Environ Sci Pollut Res
DOI 10.1007/s11356-013-1805-8
RESEARCH ARTICLE
Trace metals in Ganges soft-shell turtle (Aspideretes
gangeticus) from two barrage: Baloki and Rasul, Pakistan
Riffat Naseem Malik & Bushra Ghaffar &
Muhammad Zaffar Hashmi
Received: 14 January 2013 / Accepted: 6 May 2013
# Springer-Verlag Berlin Heidelberg 2013
Abstract The concentration of nine metals was measured in
liver, kidney, heart, muscle, plastron, and carapace of
Aspideretes gangeticus from Rasul and Baloki barrages, Pakistan. The results indicated that metal concentration were
significant different among tissues of Ganges soft-shell turtles.
However, higher concentrations of Co (5.12 μg/g) and Ni
(1.67 μg/g) in liver, Cd (0.41 μg/g) in heart, Fe
(267.45 μg/g), Cd (2.12 μg/g) and Mn (2.47 μg/g) in kidney,
Cd (0.23 μg/g), Cu (2.57 μg/g), Fe (370.25 μg/g), Mn
(5.56 μg/g), and Pb (8.23 μg/g) in muscle of A. gangeticus
were recorded at Baloki barrage than Rasul barrage. Whereas
mean concentrations of Pb (3.33 μg/g) in liver, Co
(1.63 μg/g), Cu (11.32 μg/g), Pb (4.8 μg/g) and Zn
(144.69 μg/g) in heart, Co (4.12 μg/g) in muscle, Ni
(1.31 μg/g), Pb (2.18 μg/g), and Zn (9.78 μg/g) in carapace
were recorded higher at Rasul barrage than Baloki barrage.
The metals followed the trend Fe > Zn > Ni > Cu > Mn > Pb >
Cr > Co > Cd. Metals of toxicological concern such as Cr, Pb,
and Cd were at that level which can cause harmful effects to
turtles. The results provide baseline data of heavy metals on
freshwater turtle species of Pakistan.
Keywords Bio-monitoring . Aspideretes gangeticus .
Heavy metals . Pakistan
Responsible editor: Vera Slaveykova
R. N. Malik (*) : B. Ghaffar
Department of Environmental Sciences, Faculty of Biological
Sciences, Quaid-i-Azam University, IslamabadPO 45320, Pakistan
e-mail: r_n_malik2000@yahoo.co.uk
M. Z. Hashmi
Department of Environmental Engineering, College of
Environmental and Resource Sciences of Zhejiang University,
Hangzhou 310029, People’s Republic of China
Introduction
Toxic effects of heavy metals have been reported for several
marine vertebrates (Law 1996; Franson 1996). Recent reports
have documented that marine pollution by plastic debris, tar
balls, heavy metals, and persistent organochlorine compounds
have been of great concern and that may have played a role in
declining populations of sea turtles (Godley et al. 1999). Due
to their long lifespan and high trophic level in the aquatic food
web, turtles are vulnerable to heavy metals pollution. Studies
on heavy metals or organic compounds bioaccumulation in
turtles are limited and different organs and tissues such as in
blood (Páez-Osuna et al. 2011), eggs (Páez-Osuna et al. 2010),
eggshells (Sakai et al. 1995), liver, kidney, and bone (GarcíaFernández et al. 2009) to assess and monitor heavy metal
accumulations (Sakai et al. 2000b; García-Fernández et al.
2009) and persistent organic pollutants (D’Ilio et al. 2011).
However, there is no information on toxicological effects and
detrimental threshold concentrations. Progress towards the
understanding of the possible heavy metal impact on turtle
health might be obtained with more data on accumulation and
distribution of trace elements within their body.
The Ganges soft-shelled turtle is found in the Ganges,
Indus, and Mahanadi river systems of Pakistan, northern
India, Bangladesh, and southern Nepal. This turtle inhabits
deep rivers, streams, large canals, lakes and ponds, with a
bed of mud or sand. According to the International Union
for Conservation of Nature, freshwater turtle species are
vulnerable. It tends to prefer areas where the water is turbid.
Ganges soft-shelled turtle spends more time eating aquatic
plants and a large variety of smaller animals, such as fish,
mollusks, insects, amphibians, and waterfowl (Bonin et al.
2006). Freshwater turtles have been evaluated in several
studies as biomonitors to evaluate an array of environmental
contaminants; however, most of the studies related to heavy
Environ Sci Pollut Res
metal accumulation have been carried out on marine turtles.
Illegal trade of freshwater turtle’s body parts for traditional
medicine products is amongst the major threats to freshwater turtles (Anan et al. 2009) in south Asia, in particular to
China. Many species of Asian turtles are being used to make
a popular “turtle jelly,” which endangers these species. The
threats to freshwater turtles are further magnified by changes
to their habitat resulting from human activities ranging from
logging to slash-and-burn agriculture, pollution, and the
damming and channeling of rivers (Anan et al. 2009).
The Punjab province in Pakistan is most populated and is
famous for its riverine system. A lot of barrages have been
constructed in different rivers of the Punjab province for
irrigation purposes. Rasul barrage (latitude 32º42' and longitude 73º31') was constructed on the Jehlum River at the
confluence of River Chenab which receives water from
Mangla dam, with catchments area of 24,069 Km2. The
total irrigated area of Rasul barrage is 1,967 Km2 http://
en.wikipedia.org/wiki/Rasul_Barrage. Rasul barrage has a
water discharge capacity of 24,070 cubic meters per second.
