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
waste in the form of mine tailings. Sulfide mine tailings
are of particular concern due to the biotic and abiotic
oxidation of sulfide minerals that release acidity and
metals into the environment. Revegetation can be
employed to mitigate the spread of tailings in the envi-
ronment. Revegetation often involves ameliorating tail-
ings with organic materials to promote plant growth and
improve tailings physicochemical structure. We
amended plots in the Central Manitoba Mine tailings
pond with humic substances applied at rates up to 4 g
C kg−1 through roto-tilling and seeded with Medicago
sativa and Elymus trachycaulus in 2003 and 2004. The
humic substances improved tailings fertility by increas-
ing macro aggregation, organic carbon, and macronutri-
ents but also resulted in a short-term increase in
electrical conductivity levels. In the first growing season
the humic amendment had little effect on plant yield,
except in the 2003 experiment where the yield of E.
trachycaulus decreased by 84 % with 4 g C kg−1 amend-
ment. After 7 years, the addition of humic amendment
resulted in a cover of over 38 % for M. sativa, compared
to less than 2 % in control plots. In addition, non-seeded
C. Szczerski : C. Naguit : J. Markham : S. Renault (*)
Department of Biological Sciences, University of Manitoba,
Winnipeg, Manitoba, Canada R3T 2N2
e-mail: renaults@cc.umanitoba.ca
T. B. Goh
Department of Soil Science, University of Manitoba,
Winnipeg, Manitoba, Canada R3T 2N2
species cover increased with amendment rate in the
2003 experiment but not the 2004 experiment, most
likely due to lower pH in the latter. Our results suggest
that short-term patterns of plant performance do not
reflect longer-term performance or invasion by volun-
teer plant species. Our long-term data suggest that hu-
mic amendments can be effective in establishing plant
invasion of mine tailings, although the effects vary
depending on the pH of the tailings.
Keywords Humic substances . Tailings amendment .
Medicago sativa . Elymus trachycaulus . Plant growth .
Land reclamation . Tailings structure
1 Introduction
While the economic benefits of mining are apparent,
mining also creates large amounts of processed waste
material in the form of tailings. Tailings associated
with sulfur bearing metals are particularly problematic
due to the biotic and abiotic oxidation of these sulfide
minerals releasing acidity and metals into the environ-
ment (Blowes and Ptacek 1994). This phenomena
known as acid mine drainage (AMD) has been identi-
fied as the most serious environmental problem facing
the global metal mining industry. Prior to adequate
legislation, in most jurisdictions little was required in
terms of environmental reclamation or mitigation
plans and many older mine sites became abandoned
or orphaned with little treatment.
1471, Page 2 of 14
Cost effective reclamation for acidic tailings sites
typically involves tailings stabilization to prevent con-
taminated particle movement off site by wind and
water, exposure of new oxidizable material, and to
reduce or eliminate AMD caused by water and oxygen
movements into and through the tailings impound-
ment (Ripley et al. 1996). The majority of low cost
techniques focus on the establishment of vegetation in
combination with or without chemical, biological, or
physical treatments to aid in tailings stabilization.
Selected vegetation typically consists of commercially
available, native, or metal tolerant species (Gardner et
al. 2012). In addition, revegetation can also be thought
of as a successional process where the initial vegeta-
tion consists of pioneer species that allow surrounding
native plants to colonize the site. The primary goal of
the initial stages of revegetation then is to establish
vegetative cover that requires little to no maintenance
in order to aid in the prevention of erosion and to build
up organic material in preparation for more permanent
vegetation (Ripley et al. 1996).
The successful establishment of vegetation may be
limited by the physical and chemical properties associ-
ated with sulfide containing tailings. These properties
include potentially toxic levels of heavy metals, high
salinity, low pH, macronutrient deficiencies and a low
organic carbon content contributing to relatively poor
soil structure, and a high degree of surface compaction
(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
2008). Lignite is one possible source of humic sub-
stances and has been shown to significantly improve
the carbon content and structure of mine tailings
(Ibrahim and Goh 2004). Humic substances are known
to increase water and nutrient holding capacity, water
permeability, decrease soluble metal concentrations by
complexing with metals and play a role in the formation
of soil structure including stable soil aggregation
(Stevenson 1982; MacCarthy 2001). Humic substances
also play a number of other beneficial roles in soils
including pH buffering capacity, binding of charged
toxic ions, increasing the soil cation exchange capacity,
serving as a reservoir for nutrients (nitrogen, phospho-
rus, and potassium) and retaining soil moisture
(MacCarthy 2001). Furthermore, the ability of humic
substances to remove metal ions from the soil solution
and decrease their bioavailability make them extremely
useful for reclamation of heavy metal contaminated land
Water Air Soil Pollut (2013) 224:1471
(Livens 1991). In addition, humic substances have been
shown to have direct stimulatory growth effects on
plants including increased photosynthesis and auxin like
activity (Vaughan 1969; O’Donnell 1973). However,
positive effects have varied with outcomes based on
the concentration and sources of the humic substances
used (Piccolo et al. 1993). Reduced growth has also
been shown with humic substances. It has been
suggested that high concentrations can lead to increased
heavy metal uptake causing toxicity (Whiteley and
Williams 1993) and that micronutrient deficiencies can
be caused by a tight binding of micronutrients reducing
their availability (Pertuit et al. 2001).
