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Water Air Soil Pollut (2013) 224:1471

DOI 10.1007/s11270-013-1471-y

Short- and Long-Term Effects of Modified Humic Substances


on Soil Evolution and Plant Growth in Gold Mine Tailings
Carl Szczerski & Christian Naguit & John Markham &
Tee Boon Goh & Sylvie Renault

Received: 28 August 2012 / Accepted: 4 February 2013


# Springer Science+Business Media Dordrecht 2013

Abstract Mining creates large amounts of processed species cover increased with amendment rate in the
waste in the form of mine tailings. Sulfide mine tailings 2003 experiment but not the 2004 experiment, most
are of particular concern due to the biotic and abiotic likely due to lower pH in the latter. Our results suggest
oxidation of sulfide minerals that release acidity and that short-term patterns of plant performance do not
metals into the environment. Revegetation can be reflect longer-term performance or invasion by volun-
employed to mitigate the spread of tailings in the envi- teer plant species. Our long-term data suggest that hu-
ronment. Revegetation often involves ameliorating tail- mic amendments can be effective in establishing plant
ings with organic materials to promote plant growth and invasion of mine tailings, although the effects vary
improve tailings physicochemical structure. We depending on the pH of the tailings.
amended plots in the Central Manitoba Mine tailings
pond with humic substances applied at rates up to 4 g Keywords Humic substances . Tailings amendment .
C kg−1 through roto-tilling and seeded with Medicago Medicago sativa . Elymus trachycaulus . Plant growth .
sativa and Elymus trachycaulus in 2003 and 2004. The Land reclamation . Tailings structure
humic substances improved tailings fertility by increas-
ing macro aggregation, organic carbon, and macronutri-
ents but also resulted in a short-term increase in 1 Introduction
electrical conductivity levels. In the first growing season
the humic amendment had little effect on plant yield, While the economic benefits of mining are apparent,
except in the 2003 experiment where the yield of E. mining also creates large amounts of processed waste
trachycaulus decreased by 84 % with 4 g C kg−1 amend- material in the form of tailings. Tailings associated
ment. After 7 years, the addition of humic amendment with sulfur bearing metals are particularly problematic
resulted in a cover of over 38 % for M. sativa, compared due to the biotic and abiotic oxidation of these sulfide
to less than 2 % in control plots. In addition, non-seeded minerals releasing acidity and metals into the environ-
ment (Blowes and Ptacek 1994). This phenomena
C. Szczerski : C. Naguit : J. Markham : S. Renault (*)
known as acid mine drainage (AMD) has been identi-
Department of Biological Sciences, University of Manitoba, fied as the most serious environmental problem facing
Winnipeg, Manitoba, Canada R3T 2N2 the global metal mining industry. Prior to adequate
e-mail: renaults@cc.umanitoba.ca legislation, in most jurisdictions little was required in
terms of environmental reclamation or mitigation
T. B. Goh
Department of Soil Science, University of Manitoba, plans and many older mine sites became abandoned
Winnipeg, Manitoba, Canada R3T 2N2 or orphaned with little treatment.
1471, Page 2 of 14 Water Air Soil Pollut (2013) 224:1471

Cost effective reclamation for acidic tailings sites (Livens 1991). In addition, humic substances have been
typically involves tailings stabilization to prevent con- shown to have direct stimulatory growth effects on
taminated particle movement off site by wind and plants including increased photosynthesis and auxin like
water, exposure of new oxidizable material, and to activity (Vaughan 1969; O’Donnell 1973). However,
reduce or eliminate AMD caused by water and oxygen positive effects have varied with outcomes based on
movements into and through the tailings impound- the concentration and sources of the humic substances
ment (Ripley et al. 1996). The majority of low cost used (Piccolo et al. 1993). Reduced growth has also
techniques focus on the establishment of vegetation in been shown with humic substances. It has been
combination with or without chemical, biological, or suggested that high concentrations can lead to increased
physical treatments to aid in tailings stabilization. heavy metal uptake causing toxicity (Whiteley and
Selected vegetation typically consists of commercially Williams 1993) and that micronutrient deficiencies can
available, native, or metal tolerant species (Gardner et be caused by a tight binding of micronutrients reducing
al. 2012). In addition, revegetation can also be thought their availability (Pertuit et al. 2001).
of as a successional process where the initial vegeta- The objectives of this study were: (1) to assess the
tion consists of pioneer species that allow surrounding short-term (1–2 years) and long-term (6–7 years) ef-
native plants to colonize the site. The primary goal of fects of modified humic substances on tailings physi-
the initial stages of revegetation then is to establish cochemical properties, (2) to study the growth and
vegetative cover that requires little to no maintenance elemental content of plant species seeded in tailings
in order to aid in the prevention of erosion and to build amended with modified humic substances in the short
up organic material in preparation for more permanent term, and (3) to determine if the amendment of tailings
vegetation (Ripley et al. 1996). with modified humic substances will facilitate the
The successful establishment of vegetation may be invasion of other plant species years after the initial
limited by the physical and chemical properties associ- seeding. It was hypothesized that modified humic sub-
ated with sulfide containing tailings. These properties stances would promote soil development in terms of
include potentially toxic levels of heavy metals, high aggregation, increase the carbon content of the tail-
salinity, low pH, macronutrient deficiencies and a low ings, stimulate overall growth of the selected species,
organic carbon content contributing to relatively poor and initiate succession by promoting the invasion of
soil structure, and a high degree of surface compaction other plant species.
(Ripley et al. 1996; Tordoff et al. 2000). To facilitate
plant growth, the addition of organic amendments is
often required (Tordoff et al. 2000; Green and Renault 2 Materials and Methods
2008). Lignite is one possible source of humic sub-
stances and has been shown to significantly improve 2.1 Site Description
the carbon content and structure of mine tailings
(Ibrahim and Goh 2004). Humic substances are known The Central Manitoba Gold mine tailings impoundment
to increase water and nutrient holding capacity, water is located in southeastern Manitoba, in Nopiming
permeability, decrease soluble metal concentrations by Provincial Park (50°54′16″N, 95°20′6″W). The tailings
complexing with metals and play a role in the formation cover ca. 20 ha and vary in depth from ~5 m to greater
of soil structure including stable soil aggregation than 25 m (Sherriff et al. 2009). Although the tailings
(Stevenson 1982; MacCarthy 2001). Humic substances were deposited in the 1920s–1930s, less than 0.1 % is
also play a number of other beneficial roles in soils covered with vegetation. While biotic and abiotic oxi-
including pH buffering capacity, binding of charged dation of the sulfide primary minerals pyrite (iron sul-
toxic ions, increasing the soil cation exchange capacity, fide), chalcopyrite (copper–iron sulfide) and pyrrhotite
serving as a reservoir for nutrients (nitrogen, phospho- (iron sulfide) have led to the release of metals like iron
rus, and potassium) and retaining soil moisture and copper and generate acidity, the presence of calcite
(MacCarthy 2001). Furthermore, the ability of humic in the tailings has had a buffering effect (Londry and
substances to remove metal ions from the soil solution Sherriff 2005). As a result, the tailings pH has been
and decrease their bioavailability make them extremely shown to range from 3.5 to 7 (Renault et al. 2002;
useful for reclamation of heavy metal contaminated land Londry and Sherriff 2005) with only ~20 % of the total
Water Air Soil Pollut (2013) 224:1471 Page 3 of 14, 1471