Rasul barrage receives untreated domestic and industrial
waste from the Jehlum city. While Baloki barrage (latitude
31º13' and longitude 73º31') was constructed in the Ravi
River with total irrigated area 1,965 Km2 and drainage basin
4,709 Km2. Baloki barrage receives untreated discharge
from Lahore city, Degh Nala, and Hudiara drain in upstream
region. The Hudiara drain is considered as a major source of
pollution that carries industrial, municipal, and agricultural
waste from both India and Pakistan (Farooq et al. 2011;
Hashmi et al. 2013). Both barrages are highly urbanized
with total inhabitants living in Rasul barrage at 18,374 and
208,475 people in Baloki barrage (District Census 2012).
Cotton, wheat, and rice are the main crops in the vicinity of
both barrages. Major agrochemicals used are herbicides, pesticides, and fertilizers. There is no information of heavy-metal
accumulation in freshwater turtles inhabiting in Baloki and
Rasul barrages. So, this study was conducted in these barrages
as a part of preliminary assessment of presence/absence of
freshwater soft-shell turtle species by the Pakistan Wetland
Program conducted on November 2007 (Fig. 1). Therefore,
this study aimed to determine the trace metals Cd, Co, Cr, Cu,
Fe, Mn, Ni, Pb, and Zn in six different body organs of
Fig. 1 Map of the region indicating the study sites: Baloki and Rasul barrages
Environ Sci Pollut Res
freshwater turtles (liver, kidney, heart, muscle, plastron, and
carapace) in the Baloki and Rasul barrages
Materials and methods
Sampling
A total of 12.25 km upstream right bank and approximately
1 km downstream of the Baloki and Rasul barrages was
surveyed in April–May 2008 in semi-rainy season. During
the field visits, a large number of Ganges soft-shell turtle
(Aspideretes gangeticus) was recorded. Although turtle hunting is illegal according to the Punjab Wildlife Act 1974,
revised in November 2007, illegal killing was found common
at two barrages during the study period due to poor enforcement of laws. A large group of local communities and turtle
sellers were found involved in activities such as killing and
selling of turtle plastrons during the field visits. During early
stages of the study, the turtle seller did not cooperate to sell the
turtles but later on agreed to provide the required samples if
their names and identity will not be disclosed. A total of ten
male adult turtle samples were purchased from the turtle
sellers involved in illegal trade from each barrage. The juvenile turtles were caught using fishing nets by the sellers. The
collection of sample size and further turtles handling was done
as described by Anan et al. (2009). All these turtles were
transported to the laboratory under fresh condition. In the
laboratory, the turtle’s biometric characteristics were measured and then dissected to collect tissues and organs for
heavy metal analysis. Average carapace length (cm) and width
(cm) were 30.89±5.30 and 29.19±3.01, respectively. While
plastron length (cm) was 20.11±2.22. Heart, liver, muscles,
kidneys, plastron, and carapace were separated from each
turtle. Each organ was weighed to establish wet weight,
washed with the deionized water, placed in pre-washed zipped
plastic bags with HNO3 (10 %), tagged, and refrigerated
at −20 °C before metal analyses.
Trace metal analysis
Each tissue and organ of the turtles was cut with the stainless
steel knives washed with the 10 % nitric acid solution to avoid
any contamination. The samples were cut into cubical pieces,
chopped to make the homogeneous sample for metal analysis
and oven dried at 70 °C. Dried samples measuring 1 g each
weighed and digested with 4 ml of ultra pure nitric acid
(HNO3) for microwave-assisted digestion. Digested samples
were filtered with Whatman filter paper 1 and volume increased to 25 ml with deionized water. All samples were run
in triplicate to measure Pb, Zn, Fe, Co, Cu, Cr, Mn, Ni, and Cd
using Atomic Absorption Spectrophotometer (Varian FSAA240). Procedural blanks and certified reference material, NIES
No. 1 was also prepared using identical procedures as used for
the samples. Each calibration curve was evaluated by determination of quality control standards before, during and after a
set of sample measures. The recovery rates for the studied
metals were within 90±10 %.
All the chemicals used were of analytical grade (Merck
Darmstadt, Germany). Deionized water was used throughout the analysis. Standard solutions of metals were prepared
by dilution of 1,000 ppm certified standards solutions
(Fluka Kamica Busch Switzerland) of corresponding metal.
Glass and plastic ware used in the field and laboratory
analyses was cleaned with washing detergent (Decon 90),
rinsed repeated times with deionized water, soaked in HNO3
(10 %, v/v) for 24 h, and finally rinsed with deionized water.
The level of metal accuracy with precision for studied heavy
metals was 83±2–96±3 %.
Statistical analysis
Shapiro–Wilk normality test was used to check the normal
distribution of data before analysis of variance (ANOVA).
The data was distributed normal then parametric tests were
employed for statistical analysis. One-way ANOVA was
used to examine mean differences among the organs and
sites. For all tests, P values of ≤0.05 were used to determine
significant differences. Metal concentrations are reported in
μg/g on dry weight basis. Statistical analysis was performed
using Statistica Software (Statsoft Inc. 1999).
