Assesment of Heavy Metals in Nile Tilapia (Oreochromis niloticus)

Collected from Malacca River, Malaysia

N. A. Kamarudin

, S. Z. Zulkifli

, A. Ismail

and F. Mohamat-Yusuff

Department of Biology, Faculty of Science, Universiti Putra Malaysia, 43400 UPM Serdang, Selangor, Malaysia

Department of Environmental Sciences, Faculty of Environmental Studies, Universiti Putra Malaysia, 43400 UPM

Serdang, Selangor, Malaysia

Keywords: Heavy metal, Nile tilapia, Oreochromis niloticus, Toxicology.

Abstract: Present study aimed to determine selected heavy metals (Cu, Zn, Pb) concentration in Nile tilapia

(Oreochromis niloticus) collected from Malacca River, Malaysia. Samples of O. niloticus were collected

from several locations along the Malacca River, transported back to the laboratory, treated with standard

digestion procedures and analyzed with an atomic absorption spectrophotometer (AAS). Recovery heavy

metal concentration of the reference material was satisfactorily between 98 to 102 percent. Results

demonstrate that Cu in O. niloticus from Malacca River was significantly higher than other studied metal.

Concentration of heavy metals may vary depending on locally anthropogenic activities. Concentration of

heavy metals may not be reduced after short time acclimatization. Liver was found to contain significant

amount of heavy metals. Detail studies of various aspects involving O. nilotiucs should be conducted as it

shows a potential candidate of biomonitoring agent to determine the water pollution in order to manage the

aquatic ecosystem.

1 INTRODUCTION

Since several decades ago, Malaysia has emerged as

one of the Asian countries that have rapid boost in

economy, industry, agriculture and urbanization.

However, unsustainable and improper planning for

growth and developments by authorizes and

communities could severely affect the environment.

According to [1], the accumulation of heavy metals

such as As, Cu, Pb, Zn, and Fe in the environments

are due to the anthropogenic activities or men made.

Industrial activities can lead to the severe

environmental condition from the untreated sewage

discharged. Among the characteristics of the heavy

metals are non-biodegradable, very persistence and

remain still in the environments (Zulkifli et al,

2010). The monitoring of the water sample,

sediments and the biota can be used to determine the

amount of the heavy metals in the aquatic ecosystem

such as river (Naji and Ismail, 2012).

Some heavy metals are easily enriched into

fishes including those species that are commercial

and having significantly high economic value in

markets (Ashraf et al, 2012). For example, the Nile

tilapia (Oreochromis niloticus) is an important fish

in many countries including Malaysia and

increasingly reared in aquaculture (Cogun et al,

2003). Fish is suitable for biomonitoring water

quality and easily be interpreted by the consumers in

public. Some environmental changes can be detected

by the fish and it can be used as monitor and

indicator to the pollution (Ashraf et al, 2012). Fish is

very suitable model to be used in the study of

accumulation of heavy metals in different tissues of

fish (Cogun et al, 2003). According to (Batvari et al,

2007), the present of fish in large scale, potentially

metal accumulation, long lifespan and optimum

range of size and easy to be sampled are the reasons

why fish can be used as a good bioindicator and

biomonitor. Among the advantages of using tilapia

fish are because of their tolerance to poor water

quality and the fact that they have wide range of

natural food organism such as plankton, detritus,

aquatic invertebrates and decomposing organic

matter (Popma and Masser, 1999). In addition,

tilapia is the most tolerant fish as compared to other

freshwater fish in the high salinity, high water

temperature and low dissolved oxygen.

Heavy metals can accumulate into biological

tissues through the absorption from the water body

Kamarudin, N., Zulkiﬂi, S., Ismail, A. and Mohamat-Yusuff, F.

Assessment of Heavy Metals in Nile Tilapia (Oreochromis Niloticus) Collected from Malacca River, Malaysia.

