Malaysian Journal of Sustainable Agriculture (MJSA) 3(1) (2019) 33-38
Malaysian Journal of Sustainable Agriculture (MJSA)
DOI : http://doi.org/10.26480/mjsa.01.2019.33.38
ISSN: 2521-2931 (Print)
ISSN: 2521-294X (Online)
CODEN: MJSAEJ
RESEARCH ARTICLE
EFFECTS OF HEAVY METALS (Cd, Zn and Cu) ON CARBON, NITROGEN AND IRON
MINERALIZATION IN SOIL
Falguni Akter1*, Humaira Hasan Tinni2, Parmita Banarjee2, Mohammad Zaber Hossain1
1Soil,
Water and Environment Discipline, Khulna University, Khulna, Bangladesh
fellow, Soil, Water and Environment Discipline, Khulna University, Khulna, Bangladesh
*Corresponding Author Email: falgunissku@gmail.com
2Research
This is an open access article distributed under the Creative Commons Attribution License, which permits unrestricted use, distribution, and reproduction
in any medium, provided the original work is properly cited
ARTICLE DETAILS
Article History:
Received 01 April 2019
Accepted 10 May 2019
Available online 14 May 2019
ABSTRACT
A sixty days laboratory incubation study was conducted to investigate the effect of heavy metals on soil carbon,
nitrogen and iron mineralization under aerobic condition. Sulphate salts of cadmium, zinc and copper were added
individually and in combinations to soil samples and incubated in different plastic pots. Soil organic carbon did not
change significantly throughout the incubation period. Soil microbial biomass carbon declined from 0.38mgkg-1to
0.19 mgkg-1 in Cd treated soil and 0.39 mgkg-1 to 0.28 mgkg-1 in Cu treated soil which account for about 50% and
28% reduction (p ≤0.05) in biomass carbon respectively. Cd:Zn and Cu:Cd treated soil had reduced 36.84% while
Zn:Cu had 42.11% reduction in biomass carbon.CO2-C effluxes peaked by day 15 for all the single metal amended
soil indicating that priming effects might have occurred. But in combination, metal showed some interaction for
what the respiration rates were declined for the first 15 days. Rapid ammonification with presumed immobilization
took place up to day 30. The result indicated a significant (p ≤0.05) net mineralization of nitrogen for Cd:Zn
(63.72%) and Cu:Cd (66.66%) treatments at the end of the experiment. Available iron content showed significant
changes in combined metal treatment than a single metal.
KEYWORDS
Heavy metals, microbial biomass carbon, respiration, nitrogen, iron, mineralization.
1. INTRODUCTION
Heavy metals are continuously accumulated in soil due to land disposal of
municipal and industrial wastes, emissions of automobile, use of chemical
fertilizers, pesticides and other toxic elements for agricultural purposes
[1-4]. It is clear that heavy metals commenced with compost or sewage
sludge, caused soil organic matter accumulation and decreased the
turnover rate of organic matter [5-8]. Fertilizer applications possibly
capable of influence Cd in soil which affects the movement of Cd to plant
roots in addition to Cd uptake [9]. Copper is mainly toxic to roots. High
copper interaction with iron metabolism is the causes of chlorosis as the
symptom of copper toxicity [10]. Along with this, higher concentrations of
copper can reduce the availability of zinc absorption to plant as these
micronutrients compete for the same sites of plant root [11]. Enormously
slight concentrations of some metals like copper, zinc, nickel, cadmium is
vital for the components of enzymes, pigments, structural proteins, and in
maintaining the ionic balance of cells [12]. Researchers all over the world
are concerned about heavy metals mainly due to their harmful effects on
plants, especially those on vegetative and generative parts of the plants.
They also endanger human health when the metals migrate through the
food chain [13]. Besides, heavy metals showed toxic effects on
microorganisms in various ways such as reduced litter decomposition and
nitrogen fixation, less efficient nutrient cycling [14]. There is now
considerable amount of evidence of documenting decrease in the soil
microbial biomass as a result of long-term exposure to heavy metal
contamination from past applications of sewage sludge as reviewed by
[15]. The soil organic carbon content had noteworthy positive correlations
with the dehydrogenase activity, basal soil respiration, catalase activity,
and microbial biomass-C at P<0.01 [16]. A previous researcher observed
negative relationship of basal soil respiration with heavy metal contents
[17]. Being a microbial dependent process, all those factors, which affect
microbial activities, indirectly affect N mineralization. Several studies
have shown that metals affect N mineralization, however, results are
contradictory. A researcher reported that mineralization of soil organic N,
as with microbial respiration, is unaffected at soil metal concentration at
around current EU limits [18]. This is supported by a previous researcher
who reported that inhibition of both N mineralization and nitrification are
inhibited at around 1000 mgkg-1 Zn, Cu and Ni, around 100-500 mgkg-1 Pb
and Cr and around 10-100 mgkg-1 Cd. Iron deficiency is extremely rare in
the field crops [19]. As a result, very few works deal with the iron as a
major nutrient that affects plant growth. Therefore, the present study has
been conducted to determine the effects of heavy metals (Cd, Zn and Cu)
on carbon, nitrogen and iron mineralization of an agricultural soil
artificially polluted with heavy metals. For this purpose, the rates of soil
organic carbon, nitrogen and iron mineralization, soil respiration and
microbial biomass carbon were monitored.
