Table of Content

S.No. CONTENT PAGE NUMBER

1. Abstract
2. Introduction
3. Sources of heavy metal pollution

4. Bioaccumulation of heavy metals

5. Detection of heavy metal by Histopathology

6. Hazards of heavy metals

7. Histopathological alterations due to heavy metals
pollution in fish orgains/tissues
8. Biomarkers

9. Highlights
10. Conclusion
11. Reference
TOPIC:- IMPACT OF WATER POLLUTION WITH HEAVY METALS
ON FISH HEALTH : OVERVIEW AND UPDATES

ABSTRACT
HEAVY METALS are natural trace components of the aquatic environment, but their levels
have been increasing due to industrial wastes, geochemical structure, agricultural and mining
activities.

All these sources of pollution affect the physiochemical characteristics of the water,
sediments and biological components, thus negatively affecting the quality and quantity of
fish stocks. Environmental pollution is a worldwide problem; heavy metals constitute one of
the most important pollutant challenges.

The progress of industry has led to increase emission of pollutants into ecosystem.
Environmental pollution can cause poisoning, disease and even death of fish.

Therefore, the aims of the present review article are three ways:-

 First to high lighten the impact of the bioaccumulation of heavy metals in different
organs of fish and the factor affecting their dissemination.
 Secondly to monitor the biomarkers i.e. used in the determination and diagnosis of
heavy metal toxicity and pollution.
 Finally the role that is played by the Histopathological studies on the diagnosis of the
fish disease caused due to heavy metals.
 INTRODUCTION
Water is the most important natural resource that exists on our planet and is essential for the
survival and the development of modern technology. Thus, rapid industrialization is one of
the main causes for aquatic pollution. Discharged waste water has been used in different
regions of the world for fish raising. Fish are one of the most widely distributed organisms in
the aquatic environment and being susceptible to metal contamination, may reflect the extent
of the biological effects of metal pollution in water. Metal pollution may damage aquatic
organisms (either fresh or marine water) at the cellular level and possibly affect ecological
balance. Heavy metals are dangerous because they tend to bio accumulate in the body.
Accumulation of various metals in various organs of fish may cause structural lesions and
functional disturbances.

HEAVY METAL

Heavy metals are natural components of the earth crust. They can’t be degraded or distorted.
Or we can say that the term heavy metal refers to any metallic chemical element that has a
relatively high density and is toxic or poisonous at low concentration. Heavy metals are
commonly defined as those having a specific density of more than 5g/cm3.

Since there are different definitions proposed, some based on density, some on an atomic
number or atomic weight and some on chemical properties or toxicity.

HEAVY METAL is widely distributed throughout the environment in the air, water
and land. It exists in components that may be organic or inorganic. It is highly toxic in its
inorganic form.

Some of the heavy metals are –

 Cadmium (Cd)
 Mercury (Hg)
 Arsenic (As)
 Chromium (Cd)
 Lead (Pb)
 Aluminium (Al)
 Copper (Cu)
 Iron (Fe)

Arsenic: - Arsenic reach aquatic ecosystem by a variety of sources. Arsenic compounds in

the third oxidation state (arsenites) are absorbed fairly rapidly into fish and are more toxic
than arsenic compounds in the oxidation state V (Arsenates). Arsenic is actively metabolised
in the tissue of the fish especially to organ such as liver and has tendency to accumulate in
different teleosts such as green sunfish, Tilapia massambica.
Total arsenic in marine fish, shellfish and fresh water fish tissues ranged from 0.19 to 65,
0.2 to 125.9, and 0.007 to1.46 µg g-1dw respectively.

Acute exposure can result in immediate death because of As- induction increase in mucus
production causing suffocation or direct detrimental effects on the gill epithelium.

Cadmium:- Cadmium is a naturally occurring non-essential trace element and its tendency to
bio accumulate in living organism often in hazardous levels, raises environmental concern.
Cadmium production, consumption and emission to the environment have increased
dramatically during the 20th century, due to its industrial use and consequently lead to
contamination of aquatic habitats and deformities (vertebral curvatures and yolk sac
deformities).