The objectives of this study were: (1) to assess the
short-term (1–2 years) and long-term (6–7 years) ef-
fects of modified humic substances on tailings physi-
cochemical properties, (2) to study the growth and
elemental content of plant species seeded in tailings
amended with modified humic substances in the short
term, and (3) to determine if the amendment of tailings
with modified humic substances will facilitate the
invasion of other plant species years after the initial
seeding. It was hypothesized that modified humic sub-
stances would promote soil development in terms of
aggregation, increase the carbon content of the tail-
ings, stimulate overall growth of the selected species,
and initiate succession by promoting the invasion of
other plant species.
2 Materials and Methods
2.1 Site Description
The Central Manitoba Gold mine tailings impoundment
is located in southeastern Manitoba, in Nopiming
Provincial Park (50°54′16″N, 95°20′6″W). The tailings
cover ca. 20 ha and vary in depth from ~5 m to greater
than 25 m (Sherriff et al. 2009). Although the tailings
were deposited in the 1920s–1930s, less than 0.1 % is
covered with vegetation. While biotic and abiotic oxi-
dation of the sulfide primary minerals pyrite (iron sul-
fide), chalcopyrite (copper–iron sulfide) and pyrrhotite
(iron sulfide) have led to the release of metals like iron
and copper and generate acidity, the presence of calcite
in the tailings has had a buffering effect (Londry and
Sherriff 2005). As a result, the tailings pH has been
shown to range from 3.5 to 7 (Renault et al. 2002;
Londry and Sherriff 2005) with only ~20 % of the total
Water Air Soil Pollut (2013) 224:1471
tailings area possessing a pH<4 due to the exhaustion of
calcite by a higher local load of sulfide minerals (Londry
and Sherriff 2005). Given the complexity of mine tail-
ings composition, some elements were selected for ele-
mental analysis based on a previous study of these
tailings (Green and Renault 2008). Previous analysis
indicated copper levels far exceeding the normal range
in soil while iron levels fall within the normal soil range
(Kabata-Pendias and Pendias 1992). More information
on the tailings elemental composition and their potential
impact on vegetation is provided by Green and Renault
(2008).
2.2 Experimental Design
A tailings amendment experiment was conducted twice,
once in the spring 2003 and again 2004. This repetition
was considered important since our previous work had
shown spring growing conditions vary from year to year
and have a strong impact on plant performance. The
amount of precipitation (rainfall and snowfall) during
each year varied substantially, with 512 mm in 2003
and 843 mm in 2004. The average precipitation for the
region is 558 mm. Meteorological data were obtained
from the National Climate Data and Information Archive
website (http://www.climate.weatheroffice.gc.ca).
The humic amendment was a water soluble modified
compound (Dry soluble 80™) from BlackEarth Humates
Ltd. (Edmonton, Alberta, Canada). It was processed from
a lignite (humalite) deposit. The humic substances
contained greater than 80 % humic and fulvic acids as
well as non-humic components (13.26 %K and 4.48 % P)
plus trace elements (BlackEarth Humates Ltd.) (Table 1).
Table 1 Elemental composition (μgg−1) of the gold mine tail-
ings and humic amendment
Humic Tailings Normal range
amendment in soila
Cu 3.0±0.4 3,535±21 13–24
Fe 3,878±114 31,140±431 5,000–50,000
K 157,040±1,056 7,346±1,896 3,000–26,000
P 30,519±1,002 134±18 50–1,000
Values represent the mean ± SE of three composite samples
collected prior to the treatment application and analyzed using
ICP-OES
a
Determined from Kabata-Pendias and Pendias (1992),
Kirkman et al. (1994) and Foth and Ellis (1997)
Page 3 of 14, 1471
The plots for each run (year) of the experiment were
about 10 m apart and the tailings in both locations,
appearing to be similar in colouration (i.e., composition
and oxidation state), were relatively free of surface salt
crusting present in some areas of the tailings pond, and
had no naturally established vegetation. For each run of
the experiment, four blocks (5.2×0.7 m) were
established each with three 1.4×0.7 m plots (separated
by a 0.5-m buffer strip) and amended with different
levels modified humic material (0, 7.5, and 15.0 g
amendment kg−1 tailings). These rates were used in
order to reach concentrations of 0, 2 and 4 g carbon
kg−1 tailings, respectively, and were based on prelimi-
nary experiments and results from Ibrahim and Goh
(2004). In order to calculate the rate of amendment
and fertilizer addition, density of the tailings was esti-
mated using a tapped density method in which dry
tailings were shaken in a graduated cylinder, allowed
to settle and weighed. To incorporate the amendments,
the tailings treatment blocks, including the 0 gkg−1 plot,
were first roto-tilled to a depth of 15 cm. The amend-
ment was then added to the surface of the tailings and
tilled again.
Following a 4-week incubation, two seeding subplots
(0.5×0.5 m) within each amendment plot were random-
ly selected and seeded with either Medicago sativa
(alfalfa) or Elymus trachycaulus (slender wheatgrass).
The plant species were selected based on their
established use in tailings reclamation, and tolerance to
salinity, ability to control erosion and fix nitrogen (in the
case of M. sativa). Brett Young Seeds Limited supplied
the seeds. One hundred seeds of each selected species
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
Plant Prod™ soluble fertilizer 20:20:20 (N/P/K) with
micronutrients following seeding. The rate was deter-
mined based on preliminary growth chamber experi-
ments. Garden netting was pinned down over the top
of the peat to help protect the surface from wind erosion.
Watering over the 2003 field season was conducted once
a month adding 3.1 l of water per treatment block. A
second fertilizer addition of 3.1 l (2.3 gl−1) in 2003 and
in 2004 was conducted 1 month after seeding.
