British Columbia Mine Reclamation Symposium

Reclamation of a limestone quarry to a natural plant community in the Rocky Mountains of southern Alberta Cohen-Fernández, A.C.; Naeth, M.A 2013

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  RECLAMATION OF A LIMESTONE QUARRY TO A NATURAL PLANT COMMUNITY IN THE ROCKY MOUNTAINS OF SOUTHERN ALBERTA   A.C. Cohen-Fern?ndez, PhD1 M.A. Naeth, PhD 1   1Department of Renewable Resources, Faculty of Agricultural, Life and Environmental Sciences, University of Alberta, 751 General Services Building, Edmonton, AB, T6G 2H1, Canada.  Reclamation of limestone quarries around the world is challenged by an extremely limiting environment, including steep slopes, scarce topsoil and high calcium carbonate substrates with low nutrients and water holding capacity. These issues were addressed at a limestone quarry in the Rocky Mountains of southern Alberta. Reintroduction of key components, such as vegetation and ameliorated soil were expected to speed recovery of ecosystem function. Season of seeding and soil amendment with manure mix, wood shavings and erosion control blankets were evaluated over two growing seasons to determine their effect on soil properties and native grass establishment. Season (fall, spring) of soil amending and seeding did not significantly affect revegetation or soil properties. Site characteristics such as slope, aspect, initial soil nutrients and surrounding plant communities influenced early plant community development and overall effects of soil treatments. Erosion control blankets resulted in highest seeded plant cover and lowest non-seeded plant cover despite not significantly changing soil chemical properties. Manure mix increased plant establishment, soil nutrients, microbial biomass and viable fungi and bacteria. Reclamation is postulated to be best with erosion control blankets and organic soil amendments like manure mix. Results from this work can be extrapolated to other limestone quarries or similar disturbances.   Key words: Amendments, Bacteria, Fungi, Microbial Biomass, Native Grasses, Seeding Season.    INTRODUCTION  Limestone, a sedimentary rock of mainly calcium carbonate, is widely used in a variety of industries including construction, metallurgic, pharmaceutics, food and agriculture, with an increasing worldwide production and demand. Limestone mining creates large disturbances that significantly impact soil, vegetation and fauna and result in habitat fragmentation and loss (Sort and Alca?iz 1996). Reclamation of limestone quarries, despite being an environmental necessity and mandatory in many countries, has had limited success, particularly when the goal is to achieve predisturbance conditions (Wunderle 1997).   Reclamation can succeed when soil is placed on rock spoil; however, soil for limestone quarry reclamation is scarce, coarse and nutrient deficient (Bradshaw and Chadwick 1980, Davis et al. 1985). Soil quality can be enhanced with organic amendments such as sewage sludge, biosolids and manure, which are plentiful, relatively inexpensive, readily available and reduce the need for landfill disposal. For example, sewage sludge increased plant biomass and cover in a Spanish quarry although species richness was lower than without sludge. These created substrates would be classified as anthroposols, azonal soils that have been highly modified or constructed by human activity, and are commonly built after disturbances as part of land reclamation activities (Naeth et al. 2012).  Reclamation practices can modify the composition and functioning of the soil microbial community. For example in limestone quarries, addition of sewage sludge increased biochemical and microbiological processes in the soils (Jimenez et al. 2007). Soil microorganisms are the primary agent for litter decomposition, nutrient cycling and energy flow in the soil ecosystem (Wardle 1992), with 80 to 90% of total soil metabolic activity carried out by fungi, bacteria and actinomycetes (Brady and Weil 2008). Biological diversity is used as an indicator of soil quality and ecosystem health because high species diversity may reflect high functional diversity (Brady and Weil 2008).   