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METAL AND DDT BIOACCUMULATION AND BIOMARKER RESPONSES
OF THE TIGERFISH, HYDROCYNUS VITTATUS, FROM THE JOZINI DAM AND OKAVANGO RIVER.
Miss E. Fisher, Miss Liesel Whitlow, Prof. V. Wepener & Prof. N. Smit
INTRODUCTION
• Why tigerfish?• High trophic level
(Skelton 2001)• Robbers, minnows,
sardines (Skelton 2001)• Bioaccumulate and
biomagnify (Mhlanga 2000)
• Increasing human activity.• Little known about
bioaccumulation and biomarker responses in tigerfish (Mhlanga 2000; du Preez & Steyn 1991), biomarkers (Whitlow et al. 2008)
AIM: To compare bioaccumulation of heavy
metals and DDT in tigerfish from two systems of varying impacts.
To relate biomarker responses of tigerfish from these systems, and determine whether
or not tigerfish are being stressed as a result of exposure.
HYPOTHESIS Part 1:
Tigerfish from the Jozini Dam will have higher levels of heavy metals and DDT than tigerfish
from the Okavango River.
Part 2 Tigerfish from the Jozini Dam will exhibit more
stress than tigerfish from the Okavango River as a result of the greater influx of pollutants
into this system.
STUDY OBJECTIVES
To determine the environmental partitioning of heavy metals in the Jozini Dam and Okavango River.
To compare the concentrations of heavy metals and DDT in the muscle tissue of tigerfish from these ecosystems.
To compare biomarker responses between tigerfish from the different sites, and
to relate the biomarker responses to the metal and DDT bioaccumulation in the muscle tissue of tigerfish.
SITE SELECTION
tigerfish presence (Skelton 2001)
varying degrees of impacts
Jozini Dam: slightly impacted
Okavango River: relatively unimpacted
JOZINI DAM
Photos taken from DWAF(2004)
JOZINI DAM
The Phongolo river: east of Wakkerstroom
Jozini dam wall (1972).
2492 million m3 water.
DDT sprayings Current uses (DWAF
2004).
Jozini Dam: Tigerfish
OKAVANGO RIVER
http://wpcontent.answers.com/wikipedia/commons/thumb/7/72/Okavango_RiverSign.jpg/288px-Okavango_River_Sign.jpg
OKAVANGO RIVER The Okavango river
(McCarthny 2005; Wilk et al. 2006) 10km3 + 6km3 per annum 25000km2 permanent 80000km2 seasonal 170 000 tonnes 360 000 tonnes
Threats: Boro River (Ellery & McCarthy
1998)
Potential development
http://www.cfnews13.com/uploadedImages/Site_Content/Features/Kissimmee_River/Rest1.jpg
http://www.geog.ucl.ac.uk:8080/print-version/about-the-department/people/academics/martin-todd/accord/images/map2.jpg
Un
iversity o
f Jo
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esb
urg
Zo
olo
gy
De
pa
rtme
nt
METHOD
SAMPLING Sediment & water
Acid washed polypropylene bottles
Tigerfish Caught Anesthetized &
Sacrificed Dissected Muscle frozen Biomarkers:
Hendrickson stabilization buffer
Liquid nitrogen
CONT. OF METHOD PROCESSING
Metals Dried Digestion ICP-MS ICP-OES
Pesticides Freeze dried GCMS (μg/g lipid)
Biomarkers AChE (Ellman et al. 1961)
LP (MDA) (Ohkawa et al. 1979; Uner et al.
2005)
CAT (Cohen et al. 1970)
http://www.wcaslab.com/images/icpms/plasma.jpg
CONT. OF METHOD
PC (Parvez & Raisuddin 2005; Levine et al. 1990; Fernandes
et al. 2008; Floor & Wetzel 1998)
Metallothioneins (Viarengo et al. 1997; Viarengo
et al. 1999; Atli & Canli 2008; Fernandes et al. 2008; Hubbard 2005)
CEA (de Coen & Janssen 1997; de Coen & Janssen 2003)
STATISTICAL ANALYSIS Unpaired Student t-Test PASW(18.0) Significance P<0.05
RESULTS AND DISCUSSIONSEDIMENT AND WATER COMPARISON
Cr Co Ni Cu Zn As Cd Pb0.000
0.001
0.002
0.003
0.004
0.0051
26
0.000
0.001
0.002
0.003
0.004
0.0051
26
Jozini waterJozini sediment
* * *
Jozini Metals
me
tal S
ed
[ ]
ug
/g
Me
tal w
ate
r [ ] ug
/mL
Cr Co Ni Cu Zn As Cd Pb0.000