2. MATERIALS AND METHODS
Agricultural clayey loamy soil samples were collected from a field plot at
Noapara in Jashore district. The surface soil samples were collected from
specific locations (N 23º 11.841´E: 89º 14.660´) at a depth of 0-15 cm and
were processed for the subsequent experiment and analysis. The soils
were air dried and ground to pass through a 2 mm sieve, sorted to remove
stones, plant debris and any visible soil fauna and then were mixed
thoroughly with hand trowel. The general description of the soil used for
the experiment is shown in table 1.
Cite The Article: Falguni Akter, Humaira Hasan Tinni, Parmita Banarjee, Mohammad Zaber Hossain (2019). Effects Of Heavy Metals (Cd, Zn And Cu) On Carbon,
Nitrogen And Iron Mineralization In Soil. Malaysian Journal of Sustainable Agriculture, 3(1): 33-38.
Malaysian Journal of Sustainable Agriculture (MJSA) 3(1) (2019) 33-38
Table 1: Characteristics of the soil under study
Properties
Analysis
pH
7.30
Total nitrogen (%)
0.11
Carbon (%)
1.56
Available nitrogen (mgkg-1)
25
Biomass carbon(mgkg-1)
0.38
Field capacity (%)
60.62
Soil respiration(mg of Cg-1soil day-1)
5.64
Texture
Clay loam
Total Fe (%)
3.75
CEC (cmolckg-1)
37.60
Available Fe (mgkg-1)
1.91
Approximately 0.25 kg of sieved soil samples were distributed into plastic
pots. Solutions of analytical grade sulphate (SO 4) salt each of cadmium,
zinc and copper was applied to the respective pots singly and in
Treatment Notation
combinations. Soil sample which receive no metal amendment was served
as control. All the treatments (A-G) were replicated three times. Number
of treatments and their combinations were as follows:
Table 2: Pattern of metal amendment in pre-incubated soil pots
Metal
Concentration (mgkg-1)
0
Control
--
B
Cd
A
C
Cu
3000
Zn
3000
3000
D
Cu:Cd
1500:1500
F
Zn:Cu
1500:1500
E
Cd:Zn
G
Cu:Cd:Zn
Total length of the incubation period was 60 days and the time interval for
collection of incubated soil sample was 0, 15, 30, 45 and 60 days. The
experiment was conducted aerobically. Soil pH was determined
electrochemically at soil: water ratio of 1:2.5 by using Griffin (Model 40)
glass electrode pH meter as described by [20]. Organic carbon of the soil
sample was determined by Walkly and Black wet oxidation method as
suggested by [20]. Available nitrogen and total nitrogen of the soil both
were determined by colorimetric method as suggested by [21]. Soil
microbial biomass carbon was determined by fumigation extraction
method as outlined by [22]. Total iron was determined by
spectrophotometer after digestion with HNO3: HClO4 (2:1) mixture.
1500:1500
1000:1000:1000
Available iron was determined by 1N NH4OAc (pH 7.0) as described by
[23]. Total metal content of soil before incubation was determined after
acid
digestion
(HNO3:HClO4;
2:1)
by
atomic
absorption
spectrophotometer. Soil respiration was done on 20g portion of soil
incubated in a desiccator for 24 hours along with 5ml 0.1M NaOH followed
by titration with 0.1M HCl as described in [24].
The results were analyzed by two-way analysis of variance (ANOVA) at
95% confidence interval. Differences between the control and other
treatments were assessed using least significant difference (LSD) [25].
Table 3: Metal of soil detected prior to amendment as determined by atomic absorption spectrophotometer
Metal
Concentration (mgkg-1 soil)
Cu
0.38
Pb
0.33
Cd
Zn
0.03
3.76
Cite The Article: Falguni Akter, Humaira Hasan Tinni, Parmita Banarjee, Mohammad Zaber Hossain (2019). Effects Of Heavy Metals (Cd, Zn And Cu) On Carbon,
Nitrogen And Iron Mineralization In Soil. Malaysian Journal of Sustainable Agriculture, 3(1): 33-38.