Slope and aspect of contoured embankments, depth and quality of soil covering the embankment, age of construction and surrounding vegetation are all factors affecting early ecosystem development. Thus effects of reclamation treatments must be evaluated within variable quarry structural features. The objectives of this research were to determine whether (i) fall and spring soil preparation and seeding, soil amendments and erosion control blankets would contribute to establishment of a desired plant community in limestone quarry reclamation, and whether (ii) substrate amelioration at the limestone quarry affected soil microbial biomass and number of viable fungi and bacteria colony forming units. These data could be used to determine whether ecosystem development was occurring in the reclaimed sites.   MATERIALS AND METHODS  Study Site Research was conducted at Graymont Exshaw limestone quarry near Kananaskis, Alberta, Canada (51? 07? N 115? 13? W). The quarry was located on a south facing slope, at 1,350 m elevation, below the 2000-2300 m tree line. Reclamation research treatments were established on three embankments in the quarry within 1 km of each other and one at the lime plant 7 km away. Embankments were engineered piles of limestone mine spoil, covered with clean fill (mixed topsoil and subsoil). Site New was on a newly built south facing embankment, approximately 188 m long and 7-15 m wide, with a 30 degree slope, and covered with 30 cm of clean fill. Site New HM was a 65 m long segment covered with 1-2 cm of horse manure. Site Old was on a south facing, older embankment, with 30 degree slopes of varying length and covered with 60 cm of clean fill. Site Plant was on a northwest facing berm with 15-20 degree slopes of 10-30 m lengths covered with 30 cm of clean fill. Climate is montane-subalpine. Long term mean annual precipitation is 296.2 mm as rain and 234.1 cm as snow. Mean daily temperature is 3 oC; maximum temperature is 21.9 oC in July and minimum is -14.1 oC in January.   Experimental Design In August 2007, 140 2.25 m2 (1.5 x 1.5 m) research plots were established, in a horizontal line, with 1 m buffers between plots. Half the plots at each site were randomly selected for fall and half for spring soil preparation and seeding. Within spring and fall treatments, soil treatments were randomly assigned. Each treatment, including a control, was replicated five times. New, Old and Plant sites had 2 seeding seasons x  4 soil treatments x 5 replicates = 40 plots. New HM treatments were erosion control blankets and a control for 2 seeding seasons x 2 soil treatments x 5 replicates = 20 plots.   Soil Treatments  Amendments that improved physical and chemical properties of limestone substrate in a previous greenhouse study, were locally available, and could be used without an environmental application permit, were evaluated in the field. Soil treatments including a control were used in combination with fertilizer, which improved plant establishment in the greenhouse and is a regular practice in other types of quarry reclamation when substrates have low to no nutrients. Erosion control blankets were used to physically protect soil and seeds. The erosion control blanket was Nilex SC150BN of coconut and straw. Beef manure mix was a 2 year old 6:1:1 mix of manure, waste feed and wood shavings. Wood shavings were fine screened from pine and white spruce wood. Manure was applied at 30 Mg ha-1 and wood shavings at 11.25 Mg ha-1, then incorporated into the soil at 5-10 cm with rakes and shovels. Slow release fertilizer (14-14-14 nitrogen, phosphorus, potassium) was applied at 1.1 Mg ha-1. Fall plots were amended in August 2007 and spring plots in May 2008. Fertilizer was applied at seeding and erosion control blankets after seeding.   