Macroinvertebrate community structure as an indicator of acid mine pollution

MACROINVERTEBRATE COMMUNITY STRUCTURE AS AN I N D I C A T O R OF ACID MINE P O L L U T I O N GARY DILLS* t~ DAVIDT. ROGERS,JRt

*Department of Biological Sciences, Haywood Technical Institute, Clyde, North Carolina 28721, USA tDepartment of Biology, University of Alabama, University, Alabama 35486, USA

ABSTRACT

Physicochemical parameters and benthic macroinvertebrate community structure were quantitatively related to varying degrees of acid mine drainage in a small stream system. Tributaries exposed to acid effluents were characterised by a reduction in turbidity, a decrease in pH, and an increase in mineral content. Strong positive correlation existed between hardness, iron, manganese, pH, conductivity, and sulphur. A step-wise regression analysis was performed to disclose the water parameter most closely related to species diversity ( tq) of the macroinvertebrate community. Hydrogenion concentration was highly correlated with species diversity (P < 0.001). Significant differences (P < 0"01) in species diversity existed between strongly acidic and less acidic tributaries. Stations located near areas of acid production were consistently lowest in diversity. Species diversity values for the unpolluted stations showed temporal variations with highest values occurring during late March and December. The more acidic stations showed no seasonal trends in diversity values. Varying degrees of acid mine pollution were reflected by changes in the macroinvertebrate community structure.

INTRODUCTION

During the past decade, a considerable proportion of the interest in environmental degradation has focused on the ecology of streams and rivers. The role of biologists in interpreting the effects of effluents and formulating appropriate management practices has been severely restricted by inadequate measurement techniques and a lack of fundamental data concerning the functioning of stream ecosystems (Cummins, 1969). Traditionally, stream conditions have been determined by 239 Environ. Pollut. (6) (1974)--O Applied Science Publishers Ltd, England, 1974

Printed in Great Britain

240

GARY DILLS, DAVID T. ROGERS, JR

direct chemical analysis of the water or by bioassay. The weakness of these methods is that stream conditions are indicated only at the time of sampling. Recent ecological literature shows that investigators have turned to species diversity indices to compare changes in aquatic communities caused by environmental stresses (Wilhm & Dorris, 1966; Pielov, 1966a; Reisch & Winter, 1954; Cairns et al., 1971 ; Bechtel & Copeland, 1970). As defined by Odum (1971), species diversity indices are ratios between the numbers of species and 'importance values' (biomass, numbers, productivity, etc.). Several diversity indices have been proposed (Fisher et al., 1943; Margalef, 1951; Brillouin, 1960; Preston, 1948; Good, 1953; Simpson, 1949). One of the most promising indices of diversity measures is derived from information theory (Margalef, 1956; Patten, 1962; Wilhm & Dorris, 1966; Mathis, 1968). Such measures relate to the uncertainty that exists regarding the species of an individual selected at random from a population. The greater the number of species present and/or the more evenly the individuals are apportioned among the species, the greater the uncertainty in selection, and, hence, the greater the diversity value. In estimating total community diversity, probably the most widely-used index is the Shannon-Wiener (1963) equation: A = -~;Pilog2Pi where Pi is the number of individuals in each species divided by the total individuals for all species. Where Pi is the true unknown proportion of the ith species in the whole population, Pielou (1966b) reports that the Shannon-Wiener equation never gives an unbiased approximation of diversity, but diversity is only estimated. Theoretically, /~ values range from zero to any positive number but are seldom greater than 10. The minimum A value is obtained when all individuals belong to the same species and the maximum/~ value is obtained when each species contains the same number of individuals (Wilhm, 1970). Wilhm, after sampling various habitats, reported that diversity (H) in clean water streams varied between 3 and 4 while polluted streams usually had indices of less than 1.' Diversity calculated by this method approximates normal distribution, and routine statistical inferences are thus justified (Bowman et al., 1970; Hutchenson, 1970). Generally, aquatic systems exposed to environmental stress (pollution) have fewer species and lower diversity indices than unstressed communities (Mathis, 1968; Odum, 1967; Odum, 1971). Several investigators have utilised species diversity indices to show biotic alterations in streams due to effluents. Harrel (1966) found a reduction in the species diversity of benthic macroinvertebrates below an effluent outfall and a progressive increase in diversity downstream. In Oklahoma, Wilhm & Dorris (1966), and Mathis & Dorris (1968) found similar situations in streams receiving domestic and industrial wastes, and oilfield brines. Few studies have thus far used fish diversity as a pollution indicator. This is probably due to the difficulties of sampling fish populations (Bechtel & Copeland, 1970).

MACROINVERTEBRATES AS INDICATORS OF ACID MINE POLLUTION

241

In aquatic ecosystems, the benthic macroinvertebrate community is most often investigated because of its lack of mobility and its sensitivity to physicochemical stresses. Benthic organisms provide a valuable indicator of past and present water quality conditions because of their long life histories and central position in the food chain (Mackenthum, 1966 ; Cairns & Dickson, 1971). In stream investigations, plankton are seldom utilised because they are carried long distances by currents and may reflect water quality conditions upstream rather than at the point of sampling (Mackenthum, 1966). In the present study, physicochemical conditions and the community structure of benthic macroinvertebrates were investigated from February 1970 to January 1971 in a drainage basin (Cane Creek) polluted with acid-mine drainage. A critical comparison was made of the geochemical nature of the polluted and non-polluted portions of the drainage basin. In addition, a statistical analysis was performed to show any correlation between water parameters and species diversity. The applicability of utilising the macroinvertebrate community structure to evaluate stream conditions consequent to acid flow is discussed.

