Documenti di Didattica
Documenti di Professioni
Documenti di Cultura
2449
D
Original article
T
h
Degradation of
soil cyanide by
single and mixed
cultures of
Pseudom
onas
stutzeri
and
Bacillus
subtilis
Ogbo
nnaya
Nwok
oro
and
Marie
Esther
Uju
Dibua
degradatio
n of soil
cyanide
from
0.218 mg
g-1
soil
immediate
ly with an
inoculum
concentrat
ion of 0.1
(OD600nm)
to 0.072
mg g-1 soil
after
10
days with
an
inoculum
concentrat
ion of 0.6
(OD600nm)
implying a
66.9
%
reduction.
Pseudomo
nas
stutzeri
cell
concentrat
ion of 0.1
(OD600nm)
decreased
of
0.6
(OD600nm)
(72
%
reduction)
.
The
mixed
culture of
the
two
bacteria
produced
the
best
degradatio
n of soil
cyanide
from
0.218 mg
g-1
soil
sample
with
a
combined
inoculum
concentrat
ion of 0.1
(OD600nm)
initi
ally
to
0.02
5
mg
g-1
soil
with
a
c
o
114
resistant to cyanide have been reported to remain England). Sodium dihydrogen phosphate (NaH2PO4),
active even at concentrations higher than 1 mmol L-1 Sodium hydrogen phosphate (Na2HPO4), Potassium
hydrogen phosphate (K2HPO4), Sodium chloride
(8).
The present study focuses on solving an issue of (NaCl) were purchased from Sigma-Aldrich Chemie
great ecological importance, especially in Nigeria.
Gmbh (Steinheim, Germany). Copper sulphate
The processing of cassava (Manihot esculenta) into (CuSO4x5H2O), Manganese sulphate (MnSO4x5H2O)
garri (tapioca), a commonly used food item, involves were obtained from Bio-Lab UK Ltd. (Cockspur
several operations including peeling, grating, pressing, Street, London). Sodium hydroxide (NaOH) was
and fermentation (9, 10). These processes are obtained from Nexill World Chem Chadwell Health
associated with the discharge of large amounts of (Essex, England), Potassium hydroxide (KOH) from
water, hydrocyanic acid, and organic matter. The Avondale Laboratories (Banbury, Oxon, England),
cyanide content of cassava plants varies with the plant Picric acid from Qualikems Fine Chemicals PVT. Ltd.
variety and soil conditions and may range between 75 (India), Glucose from May and Baker Ltd. (Dagenham,
and 1000 mg kg-1 of CN and large amounts of natural England), and Nutrient agar from Oxoid, Ltd. (UK).
cyanogenic glycosides found in cassava are released
during the production of starch from cassava tubers Isolation of bacteria
(11). These cyanogenic glycosides can be enzymatically
Soil samples were collected from a cassava
hydrolysed into cyanide, which is often found in the processing mill and 1 g of soil was added into a conical
waste water discharged from cassava processing flask containingmLofdistilled10
industries. The release of cyanide from industries was thoroughly shaken and 1 mL was serially diluted
worldwide has been estimated to exceed 14 million in normal saline diluents. The dilutions were plated
kg per year (12). Unused cyanide compounds are out on Nutrient agar plates and incubated for 24 h at
released in the effluent coming out from industri
35 C. Pure cultures of the isolates were obtained by
causing contamination of the soil as well as water streaking onto fresh agar plates and stored on slants
bodies affecting the biogeochemical cycle (13).
at 4 C.
Cyanide is highly toxic to humans and aquatic
organisms (14), which is why cyanide-containing
Screening isolates for their resistance to cyanide
effluents cannot be discharged into the environmen
wate
115
Development of inocula
from
the
The medium for the multiplication of
surfac
Pseudomonas stutzeri had the following
e
composition per litre of distilled water: 1.0 g of
down
K2HPO4; 0.2 g of MgSO4x7H2O; 0.01 g of
to 5
CaCl2; 0.01 g of NaCl; 2.0 mg of MnSO 4; 0.2 mg
cm
of CuSO4x5H2O; 0.2 mg of ZnSO4; 2.0 g ofdepth
glucose.
were
The medium for the multiplication of Bacillus
collect
subtilis contained the following per litre of distilled
ed
water: 1.0 g of KCl; 0.2 g of MgCl 2x2H2O; 0.25 g of
from
CaCl2x2H2O; 5.0 mg FeSO4x7H2O; 1.0 mg of
differe
MnSO4x5H2O; 5.26 g of Na2HPO4; 5.4 g of Na2HPO4. nt
The single inocula of both bacterial cultures locatio
were incubated in a Gallenkamp orbital incubator ns of
(Gallenkamp, England) for 24 h and diluted with the
sterile distilled water to optical densities of 0.1- inocul
0.6 measured in a Spectrumlab 23Aated
spectrophotometer
(Ningbo
Tianyu mappe
Optoelectronic Co., China) at 600 nm. Mixedd plot
culture inocula (1:1 ratio) was similarly grown, at
reduced to 500 mL each, combined and diluted interv
with sterile distilled water to optical densities of als of
2
0.1-0.6 measured at 600 nm.
days.