DOI: 10.5220/0009905500002480

In Proceedings of the International Conference on Natural Resources and Sustainable Development (ICNRSD 2018), pages 521-527

ISBN: 978-989-758-543-2

521

and exposed to the human via consumption from the

food web (Zulkifli et al, 2014). Bioaccumulation of

heavy metals in human body can lead to harmful

effect in human health from contamination of heavy

metals food chain. The concentration of the heavy

metals is increasing as the pass the tropic level from

lower to higher tropic level and this phenomenon is

known as biomagnifications (Zulkifli at al, 2016).

The effect on human health later can be acute and

chronic effect depends on the level exposure to the

heavy metals from the consumption of fish that

contain heavy metals such as Zn, Cu, Pb, and Fe.

Besides that, some heavy metals can cause

neurological and behavior changes in children

especially and some of them also carcinogenic,

mutagenic, teratogenic and endocrine disruptors

(Iavicoli et al, 2009). Therefore, present study aimed

to determine heavy metals (Zn, Cu, Pb)

concentration in O. niloticus collected from Malacca

River.

2 MATERIALS AND METHOD

2.1 Sample Collection

Figure 1: (I) State of Malacca (red box): (II) sampling points along Malacca River (A) Taman Merdeka; (B) Malim; (C)

BatuBerendam; (D) Bachang.

A total of samples 206 individuals of tilapia fish

(O. niloticus) were collected by using a casting net

from four different locations, namely Taman

Merdeka, Malim, BatuBerendam and Bachang

(Figure 1). The standard length (SL) range of the

collected fishwas between 18 cm to 23 cm. The fish

were divided into two condition of fresh and

acclimatized environment. The fresh samples are the

samples were immediately stored after collected

from the river, while the acclimatized samples were

kept alive in a container containing aerated tap water

for 24 hours. All samples were transported back to

the lab

2.2 Analytical Procedures

For fish samples, all samples from each station will

be pooled together. The livers and muscles were

dissected separately and the scales were removed.

Livers organ is selected due to its function as a

detoxify site of toxic chemical in the body. Only

tissues along with skin without bone were selected

for analysis. Tissues with skin were selected as

assuming that people consuming the fish together

with skin. Those samples was dried using air-

circulating oven at 60°C for 72 hours until the water

content loss and the constant weight was obtained.

ICNRSD 2018 - International Conference on Natural Resources and Sustainable Development

522

To digest biological tissues with acid, we

followed method established by [11]. In brief, about

10 ml of concentrated nitric acid (HNO

) was used to

digest 1.0g samples in a digestion tubes. The tubes

were heated in a digestion block for four hours. The

heating process was first preheating for an hour and

followed by increased gradually to 140°C for another

three hours to completely digesting the biological

samples. The temperature was increased gradually to

avoid the sample from boil vigorously and may

splatter across the tube. This may affect the total of

sample digest and alter the precision of the result.

After the digestion was completed, all the digested

samples were allowed to cool at room temperature

for 30 minutes. Then, the samples were diluted with

30 ml of distilled water to a fixed volume (40 ml).

Next, Whatman filter paper was used to filter the

digested samples and those filtrates was stored in

refrigerator until metal determination. The

concentration of Zn, Cu, Pb and Fe were analyzed by

using Flame-Atomic Absorption Spectrophotometer

(FAAS). All the data obtained was converted into

µg/g basis. The SRM 2976 - Mussel Tissue (Trace

Elements & Methylmercury) was used as the

reference material. The recovery of heavy metals was

between 98 to 102 percent.

2.3 Statistical Analysis

The data obtained from the AAS instrument were

converted into micrograms per gram (µg/g) or parts

per million (ppm) to obtain the actual data. All the

data obtained were analyzed with multivariate

analysis of variance (MANOVA) from IBM SPSS

Statistics 20.0 software. Mean data were converted

into tables and graph by using Microsoft Excel. All

the comparisons obtained were made at least at the

95% level of significance (p<0.05).