2.3 Tailings Analysis
Tailings samples (ca.500 g) were collected to a depth
of 15 cm with a Dutch auger at five different times: at
1471, Page 4 of 14
pre treatment, post treatment (4 weeks after amend-
ment application), at harvest (3 months after seeding),
1 year after treatment, and in June 2010 (6 and 7 years
post treatment for the 2003 and 2004 experiments,
respectively). The tailings were air dried and then
stored at 5 °C until further analysis. pH and conduc-
tivity measurements were made by adding distilled
water to 30 g of air dry tailings and incubated for 1 h
to form a saturated paste. Samples were filtered in
Buchner funnels with Whatman #1 filter paper under
vacuum. The filtrate was analyzed for pH using a Dual
Channel pH/Ion meter (Accumet AR25; Fisher
Scientific, Georgia, USA). Conductivity of the filtrate
was measured using a conductivity meter (Traceable,
Control Company, Texas, USA). The Walkley–Black
method was used to determine the organic carbon
content of the tailings (Kalra and Maynard 1991).
Water stable aggregation was measured based on the
method outlined by Angers and Mehuys (1993) on the
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–
0.25 mm. Next, 30 g of tailings were placed on a
wet sieving apparatus with vertical motions lasting
20 min with 32 oscillations/min. Size fractions were
collected from each sieve, dried at 105 °C, and
weighed to determine the percent of the total dry
weight for each size class. The bulk density of tailings
was determined by collecting soil cores to a depth of
15 cm with a core sampler in 2010 (Blake, 1965). The
cores were oven dried to a constant weight at 105 °C
and weighed. Elemental analysis (Cu, Fe, P and K) of
the tailings and the humic amendment was conducted
on samples of 0.1 to 1 g by inductively coupled
plasma optical emission spectrometry (ICP-OES) on
an ICP-Emission Spectrometer (Liberty 200, Varian,
USA). The available PO4−3 and inorganic N of tailings
were determined for the 2010 samples. Phosphate was
measured on bicarbonate extracts (Kalra and Maynard
1991). The microdiffusion method was used to deter-
mine available inorganic N (Kahn et al. 2000).
2.4 Plant Analysis
Ten plants per subplot were harvested at the end of the
growing season (3 months after seeding) in each run of
the experiment. Harvested plants were washed three
times in distilled water to remove tailings materials,
lyophilized, separated into roots and shoots and wieghed.
Water Air Soil Pollut (2013) 224:1471
Three of the ten harvested plants from each replicate
(subplots) were ground and pooled for elemental analysis
by ICP-OES, conducted as outlined in the “Tailings
Analysis” section. Quality control of both tailings and
plant elemental analysis included analysis of blanks,
internal replicates and standard reference materials.
In June 2010, percent cover of both seeded and
volunteer species was determined visually. Unknown
species were collected and identified with a key by
Scoggan (1957) and reference specimens at the
University of Manitoba herbarium. As the seeded spe-
cies had intermixed among the plots at the time of
sampling we did not distinguish between these
seeding subplots. Differences between treatments
were compared separately for the 2003 and 2004 run
of the experiment. Duncan’s new multiple range test
was used to determine differences between treatments
at each sampling date after performing a one-way
analysis of variance.
3 Results and Discussion
3.1 Tailings Properties
The conductivity of tailings prior to amendment was
2.11 and 3.28 dS m−1 for the 2003 and 2004 runs of
the experiment, respectively, indicating a slightly saline
soil (SPAC, 1999). This can be attributed to the oxidation
of sulfide minerals releasing ions (Cu2+, Fe2+/Fe3+ and
Na+) into the soil solution, as well as the basin like nature
of the tailings deposit, collecting runoff/dissolved min-
erals from the surrounding area. In both runs of the
experiment, following application of modified humic
substances, the conductivity of the tailings more than
doubled in the 2 gkg−1 treatment and more than quadru-
pled in the 4 gkg−1 treatment (Fig. 1). This was likely
due to the soluble nature of the modified humic amend-
ment, resulting in release of K+ and PO43− into the soil
solution (Table 1). Since conductivity values ranging
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-
sidered as high salinity (SPAC, 1999), the values of
conductivities found in amended tailings may have in-
duced some salt stress responses. By the end of the first
growing season, the conductivity values had returned to
the pre treatment values in the 2004 experiment; while in
the 2003 experiment, the effect of the amendment on the
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
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
2003 experiment
0 g C kg−1 tailings 0.60±0.09a 1.70±0.23a
2 g C kg−1 tailings 1.01±0.25a 3.21±0.29b
−1 a b
4 g C kg tailings 1.04±0.21 3.44±0.53
2004 experiment
0 g C kg−1 tailings 0.36±0.10a 1.63±0.09a
−1 ab
2 g C kg tailings 3.36±2.01 6.81±1.63
4 g C kg−1 tailings 6.57±2.32b 12.00±0.60c
Different letters represent significant differences (p<0.05) between treatments within each year and size class
through nutrient cycling, retention, and supply of nu-
trients (Sorenson et al. 2011). Soils or tailings low in
organic carbon are prone to surface erosion, possess
poor soil structure and have reduced nutrient quality
(Swift 2001). Differences between the two field sea-
sons could be attributed to moisture conditions, with
dryer conditions in 2003 than in 2004, potentially
reducing microbial activity and breakdown of the
amendment (Ibrahim and Goh 2004).