Results
The mean concentration of heavy metals in various tissues
and organs of Ganges soft-shell turtle from both sites are
presented in Table 1. Ganges soft-shell turtle samples from
Baloki barrage showed higher concentrations of metals such
as Cd, Co, Cr, Cu, Mn, Ni, and Pb. The concentrations of Cd
(P=0.00), Co (P=0.00), Cu (P=0.00), Pb (P=0.00) and Zn
(P=0.00) in heart; Fe (P=0.00), Mn (P=0.00) in kidney;
Cd (P=0.00), Co (P=0.00), Cu (P=0.00), Fe (P=0.00), Mn
(P= 0.00), Ni (P= 0.00) in muscle; Co (P= 0.00), Ni
(P=0.00), Pb (0.02) in liver; Cd, (P=0.00) Co (P=0.00),
Cr (P=0.00), Cu (P=0.00), Mn (P=0.00), Ni (0.01), Pb
(0.03), Zn (0.05) in Plastron were differed significantly.
Higher metal concentrations of Cd, Co, Cu, Mn, Pb, Fe,
and Zn were measured in kidney, liver, and heart in comparison to those concentrations measured in carapace and
plastron. Fe showed the highest concentration in all tissues.
The mean concentrations of Fe, Zn, and Pb in kidneys of
Ganges soft-shell turtles from the Baloki barrage were relatively higher (267.45, 53.06, and 2.32 μg/g) than the Rasul
barrage. The general trend of metals in kidneys was: Fe > Zn
> Pb > Cu > Co > Mn > Cd > Cr > Ni.
Environ Sci Pollut Res
Table 1 Metal concentrations (mean ± standard error, μg/g) in tissues and organs of Aspideretes gangeticus (Ganges soft-shell turtle) from the two
barrages, n=10 for each site
Specimens
Cd
Co
Cr
Cu
Fe
Mn
Ni
Pb
Zn
Heart
(BB)
Mean ± SE 0.41±0.03
0.39±0.15 0.49±0.09
8.83±0.36
1,195.8±191
1.21±0.11 4.15±1.09
0.94±0.26 121.34±4.99
Min–max
0–1.13
6.64–10.7
620–2,528
0.8–1.78
0.53–9.84
0.25–2.75
93.6–146.29
Heart
(BR)
Mean ± SE 0.09±0.02
1.63±0.14 0.41±0.11
11.32±0.48
1,291.6±133.33 1.03±0.04 3.21±0.36
1.8±0.59
144.69±5.08
Min–max
0–0.15
1.13–2.33
0.05–1.2
9.43–13.33
850–2,082
0.73–1.2
1.47–4.8
0.34–2.80
125.36–177.94
Probability
0.00**
0.00**
0.59
0.00**
0.69
0.15
0.42
0.00**
0.00**
9.47±0.3
267.45±33.4
2.47±0.27 0.57±0.14
0.23–0.56
0.05–0.89
Kidney
(BB)
Mean ± SE 2.12±0.34
3.29±0.16 1±0.08
Min–max
2.53–4.2
8.3–11.07
131–481
1.45–4.07
Kidney
(BR)
Mean ± SE 1.28±0.33
3.71±0.34 0.94±0.09
7.76±0.93
115.93±23.99
0.92±0.20 0.44±0.09
8.83±2.56 51.01±2.24
Min–max
0.07–2.83
3.15–6.68
0.55–1.63
4.575–13.77 50.5–314.5
0.03–2.52
0.05–0.9
2.5–28.5
37.22–59.01
Probability
0.13
0.28
0.65
0.1
0.00**
0.00**
0.45
0.66
0.57
0.35–3.9
0.53–1.5
2.32±1.82 53.06±2.78
0.125–1.55 0.99–3.21
45.66–75.07
Muscle
(BB)
Mean ± SE 0.23±0.03
3.71±0.04 0.94±0.06
2.57±0.11
370.25±38.60
5.56±0.63 1.02±0.16
1.23±1.40 71.99±9.27
Min–max
3.55–4.20
0.67–1.48
2.1–11.08
209.5–481
3.17–4.08
0.8–3.01
Muscle
(BR)
Mean ± SE 0.05±0.01(n=7) 4.12±0.05 0.71±0.16
1.61±0.27
164.6±25.41
2.35±0.15 1.4±0.56
1.03±0.48 60.52±4.57
Min–max
0–0.10
3.8–4.33
0.27–1.95
0.2–2.63
60.5–305.00
1.75–3.48
0.15–5.15
1.57–2.58
39.54–84.34
Probability
0.00**
0.00**
0.21
0.00**
0.00**
0.00**
0.52
0.00**
0.3
21.23±2.36
1110.8±203.32
5.79±0.71 1.67±0.15
0.07–3.90
0.03–1.55
0–75.07
Liver
(BB)
Mean ± SE 0.25±0.04
5.12±0.05 0.39±0.1
Min–max
4.9-5.45
11.03-34.65
423.5-2287
2.75-8.98
Liver
(BR)
Mean ± SE 0.18±0.05(n=8) 0.37±0.06 0.39±0.09
16.24±2.54
837.55±147.23
5.50±0.29 0.54±0.19
2.33±0.42 32.62±2.61
Min–max
(−0.05)–0.38
0.05–0.68
0.03–0.83
8.35–33.63
348–1,718.5
3.83–6.75
0.05–1.5
0.5–3.86
19.33–44.86
Probability
0.43
0.00**
0.97
0.17
0.29
0.71
0.00**
0.02*
0.19
4.02±0.1
138.3±26.72
3.09±0.51 8.58±2.12
0.03-0.45
0.03-1.08
0.25-3.75
20.47-50.37
Plastron
(BB)
Mean ± SE 0.74±0.09
3.4–4.55
45.5–306
1.17–5.87
Carapace
(BB)
Mean ± SE 0.85±0.03
0.88±0.05 1.34±0.11
3.59±0.08
110.77±18.70
1.42±0.16 0.85±0.13
0.98±0.19 7.12±0.78
Min–max
0.65–1.03
3.275–3.94
46.5–227.25
0.86–2.38
0.13–1.88
Carapace
(BR)
Mean ± SE 0.16±0.03
0.36±0.08 0.08±0.08(n=1) 0.91±0.50
128.68±3.71
0.17±0.03 1.31±0.15