0.005
0.010
Okavango sedimentOkavango water
0.11.12.13.14.15.16.1
0.000
0.005
0.0100.11.12.13.14.15.16.1
* * *
Okavango Metals
meta
l S
ed
[ ]
ug
/g
Meta
l wate
r [ ] ug
/mL
(Huntsman_Mapila et al. 2005; Huntsman Mapila et al. 2009)
RESULTS & DISCUSSIONMETALS: MUSCLE
Pongola Okavango0
2
4
6Al
Sites
Al (
g
/g d
ry w
eig
ht)
Pongola Okavango0.00
0.01
0.02
0.03
0.04
Sites
As
(g
/g d
ry w
eig
ht) As
Pongola Okavango0.000.010.020.030.040.050.060.070.080.090.100.11
Sites
Co
(
g/g
dry
we
igh
t)
Co
Pongola Okavango0.0
0.1
0.2
0.3
Sites
Cr
(
g/g
dry
wei
gh
t)
Cr
Pongola Okavango0.000.050.100.150.200.250.300.350.400.450.500.550.600.65
Sites
Pb
(g
/g d
ry w
eig
ht)
Pb
JoziniJoziniJozini
JoziniJozini
RESULTS & DISCUSSIONMETALS:MUSCLE
Coal mining and shales: Cu, Fe (Johnson et al. 2006; Huntsman Mapila et al. 2009)
Pongola Okavango0.0
0.1
0.2
0.3
0.4
0.5
0.6
*
Sites
Cu
(
g/g
dry
weig
ht)
Cu
Pongola Okavango0.00
0.05
0.10
0.15
*
Sites
Cd
(g
/g d
ry w
eig
ht)
Cd
Pongola Okavango0
25
50
75
100
*
Sites
Fe
(mg
/g d
ry w
eig
ht)
Fe
Jozini
Jozini Jozini
RESULTS & DISCUSSIONMETALS: MUSCLE
Pongola Okavango0.0
0.5
1.0
1.5
*
Sites
Mn
(
g/g
dry
wei
gh
t)
Mn
Pongola Okavango0
1
2
3
4
5
6
7
8
9
*
Sites
Zn
(g
/g d
ry w
eig
ht)
Zn
Pongola Okavango0.0
0.1
0.2
0.3
0.4
0.5
0.6
0.7
0.8
0.9
*
Sites
Ni
(g
/g d
ry w
eig
ht)
Ni
(Sawula 2004)
Jozini Jozini
Jozini
DDT EXPOSURE
DDT TOTAL
Okavango Pongola0
5001000150020002500300035004000450050005500
*
Sites
gD
DT
/g L
ipid
Jozini
RESULTS & DISCUSSIONBIOMARKERS
BIOMARKERS OF OXIDATIVE STRESS
CAT
Okavango Pongola05
10152025303540455055
*
Sites
um
olH
2O2/
min
/mg
pro
t
PC
Okavango Pongola0
10
20
30
40
*
Sites
nm
ol
carb
on
yls/
mg
pro
t
LP
Okavango Pongola0.0
0.1
0.2
0.3
0.4
0.5
0.6
*
Sites
nm
ol/
mg
pro
t
Jozini
Jozini
Jozini
RESULTS & DISCUSSIONPARAMETERS OF CEA
Lipid
Okavango Pongola0
100200300400500600700800900
100011001200
*
Sites
J/g
Protein
Okavango Pongola0
500
1000
1500
*
Sites
J/g
Glucose
Okavango Pongola0
100
200
300
*
J/g
Sites
Jozini Jozini
Jozini
RESULTS & DISCUSSIONCont. of CEA Parameters
Ea
Okavango Pongola0
100200300400500600700800900
10001100120013001400
*
Sites
J/g
Ec
Okavango Pongola0
25
50
75
100
125
150
175
Sites
J/g
CEA
Okavango Pongola0
1000
2000
3000
*
Sites
J/g
Jozini Jozini
Jozini
Okavango
RESULTS & DISCUSSIONBiomarkers of effect
MT
Okavango Pongola0.00
0.25
0.50
0.75
1.00
1.25
1.50
1.75
*
Sites
(nM
/mg
pro
t)
AChE
Okavango Pongola0.0000
0.0005
0.0010
0.0015
0.0020
0.0025
0.0030
0.0035
0.0040
0.0045
*
Sites
Ab
s/m
in/m
g p
rot
Jozini Jozini
CONCLUSIONHAVE WE ACCEPTED PART 1 OF OUR
HYPOTHESIS?
YES!
Tigerfish from the Jozini Dam are bioaccumulating higher levels of
pollutants, than tigerfish from the Okavango River.
CONCLUSIONHAVE WE ACCEPTED PART 2 OF OUR
HYPOTHESIS?
NO!
Tigerfish from the Jozini Dam are experiencing less oxidative stress than
tigerfish from the Okavango River, however they have greater responses to
exposure of toxicants.
POSSIBLE REASONS FOR STRESS
Okavango oxidative stress & lower CEA:
Barbel run Lotic environment
Jozini AChE & MT AChE: Carbamate pesticides, organophosphates, nerve
gases MT: Higher levels of metals: Cu & Cd
EXTRA CONSIDERATIONS
• NUTRIENTS• CATIONS• ANIONS• PHYSICAL WATER
PARAMETERS• GEOLOGY• SAMPLING TIME
ACKNOWLEDGEMENTSACKNOWLEDGEMENTS WATER RESOURCE COMISSION
(Resources) PROFESSOR V. WEPENER (Supervisor)
PROFESSOR N. SMIT (Co-supervisor)
MISS LIESEL WHITLOW (Data from the Okavango)
DR. H. VAN NIEKERK (SPEC-RAU)
MISS NATALIE DEGGER (Assistance with SPSS)
MR. KYLE MCHUGH (Sample Collection & Pictures)
MISS MICHELLE SOEKOE (Sample Collection)
MISS YOLANDI CLOETE (Sample Collection & other)
THANK YOU!