The total macro aggregation of the unamended
tailings was low. One month following amendment
application total aggregation was increased by
~100 % in the 2003 experiment and 250–350 % in
the 2004 experiment, depending on the amount of
amendment used (Table 2). These increases in aggre-
gation occurred in most size classes in both years of
the experiment, except that in the 2003 run of the
experiment there was no difference between non-
amended and amended treatments in the >2.0-mm size
class, whereas in the 2004 run of the experiment the
increase was 10 and 20 times in the 2 and 4 g C kg−1,
respectively. Overall, the increase in aggregation sig-
nificantly improved the tailings structure. Low levels
of soil organic carbon were likely responsible for the
reduced soil structure. A similar increase in total ag-
gregation was observed by Ibrahim and Goh (2004)
using a 4-week incubation period in mine tailings
amended with modified humic substances. Whiteley
(1993) found that materials high in sand and low in
clay were unaffected by ammonium humate amend-
ment (a similar organic salt material) and Piccolo et al.
(1997) suggested that it was the formation of clay
Water Air Soil Pollut (2013) 224:1471
0.5–1.0 mm 0.25–0.5 mm Total
2.07±0.19a 3.71±0.32a 8.08±0.72a
4.59±0.40b 7.15±0.50b 15.96±0.81b
b b
4.80±0.66 7.27±0.99 16.55±2.30b
4.42±0.78a 6.16±0.56a 12.57±1.15a
b b b
12.13±1.57 9.74±0.84 32.04±3.80b
13.78±1.24b 11.75±1.04b 44.10±1.82b
humic complexes that increased aggregate stability.
Considering the low clay content (~4 %) and high
sand content (~48 %) of the tailings reported by
Ibrahim and Goh (2004), results from this study sug-
gest that the tailings have a sufficient amount of
charged mineral surfaces (non-clay) that are bound
together by modified humic substances and lead to
the formation of water stable aggregates. The more
favorable conditions in field 2004 resulted in a
larger increase in total macro aggregation, in com-
parison to 2003.
There were no significant changes in pH following
the application of modified humic substances either
within a year after the application of the amendments,
or 6 or 7 years later. In the 2003 run of the experiment,
the pH was relatively similar among the plots regard-
less of treatment (Fig. 1), while in the 2004 run the
tailings were more acidic and much more variable
ranging from 3.0 to 7.5. Environmental conditions,
including oxidation state and water status (saturation)
in the tailings, were likely to have influenced these pH
fluctuations with varying amounts of mineral precipi-
tates being formed or the rate of sulfide minerals being
oxidized. Mine tailings also contain bacteria that are
capable of increasing the background rate of sulfide
mineral oxidation leading to the release of acidity and
metal as well as sulfate reducing bacteria that may lead
to pH increases (Johnson et al. 2002).
The amendment treatments did not affect Cu or Fe
concentrations in the soil but did result in increases in
P and K levels at the time of seeding (Table 3).
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
was no effect of amendment on plant yield in either run
of the experiment (Fig. 2). By contrast, E. trachycaulus
had a large decrease in yield with the 4 gkg−1 amend-
ment in the 2003 experiment (84 % reduction in yield
compared to controls) and no significant change in the
2004 experiment. The osmotic stress resulting from the
increase in conductivity following addition of humic
material could have also played a role in the reduction
in biomass observed in E. trachycaulus in 2003.
However, this reduction in growth was not observed in
2004, in spite of a high conductivity of the amended
tailings suggesting that other factors have contributed to
the decrease in growth. Furthermore, this difference
between species was unexpected as both species have
been shown to tolerate conductivities between 4 and
8 dS m−1 (Swift 1997). In addition to osmotic stress,
the high concentrations of soluble potassium in the
amended tailings itself could have induced nutrient de-
ficiencies due to competition for uptake within the root
cell membranes. Deficiencies of elements such as mag-
nesium and calcium (Locascio 1993) as well as iron and
zinc (Sinclair 1993) have been reported due to excess
amounts of potassium in the soil.
Studies have shown that humic substances can di-
rectly stimulate growth due to enhanced protein syn-
thesis (Bukvova and Tichy 1967), direct auxin like
activity (O’Donnell 1973), inhibition of indole-3-
acetic acid oxidase activity (Mato et al. 1971), en-
hancement of photosynthesis (Vaughan 1969) and in-
creased respiration rate (Sladky and Tichy 1959).
However, the effects of humic substances vary
depending on the concentrations, sources (peat, lig-
nite, compost, soil, or sewage sludge) and growth
substrates used (Piccolo et al. 1993; Ayuso, et al.
1996a; b). For example, humic substances extracted
2003 Experiment
2 a
a a
a 4 g C kg-1 tailings
1.5
Biomass (g)
1 1
0.5
0 0
Medicago sativa Elymus trachycaulus
Fig. 2 Plant (shoots and roots) biomass for Elymus
trachycaulus and Medicago sativa after a 3-month growth peri-
od in mine tailings amended with humic substances (mean ± SE,
Water Air Soil Pollut (2013) 224:1471
from leonardite were found to stimulate root and shoot
growth of Hordeum vulgare (barley) in an hydroponic
set up, at a maximum concentration of 5 mg C l−1
while concentrations higher than 10 mg C l−1 caused
inhibition of root and shoot growth (Ayuso et al.
1996b). A further study by Ayuso et al. (1997) carried
out using a calcareous soil with silty loam texture with
less than 3.9 gkg−1 organic content, suggested that an
addition of humic substances at an optimal rate of
50 mg C kg−1 was successful in increasing biomass
of H. vulgare (barley) while concentrations over
200 mg C kg−1 had an inhibitory effect. On the other
hand, Piccolo et al. (1993) found a stimulation of the
biomass of Lactuca sativa (lettuce) treated with humic
substances extracted from sub-bituminous coal at a
rate of 5 gl−1. The majority of authors have suggested
that humic substances should be applied in concentra-
tions of 50–300 mgl−1 in order to stimulate root and/or
shoot growth (Chen and Aviad 1990). In our study, the
concentration of modified humic substances present in
soil solution following the incubation period was not
determined, but the amendment was applied in rates of
2 g C kg−1 (7.5 gkg−1) and 4 g C kg−1 (15.0 gkg−1).