DESCRIPTION OF STUDY AREA

Cane Creek, in Walker County, Alabama, drains approximately 25 km 2 in the Warrior Basin district of the Cumberland Plateau (Fig. 1). The main channel is approximately 9 km long and has an average gradient of 3.6 m/km (Hyde, 1970). In the system of drainage classification based on the degree of branching (Horton, 1945; Strahler, 1957), Cane Creek is ranked as a fourth order stream. Riffles dominate in the upper reaches, whereas pools are common in the lower reaches. Approximately 75 ~ of the Cane Creek watershed is forest, 20 ~ consists primarily of abandoned farmland and approximately 5 ~ has been strip-mined. During the period of study there was no active coal mining in the area; however, coal was mined extensively before 1955. A small impoundment on Black Branch, a tributary of Cane Creek, afforded an opportunity to study the effects of a lake on the water quality of an acidic stream. In July 1970, the lake had an average depth of 1.5 m and encompassed approximately 3.24 ha. Emergent vegetation, consisting of cattails (Typha angustifolia L.), spike rush (Eleocharis equisetoides (Elly) Torrey), and Erianthus giganteus Walt, covered approximately one-half of the lake surface. According to the lake classification proposed by Naumann (1921), the impoundment would be classified as a 'dystrophic type'; however, many dystrophic lakes may, in fact, be considered eutrophic due to the total productivity of algae present (Ruttner, 1968). The diatoms Achanthes minutissima and Frustulia rhomboides were present in such numbers as to form thick masses on the substrate of the impoundment.

242

GARY DILLS, DAVID T. ROGERS, JR

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DESCRIPTION OF STATIONS

Based upon preliminary measurements of stream pH and conductivity, taken during November 1970, ten stations were selected within the drainage system for the collection of physicochemical and benthic macroinvertebrate samples (Fig: 1). Some physical aspects of each station are presented in Table 1. Stations 3 and 4 were found to be similar in physical characteristics but were quite different in chemical properties. Preliminary measurements showed Cane Creek at station 3 to be a nearly neutral stream and to be low in soluble ions, even though the nearby area had been extensively strip-mined previously. The chemical composition of mine drainage may vary from area to area, depending upon local hydrogeologic conditions (Colmer & Hinkle, 1947; Hyde, 1970). The determining factor in the production of acid-mine effluents is the mineral composition of the rocks associated with the coal seams. Strata containing large quantities of pyrite minerals produce acidic drainage while those with minor quantities of iron sulphide

MACROINVERTEBRATES AS INDICATORS OF ACID MINE POLLUTION

243

TABLE 1 PHYSICAL CHARACTERISTICS OF TEN STATIONS IN CANE CREEK BASIN

Station

Width (m)

Depth (cm)

1

0-5-1-5

2

0.5-2.0

3 4

3-4 5-6

5

4-6

6

5-6

7

5-7

5-10

8

1-2

4-10

9 10

7-10 5-12

10-30 10-35

Substrate

5-10 Rocks and pebbles underlain with clay. Extensive precipitate* 5-10 Rocks and pebbles underlain with clay. Extensive precipitate 5-13 Sand and gravel 5-15 Sand, gravel, and debris. Precipitate 7-14 Rocks and pebbles underlain with silt and clay. Extensive precipitate 10-20 Silt and clay Rocks and pebbles with scattered areas of clay. Precipitate Coarse sand and rock underlain with clay Sand and gravel Sand and gravel

Current Sluggish

Stream margins Mine-tailings and mixed hardwood forest

Swift

Mine-tailings and pine forest

Moderate Moderate

Small trees and shrubs Mixed pine-hardwood forest Mixed pine-hardwood forest

Swift

Sluggish Swift

Sparsely vegetated with shrubs Mixed pine-hardwood forest

Swift

Small trees and shrubs

Sluggish Sluggish

Mature hardwood forest Mature hardwood forest

* Ferric hydroxide.

minerals often result in the drainage being nearly neutral or even alkaline (Hyde, 1970). In contrast to station 3, station 4 was distinctly acidic and high in soluble ions. A single adit (subsurface mine) was the only noted source of acid effluents on the tributary at station 4. Norris Branch (station 8), an intermittent stream, drained an unmined portion of the study area. The watershed consisted primarily of old fields in early successional stages, intermingled with pine tree stands.

TOPOGRAPHY AND GEOLOGY

The land surface in Cane Creek Basin is a submaturely dissected peneplane tilted slightly to the south. Remnants of the peneplane surface are preserved as small table-like uplands that are separated by steep-sided valleys. In the northern part of the basin, the hills rise to altitudes of over 215 m above mean sea level (msl) and the valley floor lies at an altitude of about 133 m above mean sea level. In the southern part of the basin, the hills rise about 170 m and the valley floor lies at about 100 an. Cane Creek Valley has a range in width of less than 100 m in the north to more than 450 m near the confluence of Cane Creek and Lost Creek. The drainage pattern in the basin is dendritic.