2.18±0.94 9.78±0.98
Min–max
0.025–0.34
0.025–0.8
0–0.75
0.06–3.98
108.75–143.75
0.04–0.3
0.68–2.01
0.75–4
3.44–15.66
Probability
0.00**
0.00**
0.00**
0.00**
0.36
0.00**
0.03*
0.01**
0.05*
Min–max
0.17–1.2
0.7–0.99
0.99±0.15 1.99±0.64
0.75-2.10
1.93±0.32 38.63±3.53
0.15–1.7
0.7–7.5
0.76–1.71
2.97±0.80 19.59±2.31
0.63–19.72 0.25–3.25
0.24–1.39
11.35–34.37
4.08–11.12
SE standard error, Min–Max Minimum–Maximum, BB barrage Baloki, BR barrage Rasul
*P <0.05; **P <0.01
Heart samples collected from Baloki barrage exhibited
high mean concentrations of Cd, Cr, Mn and Ni whereas mean
concentration of Co, Cu, Fe, Pb and Zn were higher in Rasul
barrage. The mean concentration of metals in heart was in
order: Fe > Zn > Cu > Pb > Ni > Co > Mn > Cr > Cd. Mean
concentration of metals such as Cd, Cr, Cu, Fe, Mn, Pb and Zn
were higher in muscle at Baloki, and Co and Ni at Rasul
barrage. The trend of metal accumulation in muscle followed
the order: Fe > Zn > Pb > Mn > Co > Cu > Ni > Cr > Cd. Liver
samples from Baloki showed higher concentrations of Cd, Co,
Cu, Fe, Mn, Ni, and Zn while Pb was higher in liver samples
from Rasul barrage. The metal accumulation trend in liver
was in following order: Fe > Zn > Cu > Mn > Co > Pb
> Ni > Cr > Cd. The metals were significantly different
in plastrons samples collected from Baloki. The pattern
of metal accumulation in plastrons followed the order:
Fe > Zn > Ni > Cu > Mn > Pb > Cr > Co > Cd.
Concentrations of Cd and Cr in carapace were higher
than those in plastron, heart, liver, muscle, and kidney
(Table 1). However, mean values of Fe, Co, Pb, and Zn
were comparatively lower in carapace. The pattern of metal
levels in carapace was in the following order: Fe > Zn > Cu
> Pb > Mn > Cr > Ni > Co > Cd. The mean concentration of
Fe and Mn were significantly different in kidneys and Cu,
Pb, and Zn in heart between two sites. However, the mean
concentrations of Cd, Co, Cu, Fe, Mn, and Pb were significantly different in muscles between two sites while, the
mean concentrations of Co, Ni, and Pb were significantly
different in liver between both sites. Significant differences
between Baloki and Rasul barrages were observed for Cd,
Co, Cr, Cu, Mn, Ni, Pb, and Zn in carapace.
Discussion
The results indicated that most of the metals were significantly different between the sites and organs (Table 1).
These differences in sites and body organ may be due to
Environ Sci Pollut Res
the sources of heavy-metal pollution in both barrages and
accumulation period in the Ganges soft-shell turtles. Both
sites receive untreated, industrial, and agriculture waste
from both point sources and nonpoint sources of pollution.
The Baloki barrage receives domestic and industrial waste
from the Lahore city, Sialkot city, Faisalabad while Rasul
barrage receives from the Jehlum city (Qadir and Malik
2011; Farooq et al. 2011; Hashmi et al. 2013). However,
in the vicinity of both barrages, agriculture activities were
also recorded. The level of metal accumulation along with
other factors such as age, gender nutritional status, seasonal
and annual variations, geographic variation and trophic level
may also affect metal levels, toxicity, and elimination in
various organisms (Gardner et al. 2006). However, specific
organs have metal accumulation preferential which causes
the metal variability in the organisms. For example, Cd
accumulate preferably in kidney (Law 1996) Mn, Pb, and
Zn accumulate in calcareous tissues (bone and carapace)
(Anan et al. 2009). Food is probably the main source of
exposure to heavy metals in aquatic organisms. As turtles
feed mainly on variety of aquatic plants and other herbivorous organisms like fish, mollusks, insects, and amphibians
etc. to meet their feeding needs (Caurant et al. 1999).