tailings area possessing a pH<4 due to the exhaustion of The plots for each run (year) of the experiment were
calcite by a higher local load of sulfide minerals (Londry about 10 m apart and the tailings in both locations,
and Sherriff 2005). Given the complexity of mine tail- appearing to be similar in colouration (i.e., composition
ings composition, some elements were selected for ele- and oxidation state), were relatively free of surface salt
mental analysis based on a previous study of these crusting present in some areas of the tailings pond, and
tailings (Green and Renault 2008). Previous analysis had no naturally established vegetation. For each run of
indicated copper levels far exceeding the normal range the experiment, four blocks (5.2×0.7 m) were
in soil while iron levels fall within the normal soil range established each with three 1.4×0.7 m plots (separated
(Kabata-Pendias and Pendias 1992). More information by a 0.5-m buffer strip) and amended with different
on the tailings elemental composition and their potential levels modified humic material (0, 7.5, and 15.0 g
impact on vegetation is provided by Green and Renault amendment kg−1 tailings). These rates were used in
(2008). order to reach concentrations of 0, 2 and 4 g carbon
kg−1 tailings, respectively, and were based on prelimi-
2.2 Experimental Design nary experiments and results from Ibrahim and Goh
(2004). In order to calculate the rate of amendment
A tailings amendment experiment was conducted twice, and fertilizer addition, density of the tailings was esti-
once in the spring 2003 and again 2004. This repetition mated using a tapped density method in which dry
was considered important since our previous work had tailings were shaken in a graduated cylinder, allowed
shown spring growing conditions vary from year to year to settle and weighed. To incorporate the amendments,
and have a strong impact on plant performance. The the tailings treatment blocks, including the 0 gkg−1 plot,
amount of precipitation (rainfall and snowfall) during were first roto-tilled to a depth of 15 cm. The amend-
each year varied substantially, with 512 mm in 2003 ment was then added to the surface of the tailings and
and 843 mm in 2004. The average precipitation for the tilled again.
region is 558 mm. Meteorological data were obtained Following a 4-week incubation, two seeding subplots
from the National Climate Data and Information Archive (0.5×0.5 m) within each amendment plot were random-
website (http://www.climate.weatheroffice.gc.ca). ly selected and seeded with either Medicago sativa
The humic amendment was a water soluble modified (alfalfa) or Elymus trachycaulus (slender wheatgrass).
compound (Dry soluble 80™) from BlackEarth Humates The plant species were selected based on their
Ltd. (Edmonton, Alberta, Canada). It was processed from established use in tailings reclamation, and tolerance to
a lignite (humalite) deposit. The humic substances salinity, ability to control erosion and fix nitrogen (in the
contained greater than 80 % humic and fulvic acids as case of M. sativa). Brett Young Seeds Limited supplied
well as non-humic components (13.26 %K and 4.48 % P) the seeds. One hundred seeds of each selected species
plus trace elements (BlackEarth Humates Ltd.) (Table 1). were placed on the tailings surface of the subplots and
covered with 3 cm of moist peat to hold the seeds in
place. Each block was fertilized with 3.7 l (3.8 gl−1) of
Table 1 Elemental composition (μgg−1) of the gold mine tail- Plant Prod™ soluble fertilizer 20:20:20 (N/P/K) with
ings and humic amendment micronutrients following seeding. The rate was deter-
Humic Tailings Normal range
mined based on preliminary growth chamber experi-
amendment in soila ments. Garden netting was pinned down over the top
of the peat to help protect the surface from wind erosion.
Cu 3.0±0.4 3,535±21 13–24 Watering over the 2003 field season was conducted once
Fe 3,878±114 31,140±431 5,000–50,000 a month adding 3.1 l of water per treatment block. A
K 157,040±1,056 7,346±1,896 3,000–26,000 second fertilizer addition of 3.1 l (2.3 gl−1) in 2003 and
P 30,519±1,002 134±18 50–1,000 in 2004 was conducted 1 month after seeding.
Values represent the mean ± SE of three composite samples
collected prior to the treatment application and analyzed using
2.3 Tailings Analysis
ICP-OES
a
Determined from Kabata-Pendias and Pendias (1992), Tailings samples (ca.500 g) were collected to a depth
Kirkman et al. (1994) and Foth and Ellis (1997) of 15 cm with a Dutch auger at five different times: at
1471, Page 4 of 14 Water Air Soil Pollut (2013) 224:1471