These concentrations were selected as they had been
shown to improve tailings structure (Ibrahim and Goh
2004); such results were confirmed in our study.
Nevertheless they exceeded most of the recommended
rates of application for an experiment conducted in
soil or soil like medium and therefore the highest
concentration (4 g C kg−1) could have impacted the
growth of E. trachycaulus. Whiteley and Williams
(1993) showed that soluble extracts from lignite ap-
plied to mine tailings (~36 gkg−1) inhibited both root
and shoot growth of Agrostis capillaris (Bent grass).
Inhibition of growth has been suggested to occur due
2004 Experiment
0 g C kg-1 tailings (Control)
2 2 g C kg-1 tailings
a 1.5 a a
a a
a
a
b 0.5
Medicago sativa Elymus trachycaulus
n=4, with each replicate composed of ten subsamples). Within a
species and experiment, bars with different letters are signifi-
cantly different
Water Air Soil Pollut (2013) 224:1471
to excessive levels of humic substances that may de-
crease micronutrient uptake potentially due to excess
ligands that bind the micronutrients and make them
unavailable for plant uptake (Ayuso et al. 1996a;
Pertuit et al. 2001). Direct inhibitory effects can also
occur and involves the reduction of enzyme activity,
protein synthesis and decreases in photosynthetic and
respiratory rates (Vaughan and Malcolm 1985; Chen
and Aviad 1990).
Plant yield after the first growing season did not
correlate with plant cover 6 or 7 years later. The humic
amendment had no effect on M. sativa plant yield after
the first growing season for both runs of the experiment
(Fig. 2). However, there were significant differences in
the plant cover of M. sativa in the 2003 plots 7 years later
(Table 5). M. sativa was nearly absent in the control, but
was dominant in the 2 and 4 g C kg−1 treatments where it
accounted for nearly half of the vegetation cover. In the
2004 plots, the plant cover of M. sativa was consistent
with the biomass data 6 years earlier where no differ-
ences between treatments and control were found. The
highly variable pH in the 2004 plots could have retarded
the establishment and growth of M. sativa both in the
short and long term. Likewise, no differences were found
in E. trachycaulus biomass yield in the first growing
season of the 2004 experiment. Six and 7 years later,
there were no differences in E. trachycaulus cover be-
tween amendments and control in both runs of the ex-
periment where it accounted for less than 4 % of the
vegetation. E trachycaulus may have suffered from os-
motic stress and reduced growth in the 2003 experiment,
where its yield was reduced in the highest amendment
application. In the 2004 experiment, the low pH of some
areas of the tailings studied could have contributed to this
low vegetation cover. Nevertheless, the addition of hu-
mic substances did promote the invasion of volunteer
species over time. The most common volunteer species
in the 2004 experiment were Poa pratensis (Kentucky
bluegrass) and Salix candida (sageleaf willow) and
accounted for 6–10 % of the vegetation. Both species
may possess heavy metal tolerance since P. pratensis and
several Salix species have been reported to invade mine
tailings or mine tailings-impacted soils naturally
(McLaughlin 1988; Bourret et al. 2009). The effect of
pH on increased heavy metal availability and decreased
nutrient availability cannot be over looked and could
have contributed over time to a low vegetation cover in
the 2004 experiment. Rotor tilling and fertilizer applica-
tions, in addition to regular watering in the control
Page 9 of 14, 1471
treatments, were successful in establishing species in
the tailings in the first year of growth. The tillage may
have reduced the tailings compaction, increased water
retention capacity and improved the soil aeration.
However, the vegetation cover was relatively low in the
2003 experiment unamended tailings after 6 years com-
pared to the tailings amended with humic substances.
The beneficial effects of fertilizers on plant growth on
unamended tailings are short-lived. Our data suggest
organic amendments are more effective in promoting
plant growth long term than fertilizer alone.The remain-
der of the plant cover in both runs of the experiment was
made up of volunteer species (Appendix 1). A total of 22
species, in 13 families were found in the plots, most of
which were typical of a disturbed site. We postulate that
wind borne seeds or vegetative propagules from neigh-
boring vegetation often pioneer favorable microsites in
naturally and anthropogenically revegetated tailings
(Bagatto and Shorthouse 1999; Shu et al. 2005).
Members of Amaranthaceae, Asteraceae, Cyperaceae,
and Poaceae possess such seeds and propagules
(Bagatto and Shorthouse 1999; Szarek-Łukaszewska
2009). Woody species such as Betula spp., Populus
spp., and Salix spp. also frequently occur with the afore-
mentioned families. Rhizomatous plants such as
Equisetum spp. also play a role in colonizing mine
tailings (Shu et al. 2005). The ability of rhizomes to
translocate resources from older to younger ramets and
maintain genotypes for metal tolerance through vegeta-
tive propagation could be responsible for the ability of
Equisetum to grow on mine tailings.
At the end of the first growing season, the roots of
E. trachycaulus grown in tailings amended with humic
materials showed an increase in Cu (season 2003) and
in Fe (season 2004) levels (Table 6). This increase in
Cu occurred in parallel to a more severe decrease in
plant biomass but did not seem to be linked to the
tailings pH as the highest Cu content in roots was
found during field season one where the pH was the
highest. While humic substances have been shown to
increase macro and micro nutrient uptake (Dormaar
1975; Lee and Bartlett 1976; Rauthan and Schnitzer
1981) it has also been suggested that high instances
of mortality of non-metal tolerant cultivars of A.
capillaris grown in humic amended mine tailings was
due to increased metal uptake causing toxicity
(Whiteley and Williams 1993). On the other hand, high
concentrations of humic substances can lead to micro-
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
al. 2001). Iron content in the roots was high based on
typical levels but generally not considered excessive
with the exception of E. trachycaulus in field season
2003 in all treatments (Table 6).