Sampling site
Cyani
This experiment was conducted in the dryde
levels
season of 2013. A total of 18 plots of land (located
were
at Beach Junction, Nsukka, Nigeria) each measuring
deter
30x30 cm was mapped out around a cassava
mined
processing mill. The plots were each separately
by a modification of
inoculated with bacterial cultures. Soil samples (10
the alk by Williams
g each) starting
and Edwards (17) as
follows: various
quantit
ies of
standa
rd (50,
100,
RESULTS AND
DISCUSSION
A total of
32
bacterial
isolates were
recovered from
116
117
kinetic
properties
of
these
micr
hanisms
oorg
soil cyanide
anis
degradation
ms
by
and
cultivation
the
of
mec
of
co-
118
microorganisms in other polluted environments is 15. Dash RR, Balomajumder C, Kumar A. Cyanide removal by
recommended.
combined adsorption and biodegradation processs. Iran J
REFERENCES
1. Solomonson LP. Cyanide as a metabolic inhibitor. In:
Vennesland B, Conn EE, Knowles CJ, Westley J, Wissing F,
editors. Cyanide in biology. New York (NY): Academic
Press; 1981. p. 11-28.
2. Knowles CJ. Cyanide utilization and degradation by
microorganisms. CIBA Found Symp 1988;140:3-15.
PMID: 3073060
3. Knowles CJ. Microorganisms and cyanide. Bacteriol Rev
1976;40:652-80. PMCID: PMC413975
4.
Yu XZ, Peng XY, Wang GL. Photo induced dissociation of
ferri and ferro cyanide in hydroponic solutions. Intl J Environ
Sci Technol 2011;8:853-62. doi: 10.1007/BF03326268
5.
Smith A, Mudder TI. The Chemistry and Treatment of
Cyanidation Wastes. London: Mining Journal Books Ltd;
1991.
6. Meyers PR, Rawlings DE, Woods DR, Lindsey GG. Isolation
and characterization of a cyanide dihydratase from Bacillus
pumilus C1. J Bacteriol 1993;175:6105-12. PMCID:
PMC206703
7. Strobel GA. Cyanide utilization in soil. Soil Sci 1967;103:299302.
8. Chena SC, Liu JK. The respiratory responses to cyanide of
a cyanide-resistant Klebsiella oxytoca bacterial strain. FEMS
Microbiol Lett 1999;175:37-43. doi:
10.1111/j.1574-6968.1999.tb13599.x
9.
Ubalua AO. Cassava wastes: treatment options and value
addition alternatives. Afr J Biotechnol 2007;6:2065-73.
10. Okechi RN, Ihejirika CE, Chiegboka NA, Ibe IJ. Evaluation
of the effects of cassava
populations and physicochemical parameters at different soil
depths. Int J Biosci 2012;2:139-45.
11. Kaewkannetra P, Imai T, Garcia-Garcia FJ, Chiu TY. Cyanide
removal from cassava mill waste water using Azotobacter
vinelandii TISTR 1094 with mixed microorganisms in
activated sludge treatment system. J Harzard Mater
2009;172:224-8. doi: 10.1016/j.jhazmat.2009.06.162
12. Naveena D, Majumder CB, Mondal P, Shubha D. Biological
treatment of cyanide containing waste water. Res J Chem
Sci 2011;1:15-21.
13.
Parmar P, Soni A, Vyas K, Desai PV. Isolation and
characterization of cyanide degrading bacterial strains from
contaminated soil. Int J Environ Sci 2012;2:2006-14. doi:
10.6088/ijes.00202030084
14. Dursun AY, Aksu Z. Biodegradation kinetics of ferrous(II)
cyanide complex ions by immobilized Pseudomonas
fluorescensinapackedbedcolumn reactor. Process Biochem
2000;35:615-22. doi:
10.1016/S0032-9592(99)001107
enzmictec.2004.03.030
119
Saetak
Razgradnja cijanida u tlu s pomouPseudomonaspojedinanihstutzeri
i Bacillus subtilis
i mij
Bakterije Pseudomonas stutzeri i Bacillus subtilis odabrane su na temelju zamjetnog rasta u minimalnom
mediju s 0,8 mg mL-1 kalijeva cijanida (KCN). Tijekom ovog istra reduciranja razina cijanida u mediju s 1,5 mg mL-1
KCN. Iako su obje bakterije smanjile koncentraciju
cijanida, Pseudomonas stutzeri bila je uinkovitija. Kasnije, odabrane u razliitim koncentracijama koritene
pojedinano i
razgradnji cijanida u uzorcima tla prikupljenim u blizini mlina za preradu manioke. Bacillus subtilis je
uzrokovala razgradnju cijanida od 0,218 mg g-1
na poetku pri koncentraciji)
do 0,072 mg g-1 tla nakon deset dana pri koncentraciji od 0,6 (OD ), to je smanjenje%.Staninaod 66,
600nm
koncentracija Pseudomonas stutzeri od 0,1 (OD
) smanjila je razinu cijanida u tlu s 0,218 mg g -1
-1
600nm
600nm
na
smanjenje%. od
CORRESPONDING AUTHOR:
Ogbonnaya Nwokoro
Industrial Microbiology and Biotechnology Laboratory,
Department of Microbiology, University of Nigeria,
Nsukka, Nigeria
E-mail: ogb883@yahoo.com