pre treatment, post treatment (4 weeks after amend- Three of the ten harvested plants from each replicate
ment application), at harvest (3 months after seeding), (subplots) were ground and pooled for elemental analysis
1 year after treatment, and in June 2010 (6 and 7 years by ICP-OES, conducted as outlined in the “Tailings
post treatment for the 2003 and 2004 experiments, Analysis” section. Quality control of both tailings and
respectively). The tailings were air dried and then plant elemental analysis included analysis of blanks,
stored at 5 °C until further analysis. pH and conduc- internal replicates and standard reference materials.
tivity measurements were made by adding distilled In June 2010, percent cover of both seeded and
water to 30 g of air dry tailings and incubated for 1 h volunteer species was determined visually. Unknown
to form a saturated paste. Samples were filtered in species were collected and identified with a key by
Buchner funnels with Whatman #1 filter paper under Scoggan (1957) and reference specimens at the
vacuum. The filtrate was analyzed for pH using a Dual University of Manitoba herbarium. As the seeded spe-
Channel pH/Ion meter (Accumet AR25; Fisher cies had intermixed among the plots at the time of
Scientific, Georgia, USA). Conductivity of the filtrate sampling we did not distinguish between these
was measured using a conductivity meter (Traceable, seeding subplots. Differences between treatments
Control Company, Texas, USA). The Walkley–Black were compared separately for the 2003 and 2004 run
method was used to determine the organic carbon of the experiment. Duncan’s new multiple range test
content of the tailings (Kalra and Maynard 1991). was used to determine differences between treatments
Water stable aggregation was measured based on the at each sampling date after performing a one-way
method outlined by Angers and Mehuys (1993) on the analysis of variance.
post treatment samples both the 2003 and 2004 exper-
iments. Water-stable aggregates were measured from
four size fractions >2.0, 2.0–1.0, 1.0–0.5, and 0.5– 3 Results and Discussion
0.25 mm. Next, 30 g of tailings were placed on a
wet sieving apparatus with vertical motions lasting 3.1 Tailings Properties
20 min with 32 oscillations/min. Size fractions were
collected from each sieve, dried at 105 °C, and The conductivity of tailings prior to amendment was
weighed to determine the percent of the total dry 2.11 and 3.28 dS m−1 for the 2003 and 2004 runs of
weight for each size class. The bulk density of tailings the experiment, respectively, indicating a slightly saline
was determined by collecting soil cores to a depth of soil (SPAC, 1999). This can be attributed to the oxidation
15 cm with a core sampler in 2010 (Blake, 1965). The of sulfide minerals releasing ions (Cu2+, Fe2+/Fe3+ and
cores were oven dried to a constant weight at 105 °C Na+) into the soil solution, as well as the basin like nature
and weighed. Elemental analysis (Cu, Fe, P and K) of of the tailings deposit, collecting runoff/dissolved min-
the tailings and the humic amendment was conducted erals from the surrounding area. In both runs of the
on samples of 0.1 to 1 g by inductively coupled experiment, following application of modified humic
plasma optical emission spectrometry (ICP-OES) on substances, the conductivity of the tailings more than
an ICP-Emission Spectrometer (Liberty 200, Varian, doubled in the 2 gkg−1 treatment and more than quadru-
USA). The available PO4−3 and inorganic N of tailings pled in the 4 gkg−1 treatment (Fig. 1). This was likely
were determined for the 2010 samples. Phosphate was due to the soluble nature of the modified humic amend-
measured on bicarbonate extracts (Kalra and Maynard ment, resulting in release of K+ and PO43− into the soil
1991). The microdiffusion method was used to deter- solution (Table 1). Since conductivity values ranging
mine available inorganic N (Kahn et al. 2000). from 4.1 to 8.0 dS m−1 indicate moderately saline con-
ditions and values higher than 8 dS m−1 are often con-
2.4 Plant Analysis sidered as high salinity (SPAC, 1999), the values of
conductivities found in amended tailings may have in-
Ten plants per subplot were harvested at the end of the duced some salt stress responses. By the end of the first
growing season (3 months after seeding) in each run of growing season, the conductivity values had returned to
the experiment. Harvested plants were washed three the pre treatment values in the 2004 experiment; while in
times in distilled water to remove tailings materials, the 2003 experiment, the effect of the amendment on the
lyophilized, separated into roots and shoots and wieghed. conductivity remained visible for 1 year following the
Water Air Soil Pollut (2013) 224:1471 Page 5 of 14, 1471

16.00 2003 16.00 2004 0 g C kg-1 tailings (Control)


Experiment Experiment 2 g C kg-1 tailings
c 4 g C kg-1 tailings

12.00 12.00
Conductivity (dSm -1)

c
b

8.00 8.00
b a
b a
a
b
4.00 a 4.00 a a a a
b aa
aa a a a a a a a a a aa

0.00 0.00

2.50 b 2.50

b
2.00 2.00
Organic Carbon (%)

c
1.50 c 1.50 b
b

b b b c
1.00 a 1.00 a
a b a
b
b a a
a ab a a aa a a a
0.50 0.50 a a

0.00 0.00

9.00
9.00

b ab a 8.00
8.00 a a a a a a a
a a a a
a a a
a a a a a
a a a a a
a a
7.00
pH

7.00

6.00 6.00

5.00 5.00
Pre Post Harvest 1 Year After Pre Post Harvest 1 Year After Six
Treatment Treatment Post Seven Treatment Treatment Post Years
Treatment Years Treatment