3 RESULTS AND DISCUSSION

3.1 Accumulation of Heavy Metals in

Tilapia Fish (Oreochromis niloticus)

Figure 2 shows the distribution of metal (mean ±

standard error) Zn, Cu, and Pb in O. niloticus fish

(muscle and liver) at different sampling sites. Based

on the graph, the metal Cu concentration is highest

in Malim with mean of 1554.36 µg/g, followed by

Taman Merdeka (923.70 µg/g), Bachang (836.84

µg/g), and BatuBerendam (739.47 µg/g). While,

metal Zn concentration is highest in Malim with

mean of 124.51µg/g is far lower than metal Cu

concentration. Metal Pb concentration is highest in

Taman Merdeka (87.15µg/g) and lowest in

BatuBerendam (13.214µg/g). Multivariate test of

MANOVA shows that there are significantly

different (p<0.05) between distributions of metals at

different sampling sites.

Fishes are sometimes positioned at the top level

of a food chain. Their condition can be closely

related with the surrounding environments,

including pollution. The heavy metals concentration

in aquatic environments can be estimated from the

water, sediments and an organism. O. niloticus fish

can be one of the biomonitoring agents that can

reflex the real environment. Fish also has high

potential to bioaccumulate metals through

absorption from polluted water, food

(biomagnification) and sediment and transfer to

human via food chain (Zulkifli et al, 2016;Ahmad

and Suhaimi, 2010) . The accumulation of heavy

metals can be influenced by factors such as gender,

location, environment, season and size.

In this study, the concentration of Cu in O.

niloticus fish including the muscle and liver is the

highest in all sampling sites as the concentration of

Cu in sediments which higher at all sampling site.

However, metal Cu concentration is lower in the

river water may due to the dilution factor in rainy

season. Metals in the river water are from

environmental availability of metal in sediment

leaching out and also from direct sources of metal

influx and sediments runoff into the river. The main

source of Cu is from the industrial area and urban

development construction along the Malacca River.

Metal Cu is the common metal found in the water

and sediment and can bioaccumulate and

biomagnified in the fish from its diet (Ahmad and

Suhaimi, 2010). The metals enter the fish body

through the respiration from the gills, adsorption

through skin and detoxification in the liver (Shouta et

al, 2010).

Zn and Pb have significantly lower concentration

in the O. niloticus fish which can be related to the

predicted lower concentration of Zn and Cu in the

sediment and water samples. Other study by (Ahmad

and Suhaimi, 2010) in Lake Chini, Pahang shows

that the concentration of Zn is the highest in fish

however in this study the concentration of Zn and Pb

in fish is higher than the fish in Chini Lake due to

the a lot of human activities along Malacca River.

Assessment of Heavy Metals in Nile Tilapia (Oreochromis Niloticus) Collected from Malacca River, Malaysia

523

Figure 2: Distribution of the heavy metals concentration (mean ± standard error; µg/g dry weight) of Zn, Cu and Pb in O.

niloticus fish at four different sampling sites.(Remark: * is the actual values need to be multiply with 5).

3.2 Distribution of Heavy Metals

between Conditions

Figure 3 shows the comparison of metal

concentration (mean ± standard error) in O. niloticus

fish with different conditions of fresh and

acclimatized sample. Based on the graph, the

acclimatized sample has higher metal concentration

as compared to fresh sample with metal Cu has

highest concentration with mean of 1074.02 µg/g.

Followed by metals Zn and Pb have lowest

concentration (35.76 µg/g) in fresh sample.

According to ANOVA test, there is no significant

different (p>0.05) between the condition of fresh

and acclimatized sample in all metals.