Root phosphorus content was increased in both plant
species in all amended treatments, while only the
highest rate of humic materials (4 g C kg−1) resulted in
an increase in root potassium content (Table 6). The
presence of significant quantities of phosphate and po-
tassium in the humic amendment itself is likely to have
contributed to these increased levels. In addition, the
uptake of potassium and phosphate may have been
facilitated by the presence of the humic substances.
In both species, there was no change in shoot copper
and iron content after addition of modified humic sub-
stances (Table 7). Overall, results from the two field
seasons showed lower shoot copper contents relative to
roots with copper content higher in the E.trachycaulus
relative to M. sativa (Tables 6 and 7). Iron levels in
shoots showed more variation with higher levels than
in roots in M. sativa in 2004, lower levels in E.
trachycaulus in 2003 and similar levels for M sativa in
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
tissues are likely related to differences at the level of
uptake in the root membrane and the inherent ability to
translocate metals from their roots to their shoot portions
and the physical structural differences between the spe-
cies in trapping tailings dust (Greger 1999). Although,
the absence of visual tissue injury suggests that copper
may not have severely impaired biochemical pathways,
the levels were high enough (Table 7) to have potentially
interfered with nutrient uptake (Jones 1998) and likely
contributed to previously observed decrease in pigments
(Maksimiec and Baszynski 1996) and plant growth
(Kabata-Pendias and Pendias 1992). Iron content was
on the higher end of the typical levels found in shoot
tissues, but generally not considered excessive with the
exception of E. trachycaulus in field season 2003
(Table 7).
Following application of humic substances no
change in K occurred in the shoot tissues (Table 7).
For P, an increase was observed in both species only in
field season 2003, no change occurred in field season
2004; however, the control had a relatively high P
content compared to field season 2003. Based on the
low, normal and excessive levels of phosphate and
potassium provided in Table 7 (Jones 1998), our re-
sults suggest that deficiency may have occurred in the
plants growing in some of the un-amended and
amended tailings in both field seasons and likely
limited growth of plants on tailings despite fertilizer
application even though no difference in growth pa-
rameters were reported. While potassium deficiency
alters the plant water relations (opening and closing of
stomata and the maintenance of turgor pressure), phos-
phate deficiency typically stunts growth as phosphorus
is a critical component of some enzymes, ATP, RNA,
and DNA (Jones 1998). In our experiment, application
of the humic substances amendment containing solu-
ble phosphate and fertilizer appeared to have limited
but not completely alleviated deficiency in the
amended tailings without causing toxicity. However,
the addition of amendment did not increase plant
potassium contents. The higher levels of K and P in
tailings in the second field season reflected the higher
rate of fertilizer application, which attempted to over-
come any potential deficiencies observed in the con-
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

vided by the University of Manitoba, the Natural Sciences and
Engineering Research Council of Canada (IPS scholarship to
CS), BlackEarth Humates Ltd and Manitoba Innovation, Energy
and Mines. Thanks to Merv Fisher (BlackEarth Humates Ltd.)
for providing advice and the humic substances as well as to
Manitoba Conservation for allowing access to the site (work
permit number WPG 17623). We would also like to thank C.
Nakata, S. Green, K. Kivinen, D. Szczerski, P. Ouellette, I.
Young, R. Sheffield, J. Christensen, K. Kostyra, L. Smith, J.
Montgomery, and J. Makar for field and/or lab assistance. This
paper is dedicated to the memory of Eva Sailerova who passed
away during the study and was instrumental in getting us started
on the project.
[SPAC] Soil and Plant Analysis Council Inc. (1999). Soil anal-
ysis: handbook of reference methods. Florida: St. Lucie
Press.
Angers, D. A., & Mehuys, G. R. (1993). Aggregate stability to
water. In M. R. Carter (Ed.), Soil sampling and methods of
analysis (pp. 651–657). Florida: Lewis Publishers.
Ayuso, M., Moreno, J. L., Hernandez, T., & Garcia, C.
(1996). Effect of humic fractions from urban wastes
and other more evolved organic materials on seed
germination. Journal of the Science of Food and
Agriculture, 72, 461–468.
Water Air Soil Pollut (2013) 224:1471
Ayuso, M., Hernandez, T., Garcia, C., & Pascual, J. A. (1996).
Stimulation of barely growth and nutrient absorption by
humic substances originating from various organic mate-
rials. Biosource Technology, 57, 251–257.
Ayuso, M., Moreno, J. L., Hernandez, T., & Garcia, C. (1997).
Characterization and evaluation of humic acids from urban
waste as liquid fertilizers. Journal of the Science of Food
and Agriculture, 75, 481–488.
Bagatto, G., & Shorthouse, J. D. (1999). Biotic and abiotic
characteristics of ecosystems on acid metalliferous mine
tailings near Sudbury, Ontario. Canadian Journal of
Botany, 77, 410–425.
Blake, G. R. (1965). Bulk density. In C.A. Black (Ed.), Methods
of soil analysis, part I: Am. Soc. Agron. Monograph 9,
374–390. WT: Madison.
Blowes, D. W., & Ptacek, C. J. (1994). Acid-neutralisation in
inactive mine tailings. In J. L. Jambor & D. W. Blowes
(Eds.), Short course handbook on the environmental geo-
chemistry of sulfide mine-wastes (pp. 271–292). Ontario:
Mineralogical Association of Canada.
