Journal of Cleaner Production 236 (2019) 117709

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Journal of Cleaner Production

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Pollution indicandum and marble waste polluted ecosystem; role of

selected indicator plants in phytoremediation and determination of
pollution zones
Zeeshan Ahmad a, Shujaul Mulk Khan a, *, Muhammad Ishtiaq Ali b, Noureen Fatima b,
Shahab Ali c
a
Department of Plant Sciences, Quaid-i-Azam University Islamabad, Pakistan
b
Department of Microbiology, Quaid-i-Azam University Islamabad, Pakistan
c
Department of Botany, Hazara University Mansehra, Pakistan

a r t i c l e i n f o a b s t r a c t

Article history: The absence or presence of particular plant species indicates specific level of pollution. It was hypoth-
Received 20 August 2018 esized that the marble waste polluted ecosystem also host specific plant indicators that can grow, survive
Received in revised form and tolerate more successfully than others and can be utilized in better managements of such systems.
14 June 2019
The current research work was therefore, conducted to determine the indicators of marble polluted
Accepted 17 July 2019
Available online 18 July 2019
region of Buner, Pakistan. Ecological techniques using varying sized quadrats i.e., 1
1 m2, 5
2 m2 and
10
2 m2 for herbs, shrubs and tree species were used respectively. Standard protocols were used to
Handling Editor: Jun Bi prepare soil samples and plant parts (root, leaf and shoot) for Atomic Absorption Spectrophotometry.
Indicator i.e., Populus alba L., Arundo donax L., Erigeron canadensis L and Morus alba L. were identified via
Keywords: Indicator Species Analyses (ISA) in various polluted zones. All the collected data were put in MS Excel for
Marble waste analyses in PCORD through Cluster Analysis (CA), Two Way Cluster Analysis (TWCA) and ISA. CANOCO
Spectrophotometry software was used to examine the impact of miscellaneous environmental variables in zonation via both
Heavy metals direct and indirect gradient techniques i.e., Detrended Correspondence Analysis (DCA) and Canonical
Phytoremediation
Correspondence Analysis (CCA). A total of 102 plant species belonging to 95 genera and 48 families were
Indicator plant
recorded in the marble waste polluted ecosystem. CA and TWCA through Jaccard Distance measurements
Pollution zonation
and Wards Linkage methods gave rise to 3 major polluted zones. These zones were i) Heavily Polluted
Zone (HPZ), ii) Moderate Polluted Zone (MPZ) and iii) Less Polluted Zone (LPZ). The recorded values of
Biological Concentration Factor (BCF), Translocation Factor (TF) and Bioaccumulation Coefficient (BAC)
for analyzed heavy metals i.e. Fe, Mg, Ca, Na and Cu in root and shoot of P. alba L., A. donax L., and M. alba
L. showed that these species were significant phytostabilizers and E. canadensis L. was phytoextracters
and hence best indicandum of marble waste polluted ecosystems. It was concluded that among all the
measured environmental variables, higher phosphorus level, higher pH, moderate potassium and lower
electrical conductivity (EC) had significant effects (p < 0.05) on the functions of these indicators as
phytoremediaters. It is recommended to develop green belts of these indicators around the marble in-
dustrial areas for the better management and hazardous free environment.
© 2019 Elsevier Ltd. All rights reserved.

1. Introduction

Abbreviations: CA, Cluster Analysis; ISA, Indicator Species Analysis; TWCA, Two
Way Cluster Analysis; DCA, Detrended Correspondence Analysis; CCA, Canonical
Correspondence Analysis; TF, Translocation Factor; BAC, Bioaccumulation coeffi-
cient; BCF, Biological Concentration Factor; KP, Khyber Pakhtunkhwa; EC, Electrical
Conductivity; ST, Soil Texture; OM, Organic Matter; K, Potassium; P, Phosphorus.
* Corresponding author.
E-mail addresses: zeeshanahmad78@ymail.com (Z. Ahmad), shujaqau@gmail.
com (S.M. Khan).
Environmental pollution is a substantial issue of the modern
world, causing huge damages to the natural ecosystems. Approxi-
mately, 2.01 billion metric tons waste per annum is produced
worldwide (Bilgen et al.,2008). According to the World Bank's es-
timates, waste generation will increase up to 3.4 billion metric tons
by 2050. Only 13.5% of the today's waste is recycled and 5.5% is