Plant Species A native species mix for rapid establishment and tolerance of dry slopes, rocky sites and alkaline soils was seeded at 42.5 kg ha-1. Grasses were Poa alpina L. (alpine blue grass) 20%, Agropyron trachycaulum (Link) Maltex H.F. Lewis. (slender wheat grass) 15%, Elymus innovatus Beal. (hairy wild rye) 15%, Festuca saximontana Rydb. (rocky mountain fescue) 15%, Trisetum spicatum (L.) K. Richter variety ARC sentinel (spike trisetum) 15%, Bromus carinatus Hook. and Arn. 10% (mountain brome grass). The forb was Vicia americana Muhl. ex Willd. (american vetch) 10%. Fall plots were seeded October 8-10, 2007 and spring plots May 13-15, 2008.  Soil Property Quantification One 0-10 cm core from each of the 140 research plots was taken in May 2008 and August 2009, and analyzed individually. Samples were analyzed according to Carter (1993). Sodium adsorption ratio, soluble ions (chloride, calcium, potassium, magnesium, sodium and sulphate), pH and electrical conductivity were determined in 1:2 soil:water saturated paste. Total carbon and nitrogen were determined by combustion, total organic carbon by acid digestion then combustion, total inorganic carbon and total inorganic carbon calcium carbonate equivalent by acid digestion and total organic nitrogen by kjeldahl procedure.  Vegetation Assessment  Vegetation was assessed August 28-30, 2007, prior to plot establishment and soil treatment, since remnant vegetation from a failed seeding at Old and Plant sites and invading weeds were present in low numbers. Canopy cover by species was visually assessed in 20 x 50 cm quadrats at 10 random locations per site. After seeding, vegetation was assessed May 13-15, 2008 and 19-24, 2009 and August 11-15, 2008 and 17-24, 2009 in 0.5 x 0.5 m quadrats at the center of each of the 140 plots. Density by species and canopy and litter cover were determined. In August 2009, covers of seeded species were assessed. Density of seeded species was assessed to verify survival in August 2010 in spring plots.  Estimation of Microbial Biomass and Viable Fungi and Bacteria One soil sample from each of the 80 spring plots was collected for microbial biomass carbon and nitrogen assessments. Samples were collected with a shovel from 0-5 cm depth in August 2009. Samples were doubled bagged, stored in coolers with ice with final storage in a freezer (-20 oC). Microbial biomass carbon and nitrogen were estimated by chloroform fumigation-extraction (Vance et al. 1987). Extraction was done with 0.5 M K2SO4 solution. Analyses were performed using a total organic carbon / total organic nitrogen analyzer (Shimadzu). In August 2010, one soil sample for each spring plot was collected with a shovel from the 0-3 cm depth, stored at -20 oC until plating in September 2010. Serial dilution plate counts were used to determine total heterotrophic aerobic bacteria on plate count agar and fungi and actinomycetes on rose bengal - malt extract agar (Ottow and Glathe 1968). Four plates for each dilution of a soil sample were prepared by inoculating with 100 ?l volume of a given dilution. Plated dilutions for plate count agar were 10-5, 10-6 and 10-7. Plated dilutions for rose bengal - malt extract agar were 10-3,10-4 and 10-5. Plates were placed in a plastic bag to avoid water evaporation of the medium and incubated in the dark, at room temperature (21 oC), for 2 weeks. Colony forming units were cumulatively counted each week.   Statistical Analyses  All analyses were performed with SigmaPlot 12 (Systat Software 2011). Two way analysis of variance (ANOVA) and pair wise multiple comparisons (Holm Sidack method) were performed on soil data. Plant density and electrical conductivity data did not comply with normality and equality of variance assumptions, thus permutational ANOVA was performed with PERMANOVA v.1.6 (Anderson 2001) with season and amendment as fixed factors. Densities of seeded species due to seeding season were analyzed with Mann-Whitney rank sum test for most data and with T test for non-normal data. To identify differences in density of seeded species due to amendments and erosion control blankets at each site, fall and spring data were pooled and one way non parametric ANOVA used with amendment as the fixed factor and four levels (wood, blanket, manure, control). A posteriori pair wise comparisons with PERMANOVA were completed for treatments with significant differences. Differences in plant cover and density due to amendments and erosion control blankets were analyzed with one way ANOVA if data had normal distribution and equal variance and ANOVA on ranks otherwise.   