244

GARY DILLS, DAVID T. ROGERS, JR

The Pottsville Formation, of Pennsylvanian age and consisting of about 660 m of alternating beds of shale, sandstone and bituminous coal, surfaces at Cane Creek Basin and dips gently to the southwest, except where structural disturbances have caused minor variations. These rocks and associated coal beds often contain large quantities of iron sulphide minerals such as pyrite and marcasite (Hyde, 1970). The oxidised end-products of these minerals are the major source of acid effluents in waterways of the eastern United States (Barnes & Romberger, 1968).

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The average monthly temperatures in the Cane Creek area (Fig. 2) show wide fluctuations (US Weather Bureau, 1971). Rainfall averages 138era/year, but extended periods of drought are not uncommon. During the year of study, precipitation was unevenly distributed with monthly rainfall being greater than 8.5 cm in August and less than 3 em during November (Fig. 2).

MACROINVERTEBRATES AS INDICATORS OF ACID MINE POLLUTION

245

MATERIAL AND METHODS

Physicochemical From February 1970 to January 1971, bi-weekly water samples were collected at each station and transported to the laboratory in polyethylene containers for immediate analysis. Determination of dissolved-oxygen was made in the field, using a Precision Scientific oxygen analyser, and conductivity was determined with an Industrial Instruments conductivity bridge. Water temperature was measured with a mercury thermometer, and hydrogen-ion concentration was determined by a Sargent portable pH meter. All pH values, expressed as averages, were first converted to hydronium-ion values, averaged and then reconverted to pH values. Turbidity, phosphate, nitrate, silica, alkalinity, hardness, chromium and chloride were determined according to standard methods (American Public Health Association, 1960). Iron and manganese were measured colourimetrically in the field with a Hach Chemical Engineer's Kit. Measurements made with the Hach Kit were checked against standard laboratory methods for iron (Bipyridine Method) and manganese (Atomic Absorption Spectrophotometric Method) and were found to be accurate within +2.5 ~ and + 3 - 0 ~ , respectively. Water stage level markers were installed at each station to measure relative stream discharge. However, abnormal flooding during May 1970 destroyed the markers at three of the ten stations.

Biological A 0-1 m 2, fine-meshed Surber sampler (15 meshes/cm) was used to collect two bi-weekly macroinvertebrate benthic samples at each station from February 1970 to January 1971. The samples were collected from riffles in the same general vicinity as the water samples. Similar stream substrates were sampled and no two samples were taken from the same spot until at least two months had elapsed. Stones and gravel in the sample area were individually brushed with a paint brush to remove adhering organisms. The samples were preserved and stained in the field in a 5 oj~ formalin solution containing 100 mg/l rose bengal stain according to procedures proposed by Mason & Yevich (1966). In the laboratory the samples were washed in a No. 40 US standard soil sieve, separated from the debris, sorted into groups, identified, and enumerated. The invertebrates were then stored in vials containing 70 ~ ethanol. Taxonomic identification was made primarily from Pennak (1953) but the following publications were also used: Needham & Claassen, 1925; Curran, 1934; Ross, 1944; Needham & Westfall, 1955; Usinger, 1956; Edmondson, 1959. Verifications of invertebrate determinations were made by Herbert H. Ross, Trycoptera; Charles H. Nelson, Plecoptera; Frank N. Young, Coleoptera; Reece I. Sailer, Tabanidae; and George W. Byers, Tipulidae.

246

GARY DILLS, DAVID T. ROGERS, JR RESULTS AND DISCUSSION

Physicochemical Physicochemical data for the ten sampling stations are presented seasonally in Table 2. When the data were compared to the water quality criteria for public water supplies, as outlined by the FWCPA (1968), stations 3 and 8 were classified as 'unpolluted stations' while all other stations were classified as 'acidic stations'. Product-Moment correlation coefficients were calculated for all data for every pair of variables to assess the degree of relationship. The resulting intercorrelation matrix is shown in Table 3. By utilising the correlation matrix and seasonal data, a more meaningful discussion of these values and their geochemical significance to acid mine drainage could be presented. Maximum and minimum water temperature values occurred during August and January, respectively. The mean annual temperature values for all stations ranged from 14.5°C to 17.6°C. Station 2 had the highest average water temperature value because of the impoundment located on Black Branch. There was variability in turbidity values for both acidic and unpolluted stations. Stations 3 and 8, respectively, averaged twice and three times the turbidity values recorded at all stations except station 6. The high turbidity value at station 6 was attributed to nearby cultivated fields. Parsons (1955) ascribed low turbidity in acid streams to the precipitation of the normal silt load by the acid effluents. In addition to low turbidity, Black Branch and Shelton Branch were characterised by a hydroxide precipitate on the stream substrate. According to Barnes & Romberger (1968), the distribution of the precipitate is dependent upon the distance between where oxidation and neutralisation causes the ferric hydroxide to become stable in solution. Seasonally, turbidity values tended to be highest during summer and autumn and lowest during winter and spring (Table 2). As evidenced by the lack of measurable alkalinity, the natural buffering capacity of the acidic tributaries was probably eliminated by the acid effluents. Other workers have reported similar reductions or eradication of measurable alkalinity by acid effluents (Musser & Pickering, 1970; Parsons, 1955; Barnes & Romberger, 1968; Roback & Richardson, 1969). Station 6 did possess detectable alkalinity during the autumn and winter months. The average titratable alkalinities at stations 3 and 8 were 15.6 ppm and 3.1 ppm, respectively, with higher values measured during the summer months. The pH values for acidic tributaries varied inversely with the mineral content (r = -0.72) and showed seasonal trends; the lowest pH values were measured in summer and highest values were measured in winter. Species diversity and pH were positively correlated (r = 0.48) during the period of study (Table 3). Stream discharge, as determined by the stage level measure, was shown to be greatest during winter and spring (Table 2). Within the tributaries, pH values tended to vary directly with stream discharge while conductivity and hardness varied indirectly