Chrome, which is an essential element required in trace
quantities for normal glucose metabolism (Mertz 1969). Cr
concentration in liver (0.39 μg/g), kidney (1.0 μg/g), and
muscle (2.18 μg/g) in the present study is lower (Tables 2, 3,
and 4) than to those reported for green turtles from the south
China coast (Lam et al. 2004). Cr exposure generally affects
the liver and kidneys of the aquatic organisms (Gardner et
al. 2006). Toxic Cr concentration may cause kidney and
liver damage leading to death (D’Ilio et al. 2011).
Copper concentrations in the liver of Ganges turtles measured in present study were similar to those found in green
t ur t le s ( C h e l o n i a m y d as ) a n d h a w k s b i l l tu r t l e s
(Eretmochelys imbricata) from Yaeyama Islands, Okinawa,
Japan (Anan et al. 2009). However, Cu concentrations in
liver reported in present study, were lower to those found in
Loggerhead marine turtles from Canary Islands, Spain
(Fernandez-Turiel et al. 2003). Cu concentrations in kidney
recorded in the present study were comparable to those
found in Green turtles (8.2 μg/g) in Adriatic Sea, Italy
(Storelli et al. 2008) while lower concentrations of Cu in
kidneys were reported in Loggerhead turtles from Yaeyama
Islands, Okinawa, Japan (Sakai et al. 2000a). Cu Concentrations in muscles were similar to those found in green
turtles from south China coast (Lam et al. 2004) while lower
metal concentrations as compared to the present study were
found in Loggerhead turtles from eastern Mediterranean
Sea, Italy (Storelli et al. 2005).
Zinc concentrations in liver found in the present study
were similar to those of green turtles from south-eastern
Queensland, Australia (39.7 μg/g) (Gordon et al. 1998).
However, high concentrations in liver were reported from
South China coast in green turtles (Lam et al. 2004). The
differences in the concentration of Zn from other regions
may be due to the level of contamination, environment,
and metabolism of turtle species involved (Gardner et al.
2006).
Cadmium concentrations reported in liver (0.22 μg/g)
from Baloki barrage were similar to leatherback turtle from
UK (Davenport and Wrench 1990). Cd concentrations
recorded in the present study were very low as compared
to those found in green turtles from the South China coast
(Lam et al. 2004). The concentration of Cd (2.48 μg/g)
reported in kidney of green turtles by Lam et al. (2004)
from south China coast was similar to our study. High Cd
concentrations (142 μg/g) were found by Anan et al. (2009)
in green turtles from Japan as compared to present study.
Cd is stored in kidney and in liver (O’Shea 1999) and
distribution of metals may also influence by the duration
and concentration of exposure. Studies of freshwater turtles
have demonstrated that after a single injection (dose), the
highest concentration of Cd is found initially in liver, which
is a major site of short-term storage (Thomas et al. 1994; Rie
et al. 2001). However, tissue Cd concentrations found in
free-ranging reptiles generally indicates a shift from the liver
to kidney. During long-term exposure, Cd is redistributed
from the liver via the blood and transported as a
metallothionein-complex to the kidneys, where it is
absorbed and concentrated (Linder and Grillitsch 2000).
Concentrations of Cd were found in muscle of Loggerhead turtle from southwestern Mediterranean, Spain
(García-Fernández et al. 2009) were similar to the present
study. Higher Cd concentrations were found in green turtles
from Baja California peninsula, Mexico (Gardner et al.
2006). Mining, agricultural runoff, and wastes from metal
smelting are primary sources of Cd contamination. Cd is a
non-essential element not metabolically regulated by vertebrates and, as such, is toxic to humans and other animals
(Linder and Grillitsch 2000). Food digestion rather than
inhalation is the most likely uptake route of Cd in vertebrates. After ingestion, Cd is transported to the gut and is
excreted in the feces. The kidney is the principal storage site
for Cd, followed by the liver and other tissues (muscle, skin,
and bone) in humans and marine vertebrates (Gardner et al.
2006). Cd has a long biological half-life and accumulates
with age by binding with metallothionein within the liver
(D’Ilio et al. 2011). Cd environmental concentration such as
1 μg/g may not only affect gonadal developmental processes of freshwater turtles (Trachemys scripta) during postnatal and embryonic stages but also may disrupt reproductive processes later in life (Kitana and Callard 2008). In this
study the authors studied the effect of environmentally relevant dose of cadmium on germ cell number and oocyte
apoptosis in lab-reared T. scripta, a closely related
Table 2 Metal concentrations (mean and range μg/g) in liver of turtles from different regions
Location
Turtle species
Cd
(García-Fernández
et al. 2009)
Garcı´a-Ferna´ndez
et al. (2009)
Gardner et al. (2006)
Andalusia (Spain)
Loggerhead turtle (Caretta caretta)
5.85
Andalusia (Spain)
Loggerhead turtle (Caretta caretta)
23.38
Baja California, Mexico
Green turtle (Chelonia mydas)
3.3
Frias et al.(2006)
Storelli et al. (2005)
Torrent et al. (2004)
Kaska et al. (2004)
Lam et al. (2004)
Lam et al.(2004)
Franzellitti et al.(2004)
Anan et al. (2001)
Sakai et al. (2000a)
Sakai et al. (2000b)
Godley et al. (1999)
Godley et al. (1999)
Caurant et al. (1999)
Godley et al. (1998)
Storelli et al. (1998)
Gordon et al. (1998)
Gordon et al. (1998)
Mexico
Italy
Canary Island,Spain
Turkey
South China
South China
Italy
Japan
Japan
Okinawa, Japan
Cyprus
Cyprus
France
UK
Adriatic Sea,,Italy
Huawei
Australia
Olive Ridley turtle (Lepidochelys olivacea)
Loggerhead turtle (Caretta caretta)
Loggerhead turtle (Caretta caretta)
Loggerhead turtle (Caretta caretta)
Green turtle (Chelonia mydas) adult
Green turtle (Chelonia mydas) Juv.