Fig. 1 Conductivity, organic carbon, and pH of 2003 and 2004 amendment experiments (mean ± SE, n=4). Different letters represent
significant differences between treatments (p<0.05)

treatment, at least in the 4 g C kg−1 treatment. The greater the amount of amendment added in both runs of the
amount of precipitation in field season 2004 (843 mm experiment (Fig. 1). These increases were even more
compared to 512 mm in the previous season) was likely pronounced after 1 year in the 2003 experiments. Six
to have contributed to the more rapid decrease in or 7 years after the amendment addition, organic car-
conductivity. bon follows the same trend although the absolute
Following the addition of humic substances, the levels have decreased. Soil organic carbon plays an
amount of organic carbon, increased in proportion to important role in soil development and fertility
1471, Page 6 of 14 Water Air Soil Pollut (2013) 224:1471

Table 2 Distribution of water stable aggregates in mine tailings amended with modified humic substances at seeding time (mean ± SE,
n=4)

Aggregate distribution (%)

>2.0 mm 1.0–2.0 mm 0.5–1.0 mm 0.25–0.5 mm Total

2003 experiment
0 g C kg−1 tailings 0.60±0.09a 1.70±0.23a 2.07±0.19a 3.71±0.32a 8.08±0.72a
2 g C kg−1 tailings 1.01±0.25a 3.21±0.29b 4.59±0.40b 7.15±0.50b 15.96±0.81b
−1 a b b b
4 g C kg tailings 1.04±0.21 3.44±0.53 4.80±0.66 7.27±0.99 16.55±2.30b
2004 experiment
0 g C kg−1 tailings 0.36±0.10a 1.63±0.09a 4.42±0.78a 6.16±0.56a 12.57±1.15a
−1 ab b b b
2 g C kg tailings 3.36±2.01 6.81±1.63 12.13±1.57 9.74±0.84 32.04±3.80b
4 g C kg−1 tailings 6.57±2.32b 12.00±0.60c 13.78±1.24b 11.75±1.04b 44.10±1.82b

Different letters represent significant differences (p<0.05) between treatments within each year and size class

through nutrient cycling, retention, and supply of nu- humic complexes that increased aggregate stability.
trients (Sorenson et al. 2011). Soils or tailings low in Considering the low clay content (~4 %) and high
organic carbon are prone to surface erosion, possess sand content (~48 %) of the tailings reported by
poor soil structure and have reduced nutrient quality Ibrahim and Goh (2004), results from this study sug-
(Swift 2001). Differences between the two field sea- gest that the tailings have a sufficient amount of
sons could be attributed to moisture conditions, with charged mineral surfaces (non-clay) that are bound
dryer conditions in 2003 than in 2004, potentially together by modified humic substances and lead to
reducing microbial activity and breakdown of the the formation of water stable aggregates. The more
amendment (Ibrahim and Goh 2004). favorable conditions in field 2004 resulted in a
The total macro aggregation of the unamended larger increase in total macro aggregation, in com-
tailings was low. One month following amendment parison to 2003.
application total aggregation was increased by There were no significant changes in pH following
~100 % in the 2003 experiment and 250–350 % in the application of modified humic substances either
the 2004 experiment, depending on the amount of within a year after the application of the amendments,
amendment used (Table 2). These increases in aggre- or 6 or 7 years later. In the 2003 run of the experiment,
gation occurred in most size classes in both years of the pH was relatively similar among the plots regard-
the experiment, except that in the 2003 run of the less of treatment (Fig. 1), while in the 2004 run the
experiment there was no difference between non- tailings were more acidic and much more variable
amended and amended treatments in the >2.0-mm size ranging from 3.0 to 7.5. Environmental conditions,
class, whereas in the 2004 run of the experiment the including oxidation state and water status (saturation)
increase was 10 and 20 times in the 2 and 4 g C kg−1, in the tailings, were likely to have influenced these pH
respectively. Overall, the increase in aggregation sig- fluctuations with varying amounts of mineral precipi-
nificantly improved the tailings structure. Low levels tates being formed or the rate of sulfide minerals being
of soil organic carbon were likely responsible for the oxidized. Mine tailings also contain bacteria that are
reduced soil structure. A similar increase in total ag- capable of increasing the background rate of sulfide
gregation was observed by Ibrahim and Goh (2004) mineral oxidation leading to the release of acidity and
using a 4-week incubation period in mine tailings metal as well as sulfate reducing bacteria that may lead
amended with modified humic substances. Whiteley to pH increases (Johnson et al. 2002).
(1993) found that materials high in sand and low in The amendment treatments did not affect Cu or Fe
clay were unaffected by ammonium humate amend- concentrations in the soil but did result in increases in
ment (a similar organic salt material) and Piccolo et al. P and K levels at the time of seeding (Table 3).
(1997) suggested that it was the formation of clay Although the Cu and Fe content of the tailings was
Water Air Soil Pollut (2013) 224:1471 Page 7 of 14, 1471

