Research Journal of 
Animal, 
Veterinary and Fishery Sciences
 
__
_____________
______
____
_
_
 
ISSN 2320
 
–
 
6535
 
 
 
Vol. 
1
(
6
), 
1
-
5
, 
July
 
(201
3
)
 
Res. J. Animal, Veterinary 
and
 
Fishery
 
Sci.
 
 
 
 
International Science Congress Association
 
 
 
 
    
1
 
Impact of Tannery Effluent, Chromium on Hematological Parameters in a 
Fresh Water Fish, 
Labeo Rohita 
(Hamilton)
 
 
Praveena
 
M., 
Sandeep
 
V., Kavitha N.
 
and Jayantha Rao
 
K.*
 
Division of Toxicology, Department of Zoology, Sri Venkateswara University, Tirupati 
-
517 502, AP, INDIA
 
 
Available online at: 
www.isca.in
 
Received 
26
th
 
April
 
201
3
, revised
 
27
th
 
May 
201
3
, accepted
 
3
rd
 
July 
201
3Abstract


Chromium is one of the heavy metals present in the Tannery effluent. Chromium is kno
wn to cause various health effects. The 
health hazards associated with exposure to chromium are dependent on its oxidation state. The hexavalent form is toxic than 
trivalent form. The hematological alterations produced on exposure to sub
-
lethal concentrati
on (1/10
th
 
of LC
50
/96 hrs) of 
chromium were investigated in fresh water fish, Labeo rohita for 7days and 30 days respectively. Results revealed statically 
significant decrease in RBC, Hb, PCV, MCH, and MCHC in all the experimental animals when compared to 
the control with 
an increase in exposure days. In contrast to this, the WBC and MCV values were significantly increased. The decrease in 
hematological parameters clearly indicates that the exposed fishes have become anemic due to heavy metal exposure.Keywords: 
Tannery effluent, 
c
hromium, 
l
abeo rohita, 
h
ematological parameters.
 
 
Introduction
 
Pollution has become a serious threat and has brought drastic 
changes and ill
-
effects to the growing population as well as the 
mother Earth. Pollution of aquatic ec
o
-
systems is a major global 
problem since the past two decades. 
Rapid urbanization and 
industrialization has led to increased disposal of pollutants such 
as heavy metals, radio nuclides and various types of organics 
and inorganics into the aquatic environm
ent
. Extensive 
industrialization has measurably influences the quality of water 
of lakes, ponds and rivers all over the world. Industrial wastes 
constitute the major source of metal contamination of the 
aquatic environment
1
.
 
 
Heavy metals constitute the ma
jor contaminants. Metals are 
important pollutants, because they are not eliminated from the 
aquatic ecosystems by natural processes like organic pollutants 
and are enriched in mineral organic substances.  Metal 
contaminants are introduced into aquatic syst
em through 
smelting process, effluents, sewage and leaching of garbage 
which cause serious damage to the aquatic fauna
2,3
. The 
Tannery industry has shown tremendous expansion during the 
last 25 years and plays an important role in India’s foreign 
exchange 
earnings. However it adds pollutants to the aquatic 
environment. The tannery waste waters continue to cause 
negative effects on the aquatic organisms as they also have 
endocrine disruption effects. Tanners use large number of 
chemicals during the process, 
discharging toxic wastes into 
rivers and thereby degrading agricultural lands. The 
uncontrolled release of tannery effluents increases health risks 
to target and non
-
target organisms. 
 