Bourret, M. M., Brummer, J. E., & Leininger, W. C. (2009).
Establishment and growth of two willow species in a
riparian zone impacted by mine tailings. Journal of
Environmental Quality, 38, 693–701.
Bukvova, M., & Tichy, V. (1967). The effect of humus fractions on
the phosphorylase activity of wheat (Triticum aestivum L.).
Biologia Plantarium, 9, 401–406.
Chen, Y., & Aviad, T. (1990). Effects of humic substances on plant
growth. In P. McCarthy, C. E. Clapp, R. L. Malcolm, & P. R.
Bloom (Eds.), Humic substances in soil and crop sciences (pp.
161–181). Wisconsin: Soil Science Society of America Inc.
Dormaar, J. F. (1975). Effects of humic substances from cher-
nozemic Ah horizons on nutrient uptake by Phaseolus
vulgaris and Festuca scabrella. Canadian Journal of Soil
Science, 55, 111–118.
Foth, H. D., & Ellis, B. G. (1997). Soil Fertility (2nd ed.). Boca
Raton, FL: CRC Press Inc.
Gardner, W. C., Naeth, M. A., Broersma, K., Chanasyk, D. S., &
Jobson, A. M. (2012). Influence of biosolids and fertilizer
amendments on element concentrations and revegetation of
copper mine tailings. Canadian Journal of Soil Science, 92,
89–102.
Green, S., & Renault, S. (2008). Influence of papermill sludge
on growth of Medicago sativa, Festuca rubra and
Agropyron trachycaulum in gold mine tailings: a green-
house study. Environmental Pollution, 151, 524–531.
Greger, M. (1999). Metal availability and bioconcentration in
plants. In M. N. V. Prasad & J. Hagemeyer (Eds.), Heavy
metals stress in plants: From molecules to ecosystems (pp.
1–27). Germany: Springer Verlag.
Ibrahim, S. M., & Goh, T. B. (2004). Changes in macroaggregation
and associated characteristics in mine tailings amended with
humic substances. Communications in Soil Science and Plant
Analysis, 35, 1905–1922.
Johnson, D. B., Dziuria, M. A., Koimert, A., & Hallberg, K. B.
(2002). The microbiology of acid mine drainage: genesis and
biotreatment. South African Journal of Science, 98, 249–259.
Jones, J. B. (1998). Plant nutrition manual. Washington, D.C.:
CRC Press.
Kabata-Pendias, A., & Pendias, H. (1992). Trace elements in
soils and plants (2nd ed.). Boca Raton, FL: CRC Press Inc.
Page 13 of 14, 1471
Kahn, S. A., Mulvaney, R. L., & Hoeft, R. G. (2000). Direct-
diffusion methods for inorganic-nitrogen analysis of soil. Soil
Science Society of America Journal, 64, 1083–1089.
Kalra, Y. P., & Maynard, D. G. (1991). Methods manual for
forest soil and plant analysis. Canada: Northwest Region
Northern Forestry Centre.
Kirkman, J. H., Basker, A., Surapaneni, A., & MacGregor, A. N.
(1994). Potassium in the soils of New Zealand — a review.
New Zealand Journal of Agricultural Research, 37, 207–
227.
Lee, Y. S., & Bartlett, R. J. (1976). Stimulation of plant growth
by humic substances. Soil Science Society of America
Journal, 40, 876–879.
Livens, F. R. (1991). Chemical reactions of metals with humic
material. Environmental Pollution, 70, 183–208.
Locascio, S. J. (1993). Cucurbits: Cucumbers, Muskmelon, and
Watermelon. In W. F. Bennett (Ed.), Nutrient deficiencies
and toxicities in crop plants (pp. 123–130). Minnesota:
The American Phytopathological Society.
Londry, K. L., & Sherriff, B. L. (2005). Comparison of micro-
bial biomass, biodiversity, and biogeochemistry in three
contrasting gold mine tailings deposits. Geomicrobiology
Journal, 22, 237–247.
MacCarthy, P. (2001). The principles of humic substances. Soil
Science, 166, 738–751.
Maksimiec, W., & Baszynski, K. (1996). Chlorophyll fluores-
cence in primary leaves of excess Cu-treated runner bean
plants depends on their growth stages and duration of Cu-
action. Journal of Plant Physiology, 149, 196–201.
Mato, M. C., Fabregas, R., & Mendez, J. (1971). Inhibitory
effects of soil humic acids on indoleacetic acid oxidase.
Soil Biology and Biochemisty, 3, 285–288.
McBride, M. B. (1981). Forms and distribution of copper in
solid and solution phases of soil. In J. F. Loneragan, A. D.
Robson, & R. D. Graham (Eds.), Copper in soils and
plants (pp. 25–42). Australia: Academic Press.
McLaughlin, B. E. (1988). The distribution of Agrostis gigantea
and Poa pratensis in relation to some environmental fac-
tors on a mine-tailings area at Copper Cliff, Ontario.
Canadian Journal of Botany, 66, 2317–2322.
O’Donnell, R. W. (1973). The auxin like effect of humic prep-
arations from leonardite. Soil Science, 116, 106–112.
Pertuit, A. J., Dudley, J. B., & Toler, J. E. (2001). Leonardite
and fertilizer levels influence tomato seedling growth.
HortScience, 36, 913–915.
Piccolo, A., Celano, G., & Pietramellara, G. (1993). Effects of
fractions of coal derived humic substances on seed germi-
nation and growth of seedlings (Lactuca sativa and
Lycopersicon esculentum). Biology and Fertility of Soils,
16, 11–15.