MACROINVERTEBRATES AS INDICATORS OF ACID MINE POLLUTION

247

with stream discharge. These effects on the water quality probably reflect the dilution of the acid effluents by increased surface drainage into the streams. During the summer months, multiple factors, such as temperature, rainfall, and microbial oxidation may have produced the low pH readings. Warm summer temperatures would tend to enhance chemical and microbial oxidation of the acid-forming minerals, whereas decreased precipitation would permit the concentration of the acid effluents in surface and subsurface drainage. In addition, porous spoil-banks containing pyritic materials may have contributed to the base flow of the acidic streams by serving as ground-water reservoirs during the drier months. Corbett (1965) reported that spoil-piles from surface mining of coal in southwestern Indiana resulted in the production of significant amounts of stream flow during drought periods as compared with little or no yield from undisturbed areas. Mean annual conductance values for the acidic stations ranged from 1240micromhos/cm 2 at station 5, to 395micromhos/cm 2 at station 10 while values for the clean water stations (3 and 8) were 137 micromhos/cm 2 and 80micromhos/cm 2, respectively. Strong positive correlation existed between conductance and hardness (r = 0"96), iron (r = 0.78), manganese (r = 0.84), and sulphate (r = 0"94), while pH and conductance were strongly negatively correlated (r = - 0 . 7 2 ) . Bi-weekly measurement of conductance, reported as dissolved solids, and stream discharge were taken at stations 3 and 4 from February to June 1970. The dissolved solids and stream discharge data were related to surface area drained by each tributary and 9-2 kg/40 ha and 18"1 kg/40 ha of dissolved solids were found to be transported by the tributaries at stations 3 and 4, respectively, during the five-month study. Based upon these data, the chemical weathering of Shelton Branch drainage basin was twice as great as that of upper Cane Creek, even though the area adjacent to station 3 was the most recently mined area. The source of acid effluents in Shelton Branch originated from an extensive network of underground mines and probably from the leaching of minerals in the spoil-banks by acid effluents. Several investigators have reported similar increases in the leaching and solubility of heavy metals under low pH conditions (Decker, 1971; Cairns et al., 1971 ; Colmer & Hinkle, 1947; Pickering & Musser, 1970). Some investigators consider sulphate the best water parameter to use in identifying mine drainage because of its intimate role in pyrite oxidation, low concentration in natural waters, and relative inertness (Cairns et al., 1971 ; Brezina et al., 1970). According to Pickering & Musser (1970), the specific conductance measure may best reflect acid mine pollution in streams since it relates to both the geochemical character (ionic solute) of the water and the chemical weathering (dissolved solids) consequent to the acid drainage. As expected, dissolved-oxygen and temperature values were negatively correlated (r = - 0 . 6 8 ) . Slight variations in dissolved-oxygen measurements among the stations were attributed to differences in stream morphometry rather than to geochemical factors. Based upon the similarities in dissolved-oxygen measurements

TABLE 2

16'6 14.5 7"5 10.0

W Sp 4 S F X W Sp 5 S F ,X

30-1 29-0 25"1 28.2

18-0 16.6 11.3 14.0

30"0 29"0 25-2 28-0

W Sp 3 S F

W Sp 2 S F

W Sp 1 S F

3"8 3.6 3"2 3.3 3.4 3-9 3'6 3"3 3.5 3.6 6'6 6.7 6-3 6.6 6-5 4'3 3.8 3"0 3.2 3-3 3-8 3-4 2'8 3.0 3"1

Stage Station level p H (era)

7"6 6.8 4"9 6.1 6'3 9-2 7.9 6'2 7.3 7.6 9"8 8.3 4.4 6.3 7-2 10"4 8.8 5"8 6.7 7.9 10.8 7.6 6.7 7.9 8.3

573 945 1625 1058 1049 638 1060 1537 1107 1085 91 122 177 158 137 358 440 973 868 660 814 1085 1732 1331 1241

Dissolved Conductivity oxygen 194 291 296 312 298 238 317 423 390 342 35 45 55 54 47 152 166 321 327 241 396 309 447 411 391

0 0 0 0 0 0 0 0 0 0 10.4 9.0 23-0 20.1 15.6 0 0 0 0 0 0 0 0 0 0

Hardness Alkalinity

Water parameters

ALL VALUES ARE IN PPM, EXCEPT CONDUCTIVITY

6"4 15'2 25"7 15'0 14-5 6"8 18'9 28"8 16'0 t7'6 6-2 16.8 23-7 14"7 15-4 5'2 15"6 24"8 13.7 14"8 6"7 17"3 24'7 14.6 15"8

Temp. 19"3 27"4 22-7 20'3 22"0 17"4 18'3 5"7 14'3 13"9 11"2 11.5 11-0 11'0 11-2 19'8 24"8 37"9 33-0 28-9 20"7 26"4 30"2 27"2 26"1

Si 0"72 2"43 5'23 2"60 2"75 0"98 1"54 1'80 1"71 1"51 0"11 0-23 0"25 0-14 0"18 0"45 0"44 0'71 1"00 0"65 2"26 2'63 4'17 4"88 3'49

