent at ROW 2, ROW 4, and ROW 6 (surface ow cell efuents) varied signicantly from sites that received efuent from vertical-ow wetlands. The mean turbidity values ranged from 15 to 50 NTU among these three sites (Table 1). Flow measurements increased in the spring months, followed by a decline in the summer. The minimum, maximum, and mean ows at the mine seep were 12, 41.4, and 25.5 L/min, respectively. Although the ow rates at the mine seep were not directly affected by precipitation, the values at each of the other sampling locations varied depending on the amount of


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Table 1 Physiochemical water quality data were collected from November 2002 through December 2003 ROW 1
pH Conductivity (S) Dissolved oxygen (mg/L) Turbidity (NTU) Alkalinity (mg/L as CaCO3 ) Temperature ( C) Flow (L/min) 7.21 (0.244) A 2188 (144) A 0.18 (0.101) A 0.53 (0.284) A 50.4 (10.1) A 17.5 (0.225) A 25.5 (9.70) A

7.09 (0.272) AB 2172 (68.6) A 8.48 (2.00) D 20.7 (8.75) C 24.8 (2.44) A 19.4 (9.43) A 28.9 (11.2) A

7.09 (0.229) AB 2181 (225) A 0.43 (0.187) AB 1.10 (0.723) AB 253 (118) B 20.9 (7.23) A 26.3 (12.5) A

7.63 (0.215) C 1773 (578) B 6.22 (3.83) C 16.3 (11.6) BC 190 (71.2) B 20.1 (8.20) A 25.6 (13.8) A

6.90 (0.155) B 1795 (498) B 1.24 (0.318) AB 1.62 (0.551) AB 428 (160) C 21.5 (5.90) A 23.5 (16.9) A

7.20 (0.309) A 2173 (81.8) A 2.32 (4.92) B 50.0 (47.8) D 333 (94.6) D 20.7 (6.13) A 23.4 (17.1) A

Means and standard deviations are displayed for each metric. One-way ANOVAs were used along with Tukeys multiple comparison test to determine pair-wise differences between and within sites, = 0.05. Sampling locations with different letters are signicantly different. The sample size for each parameter is 21.

precipitation. However, no signicant difference was identied between ow measurements (Table 1). Mean electrical conductivity values remained very close to 2.2 mS/cm from ROW 1 to ROW 3, and at ROW 4 and ROW 5 fell to 1.8 mS/cm, and increased again to 2.2 mS/cm at ROW 6. The pH values remained circum-neutral throughout the treatment system. The mean pH value at the mine seep was 7.23. A larger mean pH was detected at ROW 4 compared to all other locations. Although pH values at ROW 1 and ROW 6 were signicantly greater than ROW 5, the values at ROW 2 and ROW 3 were not. The pH of efuent leaving the nal treatment cell was 7.20 (Table 1). The mean alkalinity at ROW 1 was 50.4 mg/L as CaCO3 eqiv. A signicant amount of alkalinity was produced in the vertical-ow treatment cells (Table 1). Limestone dissolution and microbial activity led to signicant increases in alkalinity within these cells. The mean alkalinity values increased from 20.7 to 254 mg/L as CaCO3 eqiv. from ROW 2 to ROW 3. Alkalinity concentrations increased from 190 to 428 mg/L as CaCO3 eqiv. on average between ROW 4 and ROW 5.

At ROW 2, nitrate was present within seven samples. No other samples collected from the remaining locations had detectable levels of nitrate, indicating that all nitrate present at ROW 2 had been converted before the waters were discharged from the vertical-ow wetland (Table 2). Two distinct differences were noted at ROW 3 compared to the previous two sampling locations. First, sulfate concentrations were signicantly lower. Sulfate concentration decreased from 1572 mg/L at ROW 2 to 1365 mg/L at ROW 3 (Table 2). Secondly, phosphate was introduced into the system. The mean phosphate concentration at ROW 3 was 2.35 mg/L compared to concentrations that were below detection limits in the previous two sampling locations. The ROW 4 mean phosphate concentration was signicantly higher than ROW 3, ROW 5, and ROW 6. However, sulfate values remained very similar to ROW 3 (Table 2). At ROW 5 a signicant decrease was observed in the mean sulfate concentration compared to ROW 4. The sulfate concentration decreased from 1433 mg/L at ROW 4 to 1118 mg/L at ROW 5. Anion concentrations remained very similar between ROW 5 and ROW 6.