https://doi.org/10.1016/j.jclepro.2019.117709
0959-6526/© 2019 Elsevier Ltd. All rights reserved.
2 Z. Ahmad et al. / Journal of Cleaner Production 236 (2019) 117709
composted. One-third or 40% of the waste is not managed appro-
priately and hence are dumped or openly burnt. The improper
running of marble industries is one of the main source of waste
generation. Marble is a metamorphic rock usually composed of
recrystallized carbonates minerals generally dolomite or calcite
€ro
(To €k et al., 2011). It usually formed in metamorphism when
limestone is exposed to pressure and heat. Marble usually contain
chemical compound MgO, CaO, SiO2, Fe2O3, TiO2, P2O5, Na2O and
Al2O3 (Knoche et al., 1995). Marble rocks have many unique prop-
erties as these are used in sculpture and architecture as building
material for beautification (Ashish, 2019; Taji et al., 2019). Chemi-
cally, it is used in pharmaceuticals and agriculture fields to decrease
acidity of the soil. It also used in paint, papers and cosmetics for its
optical properties due to low cost (Andre  et al., 2014; Li et al., 2019;
Lu et al., 2018). Though, marble industry play important role in the
socioeconomic conditions via providing employment to hundreds
of people within communities. Unfortunately, marble industry
waste approximately 70% precious minerals resources during pol-
ishing, processing and mining (Gazi et al., 2012). On the other hand
worldwide, about 40% of marbles waste generated in quarrying
operation processes in the form of rock fragments. Millions of tons
of marble waste materials are produced daily and dumped in empty
pits, river beds, roads, agriculture fields or pasture lands that lead to
wide range of environmental pollution (Aukour and Al-Qinna,
2008). It affects the composition of flora and fauna of an
ecosystem as well as other chemical and physical alterations of the
environment directly or indirectly (Yu et al., 2005). Plant and ani-
mal species in such polluted ecosystems are facing countless
environmental issues (Gaston et al., 2013). Such environmental
changes in the past had caused species extinction (Davis and Shaw,
2001). These changes may be serious for certain organisms which
cannot adjust themselves with the changing conditions especially
the one which have relatively less genetic diversity and narrow
ecological amplitude (Bano et al., 2018).
Likewise, various plant species absorb different types of toxic
pollutants from contaminated ecosystem (Paz-Alberto and Sigua,
2013; Treesubsuntorn and Thiravetyan, 2018). It acts as sinks that
reduce noxious waste concentration in the polluted ecosystem
(Prajapati and Tripathi, 2008). Some of the plants have the ability to
survive in such type of toxic environment having inimical sub-
stances (Navarro-Cano et al., 2018). This type of pollution abate-
ment function is best performed by some of the pollutant indicators
(Dyer et al., 2017; Haller et al., 2018; Kuddus et al., 2011). Pollutant
indicators are those plants that can grow on soil with concentration
of particular toxic elements where other plants are difficult to grow
and reproduce (Hellawell, 2012). These plants raise in presence of
toxic substances/heavy metals are termed as tolerant/phytostabil-
izer or hyperaccumulator of such habitat (Freeman et al., 2006). In
addition to, an indicator species is ideally a species that always
occurs under a unique set of environmental condition and disap-
pears elsewhere. Ideal indicator indicates unique set of environ-
mental conditions quite often (Burgass et al., 2017). It can signal the
change in biological state of a specific environment. These may be
utilized as proxy to diagnose health of an ecosystem. Therefore
indicator plant are the priceless gift of nature that can be utilized
wisely so that these may be preserved for future generations as well
(Kwatra et al., 2016).
In a nutshell, rapid rate of industrialization and increasing
population is demanding protection of the environment from
contamination and over exploitation. For example marble pollut-
ants had been strongly affecting the surrounding ecosystems in the
form of physical and chemical changes that in return harm the
ecosystems as well as local inhabitants depend on these systems
since long. It was hypothesized that the marble waste polluted
ecosystem has specific plant indicators that can grow, survive and
show more tolerance and contest against marble pollution and
associated heavy metals. The main focus of this research work was
to find out plant indicators via multivariate statistical techniques
and their capabilities of heavy metals accumulation. These plants,
could be a solution to this interference of environmental degra-
dation to minimize the extent of damage. These indicator species
can also be grown and use for monitoring water quality in marble
waste polluted habitats/ecosystems. These can provide a new way
of improve water quality and pollution management of Buner in
particular and other region of Pakistan in general. At the same time,
multivariate statistical techniques provide an efficient means to
reduce the complexity inherent in natural vegetation data and to
detect important environmental factors that explain this
complexity to a level of understanding (Iqbal et al., 2018). It dis-
tinguishes the main pattern in relation among species and
perceived environment. It helps in generation hypothesis about the
structure and peculiarity of indicators in a particular region, either
in terms of plant composition or environmental variables for
example pollution in our case (Beals, 1984). It reduces the
complexity of data via summarizing the ecological data with low
dimensional space by bringing plant species with akin re-
quirements together in the form of zones or communities. Zonation
or association can be distinct further on the basis of indicator values
specified by the values of various recorded environmental factors.
The methodologies adapted here could also play role as baseline for
other such studies in future. This study will also help to find out the
particular position i.e., success/failure of the regulatory measures
taken and the corrective measurements required to take up and
bring the environment to its normal and pristine stage.
2. Materials and methodology
2.1. Study area
Buner lies in Khyber Pakhtunkhwa (KP) province of Pakistan
covering an area of 1865 Km2 (720 square miles). There are 160.2
million tons of the marble reserves in Pakistan (PASDEC). Out of
these 158 million tons (98%) is present in Khyber Pakhtunkhwa and
50% of the total KP marble reserves are present in Buner (current
study area). There are more than 600 marble industries and above
100 marble industries are under construction. These cause huge
pollution that may not only affect the native fauna and flora but also
influence the human population. Therefore, the present study was
taken into consideration to quantify the current situation and
propose the future remedies based on scientific tools and
techniques.
2.2. Sampling of vegetation
The present research work was carried out around variety of
industries producing different concentration in Buner, Pakistan. A
total of 45 stations from 15 factories at a distance of 3e4 Km were
randomly selected for sampling vegetation under the influence of
marble waste processing ecosystem (Fig. 1). Quadrat quantitative
ecological techniques were used for data collection (Ahmad et al.,
2016a; Khan et al., 2013). At each station, 3 quadrats i.e., one each
from vicinity, 50 and 100 m distance were taken from each marble
factory. Phytosociological attributes such as cover, relative cover,
density, relative density, frequency, relative frequency and impor-
tance value index were measured for each plant species at each
station (Khan et al., 2017). Cover and relative cover of trees were
calculated at basal area of stem through Diameter at Breast Height.
Basal area was calculated through formula:
BA ¼ <pi>r2 (1)
 Z. Ahmad et al. / Journal of Cleaner Production 236 (2019) 117709
Where BA is Basal area; r is radius; <pi> ¼ 3.14.
The plant specimens were collected, labeled with tags, placed in
blotting papers and pressed using plant presser (Ali and Nasir,
1990; Ali and Qaiser, 1995). Ethyl Alcohol and Mercuric Chloride
solution were used to poison the plant specimens and mounted on
standard herbarium sheets having size of 11.5
17.5 inches. All the
plants were identified using Flora of Pakistan and other available
literature (Nasir et al., 1972).
2.3. Soil analyses
A composite soil samples from three sites at depth of 0.3 m were
collected from each station using soil sampling instrument and
mixed thoroughly for further use. The collected samples were put
in polythene bags, labeled, dried at room temperature and sieved.
Each sample was analyzed in three replicates for different physi-
cochemical properties i.e., pH, electrical conductivity (EC), soil
texture (ST), organic matter (OM), potassium (K) and phosphorus
(P) and its average values were taken. Soil pH was determined
following the method of (McLean, 1982). Whereas, 10 g of air dried
and well sieved soil were homogenized in 10 mL distill water via
stirring on magnetic stirrer for 60 min. The solution was filtered
through filter paper and pH was determined using pH meter
(Russel RL060P). Distilled water and soil solution was prepared in
9:1 for EC analysis. Homogenized the solution by stirring it 3 times
for 1 min with interval of 10 min and EC was determined in deci-
Siemens per meter (ds/m) (McLean, 1982). The percentage of
organic matter was determined by Walkley procedure (Nelson and
Sommers, 1996). According to it, 1 g of soil sample was taken in
conical flask (500 mL), added with K2Cr2O7 (5 mL) plus Sulphuric
acid (10 mL), and placed it for 30 min to cool down after shaking.
Fig. 1. GIS generated map of the study area representing different marble factories along with different elevation zone in the region.
3
After cooling 100e1500 mL distilled water, Phosphoric acid (3 mL)/
sodium fluoride (0.5 gm) and 5e10 drops of indicator were added
after cooling. Ferrous sulphate was added drop by drop till its color
deviate from blue-green to reddish-gray. To get the actual volume
of Ferros sulphate a blank subtract sample reading was run to
minimize Potassium dichromate. For measuring potassium 2.5 gm
soil sample was taken in 20 mL conical flask and added 50 mL
extracted reagents. The solution was shaken by placing on flat bed
recipro shaker for 30 min and filtered. The content of potassium
was determined by flame photometer using graph readings
(Mehlich, 1953) in ppm. For phosphorus, soil sample of 2.5 g was
taken, added 50 mL extracted solution, shaked it for 30 min and
filtered the solution via Whatman paper No. 42. The aliquot (5 mL)
were taken in 25 mL volumetric flask from the filtrate with the help
of pipette and color developing reagent (5 mL) was added. The
volume was raised up to mark until the bluish color was estab-
lished. Concentration of phosphorus was read at 880 nm wave-
length using spectrophotometer (VARIAN, AA240FS).
2.4. Heavy metal analysis
Selected indicators i.e., A. donax L., E. canadensis L., P. alba L. and
M. alba L. were further analyzed for their phytoremediation abil-
ity. These plants were taken from each HPZ, MPZ and LPZ of
marble polluted ecosystem. The acid digestion method was used
for samples preparation of Atomic absorption spectrophotometry
according to the protocol given by (Filgueiras et al., 2000; Zasoski
and Burau, 1977). One gram of oven dried root, shoot and soil
samples (three replicates of each) were taken and grounded via
pestle and mortar. 10 mL of Nitric acid and Perchloric acid in 3:1
were added in 50 mL conical flask and kept for 24 h. For initial
4 Z. Ahmad et al. / Journal of Cleaner Production 236 (2019) 117709
digestion, sample was placed in Fume Hood and at a temperature
up to 150  C for 1 h. The temperature was then raised up to 235  C
till white fumes appear. After cooling mixture was filtered, 40 mL
of distilled water was added. Finally, the samples were analyzed
through Atomic Absorption Spectrophotometer (VARIAN,
AA240FS) for quantification of heavy metals accumulation in mg/
kg. The standard transfer, concentration and accumulation ratio of
indicator species was evaluated by TF, BAC and BCF (Malik et al.,
2010). The aforementioned factors were examined using stan-
dard formulas as follow:
BCF ¼ (Metal) Root/ (Metal) Soil
TF ¼ (Metal) Shoot/ (Metal) Root
BAC ¼ (Metal) Shoot/ (Metal) Soil
2.5. Data analyses
Data sets were prepared in MS Excel for indicator species and
other analyses under the influence of all measured environmental
factors with special reference to marble waste polluted ecosystem