Fe CI 0'13 0'05 0'19 0 0"10 0"02 0"10 0"06 0 0"04 0 0'19 0"07 0"37 0"16 0'01 0-11 0-04 0 0-04 0'07 0'10 0-04 0 0-05

NO3 P04

3"02 4-2 0"82 4"50 3'9 0"12 5"87 4-2 0 " 0 1 5'70 4.8 0-04 4"60 4-2 0-24 2"60 3"7 0 " 1 1 4'02 3'6 0'01 5-54 3"8 0 " 0 1 4"97 4"3 0"07 4"30 3'8 0-04 0"57 4'6 0 0"43 3"7 0 0-26 4"9 0 0-30 5-7 0"10 0"38 4"7 0"03 2'48 4"6 0"09 2"13 4'3 0 " 0 1 5-88 5"0 0"01 7"17 4-9 0'02 4"40 4"7 0"03 2"48 4'1 0 " 1 1 3'60 4'0 0'04 7"32 4'0 0'19 7"65 4.4 0-46 5-26 4-2 0"20

Mn

(JTU )

401 523 813 709 611 433 524 723 759 610 44 46 43 68 50 241 225 554 582 400 454 575 927 900 714

0.07 0"01 0 0 0'02 0"04 0"02 0'01 0 0-02 0"04 0"02 0 0 0"02 0"03 0"01 0 0"01 0"01 0-03 0-01 0"01 0 0"01

S04 Cr

(W -- December-February; Sp = M a r c h - M a y ; S = J u n e - A u g u s t ; F = September-November) (micromhos/cm 2), pH, TEMPERATURE ( ° C ) , AND TURBIDITY

1970 TO

3"2 3-8 10-5 11-5 7'3 3"3 3"2 7"5 15"7 7"4 10"0 14"7 14"8 25"0 16"0 5"2 1"8 10'2 17"3 8"6 5-0 5"5 4'2 12"3 6'8

Turbidity

SEASONAL AND ANNUAL AVERAGES OF CERTAIN WATER PARAMETERS MEASURED AT TEN STATIONS IN CANE CREEK BASIN FROM FEBRUARY JANUARY 1 9 7 1

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MACROINVERTEBRATES AS INDICATORS OF ACID MINE POLLUTION

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MACROINVERTEBRATES AS INDICATORS OF ACID MINE POLLUTION

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at the acidic and non-acidic stations, dissolved-oxygen tie-up by oxidising salts associated with 'the acid effluents was not evident in this study. Concentrations of individual ions revealed both temporal and longitudinal variations along the length of the stream. During the summer and autumn, iron, manganese, and sulphate values for these ions at stations 9 and 10 were indicative of upstream precipitation of iron hydroxide and the downstream dilution of Cane Creek by the inflow of less acidic tributaries. Chloride, nitrate, phosphate, and chromium showed no seasonal trends at any station. As shown by the chemical analyses, the impoundment located between stations 1 and 2 exerted a considerable influence on the water quality of Black Branch (Table 2). Manganese, pH, chloride, sulphate, and chromium were little affected, while temperature at station 2 showed an average increase of 3.1°C when compared to station 1. The reduction in the mean annual iron content of Black Branch by 1.24 ppm between stations 1 and 2 was considered indicative of iron hydroxide precipitation in the impoundment. Probably the greatest effect upon water quality in the impoundment, as shown by the phosphate, nitrate, and silica analyses, was exerted by the biotic component. Based upon differences in silica content measured at stations 1 and 2, the depletion of silica within the lake during winter, spring, summer and autumn was 10, 30, 75, and 2 9 ~ , respectively. The high removal of silica during the summer may relate to increased metabolism in the phytoplankton in response to increased temperature. Silica depletion varied directly with temperature. Nitrate and phosphate at station 2 were reduced by 83 and 6 0 ~ , respectively, over station 1. The overall reduction may be due to the uptake of these nutrients by the phytoplankton and rooted aquatic plants present in the impoundment. Average monthly measures of dissolved-oxygen at stations 1 and 2 were 6-3 ppm and 7.7 ppm, respectively. This increase in dissolved-oxygen was attributed to the photosynthetic activity of the phytoplankton present in the impoundment. In brief, Cane Creek begins as a neutral stream in the headwaters, becomes acidic in the middle stretches, and then approaches neutrality in the lower reaches. The da~a indicate that Black Branch and Shelton Branch were the primary sources of continuous acid flow into Cane Creek. This was suggested by the proportionately greater amount of effluent ions measured at stations 1, 2, 4, and 5 and the progressive downstream decreases in the amounts of these ions at stations 7, 9, and 10. With the exception of station 3, those tributaries exposed to acid mine effluents were characterised by a lack of measurable alkalinity, a reduction in turbidity, a lower pH, and an increase in mineral content. Water parameters, measured at stations 3, 4, and 6, were compared in order to evaluate the effects of the inflow of an acidic tributary (Shelton Branch) on the water quality of a nearby neutral stream (Cane Creek). Below the inflow of the acidic stream, Cane Creek showed appreciable increases in hardness, iron, sulphate and manganese. All parameters, except pH, were highest during summer and autumn and lowest during winter and spring.