 
Tannery effluents are considered to be more dangerous than all 
the oth
er industrial wastes as pointed out by
 
Arora H.C
4
. 
The 
major constituent of tannery effluent is Chromium, a heavy 
metal which is highly toxic to aquatic fauna
5
. Chromium is an 
important pollutant from tannery effluent and causes deleterious 
effects on non
-
target organisms resulting imbalance of an 
ecosystem. Chromium compounds are used in ferrochrome 
production, pigment production, electroplating and tanning. 
Heavy metal contamination may have devastating effects on the 
ecological balance of the recipient e
nvironment and a diversity 
of aquatic organisms
6,7
. Fish is an important component of 
human nutrition, and those from contaminated sites present a 
potential risk to human health. Metals can accumulate in aquatic 
organisms including fish and persist in wate
r and sediments. 
Fishes are the simple and reliable biomarkers of pollution of 
aquatic bodies. 
When fish are exposed to elevated levels of 
metal in a polluted aquatic ecosystem, they tend to take these 
metals up from their direct environment
8
. 
The blood pa
rameters 
have been used as a sensitive indicator of stress in fish exposed 
to different water pollutants, toxicants and effluents etc. 
Hematological and biochemical profile in fish is a sensitive 
index for evaluation of fish metabolism under metallic stres
s. 
Blood is a good bio
-
 
indicator to study the problem in organ 
function. The measurement of hematological changes of fish 
under exposure to any toxicant may be used to predict its effect 
upon chronic exposure. The blood parameters get affected on 
account 
of metal toxicity.  The oxygen carrying capacity of 
blood, RBC in fishes often varies with life history, habits and 
environmental conditions. In recent years, hematological 
variables were used more when clinical diagnosis of fish 
physiology was applied to 
determine the external stressors and 
toxic substances as a result of close association between the 
circulatory system and external environment
9
. Hence the 
objective of the present investigation is to determine the 
changes in different hematological paramet
ers of the fresh water 
Research Journal of 
Animal, Veterinary and Fishery Sciences
 
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_
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_______________________________
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_
 
ISSN 2320
 
–
 
6535
 
 
Vol. 
1
(
6
), 
1
-
5
, 
July
 
(201
3
)
          
 
         
Res. J. Animal, Veterinary 
and
 
Fishery
 
Sci
.
 
  
 
International Science Congress Association
 
 
2
 
fish, 
Labeo rohita
 
exposed to sublethal concentration of 
chromium. 
 
 
Material and Methods
 
Test Animal: 
Healthy live specimens of 
Labeo rohita
 
(average 
length 10±2cms and average weight 15±2gms) were obtained 
from local ponds and tran
sported to the laboratory treated with 
0.05% KMnO
4 
solution for 2 minutes to avoid dermal infection, 
kept in large cement tanks and supplied with clean de
-
chlorinated tap water. In laboratory the fish were acclimatized 
for about 2 weeks prior to the experi
ment with a photoperiod of 
12:12 light and dark cycle with constant aeration and filtration. 
During the period of acclimatization, the fish were fed with 
commercial fish feed to satiety twice daily. The laboratory tap 
water was analyzed for physico
-
chemica
l parameters by 
adopting standard methods
10
.
 
 
Test Chemical: 
 
Analytical grade Chromium as Potassium di 
chromate supplied by BDH (India) was used as a metal toxicant 
throughout the experiment.
 
 
Experimental design: 
 
Fishes were divided into 3 groups 
contai
ning 10 fishes each with the I group serving as control  
without any treatment, the group II, III fish were exposed to 
sublethal concentration
 
(1/10
th
 
of LC
50
 
96hrs, 10ppm)
 
of 
Potassium dichromate for 7days and 30 days after determining 
LC50 value
11
. 
After
 
stipulated period, the control and 
experimental fishes were washed with tap water and dried using 
blotting paper before collecting the blood sample. Blood was 
drawn by cardiac puncture using 2cm
3
 
disposable plastic syringe 
and 21 gauge disposable needle. 
After collection the blood was 
immediately transferred to glass vials containing 1% EDTA 
solution. Blood samples were used for the measurement of RBC 
(Erythrocyte), WBC, PCV (Haematocrit), Hemoglobin (Hb) 
concentration. The methods employed for determinati
on of 
different hematological parameters were RBC 
and
 