using multivariate statistical packages (Leps and Smilauer, 2003).
Initially, Presence and absence (1, 0) data for 45 stations and 102
plant species were arranged and transposed according to software
requirements. CA and TWCA of PC-ORD version 5 were used to
identify significant polluted habitats/zones on the basis of pattern
of similarity via Jaccard Distance Measurements and Wards
Linkages method (Ahmad et al., 2016b; Greig-Smith, 1983). Indi-
cator Species Analysis (ISA) was carried to find out indicators of
each sort of habitat/zone (i.e., heavily polluted, moderate polluted
and less polluted zone). It provided the information about species
fidelity in a particular habitat to a measured set of variables.
Monte Carlo test was used for statistical significance after deter-
mination of indictor values (percentage of perfect indication
based on the combining values of the relative frequency and
relative abundance) of each indicator species using method of
(Dufre ^ne and Legendre, 1997). A threshold level of 25% indicator
value and 95% significance (p  0.05) was considered as cut off
value for determination of indicators (Dufre ^ne and Legendre,
1997; Ter Braak and Prentice, 1988). Indirect Gradient Analysis
i.e., Detrended Correspondence Analysis (DCA) using species
matrices and direct gradient analysis i.e., Canonical Correspon-
dence Analysis (CCA) using species and environmental data
matrices were performed using CANOCO software to examine the
visible relationship of distinct environmental factors with species
composition, distribution pattern and abundance in general and
indicators in particular.
3. Results
A total of 102 plant species belong to 95 genera and 48 families
were recorded from marble waste polluted ecosystem. It contains
77 herbs (75.5% of the total vegetation), 10 shrubs (9.8%) and 15 tree
species (17.7%). The family Asteraceae was the topmost dominant
family having 14 different species covering 29% of the total vege-
tation followed by Leguminosae and Poaceae families.
3.1. Results of CA and TWCA
CA and TWCA through PC-ORD version 5 classified all the plant
species and stations into 3 potential polluted zones i.e., heavily
polluted, moderate polluted and less polluted having their own
particular indicators under the influence of various measured
environmental variables and marble wastes and can be evidently
observed in the cluster dendrogram (Fig. 2). TWCA dendrogram
further characterized distribution of species at each station. The
white dots indicate the absence while black dots show presence of
plant species in the particular station (Fig. 3) of each zone.
3.2. Classification of plants into polluted zones, its associated
indicators and ecological gradients
A total of 3 major polluted zones were established under the
impact of heavy metals, soil and other climatic variables through
Jaccard Distance measurements and Wards linkage methods. The
detail description of these polluted zones and its associated indi-
cator species are as follows;-
3.2.1. Heavily polluted zone (HPZ)
A total of thirteen stations constituted HPZ encompassing 46
different types of plant species. The top most indicators of this
region were P. alba L., A. donax L and E. canadensis L. having Prob-
ability (p) Values  0.05 and Indicator Values  25% (Fig. 4). These
were the indicators of higher Phosphorus concentration in heavily
marble waste polluted region (Table 1).
Other indicators of this zone were Boerhavia procumbens Banks
ex Roxb, Cyperus rotundus L, Potentilla indica (Jacks.) Th. Wolf and
Parthenium hysterophorus L at 25% threshold level of indicator
based on Monte Carlo test of significance for observed maximum
indicator values i.e., percentage of perfect indication based on
combining values for the relative frequency and relative abundance
after ISA (Table 2).
All indicator species of HPZ were under the impact of varying
physicochemical nature of soil. It could be one of the main cogni-
tion of distinctive indicators of this zone. The soil state of this zone
perceived higher concentration of phosphorus (P) which ranges
from 4 to 8 ppm. Other associated edaphic factors constituted EC
that deviated from 1.004 to 1.654 dsm1, pH ranges 6.4e7.3, organic
matter 0.58e0.74% and potassium (K) range from 76 to 105 ppm
(Table 3).
3.2.2. Moderate polluted zone (MPZ)
Total numbers of stations segregated in this moderate polluted
Fig. 2. Cluster Analysis dendrogram via Jaccard Distance measurements (with narrow
single spaced width) of 45 stations comprehended 3 major polluted zones after Wards
linkage methods. (F ¼ factory, Q ¼ Quadrat).
 Z. Ahmad et al. / Journal of Cleaner Production 236 (2019) 117709
Fig. 3. TWCA dendrogram presenting distribution of 102 plant species in the region after Jaccard Distance Measurements with Wards Linkage Method of PCORD version 5.
(F ¼ factory, Q ¼ Quadrat).
zone were 20 escorting 90 different plant species. Top 3 uppermost
indicator species of this polluted region were M. alba L, Rosa
webbiana Wall. ex Royle and Centaurium tenuiflorum (Hoffmanns. &
Link) (Fig. 5). Higher pH together with Texture Class (silty clay
loam) soil condition was the main influencing environmental var-
iable of this zone as compared to heavily and less marble waste
polluted constituencies (Table 4). Other indicators of this zone were
Adiantum capillus-veneris L, Melilotus alba Medik, Boerhavia pro-
cumbens Banks ex Roxb and Parthenium hysterophorus L. along with
p  0.05 and IV  25% (Table 2).
When edaphic factor get changed it could sustain growth of
various indicator species. The soil pH of this region varies from 6.8
to 7.3, EC was in the range of 0.586e1.659 dsm1, organic matter
0.48e0.72%, phosphorus 3e8 ppm and potassium ambit from 49 to
113 ppm (Table 3).
3.2.3. Less polluted zone (LPZ)
This zone comprised of 12 stations and 62 different plant species
after CA and TWCA based on Jaccard Similarity Index (JSI). Topmost
indicator species of this less marble waste polluted zone were
Dalbergia sissoo DC, Dodonaea viscosa (L.) Jacq and Panicum anti-
dotale Retz (Fig. 6). These were the indicators of lower EC and
moderate potassium (K) consolidation as compared to other zones
(Table 5). Among other indicators Robinia pseudoacacia L, Cannabis
sativa L, Dysphania ambrosioides (L.) Mosyakin & Clemants,
Convolvulus arvensis L, Desmostachya bipinnata (L.) Stapf, Duchesnea
indica (Jacks.) Focke, Euphorbia chamaesyce L., and Oxalis corniculata
L are the noteworthy (Table 2).
Concentration of edaphic factor in LPZ i.e., EC range from 0.654
to 1.324 dsm1, pH ambit from 6.5 to 7.4, organic matter 0.6e0.8%,
phosphorus stretch from 4 to 7 ppm and potassium get ranges from
78 to 112 ppm that play a significant role in indicators of this region
as compared to previously mentioned zones (Table 3).
5
3.2.4. Indirect ecological gradient through Detrended
Correspondence Analysis (DCA)
Detrended Correspondence Analysis (DCA) was performed to
elucidate distribution of top most indicator species of each zone
and 45 stations along the axes of DCA plot (Fig. 7). Maximum
gradient length recoded for axis 1 was 5.264 along with 0.565
eigenvalue. The gradient length of axis 4 was 3.041 in conjunction
with 0.251 eigenvalue. Sum of all the eigenvalues/total inertia was
reported as 7.524 (Table 6).
3.2.5. Direct ecological gradient through Canonical Correspondence
Analysis (CCA)
The ordination of plant species through CCA bi-plot shows dif-
ferential and similarity indexes in plant species and the distance
between them. It was resulted that all the environmental variables
i.e., organic matter, potassium, phosphorous, pH, Soil Texture and
EC have significant effect (p  0.05) on composition and distribu-
tion pattern of indicator species (Table 7). CCA bi-plot reconfirms
the observation recorded through ISA. Like the topmost indictors of
HPZ were clustered under the impact of higher phosphorus con-
centration, organic matter, texture and potassium along with lower
pH as environmental variables. Whereas, indicator species of MPZ
were under the influence of higher pH, as denoted by long arrow as
compared to EC in addition to lower Organic Matter and Soil
Texture variables. Indicators of LPZ were assembled around the
moderate Organic Matter and lower Electrical Conductivity as
compared to the other zones mentioned previously (Fig. 8).
On the basis of aforementioned results four strongest indicator
species were further examined for their phytoremediation ability in
order to find out their role as phytoextractors and stabilizer in the
region of waste effluents. The detail descriptions of the results
obtained through Atomic absorption Spectrometry are as follows:
6 Z. Ahmad et al. / Journal of Cleaner Production 236 (2019) 117709