61.41

124.51

70.19

109.01

184.74

310.87

147.89

167.37

87.15

34.34

13.21

20.96

100

150

200

250

300

350

Taman Merdeka

Malim

Batu Berendam

Bachang

Taman Merdeka

Malim

Batu Berendam

Bachang

Taman Merdeka

Malim

Batu Berendam

Bachang

Zn Cu Pb

Metal cconcentration (µg/g)

Sampling Location

ICNRSD 2018 - International Conference on Natural Resources and Sustainable Development

524

Figure 3: Distribution of the concentration (mean ± standard error; µg/g dry weight) of Zn, Cu and Pb between different

conditions.(Remark: * is the actual values need to be multiply with 5)

The different conditions of fresh and acclimatized

could demonstrate that concentration of metal in

acclimatized fish sample is not significantly higher

as compared to the fresh sample. This is may be due

to the simple reabsorption of metal through skin or

through gills from the acclimatized water (Alex et

al, 2013). Thus, the conclusion from this parameter

is there is no significant if we consumed the fish

from acclimatized condition with the fresh fish

sample. The acclimatized condition should be

regularly checked and the water needed to be

replaced with clean water regularly. So, that the

metal leaked out in the water will not be reabsorbed

back into the body through gills or skins.

3.3 Distribution of Heavy Metals

between Organs

Figure 4 shows the comparison of metal

concentration (mean ± standard error) in O. niloticus

fish with different organ; muscle and liver. Based on

the graph, metals concentration is higher in liver

organ compare to muscle tissues. Metal Cu has

99.71% higher concentration in liver organ

compared to muscle tissues. While, metal Zn

concentration in liver has 71.78% higher in liver

organ. However, metal Pb has 42.72% higher

concentration in muscle tissues than liver. There are

significant different (p<0.05) in metal concentration

between organ muscle and liver.

Assessment of Heavy Metals in Nile Tilapia (Oreochromis Niloticus) Collected from Malacca River, Malaysia

525

Figure 4: Distribution of the concentration (mean ± standard error; µg/g dry weight) of Zn, Cu and Pb between organs of

muscle tissues and liver organs. (Remark: * is the actual values need to be multiply with 10).

In general, the level of heavy metal is lower in

muscle tissues as compared to liver organ. Similar

results have been reported with sample genus of fish

Oreochromis and other species of fish. According to

[1], the concentration of heavy metals Zn, Cu, Pb

and Ni in liver organ was higher than muscle tissues

of O.niloticus in Zambia’s lake. Also, [12] also

reported higher concentration of metals in liver

organ than muscle tissues in various species of fish.

Although muscle tissues have low concentration of

metal as compare to liver, this study is focus on the

fish muscle since it is the part which people

consumed.

Liver is an important organ with many essential

functions in storage of nutrients, metabolism

function, immunity, detoxification or transformation

of metals (Evans et al, 1993). Concentration of Zn

and Cu in liver organ at four different sampling sites

is high as related to the natural binding protein in

hepatic tissues such as metallothioneins (MT). The

MT is a ubiquitous protein that has high affinity to

essential metals, Zn and Cu and stored it for other

function such as enzymatic and metabolic demand

(Salkusak, 2012). Their synthesis is one of the best

known biochemical responses to metal exposure

where they involve in the regulation of the essential

metals and in the detoxification of non-essential

metals (Buhari and Ismail, 2017), antioxidant

activity (Atif et al, 2006) and radical scavenging

(Wright et al, 2000) and helps in metal ion

homeostasis in a cell (Shariati and Shariati, 2011).

Future studies could be conducted to study in details

the functions and behaviour of MT in various

aquatic animals.

4 CONCLUSION

In conclusion, present study determined that Cu in

O. niloticus from Malacca River was significantly

higher than other studies metal. Concentration of

heavy metals may vary depending on locally

anthropogenic activities. Concentration of heavy

metals may not reduce after short time

acclimatization. Liver contain significant amount of

heavy metals. The use of O. niloticus as a

biomonitoring agent is a good way to determine the

water pollution in order to manage the aquatic

ecosystem.

ACKNOWLEDGEMENT

This study was supported by the Fundamental

Research Grant Scheme (FRGS) (Reference No.

FRGS/1/2014/STWN01/UPM/02/4) from the

Ministry of Education Malaysia, and was performed

in research collaboration with the JSPS Core-to-

Core Program, B. Asia-Africa Science Platforms.

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