not affected the addition of modified humic substances Table 4 Available PO43− (mgkg−1) and inorganic N (mgkg−1)
in amended mine tailings in 2010 (mean ± SE, n=4)
can lead to a decrease in the soluble plant available Cu
in solution compared to untreated tailings (Senkiw and Available Inorganic N
Goh 2006). Humic substances have the ability to form PO43−
both soluble and insoluble complexes with Cu and appli-
2003
cation of organic matter is thought to be an effective
0 g C kg−1 tailings 0.37±0.03c 1.81±0.08a
mechanism of Cu retention in soils due to the strong −1 b
2 g C kg tailings 4.73±0.65 1.43±0.18a
stable complex that forms (McBride 1981; Schnitzer −1 a
and Khan 1978). Furthermore, soluble copper content 4 g C kg tailings 10.32±1.53 3.48±0.56b
might be lowered by the presence of phosphate (delivered 2004
along with the amendment) that could remove heavy 0 g C kg−1 tailings 1.19±0.77c 0.59±0.32a
−1 c
metals such as copper, cadmium, cobalt and especially 2 g C kg tailings 1.50±1.25 0.92±0.36a
−1 c
lead from solution through precipitation (Sugiyama et al. 4 g C kg tailings 5.90±2.68 1.18±0.23a
2003). The total level of P in the tailings (Table 3) was on
Different letters represent significant differences (p < 0.05)
the lower end of the range found in a mineral soil (Foth between treatments
and Ellis 1997) and the addition of modified humic sub-
stances significantly increased P content. Given the con- a high total K content as K-feldspar and mica were
centration of P in the amendment, we would expect identified by Salzsauler (2001) as the second and fourth
increases in tailings P of 225 and 450 μgg−1 in the 2 most abundant primary minerals within the tailings. Only
and 4 g C kg−1 amendment rates, respectively. These a minor amount of K is typically plant available either
values are close to what was found and, therefore, suggest free in solution or exchangeable sorbed to the surface of
little leaching of the amendment P from the tailings. charged particles (Foth and Ellis 1997). The available
These increases were reflected in the higher phosphate inorganic N of amended tailings was highly variable
levels measured in both experiments in 2010 (Table 4). between application rates and experiments and was not
Another element that was increased following amend- significantly different from unamended tailings with the
ment application was K, due to the relatively high amount exception of the 4 g treatment in the 2003 (Table 4).
of soluble K within the amendment. Given the level of K Macronutrients such as P and N are essential elements
in the amendment, we would expect the K concentration required for overall plant growth and development (Jones
in the tailings to increase by 1,180 and 2,360 μgg−1 in the 1998). The levels of available PO4−3 and inorganic N
2 and 4 g C kg−1 amendment rates, respectively. As with found in the plots seemed to be sufficient for plant growth
P, the levels found in the tailings correspond to these as other studies have found similar values on naturally
increases. Typical levels of total soil potassium range revegetated mine tailings (Wang et al. 2011).
from 3,000 and 26,000 μgg−1 in most soils types
(Kirkman et al. 1994). Mineral soils tend to fall towards 3.2 Plant Growth and Elemental Analysis
the upper end while organic soils fall on the lower end of
total soil potassium (Foth and Ellis 1997). The mine For the first growing season, the amendment showed
tailings, being mineral in nature, were expected to have no positive effect on plant yield. For M. sativa, there

Table 3 Elemental composition


(μgg−1) (field seasons 2003 and Cu Fe P K
2004) of mine tailings amended
with modified humic substances 2003
at seeding time (mean ± SE, n=4) 0 g C kg−1 tailings 3,661±197 39,308±3,984 139±20a 9,292±1056a
−1 b
2 g C kg tailings 3,702±220 38,108±3,364 443±38 11,320±1508b
−1 c
4 g C kg tailings 3,812±249 39,651±3,799 706±119 11,693±948b
2004
0 g C kg−1 tailings 4,713±446 47,685±2,614 94±23a 7,001±849a
−1 b
Different letters represent signif- 2 g C kg tailings 4,626±329 47,535±2,945 323±31 9,860±885b
icant differences (p < 0.05) −1 c
4 g C kg tailings 4,931±615 47,972±4,088 738±15 12,110±694c
between treatments
1471, Page 8 of 14 Water Air Soil Pollut (2013) 224:1471

was no effect of amendment on plant yield in either run from leonardite were found to stimulate root and shoot
of the experiment (Fig. 2). By contrast, E. trachycaulus growth of Hordeum vulgare (barley) in an hydroponic
had a large decrease in yield with the 4 gkg−1 amend- set up, at a maximum concentration of 5 mg C l−1
ment in the 2003 experiment (84 % reduction in yield while concentrations higher than 10 mg C l−1 caused
compared to controls) and no significant change in the inhibition of root and shoot growth (Ayuso et al.
2004 experiment. The osmotic stress resulting from the 1996b). A further study by Ayuso et al. (1997) carried
increase in conductivity following addition of humic out using a calcareous soil with silty loam texture with
material could have also played a role in the reduction less than 3.9 gkg−1 organic content, suggested that an
in biomass observed in E. trachycaulus in 2003. addition of humic substances at an optimal rate of
However, this reduction in growth was not observed in 50 mg C kg−1 was successful in increasing biomass
2004, in spite of a high conductivity of the amended of H. vulgare (barley) while concentrations over
tailings suggesting that other factors have contributed to 200 mg C kg−1 had an inhibitory effect. On the other
the decrease in growth. Furthermore, this difference hand, Piccolo et al. (1993) found a stimulation of the
between species was unexpected as both species have biomass of Lactuca sativa (lettuce) treated with humic
been shown to tolerate conductivities between 4 and substances extracted from sub-bituminous coal at a
8 dS m−1 (Swift 1997). In addition to osmotic stress, rate of 5 gl−1. The majority of authors have suggested
the high concentrations of soluble potassium in the that humic substances should be applied in concentra-
amended tailings itself could have induced nutrient de- tions of 50–300 mgl−1 in order to stimulate root and/or
ficiencies due to competition for uptake within the root shoot growth (Chen and Aviad 1990). In our study, the
cell membranes. Deficiencies of elements such as mag- concentration of modified humic substances present in
nesium and calcium (Locascio 1993) as well as iron and soil solution following the incubation period was not
zinc (Sinclair 1993) have been reported due to excess determined, but the amendment was applied in rates of
amounts of potassium in the soil. 2 g C kg−1 (7.5 gkg−1) and 4 g C kg−1 (15.0 gkg−1).
Studies have shown that humic substances can di- These concentrations were selected as they had been
rectly stimulate growth due to enhanced protein syn- shown to improve tailings structure (Ibrahim and Goh
thesis (Bukvova and Tichy 1967), direct auxin like 2004); such results were confirmed in our study.
activity (O’Donnell 1973), inhibition of indole-3- Nevertheless they exceeded most of the recommended
acetic acid oxidase activity (Mato et al. 1971), en- rates of application for an experiment conducted in
hancement of photosynthesis (Vaughan 1969) and in- soil or soil like medium and therefore the highest
creased respiration rate (Sladky and Tichy 1959). concentration (4 g C kg−1) could have impacted the
However, the effects of humic substances vary growth of E. trachycaulus. Whiteley and Williams
depending on the concentrations, sources (peat, lig- (1993) showed that soluble extracts from lignite ap-
nite, compost, soil, or sewage sludge) and growth plied to mine tailings (~36 gkg−1) inhibited both root
substrates used (Piccolo et al. 1993; Ayuso, et al. and shoot growth of Agrostis capillaris (Bent grass).
1996a; b). For example, humic substances extracted Inhibition of growth has been suggested to occur due