Fig. 4. Distribution plots (aec) and data attribute plots (def) of Populus alba, Arundo donax and Erigeron canadensis top 3 indicator of heavily marble waste polluted zone along with
influencing environmental factors using PCORD and CANOCO software's.

Table 1
Detail information of top 3 indicators of heavily marble waste polluted zone along with their environmental variable, Total Importance Value Index (TIVI), Indicator Values (IV)
and Probability Values (p*).

Total Number of stations were thirteen

S.NO Plant Indicators Environmental Variables IV P* Values TIVI

1 Populus alba L Phosphorus 51 0.032 261.63

2 Arundo donax Phosphorus 43 0.039 569.84
3 Erigeron canadensis L Phosphorus 39 0.044 124.60

3.3. Phytoremediation ability of the selected indicator species
Phytoremediation refers to the use of plant species and associ-
ated microbes to decrease toxic effects or concentration in the
environments. The average heavy metals concentration in soil of
less Polluted Zone varied from 33.45 to 36.15 mg/kg. Moderate
Polluted Zone from 37.95 to 42.05 mg/kg and that of Heavily
Polluted Zone from 41.05 to 47.8 mg/kg. The concentration of heavy
metals recorded in water of LPZ ranged from 12.1 to 299.05 mg/kg,
MPZ from 15.05 to 383.7 mg/kg and in HPZ varied from 15.9 to
439.45 mg/kg (Table 8).
Four indicator species i.e., P. alba, A. donax L, E. canadensis and
M. alba were further examined for their phytoremediation abilities.
Approximately, all these species had BCF>1 for Fe, Mg, Ca, Na and
Cu except for Cr, Pb and Cd metals in HPZ and MPZ. While in LPZ
these indicator bare BCF>1 for Fe, Mg and Na but less than one for
Cu, Cr, Pb and Cd heavy metals. These indicators were efficient to
take up and translocate all the measured heavy metals in HPZ and
MPZ and Fe, Mg, Ca, Na in LPZ on the basis of TF values. All the
plants screened for total metals concentration showed BAC<1 in all
zones, but comparatively a bit higher in HP and MP zone as
compared to LP region (Table 8; Fig. 9). The recorded BCF values in
root and shoot indicated that these species were the hyper accu-
mulator for Fe, Mg, Ca, Na and Cu in marble waste polluted
ecosystem. However, based on TF and BAC values these plant spe-
cies were identified as significant phytostabilizers and phytoex-
tracters of the marble waste polluted ecosystem.
4. Discussion
Competitive interactions exist among the living organisms
within ecosystem which are the key determinants of diversity and
abundance of flora as well fauna (Zhou et al., 2008). Industrial
pollution especially marble wastes affects water, air and soil that
 Z. Ahmad et al. / Journal of Cleaner Production 236 (2019) 117709 7

Table 2
Results of Indicator Species Analysis via PCORD, showing top most indicators (with bold font) of each of the 3 marble waste polluted zones (1e3) in relation to different
environmental variables at 25% threshold level of indicator based on Monte Carlo test of significance for observed maximum indicator values (% of perfect indication based on
combining values for relative frequency and relative abundance for species (p*0.05). (p* ¼ probability value; Max grp ¼ Maximum Group Value; IV¼ Indicator Value).

S.NO Botanical Names Heavily Polluted Zone Moderate Polluted Zone Less Polluted Zone defined
defined based on values of defined by the values of pH by the values of Electrical
Phosphorus Conductivity