Piccolo, A., Pietramellara, G., & Mbagwu, J. S. C. (1997). Use
of humic substances as soil conditioners to increase aggre-
gate stability. Geoderma, 75, 267–277.
Rauthan, B. S., & Schnitzer, M. (1981). Effects of soil fulvic
acid on the growth and nutrient content of cucumber
(Cucumis sativus) plants. Plant and Soil, 63, 491–495.
Renault, S., Sailerova, E., & Fedikow, M. A. F. (2002).
Phytoremediation of mine tailings and bio-ore production:
progress report on seed germination, plant growth and
metal accumulation in seedlings planted at Central
Manitoba minesite (NTS 52L13). In Manitoba Industry,
1471, Page 14 of 14
Trade and Mines, Report of activities 2002 (pp. 255–265).
Manitoba: Manitoba Industry, Trade and Mines, Manitoba
Geological Survey.
Ripley, E. A., Redmann, R. E., & Crowder, A. A. (1996).
Environmental effects of mining. Florida: St. Lucie Press.
Salzsauler, K. A. (2001). Geochemical and mineralogical in-
vestigations of abandoned mine tailings, Central Manitoba
mine site. BSc thesis. Manitoba: University of Manitoba.
Schnitzer, M., & Khan, S. U. (1978). Soil organic matter.
Amsterdam: Elsevier.
Scoggan, H. J. (1957). Flora of Manitoba: Bulletin 140. Biological
Series 47, (pp. 619). Ottawa: National Museum of Canada.
Senkiw, K. A., & Goh, T. B. (2006). Comparison of amend-
ments used to remediate acid mine tailings: environmental
and agricultural applications. In D. Langouche, & E. Van
Ranst (Eds), New Waves in Physical Land Resources (pp.
39–48). Proceedings of the Workshop for Alumni of the
M.Sc. programmes in Soil Science, Eremology and
Physical Land Resources 2006, International Centre for
Physical Land Resources, Ghent, Belgium.
Sherriff, B. L., Ferguson, I. J., Gupton, M. W., VanGulck, J. F.,
Sidenko, N., Priscu, C., et al. (2009). A geophysical and
geotechnical study to determine the hydrogeological re-
gime of the Central Manitoba gold mine tailings deposit.
Canadian Geotechnical Journal, 46, 69–80.
Shu, W. S., Ye, Z. H., Zhang, Z. Q., Lan, C. Y., & Wong, M. H.
(2005). Natural colonization of plants on five lead/zinc mine
tailings in Southern China. Restoration Ecology, 13, 49–60.
Sinclair, J. B. (1993). Soybeans. In W. F. Bennett (Ed.), Nutrient
deficiencies and toxicities in crop plants (pp. 99–104).
Minnesota: The American Phytopathological Society.
Sladky, Z., & Tichy, V. (1959). Application of humus substances
to overground organs of plants. Biologia Plantarum, 1, 9–15.
Sorenson, P. T., Quideau, S. A., MacKenzie, M. D., Landhäusser,
S. M., & Oh, S. W. (2011). Forest floor development and
biochemical properties in reconstructed boreal forest soils.
Applied Soil Ecology, 49, 139–147.
Stevenson, F. J. (1982). Humus chemistry: genesis, composition,
reactions (pp. 17–23). USA: John Wiley & Sons.
Water Air Soil Pollut (2013) 224:1471
Sugiyama, S., Ichii, T., Fujisawa, M., Kawashiro, K., Tomida, T.,
Shigemoto, N., et al. (2003). Heavy metal immobilization in
aqueous solution using calcium phosphate and calcium hy-
drogen phosphates. Journal of Colloid and Interface Science,
259, 408–410.
Swift, C. E. (1997). Salt tolerance of various temperate zone
ornamental plants. Colorado State University. Resource
Document. http://www.coopext.colostate.edu/TRA/
PLANTS/stable.shtml. Accessed 2007.
Swift, R. S. (2001). Sequestration of carbon by soil. Soil
Science, 166, 858–871.
Szarek-Łukaszewska, G. (2009). Vegetation of reclaimed and
spontaneously vegetated Zn–Pb mine wastes in Southern
Poland. Polish Journal of Environmental Studies, 18, 717–
733.
Tordoff, G. M., Baker, A. J. M., & Willis, A. J. (2000). Current
approaches to the revegetation and reclamation of metallif-
erous mine wastes. Chemosphere, 41, 219–228.
Vaughan, D. (1969). The stimulation of invertase development
in aseptic storage tissue slices by humic acid. Soil Biology
and Biochemisty, 1, 15–28.
Vaughan, D., & Malcolm, R. E. (1985). Influence of humic
substances on growth and physiological processes. In D.
Vaughan, R. E. Malcolm, & M. Nijhoff (Eds.), Soil organic
matter and biological activity (pp. 37–75). Netherlands:
Dr. Junk Publishers.
Wang, J., Zhang, C. B., Ke, S. S., & Qian, B. Y. (2011).
Different spontaneous plant communities in Sanmen Pb/
Zn mine tailing and their effects on mine tailing physico-
chemical properties. Environmental Earth Sciences, 62,
779–786.
Whiteley, G. M. (1993). Effects of colloidal lignite on the
stability of soil aggregates. Soil Technology, 63, 321–
327.
Whiteley, G. M., & Williams, S. (1993). Effects of treatment of
metalliferous mine spoil with lignite derived humic sub-
stances on the growth responses of metal tolerant and non
metal tolerant cultivars of Agrostis capillaris L. Soil
Technology, 6, 163–171.