RESULTS  Fall and spring soil preparation and seeding effects Soil chemical properties and plant density were not significantly different between spring and fall treatments in either study year. Although in 2008 total carbon at site Old (fall 6.88 ? 0.07 %, spring 6.99 ? 0.08 %) and electrical conductivity at site Plant (fall 0.29 ? 0.01 dSm-1, spring 0.48 ?0.08 dSm-1) were statistically different, values were similar and differences did not persist in 2009. Plant density was generally numerically higher in fall than spring treatments but only the Plant site had significantly more plants in fall than spring in 2008 (fall 85 ? 10 plants m-2, spring 52 ? 5 plants m-2). The statistical differences did not persist in 2009 (fall 83 ? 8 plants m-2, spring 66 ? 5 plants m-2).  Manure, wood shavings and erosion control blanket effects on soils and vegetation  Total nitrogen, carbon and organic carbon were significantly higher with manure than in controls. Total nitrogen and organic carbon were almost double that of controls in 2008 (Table 1) remaining higher in 2009 (Table 2). Electrical conductivity was more than twice that of the control in 2008, but a year later differences almost disappeared (Table 2). Total nitrogen, carbon and organic carbon did not follow a discernible trend over time after amendment with wood shavings, although total organic carbon was marginally higher at all sites than the control. Erosion control blankets had little effect on nitrogen and carbon relative to the control but concentrations were often significantly lower than with manure or wood shavings. Soil pH averaged 8, varying little with time and treatment.   Seeded plant density was generally higher with amendments than in controls (Figure 1). At the New site in 2008 and 2009 significant differences occurred, being up to twice as high with amendments as in the control. Wood shavings had less effect on plant density than manure and were often similar to controls but lower at the Old site both years. Erosion control blankets had a significant effect across sites and over time. In 2008 plant density was almost twice as high with erosion control blankets than the control at New and New HM sites; by 2009 values were over twice as high. Plant density increased in most treatments and sites from 2008 to 2009, except at the New HM site.   Soil treatment effects on soil microbial biomass, fungi and bacteria colony forming units Microbial biomass was higher in the amended soil than in the control, 15 months after amendment applications. Microbial biomass nitrogen in amended soil was numerically higher at sites New, New HM and Old but not at Plant (Figure 3). However, differences were not statistically significant. Values were similar across treatments and sites but the extreme values were found at site Old, ranging from 17.01 ug g-1 soil in the control to 47.24 ug g-1 soil with manure amendment, which often contained the higher values of microbial biomass nitrogen (Figure 3). Estimation of microbial biomass carbon in limestone soil by the total organic carbon analyzer was not reliable. The chloroform used in the procedure may have reacted with the calcium carbonate of the soil liberating carbon in the form of carbon dioxide and creating an overall reduction of carbon leading to negative values. Results are not included.   Twenty eight months after soil amendment, soil treatments had numerically higher numbers of viable fungi colony forming units at all sites compared to the unamended control (Table 3); values were not statistically different. In sites New and Old, more viable fungal colony forming units were present in soil with wood shavings; whereas in site New HM, more fungal colony forming units were present in the erosion control blanket treatment. Number of viable fungal colony forming units was similar among treatments at site Plant; however, the control counts were lower. Numbers of viable bacterial colony forming units was similar across soil treatments within the same site, but the range of values differed among sites (Table 3). Site New HM had the higher numbers regardless of soil treatment. Erosion control blankets and the control had similar mean values, around 81 x 106 colony forming units g soil-1. Sites New, Old and Plant ranged from 7 x 106 colony forming units g soil-1 to 55 x 106 g soil-1.   