Max grp IV p* Max grp IV p* Max grp IV p*

1 Acacia nilotica (L.) Delile 6 14 0.428 6 9.6 0.361 1 9.8 1.000

2 Ailanthus altissima (Mill.) Swingle 7 20.4 0.443 6 30 0.210 0 45.7 0.119
3 Broussonetia papyrifera (L.) L'Her. ex Vent. 7 19.1 0.331 6 7.2 0.938 0 13.7 0.878
4 Callistemon lanceolatus (Sm.) Sweet 7 14 0.398 6 3.7 1.000 1 2.4 1.000
5 Celtis australis L. 6 9.8 0.675 7 14.9 0.132 0 14.4 0.248
3
6 Dalbergia sissoo DC 5 4.5 1.000 6 7.4 0.505 0 50 0.006
7 Ficus carica L. 7 15.5 0.715 6 21.8 0.620 0 24.3 0.666
8 Melia azedarach L. 8 14 0.762 6 14.4 0.988 1 16.5 0.866
2
9 Morus alba L. 4 9.2 0.713 7 28.4 0.032 1 12.2 0.936
10 Morus nigra L 7 10.8 0.479 6 11.1 0.267 0 19.7 0.162
11 Pinus wallichiana A.B.Jacks. 7 14 0.398 6 2.3 1.000 1 4.9 1.000
1
12 Populus alba L 8 50.7 0.032 7 19.7 0.592 1 20.6 0.725
13 Robinia pseudoacacia L 6 7.7 0.690 7 5.6 0.406 0 25 0.091
14 Salix babylonica L 4 7.4 0.930 7 12.8 0.438 0 15.8 0.831
15 Ziziphus nummularia (Burm.f.) Wight & Arn 6 5.5 0.919 6 8.3 0.473 0 19 0.174
1
16 Arando donex 8 43.5 0.039 6 19.7 0.567 1 21.3 0.634
17 Berberis lycium Royle 7 16.7 0.186 6 3.7 1.000 1 2.4 1.000
18 Datura innoxia Mill 4 5.9 0.807 7 3 1.000 1 4.9 1.000
19 Debregeasia salicifolia (D.Don) Rendle 6 6.4 0.904 7 13.6 0.166 1 12.2 1.000
3
20 Dodonaea viscosa (L.) Jacq 7 16.7 0.180 6 3.7 1.000 0 29.4 0.048
21 Hypericum oblongifolium Choisy 7 16.7 0.186 6 3.7 1.000 1 2.4 1.000
22 Sideroxylon mascatense (A.DC.) T.D.Penn 8 24.4 0.150 6 11.1 0.259 0 15 0.254
2
23 Rosa webbiana Wall. ex Royle 6 14.3 0.643 7 33.4 0.025 0 36.4 0.158
24 Rumex hastatus D. Don 5 7.1 1.000 6 3.7 1.000 1 2.4 1.000
25 Vitex negundo L 4 7.1 0.943 6 7.9 1.000 1 17.1 0.711
26 Adenostemma lavenia (L.) Kuntze 6 7.7 0.684 7 5.6 0.388 1 2.4 1.000
27 Adiantum capillus-veneris L 4 16.7 0.480 7 41.7 0.001 1 22 0.568
28 Alternanthera pungens Kunth 6 15.4 0.230 6 2.6 1.000 1 4.9 1.000
29 Anagallis arvensis L 7 13 0.688 6 13.2 0.630 0 16.4 0.656
30 Anisomeles indica (L.) Kuntze 6 8.9 0.617 6 4.7 0.859 0 20 0.172
31 Avena sativa L 7 8.4 0.686 6 5.1 0.709 1 7.3 1.000
32 Boerhavia procumbens Banks ex Roxb 8 50 0.051 6 3.7 1.000 0 25 0.088
33 Bromus japonicus Thunb 8 26.8 0.145 7 5.3 0.794 0 15.7 0.315
34 Calendula arvensis M.Bieb 5 12 0.479 6 4.3 0.846 1 7.3 1.000
35 Cannabis sativa L 8 31.4 0.105 7 14.7 0.955 0 61.1 0.020
36 Capsella bursa-pastoris (L.) Medik 6 10.9 0.409 6 5.2 0.705 1 7.3 1.000
37 Carbenia benedicta 4 11.1 0.390 7 5.6 0.408 1 2.4 1.000
38 Cardamine impatiens L 5 4.1 1.000 6 7.4 0.507 1 4.9 1.000
39 Carthamus oxyacantha M.Bieb 6 7.7 0.679 7 5.6 0.391 1 2.4 1.000
40 Centaurea iberica Trevir. ex Spreng 6 15.4 0.308 6 3.1 1.000 1 4.9 1.000
2
41 Centaurium tenuiflorum (Hoffmanns. & Link) Fritsch 5 8.8 0.988 7 36.1 0.018 1 14.5 0.902
42 Chenopodium album L. 7 19.6 0.243 6 11.1 0.272 1 7.3 1.000
43 Dysphania ambrosioides (L.) Mosyakin & Clemants 6 7.7 0.676 6 3.7 1.000 0 25 0.086
44 Cirsium arvense (L.) Scop 8 26.1 0.147 7 6 0.847 0 16.1 0.254
45 Convolvulus arvensis L 8 20.8 0.306 7 13.7 0.512 0 61.9 0.012
1
46 Erigeron canadensis L. 8 39.4 0.044 7 34.5 0.240 1 40.1 0.354
47 Lepidium didymum L. 6 11.7 0.481 6 11.1 0.270 1 7.3 1.000
48 Cynodon dactylon (L.) Pers. 8 23.1 0.379 7 32.5 0.223 1 32.7 0.537
49 Cyperus rotundus L 6 38.5 0.051 7 9.6 0.665 1 14.6 0.869
50 Desmostachya bipinnata (L.) Stapf 4 11.2 0.873 6 19 0.304 0 39.2 0.074
51 Dichanthium annulatum (Forssk.) Stapf 8 23.8 0.222 7 14.7 0.511 1 22 0.572
52 Dicliptera chinensis (L.) Juss. 7 7.4 0.805 7 7 0.712 1 9.8 0.921
53 Digitaria sanguinalis (L.) Scop 4 5.9 0.917 7 8.7 0.464 1 7.3 1.000
54 Duchesnea indica (Jacks.) Focke 7 5.5 0.901 7 9.7 0.381 0 23.2 0.098
55 Emex spinosa (L.) Campd 6 4 1.000 7 3.3 0.761 1 4.9 1.000
56 Equisetum arvense L 5 5.6 0.919 6 7.4 0.521 1 4.9 1.000
57 Erodium cicutarium (L.) L'He r 4 6.5 0.894 6 4.3 1.000 1 9.8 0.923
58 Euphorbia granulata Forssk 6 15.4 0.315 7 11.1 0.151 1 4.9 1.000
59 Euphorbia chamaesyce L. 7 28.4 0.227 6 32.8 0.184 0 51.5 0.077
60 Filago hurdwarica (Wall. ex DC.) Wagenitz 7 16.7 0.186 6 3.7 1.000 1 2.4 1.000
61 Potentilla indica (Jacks.) Th.Wolf 8 41.2 0.051 6 7.4 0.507 0 23.5 0.088
62 Fumaria indica (Hausskn.) Pugsley 4 11.1 0.390 7 5.6 0.408 1 2.4 1.000
63 Galium aparine L. 6 15.1 0.555 7 11.6 0.668 1 19.5 0.576
64 Lathyrus aphaca L. 8 12 0.860 7 26 0.132 0 30.7 0.223
65 Lathyrus pratensis L. 5 20.9 0.306 6 11 0.457 0 15.1 0.382
66 Malva parviflora L. 7 4.5 0.919 6 4.4 0.860 1 7.3 1.000
67 Medicago polymorpha L. 5 9.3 0.810 7 20.7 0.119 1 19.5 0.575
68 Melilotus alba Medik 6 11.1 0.841 7 25.4 0.064 0 15.1 0.760
(continued on next page)
8 Z. Ahmad et al. / Journal of Cleaner Production 236 (2019) 117709

Table 2 (continued )

S.NO Botanical Names Heavily Polluted Zone Moderate Polluted Zone Less Polluted Zone defined
defined based on values of defined by the values of pH by the values of Electrical
Phosphorus Conductivity

Max grp IV p* Max grp IV p* Max grp IV p*

69 Oenothera rosea L'He r. ex Aiton 5 10.8 0.813 6 18.2 0.226 0 27.4 0.129
70 Mentha arvensis L 5 5.1 0.865 7 11.1 0.151 1 4.9 1.000
71 Mentha longifolia (L.) L 5 5.5 0.942 6 10.6 0.545 0 13.1 0.876
72 Nasturtium officinale R.Br. 7 5.8 0.915 6 5.8 0.963 1 12.2 1.000
73 Malva neglecta Wallr. 7 16.7 0.186 6 3.7 1.000 1 2.4 1.000
74 Opuntia deamii Rose 7 22.7 0.234 6 9.1 0.715 1 14.6 0.872
75 Origanum vulgare L 4 11.1 0.390 7 5.6 0.408 1 2.4 1.000
76 Oxalis corniculata L 7 15.2 0.783 6 24.3 0.588 0 74.2 0.010
3
77 Panicum antidotale Retz 5 7.1 1.000 6 3.7 1.000 0 32 0.042
78 Papaver rhoeas L. 6 8.9 0.613 6 4.2 0.853 1 7.3 1.000
79 Parthenium hysterophorus L 8 59.2 0.022 6 16.7 0.390 0 39.2 0.062
80 Paspalum distichum L 7 24.1 0.217 6 15.4 0.298 1 17.1 0.776
81 Persicaria maculosa Gray 8 12.6 0.756 7 11.7 0.925 1 13.7 0.781
82 Phalaris minor Retz 4 17.1 0.468 7 12.3 0.743 0 14.4 0.759
83 Phyla nodiflora (L.) Greene 5 11.3 0.480 6 11.1 0.275 1 7.3 1.000
84 Plantago lanceolata L 7 26.6 0.250 6 23 0.717 1 43.9 0.258
85 Persicaria barbata (L.) H.Hara 6 23.1 0.193 7 6.5 0.561 1 7.3 1.000
86 Polygonum plebeium R.Br. 7 14.5 0.307 6 2.5 1.000 1 4.9 1.000
87 Rosa webbiana Wall. ex Royle 5 4.4 1.000 6 7.4 0.511 0 23.6 0.087
88 Rumex dentatus L 7 22.7 0.308 7 20.8 0.803 0 28.3 0.531
89 Saussurea costus (Falc.) Lipsch 4 3.6 0.981 6 5 0.712 1 7.3 1.000
90 Silene conoidea L. 6 7.6 0.771 6 9.9 0.381 1 9.8 1.000
91 Silybum marianum (L.) Gaertn 5 8.2 0.734 6 7 0.678 0 9.3 1.000
92 Solanum surattense Burm. f 6 7.7 0.684 7 5.6 0.388 1 2.4 1.000
93 Sonchus asper (L.) Hill 7 16.7 0.186 6 3.7 1.000 1 2.4 1.000
94 Sonchus oleraceus (L.) L 5 18.9 0.724 6 27.4 0.671 1 29.8 0.673
95 Stellaria media (L.) Vill. 6 5 0.932 7 4.5 0.758 1 4.9 1.000
96 Taraxacum campylodes G.E.Haglund 7 5.7 0.915 7 3.4 1.000 1 7.3 1.000
97 Torilis leptophylla (L.) Rchb.f. 8 20.6 0.323 7 12.6 0.614 0 14.5 0.844
98 Verbascum thapsus L 7 8.4 0.764 7 9.7 0.528 1 12.2 1.000
99 Verbena officinalis L 4 11.6 0.499 6 8.3 0.471 1 9.8 1.000
100 Veronica biloba schreb. ex L 6 7.7 0.677 6 3.7 1.000 1 2.4 1.000
101 Vicia monantha Retz 4 7 0.805 7 8.7 0.460 1 7.3 1.000
102 Xanthium strumarium L 5 14.2 0.872 7 23.5 0.659 1 23.3 0.874