Chromium:- Chromium is an essential nutrient metal, necessary for metabolization of

carbohydrates. Chromium enter the aquatic ecosystem through effluents discharged from
leather tanneries, textiles, electroplating, metal finishing, mining, dyeing, ceramic,
photographic and pharmaceutical industries, etc. poor treatment of these effluents can lead to
the presence of Cr(VI) in the surrounding water bodies, where it is commonly found at
potentially harmful levels to fish. Hexavalent chromium (Cr6+) is considered to be toxic (i.e.
carcinogenic) because of its powerful oxidative potential and ability to cross cell membrane.

Fish assimilate Cr by ingestion or by the gill uptake tract and accumulation in fish tissues,
mainly liver occurs at higher concentration than those found in the environment.

Vick and Sharma assessed the effects of acute toxicity of Cr on fingerlings of the C. mrigala.
After 15 days of exposure significant decline in the protein and carbohydrate content of gill
was observed.

Lead (Pb):- Lead is a persistent heavy metal which has been characterized as a priority
hazardous substance. Although Pb is a naturally occurring substance, its environmental
concentrations significantly increase by anthropogenic source which include base metal
wining, battery manufacturing, etc.

Aquatic organism bio accumulates Pb from water and diet. Lead deposits in various fish
organs liver, kidneys and spleen, but also digestive tract and gills. When C. batrachus
exposed to 5ppm of lead nitrate for 150 days, it exhibited marked inhibition of gonadal
growth and showed decrease in cholesterol and lipid levels in brain, testis and ovary whereas
the liver showed an elevation of both. Acute lead toxicity is initially characterized by
damaging gill epithelium and ultimately suffocation.

Mercury (Hg):- Unpolluted water contains trace amounts which do not exceed than 0.1µg/l
of mercury. The chronic data about mercury toxicity indicated that the organic force of Hg.
Methyl mercury (MeHg+) is the most chronically toxic of the mercury compounds and it is
estimated that 70-100% of the Hg in fish is present as [MeHg+].
 Methyl mercury is generated by methylation of inorganic Hg by microorganisms such as
anaerobic sulphate-reducing bacteria, iron reducers (FeRP) and methanogens (MPA).
Simulations of ocean warning rates of 0.4 C and 1C predicted increase in the mean MeHg +
concentration of 1.7% (range 1.6-1.8%) and 4.4% (range 4.1-4.7%) respectively.

The LC50 of MeHg for fish is reported to be in the range of 0.004-0.125 mg/L depending on
the species, which is far lower than the LC50 of inorganic Hg. The adverse effect of MeHg
were first recognised in the early 1970s following the pollution of Minamata Bay, Japan and
shortly thereafter a massive human poisoning that occurred in Iraq. The primary route of
exposure for fish is through ingestion of contaminated food. Fish tissues are sensitive
indicates of aquatic pollution and have a high Hg bioaccumulation capacity for both organic
and inorganic forms.

SOURCES OF HEAVY METAL POLLUTION

The sources of heavy metals were reported by the PAD. These are as follows:-

 Mining effluents
 Industrial effluents
 Domestic effluents
 Urban storm-water
 Leaching of metals from garbage
 Solid wastes dump
 Metals inputs from rural areas
 Batteries
 Pigments
 Paints
 Glass
 Fertilizers
 Textiles
 Dental and cosmetics
 Atmospheric sources
 Petroleum industrial activates
 Moreover, heavy metal pollution can arise from many sources as smelting of Cu,
preparation of nuclear fuels, electroplating with chromium and cadmium.
 Cadmium, lead and zinc are released into tiny particles as dust from rubber tires on
road surface. These small size particles allow these toxic metals to raise on the wind
to be inhaled, or transported onto top soil or edible plants.
 Cigarette smoking is a major source of Cd exposure.
 Other sources of contamination are the manufacture and use of arsenical pesticides
and wood preservatives.
 BIOACCUMULATION OF HEAVY METALS:-
Bioaccumulation means an increase in the concentration of a chemical in biological organism
overtime, composed to the chemical’s concentration in the environmental. Fish living in
polluted waters lead to accumulate heavy metals in their tissues.