DISCUSSION The similar soil conditions and plant responses resulting from fall and spring soil preparation and seeding in our research were unexpected. Differences in soil temperature, water and nutrient availability in spring versus fall were hypothesized to significantly affect reclamation outcomes. The use of good quality amendments in fall soil preparation were expected to create better conditions for germination in spring, with increased water from snow melt being held in the organically amended substrate. Seeded species could take advantage of snow melt water to establish, before temperatures rise and soil surfaces dry in summer. Fall seeding benefits numerous species, although others are favoured by spring seeding (Kilcher 1961).   Modifications to both limiting physical and chemical properties of the substrates were important as evidenced by the favourable plant responses to manure, wood shavings and erosion control blankets. Plant density increased with treatments relative to controls, particularly with manure and erosion control blankets. Regardless of the quarry site, these physical and chemical modifications were important to reclamation success. Erosion control blankets provided a more stable substrate wherein plants could find suitable microsites for establishment on a dry, exposed surface subject to harsh temperatures, drought and wind. Nutrient uptake by the significantly higher number of established plants and decomposition by microorganisms may explain lower concentrations of total carbon, organic carbon and nitrogen in this treatment.   Manure improved substrate chemical properties, such as increased total nitrogen and organic carbon. From this and other studies, it is evident manure will improve low quality reclamation soils with low organic matter and nutrients and nutrients will remain high a few years after application. In our study, even though manure was added at a moderate rate, it increased nitrogen which remained high at the end of year 2. This becomes very important for limestone quarry reclamation, where chemical reactions among highly concentrated carbonates and high pH of the soil results in immobilized iron oxides, increased nitrification and limited available phosphorus due to phosphate adsorption to carbonate minerals or insolubilization. Nutrient limitation likely hindered vegetation in the unamended control. Beneficial results of manure increasing nutrients and plant density were also found in a greenhouse study with amended limestone materials (Cohen-Fern?ndez and Naeth in press). Manure application is well known to significantly increase electrical conductivity in soils (Hao and Chang 2003). However, it had dramatically decreased one year after application, with all values remaining below the 2 dS m-1 threshold for plant establishment (Soil Quality Criteria Working Group 1987). Although soil water input on site was relatively low, sufficient leaching took place for these salts to be flushed from the system, perhaps as a result of the water from snow melt. Plant performance with wood shavings was lower than with manure, similar to results in greenhouse experiments (Cohen-Fernandez and Naeth in press). At high rates wood products such as sawdust can reduce nitrogen mineralization and competitive ability of species with higher nitrogen requirements. This likely happened at Exshaw. Wood shavings would have lower water holding capacity than manure and in general provide less desirable sites for germination.   