2003 Experiment 2004 Experiment

0 g C kg-1 tailings (Control)


2 a 2 2 g C kg-1 tailings
a a
a 4 g C kg-1 tailings
1.5 a 1.5 a a
Biomass (g)

a a
a
1 1
a
0.5 b 0.5

0 0

Medicago sativa Elymus trachycaulus Medicago sativa Elymus trachycaulus

Fig. 2 Plant (shoots and roots) biomass for Elymus n=4, with each replicate composed of ten subsamples). Within a
trachycaulus and Medicago sativa after a 3-month growth peri- species and experiment, bars with different letters are signifi-
od in mine tailings amended with humic substances (mean ± SE, cantly different
Water Air Soil Pollut (2013) 224:1471 Page 9 of 14, 1471

to excessive levels of humic substances that may de- treatments, were successful in establishing species in
crease micronutrient uptake potentially due to excess the tailings in the first year of growth. The tillage may
ligands that bind the micronutrients and make them have reduced the tailings compaction, increased water
unavailable for plant uptake (Ayuso et al. 1996a; retention capacity and improved the soil aeration.
Pertuit et al. 2001). Direct inhibitory effects can also However, the vegetation cover was relatively low in the
occur and involves the reduction of enzyme activity, 2003 experiment unamended tailings after 6 years com-
protein synthesis and decreases in photosynthetic and pared to the tailings amended with humic substances.
respiratory rates (Vaughan and Malcolm 1985; Chen The beneficial effects of fertilizers on plant growth on
and Aviad 1990). unamended tailings are short-lived. Our data suggest
Plant yield after the first growing season did not organic amendments are more effective in promoting
correlate with plant cover 6 or 7 years later. The humic plant growth long term than fertilizer alone.The remain-
amendment had no effect on M. sativa plant yield after der of the plant cover in both runs of the experiment was
the first growing season for both runs of the experiment made up of volunteer species (Appendix 1). A total of 22
(Fig. 2). However, there were significant differences in species, in 13 families were found in the plots, most of
the plant cover of M. sativa in the 2003 plots 7 years later which were typical of a disturbed site. We postulate that
(Table 5). M. sativa was nearly absent in the control, but wind borne seeds or vegetative propagules from neigh-
was dominant in the 2 and 4 g C kg−1 treatments where it boring vegetation often pioneer favorable microsites in
accounted for nearly half of the vegetation cover. In the naturally and anthropogenically revegetated tailings
2004 plots, the plant cover of M. sativa was consistent (Bagatto and Shorthouse 1999; Shu et al. 2005).
with the biomass data 6 years earlier where no differ- Members of Amaranthaceae, Asteraceae, Cyperaceae,
ences between treatments and control were found. The and Poaceae possess such seeds and propagules
highly variable pH in the 2004 plots could have retarded (Bagatto and Shorthouse 1999; Szarek-Łukaszewska
the establishment and growth of M. sativa both in the 2009). Woody species such as Betula spp., Populus
short and long term. Likewise, no differences were found spp., and Salix spp. also frequently occur with the afore-
in E. trachycaulus biomass yield in the first growing mentioned families. Rhizomatous plants such as
season of the 2004 experiment. Six and 7 years later, Equisetum spp. also play a role in colonizing mine
there were no differences in E. trachycaulus cover be- tailings (Shu et al. 2005). The ability of rhizomes to
tween amendments and control in both runs of the ex- translocate resources from older to younger ramets and
periment where it accounted for less than 4 % of the maintain genotypes for metal tolerance through vegeta-
vegetation. E trachycaulus may have suffered from os- tive propagation could be responsible for the ability of
motic stress and reduced growth in the 2003 experiment, Equisetum to grow on mine tailings.
where its yield was reduced in the highest amendment At the end of the first growing season, the roots of
application. In the 2004 experiment, the low pH of some E. trachycaulus grown in tailings amended with humic
areas of the tailings studied could have contributed to this materials showed an increase in Cu (season 2003) and
low vegetation cover. Nevertheless, the addition of hu- in Fe (season 2004) levels (Table 6). This increase in
mic substances did promote the invasion of volunteer Cu occurred in parallel to a more severe decrease in
species over time. The most common volunteer species plant biomass but did not seem to be linked to the
in the 2004 experiment were Poa pratensis (Kentucky tailings pH as the highest Cu content in roots was
bluegrass) and Salix candida (sageleaf willow) and found during field season one where the pH was the
accounted for 6–10 % of the vegetation. Both species highest. While humic substances have been shown to
may possess heavy metal tolerance since P. pratensis and increase macro and micro nutrient uptake (Dormaar
several Salix species have been reported to invade mine 1975; Lee and Bartlett 1976; Rauthan and Schnitzer
tailings or mine tailings-impacted soils naturally 1981) it has also been suggested that high instances
(McLaughlin 1988; Bourret et al. 2009). The effect of of mortality of non-metal tolerant cultivars of A.
pH on increased heavy metal availability and decreased capillaris grown in humic amended mine tailings was
nutrient availability cannot be over looked and could due to increased metal uptake causing toxicity
have contributed over time to a low vegetation cover in (Whiteley and Williams 1993). On the other hand, high
the 2004 experiment. Rotor tilling and fertilizer applica- concentrations of humic substances can lead to micro-
tions, in addition to regular watering in the control nutrient deficiency due to excessive ligand binding to
1471, Page 10 of 14 Water Air Soil Pollut (2013) 224:1471