252

G A R Y DILLS, D A V I D T. ROGERS, JR

B E N T H I C M A C R O I N V E R T E B R A T E SPECIES

Seventy-five species in 72 genera were collected during this investigation. Diptera (23 species, 31 ~ ) and Coleoptera (10 species, 14 ~ ) dominated the numbers of species, while Plecoptera, Trycoptera and Ephemeroptera each contained 7 species (10 ~). The unpolluted stations (3 and 8) yielded the greatest number of species--61 and 49, respectively. Stations nearest the source of the effluents (1, 2, 4, 5, 6 and 7) averaged only 24 species. Stations 1 and 2, respectively, possessed 24 and 22 species. The lake, located between stations 1 and 2, did not appear greatly to affect the total number of species present, although alterations in the water quality of the stream were noted. Station 5 was lowest in total species present with only 20 species. The downstream stations (9 and 10) possessed relatively more species than the polluted upstream tributaries with 28 and 33 species, respectively. Station 4, considered to be near an acidic effluent source, possessed a total of 33 species. The disproportionate number of species at station 4, as compared to other acidic stations (24 species), may be related to the total amount of effluent ions present and not to the pH measure. The dissolved load at station 4, as determined by the mean annual conductivity measure, was approximately 3 9 ~ less than that measured at stations 1 and 2, and 50 ~ less than that at station 5, while pH values at these stations showed considerable similarities (Table 2). The Student t-test showed conductivity values measured at station 4 to be significantly different from those recorded at acidic station 1 (t c = 2.10; P < 0.05), station 2 (to = 3.40; P < 0.05) and station 5 (tc = 3.18; P < 0.05). Species diversity values showed little variation at the acidic stations (Table 4). TABLE 4 A COMPARISON OF TOTAL NUMBER OF SPECIES, SPECIES p n , AND CONDUCTIVITY VALUES MEASURED AT STATIONS 1, 2, 4, AND 5 IN CANE CREEK BASIN DURING FEBRUARY 1970 TO JANUARY 1971

Annual pH (X)

Site 1 Site 2 Site 4 Site 5

3.5 ± 3"6 ~ 3.6 ± 3.3 i

0.9 0.7 0.8 0.7

Annual conductivity (X)

1049 ± 1085 ± 660 -1241 ±

496 456 380 165

Species diversity (X)

Total No. of species

1"75 1.64 1"89 1"65

24 22 33 20

The previous comparison of pH and ionic content of the acidic streams lends credulity to the supposition that the ionic solute, rather than the pH, may exert the greatest influence upon the macroinvertebrate community composition. An attempt was made to discern a relationship between conductivity and total number of species, but no such conclusion could be drawn from the scatter diagrams constructed for this purpose. Warnick & Bell (1969) found heavy metals to be the most important parameter influencing the mortality of different species of aquatic

MACROINVERTEBRATESAS INDICATORSOF ACID MINE POLLUTION

253

insects. These workers further stated that analysis of the killed insects revealed that they had absorbed significant amounts of metal ions. There has been a tendency to ascribe great significance to the pH parameter in aquatic investigations (Hynes, 1970). Alone, pH reveals little of the chemistry of the water, but some dissolved elements may be directly harmful to aquatic life (Warren, 1971). Hynes (1970) further reports that, based upon field studies, it is difficult to distinguish the effects of high hydronium-ion concentration from other factors and most investigations lack any proof that acidity is the controlling factor. Regardless of these facts, many workers have attributed the composition and distribution of stream invertebrate communities to the effects of low pH (Agnew, 1962; Hall, 1951 ; Tarzwell & G aufin, 1953). The species composition was distinctly different at the polluted and non-polluted stations. Excluding stations 9 and 10, 17 species present at either stations 3 or 8 were absent at the polluted stations while only 5 species present at the acidic stations were found at stations 3 and 8. Fifteen species found at station 3 were not collected at station 8 while only 4 species found at station 8 were absent at station 3. Lirceus (43 ~o) formed large populations at stations 3 and 8; while dipteran populations (Tendipedidae) were proportionately greater at the acidic stations. According to Pennak (1953), two or more species of isopods are rarely found within the same

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Fig. 3. Statistics of species numbers for sampling stations in Cane Creek Basin. The solid line = range of values; the unshaded bars = standard deviation; the black bars = two standard errors to each side of the mean (vertical lines) ; and the numerical values = sample size.

254

GARY DILLS, DAVID T. ROGERS, JR

habitat; however, Lirceusfrontinalis (Raf) and Asellus militaris (Say) were collected in Norris Branch (station 8) during the spring of 1970. Generally, the unpolluted stations were characterised by a large number of species and individuals while the stations exposed to acid effluents had a relatively smaller number of species and individuals (Fig. 3). It is assumed that these differences are attributable, in part, to the presence of the effluent ions in the acidic streams. However, Hynes (1970) feels that the presence of iron hydroxide precipitate in acid streams is as important in eliminating aquatic species as pH. This effect was not measured in the present study. The low numbers of species and individuals in the acidic tributaries are characteristic of physically stressed communities (Odum, 1971). The benthic fauna at the acidic stations was dominated by chironomids, megalopterans, and Ceratopogonidae. Contrary to the results reported by Roback & Richardson (1969), Odonata, Plecoptera and Ephemeroptera benthic, insects were present under various acid-mine drainage conditions.

Species diversity Four stream bottom samples at each station, representing a sampled area of 0-4 m 2, were pooled to compute monthly species diversity indices for all stations in Cane Creek (Fig. 4). Monthly diversity (/~) values ranged from 0.79 at station 6

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Statistics of species diversity values for sampling stations in Cane Creek Basin. Explanation as in Fig. 3.