Nutrient analysis 3.3. Metals analysis

The results from the anions analysis indicated a signicant decrease in sulfate concentrations and an introduction of nitrate and phosphate within the system. No detectable levels of nitrite, nitrate, or phosphate were identied at ROW 1 (Table 2).

The results of metals analysis indicated that few metals varied signicantly between sampling locations. Concentrations of all analyzed metals at each sampling location are identied in Table 2. Copper concentrations were below detection

Table 2 Water quality data were collected from January 2003 to December 2003 ROW 1
Cl (mg/L) NO3 (mg/L) PO4 (mg/L) SO4 (mg/L) Mn (mg/L) Fe (mg/L) S- (mg/L) BOD (mg/L) CHLa (mg/L) 8.91 (0.799) B BDL BDL 1538 (93.1) CD 0.89 (0.296) BC 19.01 (7.19) A 0.026 (0.024) A 10.2 (7.78) A 0.75 (1.58) A

8.94 (0.915) B 1.31 (0.29) A BDL 1573 (106) D 0.60 (0.228) C 1.62 (0.531) B 0.015 (0.009) A 9.77 (6.71) A 1.81 (1.42) A

6.82 (1.23) A BDL 2.35 (0.453) B 1364 (236) B 1.35 (1.33) BC 7.10 (11.5) B 16.71 (3.46) C 88.5 (20.5) C 1.94 (2.72) A

7.03 (1.07) A BDL 3.30 (0.314) A 1433 (68.7) BC 1.30 (0.747) BC 1.01 (0.806) B 0.011 (0.005) A 12.2 (6.89) AB 10.5 (7.40) AB

7.28 (1.62) A BDL 1.69 (0.228) B 1118 (210) A 1.66 (1.61) AB 5.25 (15.8) B 30.25 7.97 D 147 (50.3) D 58.8 (55.4) BC

7.12 (1.72) A BDL 1.78 (0.741) B 1192 (149) A 2.52 (1.48) A 0.95 (1.02) B 9.31 2.88 B 43.1 (10.7) B 60.6 (73.1) C

Sulde (S-), biochemical oxygen demand (BOD) and chlorophyll a (CHLa) values were collected at each sampling location. CHLa samples from cell 2 (ROW 3) and cell 4 (ROW 5) were collected at the east and north surfaces, respectively. Means and standard deviations are displayed for each metric. One-way ANOVAs were used along with Tukeys multiple comparison test to determine pair-wise differences between and within sites, = 0.05. Sampling locations with different letters are signicantly different.

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limits. Nickel concentrations also did not vary signicantly and ranged from 0.10 to 0.12 mg/L. Zinc concentrations did not vary signicantly between any of the sampling locations, and means ranged between 0.02 and 0.04 mg/L. No signicant difference was observed for calcium concentrations between sampling locations. The mean concentrations at ROW 1 through ROW 6 ranged from 640 to 687 mg/L. Magnesium concentrations, like calcium, were not signicantly different among sampling locations. Manganese and iron were the only two metals that were signicantly different between sampling locations. The results of the manganese and iron analyses are shown in Table 2.


Sulde analysis
Fig. 2 Chlorophyll a values measured at ROW 4, north end of cell 4 and at ROW 6 during the summer collection period. The large decrease in September is very likely due to a recent storm event prior to sampling.

Sulde concentrations were relatively high at ROW 3, ROW 5, and ROW 6. ROW 5 sulde concentrations were signicantly greater than all other sites, and ROW 3 concentrations were signicantly greater than ROW 1, ROW 2, ROW 4, and ROW 6 (Table 2). The mean sulde concentrations at ROW 3, ROW 5, and ROW 6 were 16.7, 30.3, and 9.3 mg/L, respectively. A temporal trend in sulde concentration was observed at these locations. Sulde concentrations, like sulfate, are dependent on microbial activity, which is regulated by the ambient temperature. The decrease in sulde concentrations observed was expected. The mean sulde concentrations at ROW 1, ROW 2, and ROW 4 were 26, 15, and 11 g/L, respectively (Table 2). No temporal trend was observed with the sulde concentrations at these sites.


Macroinvertebrate community structure


Biochemical oxygen demand analysis

Biochemical oxygen demand analysis indicated signicant oxygen demand from the efuent of the vertical-ow wetlands. BOD values at each sampling location are identied in Table 2. The mean BOD values at ROW 3, ROW 5, and ROW 6 were 88.5, 147.3, and 43.1 mg/L, respectively. ROW 5 BOD concentrations were signicantly greater than all other sites, and ROW 3 concentrations were signicantly greater than ROW 1, ROW 2, ROW 4, and ROW 6. The BOD concentrations also decreased over time at the majority of the sampling locations.