Table 5 Percent cover


of the vegetation in the Percent cover (%)
2003 and 2004 experiments in
2010 (mean ± SE, n=4) M. sativa E. trachycaulus Volunteer species

2003
0 g C kg−1 tailings 1.67±0.68b 0.83±0.48a 6.58±2.42a
−1 a a
2 g C kg tailings 38.75±4.88 2.17±1.54 33.33±7.28b
4 g C kg−1 tailings 48.42±13.52a 0±0a 64.01±10.25c
2004
0 g C kg−1 tailings 3.33±3.33b 2.17±1.21a 36.58±9.56a
−1 b a
Different letters represent signif- 2 g C kg tailings 2.25±2.03 2.92±2.92 27.86±5.10a
icant differences (p<0.05) −1 b a
4 g C kg tailings 2.25±1.27 0.58±0.37 25.51±11.60a
between treatments

various metallic nutrients (Ayuso et al. 1996b; Pertuit et uptake of potassium and phosphate may have been
al. 2001). Iron content in the roots was high based on facilitated by the presence of the humic substances.
typical levels but generally not considered excessive In both species, there was no change in shoot copper
with the exception of E. trachycaulus in field season and iron content after addition of modified humic sub-
2003 in all treatments (Table 6). stances (Table 7). Overall, results from the two field
Root phosphorus content was increased in both plant seasons showed lower shoot copper contents relative to
species in all amended treatments, while only the roots with copper content higher in the E.trachycaulus
highest rate of humic materials (4 g C kg−1) resulted in relative to M. sativa (Tables 6 and 7). Iron levels in
an increase in root potassium content (Table 6). The shoots showed more variation with higher levels than
presence of significant quantities of phosphate and po- in roots in M. sativa in 2004, lower levels in E.
tassium in the humic amendment itself is likely to have trachycaulus in 2003 and similar levels for M sativa in
contributed to these increased levels. In addition, the 2003 and E. trachycaulus in 2004. Differences between

Table 6 Copper, iron, potassium


and phosphorus content Cu Fe K P
(μgg−1 dry weight) of Medicago
sativa and Elymus trachycaulus M. sativa
roots after a 3-month period of 2003
growth in mine tailings amended
0 g C kg−1 tailings 134±45a 313±87a 8,957±900a 703±89a
with modified humic substances −1 a a ab
(mean ± SE, n=4) 2 g C kg tailings 134±19 391±85 12,535±822 2,769±150b
−1 a a b
4 g C kg tailings 107±16 407±154 14,414±2055 2,702±222b
2004
0 g C kg−1 tailings 120±19a 190±31a 11,621±207a 1,440±107a
−1 a a a
2 g C kg tailings 156±30 247±33 13,859±1220 2,892±344b
−1 a a a
4 g C kg tailings 106±15 208±72 13,595±846 2,404±113b
E. trachycaulus
2003
0 g C kg−1 tailings 575±43a 1,557±281a 12,015±473a 608±80a
−1 b a a
2 g C kg tailings 951±162 2,328±474 13,516±233 2,468±402b
−1 b a b
4 g C kg tailings 1,180±191 1,909±368 29,444±1875 3,352±340c
2004
0 g C kg−1 tailings 545±141a 290±20a 16,840±1,909a 1,186±76a
−1 a b a
Different letters represent signif- 2 g C kg tailings 672±256 478±33 16,773±2,411 1,714±128b
icant differences (p < 0.05) −1 a a a
4 g C kg tailings 573±220 314±95 17,514±2,425 1,800±249b
between treatments
Water Air Soil Pollut (2013) 224:1471 Page 11 of 14, 1471

the species iron and copper contents in the shoot and root low, normal and excessive levels of phosphate and
tissues are likely related to differences at the level of potassium provided in Table 7 (Jones 1998), our re-
uptake in the root membrane and the inherent ability to sults suggest that deficiency may have occurred in the
translocate metals from their roots to their shoot portions plants growing in some of the un-amended and
and the physical structural differences between the spe- amended tailings in both field seasons and likely
cies in trapping tailings dust (Greger 1999). Although, limited growth of plants on tailings despite fertilizer
the absence of visual tissue injury suggests that copper application even though no difference in growth pa-
may not have severely impaired biochemical pathways, rameters were reported. While potassium deficiency
the levels were high enough (Table 7) to have potentially alters the plant water relations (opening and closing of
interfered with nutrient uptake (Jones 1998) and likely stomata and the maintenance of turgor pressure), phos-
contributed to previously observed decrease in pigments phate deficiency typically stunts growth as phosphorus
(Maksimiec and Baszynski 1996) and plant growth is a critical component of some enzymes, ATP, RNA,
(Kabata-Pendias and Pendias 1992). Iron content was and DNA (Jones 1998). In our experiment, application
on the higher end of the typical levels found in shoot of the humic substances amendment containing solu-
tissues, but generally not considered excessive with the ble phosphate and fertilizer appeared to have limited
exception of E. trachycaulus in field season 2003 but not completely alleviated deficiency in the
(Table 7). amended tailings without causing toxicity. However,
Following application of humic substances no the addition of amendment did not increase plant
change in K occurred in the shoot tissues (Table 7). potassium contents. The higher levels of K and P in
For P, an increase was observed in both species only in tailings in the second field season reflected the higher
field season 2003, no change occurred in field season rate of fertilizer application, which attempted to over-
2004; however, the control had a relatively high P come any potential deficiencies observed in the con-
content compared to field season 2003. Based on the trol treatments of the first field season.