Malaysian Journal of Sustainable Agriculture (MJSA) 3(1) (2019) 33-38
3. RESULTS AND DISCUSSION
Treatment
Days
0
Control
45
15
Table 4: Effects of heavy metals on changes of organic carbon in soil (%)
Cd
Zn
Cu
Cd:Zn
Zn:Cu
Cu:Cd
Cd:Zn:Cu
0.89
0.90
0.89
0.89
0.90
0.89
0.90
0.89
0.88
0.87
0.90
0.86
0.89
0.87
0.86
0.87
0.89
30
0.90
0.88
60
0.89
0.88
0.87
0.88
0.89
0.85
0.90
0.88
0.90
Organic carbon did not change significantly due to the application of heavy
metals during the entire incubation period (Table 4). Among the
treatments, Zn accumulated a little amount of carbon until 45 days while
Cu: Cd decreased more carbon than any other treatments. From the study,
it was observed that soil organic carbon was unaffected due to heavy
0.85
0.88
0.90
0.88
0.87
0.89
0.89
0.88
0.85
0.89
0.84
0.87
metals application. Less incubation time may be the reason for not getting
significant changes in the soil organic carbon content. A group of
researchers has worked on effects of heavy metals in acid and calcareous
soils for mineralization of organic carbon in incubation and found low
mineralization rate of organic-C after 28 weeks of incubation [26].
Table 5: Effects of heavy metals on changes of microbial biomass carbon in soil (mg kg-1)
Treatment
Days
0
Control
Cd
Zn
Cu
Cd:Zn
Zn:Cu
Cu:Cd
Cd:Zn:Cu
0.38
0.38
0.39
0.39
0.38
0.38
0.38
0.38
45
0.38
0.24*
0.37
0.31
0.30
0.30
0.32
0.38
15
30
60
0.38
0.35
0.38
0.39
0.30
0.37
0.35
0.37
0.19*
0.36
0.33
0.37
*Asterisks indicate significant difference from the control treatment
(N=8, P≤0.05).
0.36
0.31
0.28*
0.36
0.34
0.24*
0.38
0.31
0.22*
0.38
0.24*
0.37
their combinations along with Zn showed significant (p≤0.05) decreases
in soil MBC at day 60 in spite of the unusually very high levels of other
metals [27]. The availability and toxicity of a given metal ion is always
much lower in soil than in water solution as a result of adsorption of the
metal ions by soil organic and inorganic colloids and microorganisms can
overcome from their adverse effects. Interactions of metals used in
combination may be the result of simple competition, antagonism,
additive action or synergism [28, 29]. The finding is in agreement with a
scholar who observed diminished microbial biomass C in cadmiumcontaminated sewage sludge compost to the soil [30].
The table 5 showed that microbial biomass carbon (MBC) declined due to
the application of heavy metals with incubation time. Cd has decreased
more than any other treatments. The toxic effects of metals imposed on
soil in the following order: Cd>Cu>Zn. This order probably explains the
toxicity experienced by the soil microbes by the data on soil microbial
biomass carbon. With respect to the MBC the result indicates that due to
application of different metals and their combinations, the MBC didn’t
show any significant responses until day 45 except Cd. Cd, Cu itself and
Table 6: Effects of heavy metals on changes of respiration in soil (mg C g-1 soil day-1)
Treatment
Days
0
Control
Cd
Zn
Cu
Cd:Zn
Zn:Cu
Cu:Cd
Cd:Zn:Cu
5.64
5.40
6.82
4.70
6.30
5.20
6.20
5.90
45
8.40
7.80
7.38
7.87
5.88*
5.91
5.55*
15
30
9.20
8.52*
7.26
7.50
9.15
60
7.90
7.98
8.85*
7.38
7.44
7.39
*Asterisks indicate significant difference from the control treatment
(N=8, P≤0.05).
8.24
Initially respiration was increased and then became slowing due to
application of heavy metals during the entire incubation period (Table 6).
The maximum values for respiration were found from day 0 to day 15. The
result indicated that organic carbon release occurred at this time, showing
Sample
A
Treatment days
Control
0
15
30
45
60
82.02
93.16
118.11
91.04
46.75
6.40
5.50*
5.29*
5.88
5.40
4.68
6.01
5.51
5.30
5.18
5.72
5.73
4.02
the priming effect of the added organic materials. This finding is an
agreement with the study of [31]. It was also found that soil respiration
and especially the respiration per unit biomass (qCO2) increased with
increasing amounts of heavy metals due to the contribution of fungi to soil
respiration increased [32].