Loggerhead turtle (Caretta caretta)
Green turtle (Chelonia mydas)
Loggerhead turtle (Caretta caretta)
Green turtle (Chelonia mydas)
Loggerhead turtle (Caretta caretta)
Green turtle (Chelonia mydas)
Loggerhead turtle (Caretta caretta)
Leatherback Turtle (Dermochelys coriacea)
Loggerhead turtle (Caretta caretta)
Green turtle (Chelonia mydas)
Green turtle (Chelonia mydas)
3.28
3.36
2.53
10.8
1.4
1.098
2.84
18.2
9.74
3.90
8.64
5.89
2.58
0.22-88.0
2.24
Sakai et al. (1995)
Japan
Loggerhead turtle (Caretta caretta)
12.5
9.29
Aguirre et al. (1994)
Davenport and
Wrench (1990)
Present study
Present study
Hawaiian Islands
British Island
Green turtle (Chelonia mydas)
Leatherback turtle (Dermochelys coriacea)
9.3
0.22
Baloki barrage
Rasul barrage
Ganges soft-shell turtle (Aspideretes gangeticus)
Ganges soft-shell turtle (Aspideretes gangeticus)
0.25
0.18
BDL below detection limit, NA not available.
Co
Cr
0.304
0.585
0.847
BDL
0.25
2.2
<0.03
Cu
Fe
Pb
Zn
5.40
0.69
26.82
21.60
2.75
107.3
8.2
7.69
15.02
2.98
9.168
133
7.4
139
17.7
8.73
3.32
0.16
2.94
3.55
0.862
0.152
13.48
NA
211.6
128.9
604
126
Mn
Ni
10.5
16.27
0.712
0.27
4.74
2.18
1.86
<0.03
0.059
8.25
0.507
0.08
0.12
BDL
BDL
NA
0.02-14.0
NA
<0.0
5.12
0.37
0.39
0.39
87.2
28.1
59.5
25
NA
31.9
17.9
649
2.0
87.6
0.15
1170
NA
1.6
NA
21.23
16.24
1110.8
837.55
5.79
5.50
<0.03
1.67
0.54
BDL
39.7
27.9
BDL
30.6
2.62
1.93
3.33
38.63
32.62
Environ Sci Pollut Res
Reference
Environ Sci Pollut Res
Table 3 Metal concentrations (mean and range, μg/g) in kidney of turtles from different regions
Reference
Location
Turtle species
Cd
Garcı´a-Ferna´ndez
et al. (2009)
Garcı´a-Ferna´ndez
et al. (2009)
Gardner et al. (2006)
Frias et al.(2006)
Maffucci et al. (2005)
Storelli et al. (2005)
Torrent et al. (2004)
Kaska et al. (2004)
Lam et al.(2004)
Franzellitti et al.(2004)
Anan et al. (2001)
Sakai et al. (2000a)
Sakai et al. (2000b)
Godley et al. (1999)
Godley et al. (1999)
Andalusia (Spain)
Loggerhead turtle (Caretta caretta)
10.49
Andalusia (Spain)
Loggerhead turtle (Caretta caretta)
Baja California, Mexico
Mexico
Italy
Italy
Canary Island,Spain
Turkey
South China
Italy
Japan
Japan
Okinawa, Japan
Cyprus
Cyprus
Caurant et al. (1999)
Storelli et al. (1998)
Gordon et al. (1998)
Sakai et al. (1995)
Aguirre et al. (1994)
Present study
Present study
France
Adriatic Sea,,Italy
Australia
Japan
Hawaiian Islands
Balloki barrage
Rasul barrage
Co
Pb
Zn
1.26
0.17
9.29
31.47
3.77
0.52
27.88
Green turtle (Chelonia mydas)
Olive Ridley turtle (Lepidochelys olivacea)
Loggerhead turtle (Caretta caretta)
Loggerhead turtle (Caretta caretta)
Loggerhead turtle (Caretta caretta)
Loggerhead turtle (Caretta caretta)
Green turtle (Chelonia mydas) Juv.