Chlorophyll a analysis

Chlorophyll a concentrations within the different treatment cells varied signicantly. Chlorophyll a concentrations from each sampling location are identied in Table 2. An increase in concentration was observed within the treatment cell that received efuent at ROW 3 and all subsequent sites. ROW 6 chlorophyll a concentrations were signicantly greater than ROW 1 through ROW 4, and ROW 5 concentrations were signicantly greater than ROW 1 through ROW 3. A temporal trend was observed in chlorophyll a concentrations within the highly productive treatment cells. The values increased in the summer months and followed with a general decrease in the autumn (Fig. 2).

Differing community structure was identied within each treatment cell. All invertebrate taxa collected within each treatment cell are shown in Table 3. No individual were collected within cell 5. Three collection periods were included in the study: JuneAugust 2003, AugustSeptember 2003 and SeptemberOctober 2003. Most of the eight community metrics varied among treatment cells during each collection period. Invertebrate densities ranged from 20 to 843 individuals in cell 3 (Table 4). Invertebrate densities increased in cell 3 and cell 4 compared to cell 1 and cell 2. Although the number of individuals increased, the contribution of the dominant taxa also increased greatly. The contribution of the dominant taxa ranged from 0.450 to 0.994. The dominant taxon in all treatment cells was Chironomidae, a typically tolerant taxon, with exception to Coleoptera in cell 1 during the second collection period. The only EPT taxa collected were Ephemeroptera, and only two different families of Ephemeroptera were identied, Caenidae and Baetidae. The ratio of EPT/Chironomidae abundance ranged from 0 to 0.354. EPT index values indicated the presence of EPT taxa in cell 2cell 4. The ratio of scraper and collector functional feeding groups ranged from 0.000 to 0.994. Three major functional groups were observed within the different treatment cells. During the rst collection period, 40% of the individuals collected were engulfers and 60% were collectors in cell 1. The distribution in cell 2 was more diverse, with 46% of all individuals as collectors, 34% as scrapers and 20% as engulfers. Collectors became dominant in cell 3 and cell 4, comprising 78% and 90% of the individuals, respectively. During the second collection period, 100% of the individuals collected were engulfers in cell 1. This distribution of functional groups is very likely due to the small number of individuals collected. The distribution in cell 2 was once again more diverse, with 20% of all individuals as collectors, 30% as scrapers and 50% as engulfers. Collectors again were dominant in cell 3 and cell 4, comprising 94% and 86% of the


Table 3 Invertebrate assemblages were evaluated at each sampling location Order Sub-order Family Genus C1
Coleoptera Hydrophilidae Gyrinidae Dytiscidae Haliplidae Anisoptera Gomphidae Cordolidae Libellulidae Coenagrionidae Protoneuridae Chironomidae Chironomidae Ceratopogonidae Caenidae Baetidae Physidae X X X X X X X

Cell 1 collections C2

Cell 2 collections C1

Cell 3 collections C1

Cell 4 collections C1







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Zygoptera Diptera

Chironomus Ablabesmyia



Samples were collected from multi-plate articial substrate samplers with 0.6 m2 surface area. Three collection periods were evaluated (C1, C2, and C3). Presence (X) and absence () were indicated for each collection.

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Samples collected from multi-plate articial substrate samplers with 0.6 m2 surface area. No individual were collected in cell 5. Three collection periods were evaluated (C1, C2, and C3).

Table 4 Macroinvertebrate community structure was evaluated for each assemblage collected at all sampling locations

