1. BioReLab.

, Tehri--249199 (UK)
BioReLab., Department of Zoology/Entomology, HNBGU (A Central University), Campus Badshahithaul Tehri
2. Department of Zoology, Government PG College, Kotdwara Garhwal (UK) INDIA
1.
Corresponding author: asokumr@gmail.com Cont: +919450501471
 I NTRODUCTION

In recent years, Uttarakhand has emerged as one of the most attractive

industrial destinations in India.

The Government is encouraging private participation in all industrial activities
and as a result big players such as HLL and Dabur have set up units in the state.

The New Industrial Policy announced in 2003 by the state Government puts in
place the regulatory framework for Uttarakhand's industrialization.

The New Industrial Policy indicates that private resources may be tapped while
promoting integrated Industrial Estates in Uttarakhand.

The industrial sector of the Uttarakhand state growing day by day and it is very
good for the development of state and country also, but we should remember
the second phage of industrialization.

It is the industrial effluent, disposed by the industries. The industrial effluents
have many pollutants including heavy metals.
 H EAVY M ETAL P OLLUTION

In early days of abundant resources and negligible development pressures, little
attention was paid to environmental issue, although some environment related
legislation pertaining to different sectors was authorized.

Rapid economic changes have resulted in elevated level of toxic heavy metals
and radionuclides entering the biosphere.

Solid and/or liquid wastes containing toxic heavy metals may be generated in
various industrial processes such as chemical manufacturing, electric power
generating, coal and ore mining, smelling and metal refining, metal plating and
others.

Continuous monitoring of heavy metals level in the environment is very
important since it cannot be degraded and becoming public health problem
when increased above acceptance level.

Health problem due to heavy metals pollution include nausea, vomiting, bone
complications, nervous system impairments and even death become a major
problem throughout many countries when metal ions concentration in the
environment exceeded the admissible limits.
 M ETHODOLOGY
Sample Collection and Processing:
The liquid and solid wastes were collected from different sites of Uttarakhand state including Sevage and
Municipality (APHA 1998). The leachate from solid waste was prepared according to the method described
by French Standard method (Ferrari et al., 1999, Srivastava et al., 2005 and Savitha et al., 2010). The metal
concentrations analysed by ICP-MS (Ashok et al 2010).

Microbial strain and Heavy metal Training:

Streptomyces sp. strains were maintained in agar slants (Holt et al 1994) containing nutrient broth, which was
isolated from sludge contaminated with heavy metal. Individual stock metal ion solutions of different
concentration of Cu from CuSO4 .5H2O, Cd from 3CdSO4.8H2O, Cr from K2Cr2O7, Ni from NiCl.6H2O, Pb from
Pb acetate and Zn from ZnSO4 respectively was prepared. (Hammaini et al 2006).Microbial strains were
allowed to grow in the supplemented media with metal concentration, which was increased 0 µg/ml to 350
µg/ml (Kumar et al 2010b).

Biosorption Experiment:
Experiments of heavy metals biosorption were done in Erlenmeyer flasks containing 150 ml and 15.0 ± 1.0 mg of
cells. To ensure equilibrium, cells and waste were maintained in contact for 48 hours, under constant
agitation, at 30-350C ± 2ºC. After 48 hours, cells were separated from the medium and residual metal
concentrations were monitored by ICP-MS. The optimum pH and temperature maintained for the growth of
microorganisms in the batch culture. (Chatarzee et al 2001; Cybulski et al, 2003; Kumar et al 2010b).
 R ESULTS
Tolerance of Microbes
The cultures conditions: Temperature (28–30oC), pH of 6.8 ± 0.2.
The metal concentration-0 mg/l (control) to 300mg/l
Streptomyces sp showed the tolerance for Cr and co-tolerance for Pb and Cu.
The control and optimization of bioremediation processes is a complex system of
many factors. They are
Existence of a microbial sps. capable of degrading the pollutants
availability of contaminants to the microbial population
Environment factors such as temperature
pH
Presence of oxygen or other electron acceptors
Nutrients
Microbes were allowed to grown in the media containing heavy metals
concentration and they were adopted for the concerned metals. Understanding
metal–microbe relationships has led to advances in bioremediation (Malik 2004;
Bruins et al. 2000).
Biosorption
capacities
depend on - M ETAL TOLERANCE
Metal
concentration
pH of the
solution Streptomyces sp.
Contact time
80
Ionic strength
Presence of 70
competitive ions.
Morley and Gadd 60
(1995) concluded 50
for fungal
biomass that the 40
different cell wall 70
polymers have 30
55

No. of Colonies
various functional
20
groups and
differing charge 10 25
distributions and 15 10 5
therefore 0
different metal- C 25 50 100 200 250
binding capacities
and affinities. Concentration of metal (mg/l)
 B IOREMEDIATION A CTIVITY
Three metal Cu, Cr and Pd were sorbed by Streptomyces sp.