Continued decomposition of organic matter and dead microorganisms likely resulted in mineralization and an increase of available nitrogen. Manure treatment resulted in higher plant density and more vigorous plants. Even after 15 months, higher microbial biomass nitrogen and significantly higher total nitrogen and total organic carbon was found in manure treatments. High microbial biomass nitrogen at site Plant in wood shavings treatments was probably associated with high decomposition rates due to more soil water because of its north facing position. North facing slopes commonly have more water (Hanna et al. 1982)  which is important for microbial decomposition of organic matter and thus increases microbial population and biomass (Kieft et al. 1987). Microbial activity at the time of measurements had likely already decomposed the more readily decomposable fraction of the amendments and mineralized some nutrients which were utilized by vegetation. This is supported by the more lush and dense vegetation at Plant site.   The number of organisms in the quarry soil, despite being highly disturbed, corresponds to a range described by other researchers (104 to 105) (Beck et al. 2005). Although only a small number of bacteria in soil can be cultured by standard isolation techniques it gives an indication of differences in detectable microorganisms caused by treatment. High pH and calcium carbonate of a limestone quarry site are more favourable for bacteria than fungi. This partially explains the higher bacterial (colony forming units) counts compared to fungal counts at all locations within the overall site.  Wood shavings in the soil favoured the fungal community at south facing, exposed sites. The cultured organisms were predominantly molds, because wood shavings have lignocellulose as one component and various fungi are able to degrade this material (Rodriguez et al. 1996). Erosion control blankets favoured fungi likely because of straw and shade. Manure treatments did not cause a significant increase in colony forming units. Site New HM had conditions for more bacteria colony forming units. This could result from bacteria inherent in the horse manure or by more organic matter and better chemical and physical conditions for local bacteria to establish.  Variability in number of viable colony forming units of bacteria and fungi increased under more exposed conditions. More favourable north facing sites resulted in more stable (similar numbers) fungi and bacteria colony forming units across amended treatments and controls. More than a third of the morphotypes were actinomycetes. They may become more dominant during later stages of decay when easily metabolized substrate has been used (Brady and Weil 2008). Soil at Plant site had higher water content regardless of soil treatment and there was a small wetland at the bottom of the berm. Wetter conditions at the site and addition of amendments may have resulted in an increase of bacteria populations. More rapid mineralization of nutrients in amendments and dead microorganisms were used by plants and therefore removed from the soil.  REFERENCES  Anderson, M.J., 2001, A new method for non-parametric multivariate analysis of variance, Austral Ecology, 26, 32-46. Beck, L., J. R?mbke, A.M. Breure and C. Mulder, 2005, Considerations for the use of soil ecological classification and assessment concepts in soil protection. Ecotoxicology and environmental safety,. 62, 189-200.  Brady, N.C. and R.R. Weil, 2008, The nature and properties of soils. Fourteenth edition. Pearson Prentice Hall. Upper Saddle River, New Jersey. 975 pp.  Bradshaw, A.D. and Chadwick, M.J., 1980, The restoration of land: The ecology and reclamation of derelict and degraded land, University of California Press, Berkeley, California, 317 pp.   Carter, M. R., 1993, Soil sampling and methods of analysis, Lewis Publisher, Boca Raton, Florida. Cohen-Fernandez, A.C. and Naeth, M.A. In press. Anthroposol development from limestone quarry substrates, Canadian Journal of Soil Sciences. Cohen-Fernandez, A.C. and Naeth, M.A., 2013, Erosion control blankets, organic amendments and site variability influenced the initial plant community at a limestone quarry in the Canadian Rocky Mountains. Biogeosciences Discussion, 10, 3009?3037. Davis, B.N.K., Lakhani, K.H., Brown, M.C. and Park, D.G., 1985, Early seral communities in a limestone quarry: an experimental study of treatment effects on cover and richness of vegetation, Journal of Applied Ecology, 22, 473-490.   