Cell 2 collections

88 6 0.455 0.354 1 0.796 0.946 0.528


20 3 0.450 0.316 1 0.500 0.873 0.795

individuals, respectively. In the nal collection period, 83% of the individuals were collectors and 17% were engulfers within cell 1. The distribution in cell 2 was once again more diverse, with 27% of all individuals as collectors, 41% were scrapers and 33% were engulfers. Collectors again were dominant in cell 3 and cell 4, comprising over 99% of the individuals in both collections. Shannons diversity values ranged from 0.042 to 1.244. Diversity values were much greater in cell 1 and cell 2 compared to cell 3 and cell 4. Hills evenness values ranged from 0.030 to 1.748. Similar to Shannons diversity, Hills evenness values were much greater in cell 1 and cell 2 compared to cell 3 and cell 4 (Table 4). Values from individual community metrics varied among treatment cells and among collection periods. Richness decreased during the second collection period and rebounded during the third period for cell 1cell 3. Richness values at cell 4 increased from 4 to 6, but decreased to 4 in the third collection. The contribution of the dominant taxa increased over all collection periods for cell 1cell 4. The values at cell 2 decreased from one collection to the next. Hills evenness values decreased over all collection periods for cell 1cell 3. In cell 4 the value increased from 0.425 to 0.432 and then decreased to 0.043. Shannons diversity decreased over all collection periods for cell 1 and cell 4. Cell 3 diversity values increased from 0.277 to 0.319 and then decreased to 0.042. Cell 2 diversity values increased from 0.528 to 0.795 and nally 1.244.

C3 C2 C3 C2 C3 C2 C1 C1

Cell 4 collections

Cell 3 collections

37 5 0.270 0.269 1 0.459 0.773 1.244

843 7 0.866 0.031 2 0.804 0.539 0.277

52 2 0.942 0.000 0 0.942 0.221 0.319

501 4 0.994 0.000 0 0.994 0.030 0.042

218 4 0.894 0.005 1 0.959 0.425 0.307

211 6 0.858 0.005 1 0.906 0.432 0.241

320 4 0.991 0.003 1 0.994 0.043 0.060



The RAPS built in Red Oak, Oklahoma, was designed and built to treat AMD; however, the injection of FBA into the mine void spaces, signicantly changed post-injection seep water quality. The water was circum-neutral pH and had relatively low concentrations of many metals (Tables 1 and 2). Therefore, observable ecosystem functions within the treatment system were not inuenced by typical AMD. Dissolved oxygen concentrations at ROW 4 decreased in the late summer months. The constant addition of low dissolved oxygen, high sulde, and high phosphate waters into the third treatment cell, along with high ambient temperatures, could have led to a signicant increase in microbial proliferation and a decrease in dissolved oxygen concentrations within the water column. The dissolved oxygen concentrations were below recommended criteria for warm water aquatic communities (EPA, 2002). The conditions in cell 5 became very poor in a short-time period after the introduction of efuent from cell 4. The dissolved oxygen concentrations increased significantly above saturation and then dropped below 1 mg/L and remained low through out the sampling period. The sulde concentrations at ROW 6 remained high unlike ROW 4 where sulde concentrations from ROW 3 volatilized relatively rapidly (Table 4). Prior to this study, after the initial introduction of mine drainage into the treatment system, a containment wall in cell 2 was reconstructed due to a previous failure. Flow into the system was halted and mine drainage present in the anoxic column of cell 2 and cell 4 was released directly into cell 5 after ows were stopped. This water was saturated with organic material from the two vertical-ow

C3 C2 C1

Cell 1 collections

Invertebrate density Taxa richness Contribution of dominant taxa Ratio of EPT/Chironomidae abundance EPT index Ratio of scraper and collector functional feeding groups Hills evenness Shannons diversity