February March April May June July Time August September October November December 1971 January

1970

TABLE 5

1.18 0-89 1.44 2.11 1'79 2.51 1-60 1"75 1.47 1'75 1.53 1.64

1'64

1'75

2

1.93 1.67 1.48 1.04 1.54 1-80 1 "38 2.01 2'23 1.98 1.80 2'13

1

3-11

3'88 4.13 2.67 2"93 3.55 2.45 3.28 2.18 1.62 3.17 3.44 4.02

3

1.89

2.61 1"50 1.81 2.86 1.97 0.85 1"44 1.73 1-92 1.54 2-15 2-34

4

1.65

1.46 2"26 1.93 1.25 1.39 2.32 1-60 1.43 1.71 1.12 1.57 1.70

5

2.05

2-26 2-79 2.69 2.91 2.30 1.55 0-79 1.01 1-88 1.87 2-04 2.46

6

Stations

1.94

1.91 2"01 2.75 2.73 2.25 1-70 1-86 1.26 1.26 1.67 1.79 2.03

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2.68 3.44 2'49 2.61 3.08 2.79 1-98 dry 1.96 2.73 3.00 2-26

8

2-01

2-15 1-29 2.65 2"24 2.13 1-29 1-49 1-36 2.14 1.93 2.22 2.45

9

MONTHLY AND ANNUAL SPECIES DIVERSITY VALUES CALCULATED FOR TEN STATIONS IN C A N E CREEK BASIN

2.22

1.95 1.72 3'05 3.19 2"90 2.37 2-22 0.96 1.52 1.50 2-48 2.74

10

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256

GARY DILLS, DAVID T. ROGERS, JR

to 4.13 at station 3. Stations located near the areas of acid production were consistently lowest in diversity. Cairns et al. (1971) reported similar values when diversity measurements of a stream polluted by acid drainage were compared to 'healthy' mountain streams. The lowest mean annual diversity was recorded at station 2. On the basis of a t-test performed on species diversity values calculated for stations 1 and 2, the impoundment did not have a significant effect upon the community structure of the benthic macroinvertebrates (tc = 0.70; P < 0.25). Diversity values at stations 9 and 10 may have reflected a recovery response to the acid pollution in the downstream reaches. However, macroinvertebrate diversity tends to increase with increasing stream order (Harrel & Dorris, 1968) and the effects of self-purification of the acidic streams, and natural succession of species diversity were not identified. Stations 3 and 8 were previously classified as unpolluted stations; however, diversity indices of these stations revealed considerable differences in community complexity (Table 5). A t-test, performed on species diversity indices calculated for stations 3 and 8, showed these sites to be significantly different at the 5 % level (Fig. 4). The lower diversity at station 8 may be attributed to the damage to certain macroinvertebrate species by drought conditions during summer and autumn. Harrel & Dorris (1968) reported that drought conditions tended to upset and reduce community diversity in an Oklahoma stream. Other investigators of small streams found certain invertebrate species to be totally eliminated by drought (Hynes, 1941 ; Sprules, 1941 ; Hynes, 1958). Species diversity indices for station 8 tend to support the contention by Patrick (1970) that the rigorous and unpredictable stream environment is largely responsible for low equatibility and low diversity in benthic communities. The mean annual diversity at station 3 was indicative of a 'cleanwater' station (Wilhm, 1970). Monthly species diversity values for stations 3 and 8 showed temporal variations with highest values occurring during late March and early June (Table 5). Low diversity at these stations during early spring and autumn corresponded with the emergence pattern of many aquatic insects (Hynes, 1970; Mackay & Kaliff, 1968). Seasonal changes in species diversity in natural streams, such as Norris Branch, may be affected by variations in stream flow, the input of detrital material, or they may reflect life history patterns of the invertebrates (Clifford, 1969; Mackay & Kaliff, 1968). During autumn, the stream macroinvertebrate community may be low due to the hatching of eggs of those species that grow during the winter (Hynes, 1970). Species diversity values for the acidic streams showed no seasonal response, while the unpolluted stations showed lowest diversity values to occur during late summer (Fig. 5). In relation to community structure, systems of low diversity are usually less predictable than systems of high diversity (Margalef, 1969). Based upon statistical correlations and interpretations of geochemical associations, such as conductivity and manganese, hardness and pH, certain water parameters were suspected to be changing together and not to be totally independent of

257

MACROINVERTEBRATES AS I N D I C A T O R S OF A C I D MINE P O L L U T I O N

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Fig. 5. Monthly species diversity values determined for stations 3 and 5 from February 1970 to January 1971. Values presented on a 3-month period centred moving average as described by Croxton & Cowden (1960). each other. Therefore, to disclose the p a r a m e t e r s a n d / o r p a r a m e t e r interactions t h a t m o s t affected benthic diversity further, a step-wise regression analysis was p e r f o r m e d on the data. I n d e p e n d e n t variables were specified to be dissolvedoxygen, alkalinity, t e m p e r a t u r e , chloride, nitrate, p h o s p h a t e , p H , a n d t u r b i d i t y while species diversity was specified to be the d e p e n d e n t variable. A s s h o w n in T a b l e 6, p H , alkalinity, a n d p h o s p h a t e were highly c o r r e l a t e d ( P < 0.001, P < 0.01, P < 0.625, respectively) with species diversity; t h a t is these factors acting TABLE 6 ANALYSIS OF VARIANCE OF REGRESSION OF THE EFFECTS OF p~-{, ALKALINITY, AND PHOSPHATE ON SPECIES DIVERSITY

Source

Total Reg. (pH. alkalinity, phosphate) pH Alkalinity Phosphate Residual

DF

SS

219 3 1 1 1 216

134'75 49.51 18.17" 4.33* 2.60* 85.24

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16.51 I8.17 4.33 2,60 0'39

F

41.82 46.01 10.95 6.57

P

<0.001 <0.001 <0.01 <0.025

* Computed by step-wise regression; hence sum of squares for each variable was computed as if that variable were in the last position and, because of intercorrelations among these variables, the sum of squares for the parts will not total to the sum of squares for regression.