Accumulation depends on –

 Metal concentration
 Time of exposure
 Way of metal uptake
 Environmental conditions
 Water temperature
 PH
 Hardness
 Salinity
 Intrinsic factors-
 Fish age
 Feeding habits
 Most of these metals accumulate mainly in liver, kidney and gills.
 Metals in natural water occur in particulates or in soluble form, including labile and
non-labile fractions.
 The labile metal compounds are most dangerous to fish. They include various ionic
forms of different availability to fish.
 The results of many field studies of metal accumulation in fish living in polluted
water show that considerable amount of various metals may deposited in fish tissue
without causing mortality.
 Various metals are accumulated in fish body in different amounts. Generally, the
higher metal concentration in the environment the more it may be taken up and
accumulated by fish.

Metals levels in the line fish usually follow the ranking-

Fe > Zn > Pb > Cu > Cd > Hg

 The levels of Zn may be very high up to over 300 µg/g dw.

 In India, various studies have been conducted on heavy metal bioaccumulation in the
muscle tissue of fish collected from different fresh water aquatic systems in relation to
their concentration in water.
 Recently, Mahino and Nazura collected water samples from 22 km segment II of
Yamuna River from Okhla barrage, India.
  The water samples were used to determine the presence of Cr, Ni and Pb. They found
that concentrations of heavy metals were much above the maximum permissible
limits set by WHO.
 Bioaccumulation factor used to evaluate the heavy metal concentration in different
fish tissues and calculated as the ratio of concentration of pollutant accumulated in the
tissue of organism with respect to the concentration of that pollutant using the
formula:-

Concentration of heavy metal in fish tissue (µg g-1)

Concentration
of heavy metals in surrounding water (µg L-1)

Adverse effects of heavy metals on fish are not only related with material accumulation but
also to cumulative toxic effects. Lethal disturbances are resulted due to exceeding certain
values of heavy metals concentration in fish.

DETECTION OF HEAVY METALS BY HISTOPATHOLOGY:-

The effect of heavy metals can be detected through the histopathology techniques. The
histological analysis of fish organs i.e. liver, kidney, gilt, skin, muscles is done.

Method:-

 Tissue samples are put in a series of ascending grades of ethyl alcohol (70-100%) for
dehydration.
 Clarification is done in Xylol or chloroform.
 The treated samples are embedded in paraffin wax (45oC).
 The paraffin blocks are prepared and with a microtome cuts at thickness of 0.6 µm are
done.
 The tissue sections are stained with haematoxylin and eosin.
 For long-term examination, the stained slides are covered using Canada balsam and
examined with a light microscope.
 HAZARDS OF HEAVY METALS
Mechanism of pathogenicity:-

 Lead and cadmium are industrial pollutants which have strong negative effect on
animal and human health.
 These metals are accumulated in the organism mainly in liver and kidneys.
 Lead exerts its effect, physiologically and biochemically as a mimetic agent
substituting for essential elements participating in metabolism such as calcium, iron
and zinc.
 The maximum Cd and Pb levels permitted for sea fish are 0.1 and 0.4 mg/g
respectively.
 The levels of Cd and Pb in the underground water were reported as 0.003 and 0.01
mg/L respectively.
 WHO/FAO reported that Cd in surface water is usually found together with zinc but
at much lower concentration. The Cd present in the surface water may be either
dissolved or insoluble of the dissolved forms, those which may be poisonous to fish,
include the sample and various inorganic & organic complex ions. Its acute toxic
action damage to central nervous system and parenchymatous organs, very small
concentration of Cd may produce specific effects after a long exposure period.
 The acute lethal concentration of Cd for different species of fish ranges from 2-20
mg/L.
 The gill is an important site for the entry of the heavy metals and is the first target
organ for exposure in fish.
 The high concentration of metals in the gills of P. fluvidraco and C. carpio is due to
the metals complexation with the mucus.
 The concentration of metals in the gill reflects the level of metals in the water where
the fish live, whereas the concentration in liver and kidney represent the storage of
metals. Thus, the gills in fish are more often recommended as environment indicator
organ of water pollution than any other fish organs.
 In liver, higher concentration of Pb were recorded in P. fluvidraco (0.76 µg/g) than in
C. carpio (0.63 µg/g) respectively.
 The increased metal bioaccumulation (Cu, Cr, Cd, Pb) in the different tissues of two
fishes significantly different for during summer and winter season. And the variation
in the level of heavy metals among different species depends upon its feeding habit,
age, size and length of the fish and their habitats.
 Dissolved form of Cd in water surface may be poisonous to fish. Very small
concentration of Cd may produce specific effects after a long exposure period.
 The Cd is deposited in soft tissues of the body with 50-70% accumulation in both
kidney and liver.
 HAZARDS/EFFECT OF HEAVY METELS
There are so many health problems occurring due to heavy metals. Some of them are as
following:-