ultimately disturb the surrounding ecosystems. (Smithson et al.,
2013). Studies ranging from baseline to specialized one are
imperative to come up with a reasonable solution for such deteri-
orated natural systems.
Current study acknowledged a total of 102 plant species belong
to 95 genera and 48 families in the marble waste polluted
ecosystem of Buner region of Pakistan. The family Asteraceae was
the topmost dominant family having 14 different plants covering
29% of the total plant species. They showed significance with higher
Phosphorus concentration, higher pH, moderate potassium and
lower electrical conductivity environmental variables. Extensive
review of literature showed that there is no reasonable data on
vegetation of marble waste polluted habitats for comparison as
well as foundation purposes. However (Kabir et al., 2010), reported
eighty plants in a cement polluted ecosystem from Karachi,
Pakistan with higher concentration of Calcium carbonate, EC, pH,
TDS, exchangeable Potassium, Sodium and low quantity of Organic
Matter concentration. In addition, they documented the industrial
soil was mostly of porous type with considerable amount of water
holding capacity. More such studies can be great importance for
better management of such regions and systems.
One of the important applications of present research work was
the use of multivariate statistical approach. It allows the researcher
to compare the multiple classifications of the same sites/zones for
substantial information resulting from the analysis till inferences
(Khan et al., 2016). In present research work CA and TWCA of PC-
ORD version 5 for the identification of significant polluted zones
on the basis of similarity index resulted 3 major zones i.e., HPZ
devising higher phosphorus concentration, MPZ with higher pH
and silty clay loam soil condition and LPZ accompanied by lower
Electrical Conductivity and Potassium alliance. Indicator species of
each sort of zone were find out using ISA that provided the infor-
mation about species fidelity in a particular habitat (Dufre ^ne and
Legendre, 1997). A threshold level of 25% along with 95% signifi-
cance (p  0.05) was considered as cut off value for determination
of these indictors (Dufre ^ne and Legendre, 1997; Ter Braak and
Prentice, 1988). Indicators of the HPZ were P. alba, A. donax L and
E. canadensis. Similarly M. alba, R. webbiana and C. tenuiflorum were
the indicator of MPZ. While that of LPZ were D. sissoo, D. viscosa and
P. antidotale. The aforementioned indicators of each zone unique
due to presence of more concentration of marble waste in the re-
gion and variation edaphic factors. Furthermore, DCA and CCA
determined the impact of marble waste pollution and other
edaphic factors on species in general and indicator species in
particular. Moreover, DCA and CCA reconfirm the finding regarding
indicator species. The aforementioned statistical techniques were
quite often used by different researcher in classification, identifi-
cation of indicator species and impact of different environmental
gradient in normal ecosystem and vegetation studies (Iqbal et al.,
2015; Khan et al., 2012). These techniques have rarely been taken
into consideration for such polluted system and one of its pioneer
in nature in our current study.
Industrial effluents encompass heavy metals and different types
of nutrients that effect plant species and physicochemical proper-
ties of soil in a variety of ways (Akbar et al., 2007). Majority of these
effluents were disposed without any treatment in water or soil that
 Z. Ahmad et al. / Journal of Cleaner Production 236 (2019) 117709 9