258

G A R Y D I L L S , D A V I D T. R O G E R S , J R

together may have some effect on species diversity. Due to the sampling regime, these data are probably correlated with each other through time and differences may not be as strong as they appear in analysis. An analysis of variance was performed on species diversity values for stations 3, 4, 6 and 7 to show possible similarities among the stations (Table 7). Station 3 was previously classified as an unpolluted station while 4, 6 and 7 were stations variously affected by acid effluents. Based upon this analysis, it was concluded that the means of the samples taken at stations 3, 4, 6 and 7 were meaningfully different at the 1 ~ level of significance. To show the stations that were significantly different from each other, the Least Significant Difference (LSD) test was performed on the means. On the basis of a 5 ~ level of significance, station 3 was found to differ from stations 4, 6 and 7 while stations 4, 6 and 7 were not significantly different TABLE 7 ANALYSIS OF VARIANCE AMONG SPECIES DIVERSITY VALUES MEASURED AT STATIONS CANE CREEK BASIN

3, 4, 6,

AND

Source o f variation

SOS

df

Mean square

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F.99

Total Between means Within samples (error)

21.85 12.25 9"16

47 3 44

4.08 0.22

16'75

4"31

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Annual mean values for species diversity (H) and mean number of species for stations 3, 6, 7, 9 and 10 in Cane Creek Basin from February 1970 to January 1971.

MACROINVERTEBRATES AS INDICATORS OF ACID MINE POLLUTION

259

from each other. The species diversity measure was thereby shown effectively to differentiate polluted from non-polluted stations. Diversity values and number of species for stations 3 and 6 best reflect the devastating impact of ionic concentrations (Shelton Branch) on the benthic community of a neutral stream (Fig. 6). Below the confluence of Shelton Branch with Cane Creek, species diversity and the total number of species were reduced by 53 and 68 %, respectively, when compared to the upstream diversity. Several critiques have recently been written concerning the widespread use of species diversity indices in lieu of species-number relationships in community ecology investigations (Warren, 1971; Hurlbert, 1971). A comparison of Figs. 3 and 4 shows the total number of species value most clearly to delineate the polluted

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260

GARY DILLS, DAVID T. ROGERS, JR

f r o m the n o n - p o l l u t e d stations. However, a mere c o m p i l a t i o n o f the n u m b e r o f species present in the acidic a n d non-acidic streams w o u l d have revealed little o f the seasonal v a r i a t i o n s in the a p p o r t i o n m e n t o f the individuals a m o n g the species. A s shown by Fig. 7, the n u m b e r s o f species at stations 3, 4 a n d 6 were consistent in value while the total n u m b e r o f individuals a n d the diversity measure (/~) showed seasonal variations. A s r e p o r t e d by Patrick (1970), the n u m b e r o f benthic rnacroinvertebrate species in streams often remains consistent even t h o u g h the n u m b e r o f individuals m a y vary considerably. Based u p o n evidence derived from this study, the a u t h o r s suggest that the diversity index (/~) should c o m p l e m e n t t a b u l a r analysis when an intensive investigation o f the aquatic c o m m u n i t y is in order. In addition, the diversity indices w o u l d be o f considerable value when a c o m p a r i s o n can be m a d e o f stressed a n d unstressed p o r t i o n s o f an aquatic system.

ACKNOWLEDGEMENTS

This research was s u p p o r t e d in p a r t by a research g r a n t from the Society o f Sigma Xi. T h e a u t h o r s express their t h a n k s to Joyce Dills and R o b e r t M a s o n who h e l p e d in s a m p l e p r e p a r a t i o n .

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CA1RNS, JOHN, JR, CROSSMAN,J. S., DICKSON, K. L. • HERRICKS,E. E. (1971). The recovery of damaged streams. A S B Bull., 18, 79-106.

CAmNS, JOHN, JR &DIcKSON,K. L. (1971). A simple method for the biological assessment of the effects of waste discharge on the aquatic bottom-dwelling organisms. J. 14~at. Pollut. Control Fed., 43, 755-72. CLIFFORD, H. F. (1969). Limnological features of a northern brown-water stream, with special reference to the life histories of the aquatic insects. Am. Midl. Nat., 82, 578-97. COLMER,ARTHURR. & H1NKLE, M. E. (1947). The role of micro-organisms in acid mine drainage: A preliminary report. Science, N. Y., 106, 253-8. CORBETT, DON M. (1965). Water supplied by coal surface mines, Pike County, Indiana. Water Resources Research Center Report o f Investigations, No. 1. CROXTON, F. E. & COWDEN,D. J. (1960). Practical business statistics. New York, Prentice-Halh CUMMINS, K. W. (1969). The stream ecosystem. Tech. Rep. Am. Ass. Adt~mt Sci., No. 7.

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