33 6 0.606 0.000 0 0.606 1.748 0.976

4 2 0.750 0.000 0 0.000 0.563 0.812

54 4 0.833 0.000 0 0.833 0.382 0.530


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cells and carried many nutrients and other compounds into cell 5. The signicant introduction of these compounds into cell 5 facilitated the present conditions in cell 5. The poor construction of the cell was remediated prior to the initiation of this study, yet the impacts to cell 5 remained. Data collected from 5-day BOD tests indicated that efuents leaving the vertical-ow wetlands had a signicant oxygen requirement. The oxygen was utilized in the biochemical degradation of organic materials or the oxidation of suldes and ferrous iron. Due to the suspected limited amount of ferrous iron in the efuent, the oxygen consumed by processes occurring in the efuent were limited to oxidation of suldes and biochemical degradation of organic materials. Odum (1969) identied the presence of organic matter as an indicator of successional stages in an ecosystem. Although organic matter is important in ecosystem development, enrichment (or over abundance) of organic matter can lead to anoxic conditions and limit community structure (Chandler, 1970; Hilsenhoff, 1987; Thomas and Daldorph, 1994; Kadlec and Knight, 1996). Signs of enrichment were present in cell 3 and cell 4 based on chlorophyll a concentrations and macroinvertebrate community structure. Chlorophyll a extractions identied oligotrophic through hypertrophic status within the treatment system cells. Chlorophyll a concentrations at ROW 1 through ROW 3 were oligotrophic, <3.5 mg/m3 , in all but a few samples which were mesotrophic, 3.59 mg/m3 . Chlorophyll a concentrations at ROW 4 were primarily eutrophic, 925 mg/m3 , and chlorophyll a concentrations at ROW 5 and ROW 6 were primarily hypertrophic, >25 mg/m3 (Nurnberg, 1996). The majority of evidence suggests that phytoplankton growth is limited by phosphorus, and phosphate was introduced into the system at ROW 3 (Schindler, 1977; Table 2). The excessive chlorophyll a concentrations were linked to the signicant increase in phosphate concentrations. Chlorophyll has not typically correlated well with other methods of measuring productivity in wetland ecosystems, but is useful when identifying top-down effects in shallow water bodies (Hall and Moll, 1975; Lammens, 1988; Berger, 1989; Havens, 1993; Fennessy et al., 1994). A diverse range in chlorophyll a concentrations was identied among the treatment cells. The presence of producers is extremely important in healthy functioning ecosystems, but balance is required to maintain homeostasis. Population density, biotic control of grazing, and nutrient cycling provides the chief positive-feedback mechanisms that contribute to stability and preventing destructive oscillations (Odum, 1969). Impacts to consumers become increasingly obvious as populations of producers grow exponentially due to nutrient cycling that is unstable. Chlorophyll a was used as an indirect measure of productivity in this study. Data collected in cell 1 and cell 2 indicated oligotrophic status. The limited productivity will likely impact other ecosystem functions as succession continues within the treatment system. Although productivity is not limited in cell 3 and cell 4, nutrient enrichment and large algal growths limit ecosystem functions. Macroinvertebrate community structure varied signicantly among treatment cells. Shannons diversity index and Hills evenness values indicated that more diverse and evenly distributed communities were collected in cell 1 and cell 2 compared to cell 3 and cell 4. Although the number of individu-

als increased dramatically in cell 3 and cell 4, the contribution of the dominant taxa increased dramatically as well. The family Chironomidae, a typically tolerant family, heavily dominated cell 3 and cell 4 communities. A more diverse number of functional groups were also observed in cell 2 collections. Cell 3 and cell 4 samples were also almost entirely composed of collectors. Species diversity, equitability, and complexity of food chains are all important ecosystem attributes. Odum (1969) used these, as well as many others, to dene successional stages in ecosystem development. A linear and predominantly grazing food chain would describe early or developmental stages and web-like food chain that are dominated by detritivores would be characteristic of mature stages. Low diversity and equitability would be found in developmental stages and high equitability and diversity would characterize mature stages in ecological succession. Many of these attributes described the communities that were collected within the different treatment cells. The invertebrate community in cell 1 was limited in the number of individuals but had relatively good diversity and evenness. After most of the iron was removed in the rst treatment cell the community structure improved. Iron precipitant within aquatic ecosystems has adversely affected macroinvertebrates in previous studies. The precipitant as a physical structure negatively affects habitat and normal invertebrate function (DeNicola and Stapleton, 2002; Batty et al., 2005). The toxicity of iron is not typically a concern. High sulde and phosphate loading into cell 3 led to the decline in the quality of the invertebrate community in the following treatment cells. Resh and Rosenberg (1984) noted that changes in invertebrate communities could be attributed to changes in water quality constituents such as sulde and phosphate. Aquatic organisms are dependent on the presence of suitable habitat. Pioneering aquatic species were observed along the littoral fringe of each treatment cell. The macroinvertebrate communities would improve with further improvement of water quality and additional development of suitable habitat.



Many ecosystem functions were identied in this study. Productivity varied among different treatment cells. Nutrient cycling was very rapid to conservative, and macroinvertebrate community diversity, evenness, and trophic structure were relatively high at different sampling locations. The RAPS considered in this study could be dened as an ecosystem in early successional stages based on metrics presented by Odum (1969). Trend in destructive oscillations were observed, and continued discharge of low oxygenated, high phosphate water into the later cell will prohibit a trend toward homeostasis. The data collected in this study are not typical of all reducing and alkalinity producing systems. The presence of more toxic metals and acidic efuents would likely decrease the quality of the communities within the primary treatment cells. Improvements in water quality within the nal cells would produce more diverse communities, and contribute to more quality ecosystem functions. More extensive research is needed to construct a more dened understanding of ecosystem func-

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