Loggerhead turtle (Caretta caretta)
Green turtle (Chelonia mydas)
Loggerhead turtle (Caretta caretta)
Green turtle (Chelonia mydas)
Loggerhead turtle (Caretta caretta)
Green turtle (Chelonia mydas)
73.10
5.28
57.20
8.35
5.01
16.96
2.48
8.35
142
38.3
38.5
30.5
3.46
4.35
6.4
2.60
1.21
4.60
2.08
0.03
4.46
32.47
Loggerhead turtle (Caretta caretta)
Loggerhead turtle (Caretta caretta)
Green turtle (Chelonia mydas)
Loggerhead turtle (Caretta caretta)
Green turtle (Chelonia mydas)
Ganges soft-shell turtle (Aspideretes gangeticus)
Ganges soft-shell turtle (Aspideretes gangeticus)
13.3
7.52
117.9
39.4
26
2.12
1.28
11.39
Cr
Cu
Fe
15.2
Mn
11.59
Ni
0.2
1.21
2.2
8.27
1.3
2.2
30
5.6
1.5
0.21
0.12
2.44
3.99
0.31
0.12
0.81
0.16
97.0
23.10
9.09
NA
143.5
169
25.4
29.6
2.45
1.81
3.29
3.71
1.00
0.94
2.21
NA
ND
1.30
3.6
9.47
7.76
0.21
35.9
1.57
267.45
115.93
2.47
0.92
0.57
0.44
BDL
BDL
10.23
8.83
NA
76.3
25.8
53.06
51.01
Table 4 Metal concentrations (mean and range, μg/g) in muscle of turtles from different regions
Reference
Location
Turtle species
Cd
Garcı´a-Ferna´ndez et al. (2009)
Garcı´a-Ferna´ndez et al. (2009)
Gardner et al. (2006)
Frias et al.(2006)
Maffucci et al. (2005)
Storelli et al. (2005)
Torrent et al. (2004)
Andalusia (Spain)
Andalusia (Spain)
Baja California, Mexico
Mexico
Italy
Italy
Canary Island,Spain
0.04
0.20
31.1
2.6
107.0
0.07
1.14
Kaska et al. (2004)
Lam et al. (2004)
Lam et al.(2004)
Franzellitti et al.(2004)
Turkey
South China
South China
Italy
Loggerhead turtle(Caretta caretta)
Loggerhead turtle (Caretta caretta)
Green turtle (Chelonia mydas)
Olive Ridley turtle (Lepidochelys olivacea)
Loggerhead turtle (Caretta caretta)
Loggerhead turtle (Caretta caretta)
Loggerhead turtle
(Caretta caretta)
Loggerhead turtle (Caretta caretta)
Green turtle (Chelonia mydas) Adult
Green turtle (Chelonia mydas) Juv.
Loggerhead turtle (Caretta caretta)
3.57
0.17
BDL
0.36
Sakai et al. (2000a)
Godley et al. (1999)
Godley et al. (1999)
Caurant et al. (1999)
Storelli et al. (1998)
Sakai et al. (1995)
Davenport and Wrench (1990)
Present study
Present study
Japan
Cyprus
Cyprus
France
Adriatic Sea,,Italy
Japan
British Island
Balloki barrage
Rasul barrage
Loggerhead turtle (Caretta caretta)
Loggerhead turtle (Caretta caretta)
Green turtle (Chelonia mydas)
Loggerhead turtle (Caretta caretta)
Loggerhead turtle (Caretta caretta)
Loggerhead turtle (Caretta caretta)
Leatherback turtle (Dermochelys coriacea)
Ganges soft-shell turtle (Aspideretes gangeticus)
Ganges soft-shell turtle (Aspideretes gangeticus)
0.06
0.57
0.37
0.08
0.14
0.06
0.06
0.23
0.05
Co
Cr
Cu
Fe
Mn
Ni
1.01
5.04
0.10
3.1
0.20
0.59
2.85
0.11
0.08
2.18
2.71
1.55
1.56
3.74
1.50
0.81
19.8
5.03
1.27
1.07
0.21
0.28
0.08
Pb
Zn
0.06
0.26
0.01
1.78
NA
0.04
2.26
13.08
65.39
0.41
2.7
27.90
6.70
2.42
0.26
0.08
NA
238.7
147.7
30.90
0.02
2.46
BDL
25.0
0.73
19.60
0.13
3.71
4.12
0.94
0.71
0.83
0.26
2.57
1.61
20.1
0.30
370.25
164.6
5.56
2.35
1.02
1.4
8.23
3.03
24.2
1.89
71.99
60.52
Environ Sci Pollut Res
Environ Sci Pollut Res
freshwater turtle species. They found that Cd dose at 1 μg/g
reduced the germ cell numbers in T. scripta but oocytes
apoptosis increased with Cd concentrations. Furthermore,
they also found that the effects of Cd on turtle gonadal
development was extended up to 3 months post hatch. In
contrast, our study showed the higher environmental levels
of Cd in kidney from the Baloki barrage (2.12 μg/g) and
Rasul barrage (1.28 μg/g), suggesting Cd may pose threats
to reproductive success of freshwater turtles.