Copper reduction ranged from 0.288-1.129mg/l, chromium reduction ranged

from 0.386-6.42mg/l, which showed maximum reduction in chromium by
Streptomyces sp and reduction in lead ranged from 0.063-0.286mg/l.

Sharma and Loyal (2009) studied the chromium (III) removal by

Streptomyces sp from tannery effluent and found the similar results as in
case of chromium removal.

Puranik, et al. (1995) found Streptomyces pimprina was potent biosorbent

for cadmium in his study.

Streptomyces rimosus biomass used for Cu, Zn and Cr biosorption by many

investigators (Addour, et al. 1999; Chergui , et al. 2007; Ashwini, et al.
2009).
 A CTIVITY OF S TREPTOMYCES SP

Table 1: Metal Site Sample CM (Cu) CMF (Cu) CM (Cr) CMF (Cr) CM (Pb) CMF (Pb)
conc. in the waste
samples (CM and Solid* ND ND 2.09 0.564 ND ND
HR
CMF). Liquid 3.90 3.198 8.56 2.140 0.858 0.582
Optimal
Solid ND ND 3.34 0.935 ND ND
conditions: DD
Temp. 300C ± Liquid 4.78 3.872 1.85 0.536 0.560 0.380
30C and pH 6.8
Solid ND ND 0.56 0.174 ND ND
±0.2. MR
Liquid 5.95 4.821 1.05 0.294 0.202 0.139
Sample were of Solid ND ND 0.96 0.288 ND ND
six cities: RK
HR: Haridwar Liquid 1.80 1.512 1.45 0.405 0.865 0.579
DD: Dehradun Solid ND ND 3.12 0.845 ND ND
MR: Mossorrie KR
Liquid ND ND 2.15 0.602 0.564 0.383
RK: Rishikesh
KR: Kashipur Solid ND ND 3.45 0.935 ND ND
RR: Roorkee
RR
Liquid 4.78 3.858 ND ND 0.562 0.382
 Ashok
Kumar B IOACCUMULATION
Graph 2: Cu
Bioaccumulatio
n (mg/g of Cr
8
Biomass) by Pb
Streptomyces sp
6
(multi resistant).
4
They reduced
three metals
2
viz. Cr, Cu and
Pb. 0
Max. reduction
recorded by
acclimated
species was
chromium, it
was 72%.
 Ashok
Kumar
D ISCUSSION

Microorganisms are intimately involved in metal biogeochemistry with a
variety of processes determining mobility and therefore, bioavailability.

The balance between mobilization and immobilization varies depending

on the organisms involved, their environment and physicochemical
conditions.

Metal mobilization can arise from a variety of leaching

mechanisms, complexation by metabolites and siderophores and
methylation, where this results in volatilization. In addition, reduction of
higher valency species may effect mobilization, e.g., Cr(VI) to Cr(III).

In the context of bioremediation, solubilization of metal contaminants

provides a means of removal from solid matrices, such as
soils, sediments, dumps and other solid industrial wastes (Kumar et al
2010).
 C ONCLUSION
The tolerance test indicated that among experimental heavy metals
maximum tolerance was shown to Cr, Pb and Cd, showing the
growth of Streptomyces sp up to 200-250 ug/ml.
The heavy metals reduced by filamentous bacteria Streptomyces sp
are copper, chromium and lead.
The average Cu reduction by Streptomyces sp was 18%, Cr reduction
was recorded as 72% and Pb reduction was 32%.
The gram positive bacteria Streptomyces sp was proved as strong
biosorbent for chromium heavy metal.
Further studies should be carried out to establish the quantity or level
of bioremediation which affects the treatment efficiency.
Acknowle
dgements

Authors are thankful to Prof. J.P. Bhatt, HOD

Biotechnology and Zoology, HNBGU (A
central University) Srinagar Garhwal
(Uttarakhand) INDIA for his valuable
suggestions and Dr. Gaurav Gupta, Director
Himachal Institute of Life Sciences, Panonta
Sahib (HP) INDIA for providing the lab
facility.

The authors are also grateful to Dr. A.K
Singh, Scientist C, Wadia Institute of
Himalayan Geology for valuable comments
and technical support.
S NAPSHOTS ……..
 Ashok
Kumar

T Y
f y e
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