Table 3 phytoremediation.
Physicochemical analysis of heavily, moderate and less polluted zones of marble Current study focused partly on identification of plant indicators
waste polluted ecosystem.
that tolerate marble wastes and reduce its toxicity. Concentrations
Station No. pH EC dsm-1 OM % P (ppm) K (ppm) Texture of marble wastes not only influence indicator species but also bring
Heavily polluted zone about changes in the soil conditions. The phytoremediation ability
F1Q1 7.2 1.222 0.7 6 101 Sandy clay loam of selected plants i.e., P. alba, A. donax L, E. canadensis and M. alba,
F2Q1 6.7 1.023 0.7 5 102 Clay loam soil and water samples were determined. It discloses
F3Q1 6.8 1.004 0.7 7 83 Clay loam
33.45e36.15 mg/kg, 37.95e42.05 mg/kg and 41.05e47.8 mg/kg
F4Q1 7.3 1.654 0.6 4 78 Silty clay loam
F5Q1 6.5 1.365 0.6 5 105 Sandy clay loam concentration of heavy metals in less, moderate and heavily
F6Q1 7.2 1.365 0.7 5 97 Clay loam polluted zone soil, respectively. The concentration of heavy metals
F7Q1 6.8 1.001 0.7 7 96 Loam recorded in water of LPZ ranged from 12.1 to 299.05 mg/kg, Mod-
F9Q1 6.8 1.024 0.7 5 78 Loam
erate polluted zone stretch from 15.05 to 383.7 mg/kg and in
F10Q1 6.4 1.324 0.6 8 96 Silty clay loam
F11Q1 7.3 1.332 0.6 4 78 Silty clay loam
heavily polluted zone varied from 15.9 to 439.45 mg/kg. Similarly
F12Q1 7.1 1.145 0.7 4 92 Loam (Malik et al., 2010) worked on 16 plant species to assess 6 toxic
F13Q1 6.8 1.221 0.7 4 76 Silty clay loam metal (Cu, Co, Cr, Pb, Zn and Ni) contents collected from industrial
F14Q1 6.9 0.658 0.7 6 83 Sandy clay loam zone of Islamabad, Pakistan. Where, total metal concentrations
Moderate polluted zone
varied between 2 and 927.2 mg/kg in soil. Edaphic factors of HPZ
F1Q2 7.1 1.024 0.6 6 88 Sandy clay loam
F2Q2 7.2 1.001 0.7 6 113 Clay loam i.e., Phosphorus varied from 4 to 8 ppm, pH 6.4e7.3, organic matter
F2Q3 7.1 1.258 0.7 4 94 Sandy clay loam 0.58e0.74% and K 76e105 ppm. Soil pH of MPZ various from 6.8 to
F3Q2 7.1 1.002 0.7 5 90 Sandy clay loam 7.3, organic matter 0.48e0.72%, phosphorus 3e8 ppm and potas-
F4Q2 6.9 1.532 0.5 3 49 Sandy loam sium 49e113 ppm. Whereas, pH ranges from 6.5 to 7.4, organic
F5Q2 7.2 1.001 0.7 5 89 Silty clay loam
F6Q2 6.8 1.004 0.6 5 83 Loam
matter 0.6e0.8%, Phosphorus 4e7 ppm and potassium 78e112 ppm
F6Q3 6.9 1.221 0.7 6 88 Loam in LPZ that plays a significant role in heavy metal accumulation of
F7Q2 7.3 1.659 0.5 5 89 Sandy clay loam the aforementioned plants. Comparing (Malik et al., 2010) that
F7Q3 7.3 0.922 0.7 6 112 Loam indicated accumulation of heavy metals varied greatly among
F8Q1 7.2 1.256 0.6 4 104 Clay loam
plants. Primarily its uptake dependent on plant species, inherent
F8Q2 7.2 1.584 0.7 5 93 Sandy clay loam
F9Q2 6.8 1.11 0.7 7 94 Clay loam control and soil condition as well. Huge number of factors controls
F10Q2 6.9 1.254 0.7 4 104 Loam the accumulation and bioavailability associated with climatic, soil
F11Q2 7.3 1.114 0.7 6 95 Clay loam conditions and plant genotype counting passive or active transfer
F12Q2 6.8 1.002 0.7 4 91 Sandy clay progressions, speciation and sequestration, redox conditions, type
F13Q2 6.9 1.251 0.5 7 91 Loam
F14Q2 6.8 1.021 0.7 6 103 Loam
of plant root system and the response of plant species to elements
F15Q1 6.9 1.112 0.7 6 105 Loam along with seasonal cycles. The positively charged ions i.e., cations
F15Q2 6.9 0.586 0.6 8 85 Sandy clay loam of heavy metal formed strong bond with the negatively charged
Less polluted zone carboxyl group of protein and often disrupt ionic bond that may
F1Q3 6.8 0.654 0.7 5 98 Loam
ultimately leads to denaturation of proteins. In addition to, higher
F3Q3 6.8 1.231 0.7 6 94 Loam
F4Q3 6.7 1.006 0.7 7 78 Sandy clay loam pH concentration resulted greater retention of heavy metals and
F5Q3 6.9 1.003 0.7 5 79 Sandy clay loam their low solubility in soil. Higher pH application stabilized toxic
F8Q3 7.4 1.259 0.6 6 88 Silty clay loam elements that results decreased leaching effect of soil toxic ele-
F9Q3 6.9 1.11 0.6 5 89 Loam ments. This confined absorbability of metals from soil solution and
F10Q3 6.9 1 0.6 6 96 Sandy clay loam
F11Q3 7.2 1.324 0.8 4 84 Clay loam
its translocation into tissue of plants (Liu et al., 2005).
F12Q3 7.1 1.024 0.7 5 91 Loam The recorded BCF, TF and BAC values of P. alba, A. donax L, E.
F13Q3 6.5 1.033 0.7 5 89 Sandy clay loam canadensis and M. alba in root and shoot indicated that these spe-
F14Q3 6.9 1.112 0.6 7 112 Clay loam cies are identified as a significant phytostabilizers and phytoex-
F15Q3 6.7 1.004 0.6 6 88 Clay loam
tracters of marble waste polluted ecosystem (Mulk et al., 2015).
OM ¼ organic matter, P ¼ phosphorus, K¼ Potassium, EC ¼ electrical conductivity, evaluated the impact of marble industry effluents on water and
F ¼ Marble factory, Q ¼ Quadrat/station. sediments quality and concluded that heavy metals concentration
both in sediments and water significantly increased due to marble
wastewater (Zornoza et al., 2013). appraised effect of pig slurry
may lead to deteriorated river water. These water are being used for
alone and combined with marble waste in terms of heavy metal
many activities like agriculture as well as drinking. Such soils are
stabilization, organic matter mineralization and microbial activ-
also cultivated for economic crops, vegetables and fruits. As a result
ities. They concluded that the combined pig slurry and marble
of these pollutants neither water nor soil remained safer for use by
waste were the most significant treatment along highest general
both plants and animals. The removal of pollutants from industrial
reduction in metal availability and increases in microbial size and
waste has been tried by different ways. It included ion exchange
activity. It helps to stabilize organic matter and decrease carbon
method, chemical extract, polymer micro encapsulation, excava-
loss. The application of Pig slurry increased the accumulation of
tion, precipitation and electrolytic techniques etc., (Antunes et al.,
heavy metals in shoot. While, pig slurry along with marble waste
1998; Waltner-Toews, 2001). The bigger challenge in this regard
decreased this accumulation (Kabas et al., 2012). In addition to
is the ecofriendly management of these pollutants. Phytor-
(Kumar and Thambavani, 2012), worked on the biological moni-
emediation is one of the most rapidly developing field using plants
toring of roadside plants exposed to vehicular pollution and indi-
to decrease the harmful effect and detoxify various pollutants
cated that Azadirachta indica, Pongamia pinnata, Delonix regia,
(Mani and Kumar, 2014). It leads to preserve biodiversity of an
Polyalthia longifolia and Ficus religiosa were indicators of urban air
ecological ecosystem and certify to sustain life beyond these floras
pollution. Whereas (Akbar et al., 2007), studied the effect of marble
(Srivastava and Vellend, 2005). The current study is one of such
waste effluent on development and growth of Maize plants and
attempt recognizing the evaluation of vegetation of marble waste
indicated that increase in concentration of such waste increase pH
polluted habitat and role of selected indicator plants in
of soil and can be used as a best source (in low amount) for soil with
10 Z. Ahmad et al. / Journal of Cleaner Production 236 (2019) 117709

Fig. 5. Distribution plots (aec) and data attribute (def) of Morus alba (1st indicator species), Rosa webbiana (2nd indicator) and Centaurium tenuiflorum (3rd indicator) of moderate
polluted zone in relation to various measured environmental variables after species distribution analysis of PCORD and Canonical correspondence analysis of CANOCO software.

Table 4
The top 3 indicator species of moderate marble waste polluted zone together with their respective environmental variables and indicator values after ISA.

Total number of stations were twenty

S. No Indicator plants Environmental Variables IV P* Values TIVI

1 Morus alba L pH 28 0.032 172.37

Texture class 27 0.029
2 Rosa webbiana Wall. ex Royle pH 33 0.025 508.47
3 Centaurium tenuiflorum (Hoffmanns. & Link) pH 36 0.018 74.19

Fig. 6. Distribution plots (aec) and data attributes (def) of topmost 3 indicator species i.e., Dalbergia sissoo, Dodonaea viscosa and Panicum antidotale of less marble waste polluted
zone together with respective environmental variables.
 Z. Ahmad et al. / Journal of Cleaner Production 236 (2019) 117709 11

Table 5
Detail information of uppermost indicators of less marble waste polluted region in conjunction with corresponding environmental variables, Total Importance Index (TIVI),
Indicator Values (IV) and Probability (p*) Values after ISA.

Total Number of stations were twelve

S.NO Indicator plants Environmental Variables IV P* Values TIVI

1 Dalbergia sissoo DC Electrical Conductivity 50 0.006 22.02

2 Dodonaea viscosa L. Electrical Conductivity 29 0.048 100.00
3 Panicum antidotale Retz Potassium 32 0.042 05.36

Fig. 7. DCA diagram presenting distribution of stations (A) and indicator species (B) into 3 polluted zones via CANOCO software. (F ¼ factory, Q ¼ Quadrat).

Table 6
DCA summary of topmost indicators and stations of each polluted zone along eigenvalues with different gradient lengths of all four axes.