Table 7 Copper, iron, potassium


and phosphorus content Cu Fe K P
(μgg−1 dry weight) of Medicago
sativa and Elymus trachycaulus M. sativa
shoots after a 3-month period of 2003
growth in mine tailings amended
0 g C kg−1 tailings 74±6a 409±64a 14,330±270ab 1,170±70a
with modified humic substances
(mean ± SE, n=4) 2 g C kg−1 tailings 95±7a 501±47a 14,871±515b 2,163±136b
−1 a a a
4 g C kg tailings 77±5 470±90 13,178±109 2,435±273b
2004
0 g C kg−1 tailings 76±15a 360±38a 16,508±2,045a 1,840±190a
−1 a a a
2 g C kg tailings 62±117 441±112 21,687±1,168 2,410±236a
4 g C kg−1 tailings 79±26a 375±114a 20,521±1,487a 2,168±205a
E. trachycaulus
2003
0 g C kg−1 tailings 209±75a 938±330a 11,737±701a 608±80a
−1 a a a
2 g C kg tailings 195±18 1,016±139 13,373±114 2,468±402b
4 g C kg−1 tailings 173±29a 753±152a 16,226±2,745a 3,352±340c
2004
0 g C kg−1 tailings 145±61a 293±25a 16,061±1,923a 1,557±174a
−1 a a a
2 g C kg tailings 142±54 402±90 15,280±1,761 1,977±184a
−1 a a a
a
Levels proposed by Jones 4 g C kg tailings 130±53 256±90 16,917±2,121 2,049±352a
(1998) Low levelsa 2–5 <50 <1,500 500–2,000
Different letters represent signif- Normal levelsa 5–30 100–500 15,000–30,000 2,000–8,000
icant differences (p < 0.05) Excessive levelsa 20–100 500–1,000 >50,000 >10,000
between treatments
1471, Page 12 of 14 Water Air Soil Pollut (2013) 224:1471

4 Conclusion Appendix

Our short term data from field seasons 2003 and 2004
suggest that tailings amended with humic substances Table 8 Species list of plants growing on tailings amended with
modified humic substances at the Central Manitoba Mine
improved the physical properties of the tailings by
Tailings
increasing macro aggregation, organic carbon, and
macronutrients but also increased soil electrical con- Family Genus Species
ductivity to levels stressful for plants. Plant growth
Amaranthaceae Rumex salicifolius Weinm.
varied depending on the species and the year of plant-
Suaeda maritima (L.) Dumort.
ing. The inhibition of E. trachycaulus may be attrib-
Asteraceae Anaphalis margaritaceae (L.) Benth.
uted to a high concentration of humic substances that
are directly or indirectly impairing the growth and/or Sonchus arvensis L.
the osmotic stress induced by high concentrations of Taraxacum officinale F.H. Wigg.
ions in the amended tailings. On the other hand, our Betulaceae Betula papyrifera Marshall
results suggest that M. sativa is capable of tolerating Bryaceae Bryum caespiticium Hedw.
the high conductivities of the amended tailings. Our Cyperaceae Carex aurea Nutt.
results also showed that roto-tilling, fertilizer applica- Ditrichaceae Ceratodon purpureus (Hedw.) Brid.
tions and regular watering over the growing season Equisetaceae Equisetum variegatum Schleich. ex F.
Weber & D.M.H. Mohr
(depending on the precipitations) were successful in
establishing M. sativa in the neutral tailings with and Fabaceae Medicago sativa L.
without humic substances amendment. However, the Melilotus albus Medik.
effects of fertilizers without organic amendment are Onagraceae Chamerion angustifolium ssp.
angustifolium (L.) Holub
short-lived. Our long term data from field season 2010
Pinaceae Larix laricina (Du Roi) K. Koch
suggest that the humic amendment in addition to fer-
Poaceae Agropyron trachycaulum (Link)
tilizer can result in a permanent plant cover. As a Malte ex H.F. Lewis
consequence, 22 volunteer species, 17 of which are
Poa palustris L.
native, were able to colonize the amended tailings,
pratensis L.
which were nearly void of vegetation prior to amend-
Sphenopholis intermedia (Rydb.) Rydb.
ment application. Overall, near neutral pH, adequate
Thinopyrum ponticum (Host)
nutrients, favorable climatic conditions, and inherent D.R. Dewey
plant tolerance to high conductivity resulted in the Rosaceae Potentilla norvegica L.
highest biomass yield and vegetation cover. Careful Salicaceae Populus balsamifera L.
selection of site, plant species, amendment and appli- tremuloides Michx.
cation rate seem to be key in revegetation since pH
Salix candida Flüggé ex Willd.
and elemental composition can vary considerably
lucida Muhl.
within a mine tailings site.

Acknowledgements Research funds for this project were pro- References


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