Table 7: Effects of heavy metals on changes of available N in soil(mg kg-1)
B
C
D
E
F
G
H
Cd
Zn
Cu
Cd:Zn
Zn:Cu
Cu:Cd
Cd:Zn:Cu
86.12
71.36*
127.13
93.50
46.75
95.14
82.84
125.49
100.07
40.19
61.52
49.21*
117.29
104.17
41.83
83.66
58.23*
113.19
87.76
30.35*
77.92
65.62*
107.45
63.98*
43.47
66.44
54.92*
108.27
104.17
22.15*
83.66
63.98*
101.71*
77.10
44.29
Cite The Article: Falguni Akter, Humaira Hasan Tinni, Parmita Banarjee, Mohammad Zaber Hossain (2019). Effects Of Heavy Metals (Cd, Zn And Cu) On Carbon,
Nitrogen And Iron Mineralization In Soil. Malaysian Journal of Sustainable Agriculture, 3(1): 33-38.
Malaysian Journal of Sustainable Agriculture (MJSA) 3(1) (2019) 33-38
*Asterisks indicate significant difference from the control treatment (N=8,
P≤0.05).
(117.29 mg kg-1). The probable reason for the decrement of available N in
soil after 30 days of incubation was due to ammonia volatilization and/or
immobilization during large heterotrophic microbial activities which
result in similar or lesser N to be mineralized in other treatments. Besides
this, the recalcitrant organic fractions might slow down N mineralization
over time. A scholar worked on nitrous oxide production from an ultisol
treated with different nitrogen sources and moisture regimes and found
similar results [34].
Net N mineralization over the 60-day incubation period differed
significantly (p≤0.05) among the treatments (Table 7). Minearalization
and immobilization of N were closely associated in all treatments and were
predominant until day 15 of the incubation period. This finding is in
agreement with [33]. The highest rate for Cd (127.13mg kg-1) displayed a
high degree of ammonification and also for Zn (125.49 mg kg-1) and Cu
Table 8: Effects of heavy metals on changes of total N in soil (%)
Sample
A
Treatment days
Control
30
0.46
0
15
60
C
D
E
F
G
H
Zn
Cu
Cd:Zn
Zn:Cu
Cu:Cd
Cd:Zn:Cu
0.44
0.52
0.46
0.50
0.43
0.46
0.53
0.45
0.45
0.38
0.20*
0.36
0.37
0.46
45
B
Cd
0.40
0.39
0.44
0.42
0.38
0.19
0.21*
0.48
0.25*
O
Treatment days
Control
2.08
2.02
1.10
0.96
1.14
O
Control
3.63
3.24
4.68
3.02
2.36
0.46
0.21*
0.39
0.22*
0.21*
B
C
D
E
F
G
Cd
Zn
Cu
Cd:Zn
Zn:Cu
Cu:Cd
Cd:Zn:Cu
0.37
0.30
1.49
0.49
0.60
0.36
0.18
1.77
0.89
0.53
2.59
2.02
1.06
1.17
1.17
2.44
0.95
1.03
1.31
0.92
2.50
1.33
0.96
0.89
0.85
2.56
6.37**
5.46**
0.78
0.50
2.46
7.32**
1.21
0.85
0.57
maximum mineralization (6.37 mgkg-1) was found at day 15 and day 30
the result was 5.46 mgkg-1 due to the synergistic interaction between them
but after that the value decreased drastically due to antagonistic
interaction between them. Kabata-Pendias [36] found both antagonistic
and synergistic relation between Cu and Cd.
From the Table 9, we found that as compared with control treatment there
was no significant relationship among the heavy metal treatments when
they are applied singly, but in combination a significant (p≤0.01)
relationship exist in Cu:Cd and Cd:Zn:Cu treatments. In case of Cu:Cd,
Sample
0.33*
A
**Asterisks indicate significant difference from the control treatment
(N=8, P≤0.01)
Treatment days
0.41
0.29*
0.32
Table 9: Effects of heavy metals on changes of available iron in soil (mg kg 1)
Sample
0.34
0.26*
[35]. Cd toxicity increased which affect N mineralization in soil [36].
Worked on cadmium and zinc uptake in wheat and found similar results.