Axes 1 2 3 4 Total inertia

Eigenvalues 0.565 0.450 0.360 0.251 7.524

Length of gradient 5.264 3.702 3.420 3.041
Cumulative % variance of species data 7.5 13.5 18.3 21.6

Table 7
Summary of Canonical corresponding analyses of all indicator species in conjunction with various measured environmental variables of the marble waste polluted ecosystem.

Axes 1 2 3 4 Total inertia

Eigenvalues 0.277 0.183 0.175 0.150 7.524

Species environment correlations 0.863 0.787 0.768 0.784
Cumulative % variance of species data 3.7 6.1 8.5 10.4
Cumulative % variance of species environmental relation 27.5 45.6 63 77.8

Summary of Monte Carlo test

Test of significance of first canonical axis Test of significance of all canonical axes

Eigenvalue 0.277 Trace 1.009

F-ratio 1.454 F-ratio 0.981
P value 0.354 P-value 0.048

acidic nature because of higher calcium contents. The root length
and stem girth enhanced at 40% concentration of effluent while
decreases at higher concentration.
5. Conclusions
It was concluded that marble waste polluted ecosystem had
unique indicator species that performs well than others. P. alba L,
A. donax L and E. canadensis L. were the indicators of HPZ along with
higher Phosphorus concentration. Morus alba, Rosa webbiana and
Centaurium tenuiflorum were indicators of MPZ in conjunction with
higher pH and silt clay loam soil condition. Similarly, Dalbergia
sissoo, Dodonaea viscosa and Panicum antidotale were topmost
indicators of LPZ along with lower electrical conductivity and
moderate potassium concentration. Furthermore, atomic absorp-
tion spectrophotometry analysis identified Populus alba, Arundo
donax L, Erigeron canadensis and Morus alba as a significant phy-
tostabilization and phytoextraction of marble waste polluted
ecosystem on the bases of BCF, TF and BAC values of these plant
species. These indicator species grow, survive and showed more
tolerance and contest against marble pollution and its associated
heavy metals in a better way and therefore, can be used for soil and
water quality monitoring in marble waste polluted zones. It is
recommended to examine the protein contents of Morus alba fruit
to analyze how much marble waste polluted ecosystem affect the at
constituents level? Further study could be done to examine use of
12 Z. Ahmad et al. / Journal of Cleaner Production 236 (2019) 117709

Fig. 8. CCA bi-plot of CANOCO software representing A) all stations and B) top most indicators of each polluted zone under the influence of various measured environmental
variables. (F ¼ factory, Q ¼ Quadrat).

Table 8
Translocation factor (TF), Bioaccumulation coefficient (BAC) and Biological concentration factor (BCF) (in mg/kg) of selected indicator species of Heavily Polluted Zone (HPZ),
Moderate Polluted Zone (MPZ) and Less Polluted Zone (LPZ) of marble waste polluted ecosystem.

Plant species Factors Fe Mg Ca Na Cu Cr Pb Cd

Arundo donax BCF- HPZ 1.008 1.005 0.969 1.011 0.948 0.890 0.861 0.865
TF-HPZ 0.833 0.839 0.872 0.852 0.860 0.871 0.875 0.867
BAC-HPZ 0.839 0.843 0.845 0.862 0.815 0.775 0.753 0.750
BCF-MPZ 1.022 1.049 1.033 0.969 0.976 0.961 0.836 0.861
TF-MPZ 0.880 0.798 0.889 0.847 0.788 0.864 0.863 0.857
BAC-MPZ 0.899 0.837 0.918 0.821 0.770 0.831 0.722 0.738
BCF-LPZ 1.006 0.990 0.968 0.962 0.871 0.805 0.810 0.823
TF-LPZ 0.844 0.856 0.830 0.870 0.800 0.782 0.703 0.792
BAC-LPZ 0.849 0.848 0.803 0.838 0.696 0.630 0.569 0.651
Erigeron canadensis BCF-HPZ 1.000 0.980 1.001 1.025 0.998 0.825 0.816 0.845
TF-HPZ 0.840 0.861 0.871 0.841 0.868 0.860 0.860 0.865
BAC-HPZ 0.840 0.843 0.872 0.862 0.866 0.709 0.702 0.731
BCF-MPZ 0.982 1.001 1.045 1.010 0.982 0.872 0.882 0.900
TF-MPZ 0.847 0.864 0.844 0.889 0.863 0.788 0.858 0.833
BAC-MPZ 0.832 0.865 0.881 0.898 0.848 0.687 0.757 0.749
BCF-LPZ 1.006 1.043 0.939 0.962 0.869 0.804 0.802 0.809
TF-LPZ 0.830 0.686 0.839 0.870 0.834 0.690 0.782 0.703
BAC-LPZ 0.834 0.716 0.788 0.838 0.725 0.555 0.627 0.568
Populus alba BCF-HPZ 0.986 0.980 1.006 1.002 1.009 0.873 0.870 0.876
TF-HPZ 0.863 0.868 0.860 0.860 0.848 0.872 0.846 0.857
BAC-HPZ 0.851 0.850 0.864 0.862 0.855 0.761 0.736 0.750
BCF-MPZ 1.038 1.029 1.040 1.027 1.010 0.908 0.886 0.883
TF-MPZ 0.852 0.851 0.834 0.852 0.757 0.756 0.801 0.759
BAC-MPZ 0.884 0.875 0.867 0.875 0.764 0.687 0.710 0.670
BCF-LPZ 1.006 1.013 0.928 0.973 0.846 0.822 0.814 0.823
TF-LPZ 0.830 0.804 0.819 0.829 0.811 0.814 0.959 0.792
BAC-LPZ 0.834 0.815 0.760 0.807 0.686 0.669 0.781 0.651
Morus alba BCF-HPZ 1.002 1.002 1.016 1.014 1.021 0.894 0.889 0.877
TF-HPZ 0.860 0.858 0.859 0.867 0.865 0.861 0.842 0.846
BAC-HPZ 0.862 0.860 0.872 0.879 0.883 0.770 0.748 0.742
BCF-MPZ 1.008 1.000 1.033 1.030 0.976 0.872 0.886 0.840
TF-MPZ 0.838 0.849 0.838 0.874 0.803 0.857 0.842 0.863
BAC-MPZ 0.845 0.849 0.866 0.899 0.784 0.747 0.746 0.725
BCF-LPZ 0.991 1.019 0.969 0.978 0.836 0.796 0.816 0.805
TF-LPZ 0.842 0.805 0.798 0.811 0.750 0.785 0.728 0.761
BAC-LPZ 0.834 0.821 0.774 0.793 0.626 0.624 0.594 0.613
Soil samples HPZ 0.871 0.879 0.878 0.877 0.821 0.943 0.953 0.956
MPZ 0.766 0.762 0.759 0.776 0.716 0.827 0.841 0.837
LPZ 0.694 0.669 0.721 0.714 0.688 0.719 0.722 0.723
Marble waste water HPZ 2.172 1.095 8.789 2.377 0.622 0.740 0.690 0.318
MPZ 1.895 1.013 7.674 2.176 0.431 0.380 0.510 0.301
LPZ 0.963 0.796 5.981 1.564 0.397 0.290 0.450 0.242
 Z. Ahmad et al. / Journal of Cleaner Production 236 (2019) 117709
Fig. 9. Bioaccumulation coefficient (BAC), Translocation factor (TF) and Biological concentration factor (BCF) of some of the Arundo donax, Erigeron canadensis, Populus alba and
Morus alba indicator species.
these plants for other polluted systems to see if these plants could
be used as phytostabilizers and phytoextractors at broader scale.
Acknowledgement
The authors would like to extend their heartiest thanks to the
owners of marble factories and people living in the surroundings
for their support in one way or the other. We are also thankful to all
the members of Environmental Microbology Lab. QAU, Islamabad
for their technical assistance and facilitation.
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