Hanna, A.Y., Harlan, P.W. and Lewis, D.T., 1982, Soil available water as influenced by landscape position and aspect, Agronomy Journal, 74, 999-1004.  Hao, X. and Chang, C., 2003, Does long-term heavy cattle manure application increase salinity of a clay loam soil in semi-arid southern Alberta?, Agriculture, Ecosystems & Environment, 94, 89-103. Jimenez, P., O. Ortiz, D. Tarrason, M. Ginovart and M. Bonmati, 2007, Effect of differently post-treated dewatered sewage sludge on B-glucosidase activity, microbial biomass carbon, basal respiration and carbohydrates contents of soils from limestone quarries, Biology and Fertility of Soils 44, 393-398.  Kilcher, M.R., 1961. Fall seeding versus spring seeding in the establishment of five grasses and one alfalfa in southern Saskatchewan, Journal of Range Management, 14, 320-322.  Kieft, T.L., E. Soroker and M.K. Firestone, 1987, Microbial biomass response to a rapid increase in water potential when dry soil is wetted. Soil Biology and Biochemistry, 19, 119-126.  Larney, F.J., Buckley, K.E., Hao, X. and McCaughey, W.P., 2006, Fresh, stockpiled, and composted beef cattle feedlot manure, Journal of Environmental Quality, 35, 1844-1854.  Naeth, M. A., Archibald, H. A., Nemirsky, C. L., Leskiw, L. A., Brierley, J. A., Bock, M. D., VandenBygaart, A. J., and Chanasyk, D. S., 2012, Proposed classification for human modified soils in Canada: Anthroposolic order. Canadian Journal of Soil Sciences, 92, 7-18. Ottow, J.C. and H. Glathe., 1968, Rose bengal-malt extract-agar, a simple medium for the simultaneous isolation and enumeration of fungi and actinomycetes from soil. Applied Microbiology, 16 ,170-171.  Rodriguez, A., F. Perestelo, A. Carnicero, V. Regalado, R. Perez, G. de la Fuente and M. Falcon., 1996, Degradation of natural lignins and lignocellulosic substrates by soil-inhabiting fungi imperfecti, FEMS Microbiology Ecology, 21, 213-219.  Saviozzi, A., Levi-Minzi, R., Riffaldi, R. and Vanni, G., 1997, Laboratory studies on the application of wheat straw and pig slurry to soil and the resulting environmental implications, Agriculture,  Ecosystems and Environment, 61, 35-43. Soil Quality Criteria Working Group., 1987, Soil quality criteria relative to disturbance and reclamation, Alberta Agriculture, Food and Rural Development, Edmonton, Alberta.   Sort, X. and Alca?iz, J.M., 1996, Contribution of sewage sludge to erosion control in the rehabilitation of limestone quarries, Land Degradation & Development, 7, 69-76. Systat Software., 2011, SigmaPlot for Windows version 12, Systat Software Inc. Chicago, Illinois. Vance, E.D., P.C. Brookes and D.S. Jenkinson, 1987, An extraction method for measuring soil microbial biomass C. Soil Biology and Biochemistry, 19, 703-707.  Wardle, D.A., 1992, A comparative assessment of factors which influence microbial biomass carbon and nitrogen levels in soil. Biological Reviews, 67, 321-358.    Table 1. Mean measured soil properties in soil treatments in 2008. Site Treatment Total Carbon (%) Total Organic Carbon (%) Total Nitrogen (%) Hydrogen Ion Activity (pH) Electrical Conductivity         (dS m-1) New Control 6.81 ? 0.05 b 0.68 ? 0.02 b 0.06 ? 0.00 b 8.17 ? 0.04 0.18 ? 0.01 b  Blanket 6.74 ? 0.03 b 0.61 ? 0.04 b 0.05 ? 0.00 b 8.11 ? 0.02 0.22 ? 0.02 b  Manure 7.25 ? 0.09 a 1.18 ? 0.09 a 0.10 ? 0.01 a 8.17 ? 0.02 0.50 ? 0.11 a  Wood 7.10 ? 0.10 a 1.01 ? 0.08 a 0.06 ? 0.00 b 8.14 ? 0.01 0.17 ? 0.01 b New HM Control 7.73 ? 0.15 1.45 ? 0.17 a 0.18 ? 0.02 a 8.25 ? 0.03 0.48 ? 0.03   Blanket 7.23 ? 0.13 0.83 ? 0.11 b 0.10 ? 0.01 b 8.22 ? 0.02 0.38 ? 0.05 Old Control 6.73 ? 0.10 b 0.74 ? 0.09 b 0.06 ? 0.00 b 8.16 ? 0.02 0.18 ? 0.00 c  Blanket 6.81 ? 0.09 ab 0.70 ? 0.09 b 0.07 ? 0.00 b 8.16 ? 0.03 0.25 ? 0.02 b  Manure 7.13 ? 0.09 a 1.16 ? 0.08 a 0.11 ? 0.01 a 8.21 ? 0.03 0.56 ? 0.11 a  Wood 7.05 ? 0.10 ab 1.09 ? 0.08 a 0.07 ? 0.00 b 8.24 ? 0.02 0.19 ? 0.00 c Plant Control 7.69 ? 0.21 1.46 ? 0.13 b 0.10 ? 0.01 b 8.29 ? 