 External organs are affected.

 Causing loss of equilibrium.
 Increase opercular movement.
 Irregular vertical movements.
 Leading to death.
 Renal damage
 Nervous system damage
 Gill damage
 Severe destructive pathological changes i.e. structural lesions

HISTOPATHOLOGICAL ALTERATIONS DUE TO HEAVY METAL

POLLUTION IN FISH ORGAN/TISSUE

 Histopathology deals with the study of pathological changes of the microscopic

structure of the body tissue. Any peculiar type of alteration of cells may indicate the
presence of the disease or the effect of toxic substances.
 There are various studies done on the pathological changes. And we all only give the
reviews on this.
 Histopathological studies give us useful data concerning tissue prior to external
manifestation.
 Various changes found in the fish organ and tissues. For example:-
Skin – Tilapia fish was adversely affected by heavy metals pollution showing hyper
activation of goblet cells and dermal melanosis and dermal granuloma.
Kidneys – In the polluted kidneys of carp fish there appears hyaline casts, interstitial
nephritis and renal necrosis and mononuclear cells infiltration.
Brain – Sections of polluted carp brains exhibited symptoms of meningitis and
gliosis.
 BIOMARKERS
 Fish species were recently suggested as environmental biomarkers.
 Fishes are considered as early warning for the degradation of environmental quality,
but also specific measures of mutagenic compounds in the biological materials.
 The gill, liver, and kidneys are commonly the primary target organs for pollution.
 Histopathological lesions and increase in size were reported in various fish exposed to
heavy metals.
 Riji and George reported that the diagnostic tools used in the diagnosis of fish disease
should be as follows-
 Case history –
 Water quality
 Feeding percentage
 Feed intake
 Fertilization
 Time of last algal bloom
 Living details
 Treatment details
 Source of seed
 Stocking details
 Microscopic diagnosis (including light and electron microscope slides preparations, H
& E stain and special stains) for histological diagnoses.
 Immunological and molecular diagnoses.
 Microbiological diagnoses.
 Wet digestion-
Micro digestion
 Biomarkers
 CONCLUSION
This review highlights the causes and consequences of heavy metal contamination on fish
organ. There is a strong evidence of a co-relation between heavy metals concentration in
different fish tissue and those of the surface water of the polluted area.

Industrial wastes are potential source of heavy metal pollution in aquatic

environment. In recent years, the anthropogenic pollution of aquatic ecosystems increased the
need for studies to identify the impact of heavy metals on the species living there.

Monitoring programs for bioaccumulation measurements serve as a biomarker for fish from
contaminated places and provide information about the environmental conditions.
Histopathological changes are more sensitive and occur earlier than any other evidence. They
provide a better assessment technique of fish health and to the effects of pollution on each
biochemical parameter. Metal pollution may damage aquatic organism at the cellular level
and possibly affect the ecological balance.

Monitoring of environmental parameters is a key activity in managing the environment

restoring polluted environments and anticipating the effects of man- made changes on the
environment.