Lead is non-essential metal and poses toxic effects to
various organisms at elevated concentrations. Comparison of
Pb concentration in liver samples revealed that it was significantly different at both sites. Pb concentrations in turtle liver
samples from Rasul barrage were higher (2.33 μg/g) than the
Baloki barrage (1.93 μg/g). Pb concentrations reported in liver
in the present study were similar to Olive Ridley turtles in
Mexico (Frías-Espericueta et al. 2006). However, as compared to the present study, higher Pb concentrations
(2.75 μg/g, dry wt.; 2.94 μg/g, wet wt. respectively) in liver
were found for Loggerhead turtles from southwestern Mediterranean, Spain (García-Fernández et al. 2009). The concentrations of Pb were greater in kidneys at Baloki barrage
(2.32 μg/g) as compared to the Rasul barrage (1.93 μg/g).
Comparison of Pb with previous studies revealed that it was
higher in the present study than reported in Loggerhead marine turtles from Canary Islands, Spain (Torrent et al. 2004).
At Baloki barrage, A. gangeticus showed higher Pb
concentrations (1.23 μg/g) in muscle as compared to
those measured at Rasul barrage (1.03 μg/g). The concentrations of Pb were higher in the present study as
compared to the Loggerhead turtles from southwestern
Mediterranean, Spain (García-Fernández et al. 2009)
South China coast (Lam et al. 2004). Significantly high
Pb concentrations were observed in heart tissue of Ganges soft-shell turtle between both sites.
High Co concentrations in liver were reported in the
present study as compared to hawksbill turtles (E.
imbricata) and green turtles (C. mydas) from Yaeyama
Islands, Japan (Anan et al. 2009). Co concentrations in
liver at Rasul barrage was within range reported by
Lam et al. (2004) in Green turtles (0.58 μg/g) as compared to the present study. Higher Co concentrations in
kidney (11.39 μg/g) were reported by Lam et al. (2004)
in green turtles from the South China coast as compared
to the current study. While lower concentrations
(2.2 μg/g) in green turtles’ kidney were reported by
Anan et al. (2009) from Japan as compared to the
current study. The Co concentrations were measured
higher as compared to those of muscle of green turtles
from South China coast (Lam et al. 2004).
These concentrations were low as compared to those
reported in green turtles from South China coast (Lam
et a l. 20 04). The pr esent study showed h igh
concentration of Ni in turtle’s kidneys as compared
those found by Lam et al. (Lam et al. 2004) in green
turtles from South China coast and by Sakai et al.
(2000a) in Loggerhead marine turtles from Japan. Similar concentrations of Ni in muscle (1.07 μg/g) were
measured in green turtles from south China coast (Lam
et al. 2004) while lower concentrations of Ni were
observed in Loggerhead turtles from Yaeyama Islands,
Okinawa, Japan (Kszos et al. 1992).
Iron is the fourth most abundant element in the
Earth’s crust and occurs in most rocks and soils. The
highest concentration of Fe (1,170 μg/g) was reported
by Alonso Aguire et al. (Alonso Aguirre et al. 1994) in
liver of green turtles from Hawaiian Islands than the
present study. Comparatively, low concentrations of Fe
in liver were observed in Loggerhead turtles from
Yaeyama Islands, Okinawa, Japan (Sakai et al. 2000a).
The concentrations in turtle kidney found in the present
study were high as compared to those reported in Loggerhead turtles in Japan (Sakai et al. 2000a). Low
concentrations in comparison with present study were
found in muscles of Loggerhead turtles in Japan (Sakai
et al. 2000a). Lam et al. (2004) reported high concentrations of Fe in livers of green turtles from South
China as compared to those found in the present study.
Manganese is essential as a cofactor for some enzymes. Comparatively high concentrations of Mn were
observed in green turtle kidney from South China (Lam
et al. 2004) while low concentrations were observed in
Loggerhead turtles from Japan (Sakai et al. 2000a). Low
concentrations of Mn were reported in green turtles and
Loggerhead turtles from South China and Japan, respectively (Lam et al. 2004).
Conclusion
The A. gangeticus samples (liver, kidney, heart, muscle,
plastron, and carapace) collected from Baloki barrage
showed higher metal concentrations i.e., of Cd, Co, Cr,
Cu, Mn, Ni, and Pb than the turtle samples collected from
Rasul barrage. Among tissues and organs, the kidney revealed high concentrations of Fe, Zn, and Pb from Baloki.
Heart samples collected from Baloki barrage showed higher
concentrations of Cd, Cr, Mn, and Ni as compared to those
found in Rasul barrage. Cd, Cr, Cu, Fe, Mn, Pb, and Zn
were higher in muscle tissues of A. gangeticus from Baloki
while Co and Ni were high from Rasul barrage. High
concentrations of Cd, Co, Cu, Fe, Mn, Ni, and Zn were found
in livers of A. gangeticus at Baloki, while Pb was found in
greater concentration in Rasul barrage samples. Among the
external tissues plastron from Baloki showed a high concentration of Fe, Zn, Ni and Cu. Carapace tissue of A. gangeticus
Environ Sci Pollut Res
showed greatest differences between Baloki and Rasul barrage in Cd, Co, Cr, Cu, Mn, Ni, Pb, and Zn. Based on the
metals accumulation in different tissues, it was found that the
Baloki barrage was more polluted than the Rasul barrage.
However, there is more need to investigate the environmental
contamination impact on the freshwater turtle population for
better conservation of freshwater turtles.
Acknowledgments The authors are grateful to Pakistan Wetlands
Program (PWP) for providing transport during sampling and the first
author especially acknowledges the support of Mr Richard Garstang
toward this study.
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