The existence of antagonistic relationship between Cd and Zn prevails
throughout the incubation time. This phenomenon enhances the
nitrification by which nitrogen may be lost from the system. As a result,
values of total nitrogen obtained were minimized. Another researcher
found an antagonistic relationship between zinc and cadmium [36]. On the
contrary, it is believed that an additive action or synergistic effect between
Cu and Zn could have been responsible for the enhanced toxicity of Cu:Zn
in the soil sample and was reported by [37]. The combination of Cd:Zn:Cu
was less effective on the total nitrogen change. Based on a study, the
addition of heavy metals reduced N mineralization in soil [38]. This
phenomenon results from a decrease in nitrifying bacteria activity.
However, metal variables revealed positive interactions with parameters
and N-related indices such as urease, N-acteyl-glucosaminidase, total N
mineralization, ammonification and metabolic quotient [39].
Results indicated that net N mineralization was significantly (P≤0.05)
different among the treatments over the 60-day incubation period (Table
8). The total nitrogen content decreased throughout the incubation period.
It indicates mineralization and immobilization of N were closely
associated in all soils and were predominant until day 15 of the incubation
period. These findings are in agreement with [34]. The highest value for
Cd:Zn (0.52%) at day 30 displayed a high degree of ammonification
followed by Cu: Cd which was 0.50%and the lowest value for Cd (0.19%)
was found at day 60. Probably this is a result of small mineralization or
rapid remineralization of nitrogen over a period of time. Ammonia
volatilization
and/or
immobilization
by
the
heterotrophic
microorganisms could result in similar or lesser N to be mineralized in
other treatments. The recalcitrant organic fractions might slow down N
mineralization over time. Cd and Zn interaction may change the pH of soil
0
15
30
45
60
0.40
0.40
0.22*
*Asterisks indicate significant difference from the control treatment (N=8,
P≤0.05).
0
15
30
45
60
0.49
0.40
Table 10: Effects of heavy metals on changes of total iron in soil (%)
A
B
C
D
E
F
G
Cd
Zn
Cu
Cd:Zn
Zn:Cu
Cu:Cd
Cd:Zn:Cu
4.11
3.60
1.42**
0.88*
0.53
2.14
2.98
1.17**
0.79*
0.84
*Asterisks indicate significant difference from the control treatment (N=8,
P≤0.05); **Asterisks indicate significant difference from the control
treatment (N=8, P≤0.01)
1.85
3.40
0.14**
0.30
0.15
2.14
3.45
1.95*
0.36**
0.19
1.96
3.02
5.74
0.36**
0.16
1.85
3.32
1.38**
0.70*
0.32
3.63
3.22
1.60**
1.61
1.95
Initial mineralization took place in all the treatments until 15 days of the
incubation together with the probability of Fe immobilization (Table 10).
As compared to control, at day 30 except Zn:Cu, all treatments varied
significantly (p≤0.05) over the incubation period. At day 45, all treatments
Cite The Article: Falguni Akter, Humaira Hasan Tinni, Parmita Banarjee, Mohammad Zaber Hossain (2019). Effects Of Heavy Metals (Cd, Zn And Cu) On Carbon,
Nitrogen And Iron Mineralization In Soil. Malaysian Journal of Sustainable Agriculture, 3(1): 33-38.
Malaysian Journal of Sustainable Agriculture (MJSA) 3(1) (2019) 33-38
showed the decreasing pattern and varied significantly except Cu and
Cd:Zn:Cu. This may due to higher concentration of single metal along with
the antagonistic interaction among the metals used helps the release of
iron from exchange position. A scholar observed an antagonistic
interaction between Cu and Zn. A scholar found an antagonism relation
between Cd and Zn.Cd:Zn:Cu showed increment in total Fe at day 45 along
with at the end of the incubation period [40]. This may due to the organism
cell lyses. However, at the end of the incubation period no significant
change in total Fe observed.
4. CONCLUSIONS
The effect of heavy metals applied in soil in single and in combination on
the changes in carbon, nitrogen and iron mineralization revealed that the
single metal treatment showed more effect on microbial activities than in
combination. The combined effect showed interactions among the metals
which was antagonistic in nature.
ACKNOWLEDGEMENT
The authors thank to all staff of Soil, Water and Environment discipline,
Khulna University, Khulna, Bangladesh.
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Nitrogen And Iron Mineralization In Soil. Malaysian Journal of Sustainable Agriculture, 3(1): 33-38.
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Cite The Article: Falguni Akter, Humaira Hasan Tinni, Parmita Banarjee, Mohammad Zaber Hossain (2019). Effects Of Heavy Metals (Cd, Zn And Cu) On Carbon,
Nitrogen And Iron Mineralization In Soil. Malaysian Journal of Sustainable Agriculture, 3(1): 33-38.