0.01 0.30 ? 0.02 b  Blanket 7.68 ? 0.19 1.41 ? 0.12 b 0.08 ? 0.01 b 8.27 ? 0.01 0.30 ? 0.03 b  Manure 8.06 ? 0.25 2.59 ? 0.46 a 0.16 ? 0.01 a 8.27 ? 0.01 0.70 ? 0.14 a  Wood 7.67 ? 0.18 1.57 ? 0.14 b 0.10 ? 0.01 b 8.31 ? 0.01 0.26 ? 0.02 b Values are mean ? standard error. Values of ? 0.00 in the table were <0.006. Letters indicate statistically significant differences among treatments in a site. Absence of letters indicates no statistical differences. Table 2. Mean measured soil properties in soil treatments in 2009. Site Treatment Total Carbon (%) Total Organic Carbon (%) Total Nitrogen (%) Hydrogen Ion Activity (pH) Electrical Conductivity         (dS m-1) New Control 7.15 ? 0.07 b 0.93 ? 0.03 b 0.07 ? 0.00 b 8.18 ? 0.07 0.33 ? 0.04  Blanket 7.10 ? 0.07 b 0.90 ? 0.02 b 0.08 ? 0.01 b 8.19 ? 0.05 0.33 ? 0.05  Manure 7.53 ? 0.11 a 1.22 ? 0.07 a 0.11 ? 0.01 a 8.07 ? 0.05 0.38 ? 0.04  Wood 7.45 ? 0.10 a 1.05 ? 0.04 ab 0.09 ? 0.01 b 8.02 ? 0.05 0.39 ? 0.04 New HM Control 7.49 ? 0.18 1.25 ? 0.10 0.11 ? 0.01 8.1 ? 0.08 0.34 ? 0.03  Blanket 7.39 ? 0.22 1.14 ? 0.11 0.12 ? 0.01 8.06 ? 0.06 0.34 ? 0.02 Old Control 6.96 ? 0.05 0.86 ? 0.03 b 0.07 ? 0.00 8.2 ? 0.06 0.32 ? 0.03  Blanket 7.31 ? 0.21 0.99 ? 0.04 b 0.07 ? 0.01 8.03 ? 0.06 0.36 ? 0.06  Manure 7.28 ? 0.17 1.16 ? 0.10 a 0.10 ? 0.01 8.15 ? 0.03 0.32 ? 0.04  Wood 7.28 ? 0.05 1.08 ? 0.03 ab 0.08 ? 0.01 8.1 ? 0.04 0.31 ? 0.04 Plant Control 8.56 ? 0.22 1.26 ? 0.07 0.07 ? 0.00 b 8.18 ? 0.07 0.30 ? 0.02  Blanket 9.08 ? 0.33 1.26 ? 0.05 0.08 ? 0.00 b 8.16 ? 0.04 0.32 ? 0.04  Manure 9.60 ? 0.42 1.41 ? 0.09 0.11 ? 0.01 a 7.97 ? 0.09 0.39 ? 0.05  Wood 8.96 ? 0.40 1.33 ? 0.11 0.08 ? 0.00 b 7.99 ? 0.08 0.40 ? 0.07 Values are mean ? standard error. Values of ? 0.00 in the table were <0.006. Letters indicate statistically significant differences among treatments in a site. Absence of letters indicates no statistical differences. .   Table 3. Colony forming units (CFU) of fungi and bacteria in amended soil in August 2010.    Site    Soil Treatment New  New HM  Old  Plant  x 10 4 Fungi CFU g-1 soil  Control  14.61 ? 5.73 12.73 ? 5.77  4.66 ? 1.63 14.34 ? 3.65 Blanket  15.45 ? 14.54 23.7 ? 1.6   9.82 ? 5.84 17.4 ? 4.43 Manure  19.16 ? 3.01   13.94 ? 3.29 20.22 ? 8.7 Wood 44.49 ? 16.69   40.07 ? 15.29 18.79 ? 14.91   x 10 6  Bacteria CFU g-1 soil  Control  32.81 ? 31.06 81.47 ? 41.53 10.9 ? 0.83 39.91 ? 18.07 Blanket  11.52 ? 10.01 82.88 ? 9.11 25.7 ? 11.77 49.26 ? 10.99 Manure  7.34 ? 7.01   41.76 ? 6.83 55.18 ? 15.49 Wood 16.79 ? 13.87   45.9 ? 31.68 39.17 ? 26.53 Values are mean ? standard error.  Different letters indicate statistically significant differences among soil treatments of the same site. Absence of letters indicates no statistical differences.  Site New0204060801001202008 2009 Site New HMSite OldControl Blanket Manure Wood020406080100120 Site PlantControl Blanket Manure WoodabaaA abbcAbabbSoil TreatmentBABAABBbAbcAAaAA aAASeeded plant density (plants per square meter) Figure 1. Mean seeded plant density at each soil treatment and site in 2008 and 2009. Letters indicate significant differences among soil treatments in the same year; upper case letters for 2008 and lower case letters for 2009. Bars indicate standard error.    Soil Treatment Plant cover (%)0102030405020082009Site New Site New HMSite BControl Blanket Manure Wood01020304050Site OldControl Blanket Manure WoodSite PlantbbbAabaaaABABBABAAAAAAaaa aaaaaaa Figure 2. Mean plant cover at each soil treatment and site in 2008 and 2009. Letters indicate significant differences among soil treatments in the same year; upper case letters for 2008 and lower case letters for 2009. Bars indicate standard error.  Microbial biomass nitrogen (ug g soil-1 )010203040506070Soil TreatmentControl Blanket Manure Wood010203040506070Control Blanket Manure WoodSite New Site New HMSite Old Site Plant  Figure 3. Mean microbial biomass nitrogen in soil samples from spring plots at the quarry and plant sites in samples collected in August 2009. Bars indicate standard error. Absence of letters indicates no statistical differences. 

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