WBC count 
by Neubauer’s improved Hemocytometer using diluting fluids
12
, 
PCV by Wintrobe’s method and Hb using Sahli’s 
hemoglobinometer. The haematological Indices, Mean 
Corpuscular
 
Volume (MCV), Mean 
Co
rpuscular
 
Haemoglobin 
(MCH) and Mean
 
Corpuscular
 
Haemoglobin Concentration 
(MCHC) were calculated using the formula of
 
Baker F.J. and 
Silverton R.E
.
 
given below
13
 
given below:
 
 
Mean Corpuscular Volume (MCV) μm = 
Haematocrit (%) x 10 
 
                      
                                      
Erythrocyte count (mm3) 
 
 
 
Mean Corpuscular Haemoglobin (MCH) pg = 
 
Haemoglobin (g%) x 10 
 
                                           
  
Erythrocyte count (mm3) 
 
 
 
Mean Corpuscular Haemoglobin Concentration 
 
(MCHC) % = 
Haemoglobin (g%) x 100
 
Haematocrit (%)
 
 
Statistical Analysis: 
All measurements were performed in 
average of three replicates. Data obtained was analyzed using 
the SPSS/PC+ Statistical package (ver.11.5). Significant 
difference between control and experimen
tal groups were 
determined using Duncan’s test for multiple range 
comparisons
14
. Results were considered as statistically 
significant at 95% confidence level (p.05).
 
 
Results and Discussion
 
In the present study the Physico
-
chemical parameters of water 
su
ch as pH, Dissolved Oxygen, and Alkalinity etc., were 
presented in table
-
1. The exposure of fish, 
Labeo rohita
 
to 
sublethal concentration of Potassium dichromate for 7days and 
30 days caused significant alterations in hematological 
parameters are represent
ed in table
-
2. The results reveal that the 
Red blood corpuscels (RBC), Haemoglobin (Hb), Haematocrit 
(PCV), MCH, and MCHC values were significantly decreased 
after 7days and 30days exposure periods when compared to 
control, being statistically significant 
(p.05). On contrast to 
this, the White blood cell (WBC), and MCV values were found 
statistically significant increase (p.05) in experimental fish 
when compared with control.
 
 
Table
-
1
 
Physico
-
chemical parameters of Tap water used in the 
Laboratory for s
ub lethal tests
 
Parameter
 
Values
 
pH
 
7.6±0.1
 
Temperature
 
27±2ºC
 
Dissolved Oxygen
 
6.52±0.2 mg/lit
 
Alkalinity
 
32.5±1.5 mg/lit
 
Turbidity
 
0.22±0.02 mg/lit
 
Salinity
 
0.25±0.05 mg/lit
 
Free Carbondioxide
 
2.35±0.04 mg/lit
 
 
The fishes exposed to sublethal con
centrations of heavy metal 
chromium in the present investigation showed remarkable 
hematological alterations.  Hematology is used as an index of 
fish health status in number of fish species to detect different 
stress conditions like diseases, hypoxia, 
and 
exposure
 
to metals 
and pollutants etc.
15,16
. The statistically significant decrease 
(p.05) is not uncommon in fish exposed to sublethal 
concentrations of metals, toxicants and therapeutic agents. The 
general reduction in blood parameters is an indication
 
of 
anaemia. 
 
 
The heavy metal induced significant decrease in RBC, Hb and 
PCV. The RBC count coupled with low haemoglobin content 
may be due to destructive action of pollutants on erythrocytes. 
Our results are in good agreement with the earlier works 
repo
rted
17,18
. The decrease in haemoglobin concentration 
indicates the fish inability to provide sufficient oxygen to the 
tissues
19
. In support to our present work, 
it
 
is found that there 
was a decline in the values of RBC, Hb and PCV in fishes 
exposed to comb
ined metal solution
20
. Significant decrease in 
Research Journal of 
Animal, Veterinary and Fishery Sciences
 
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ISSN 2320
 
–
 
6535
 
 
Vol. 
1
(
6
), 
1
-
5
, 
July
 
(201
3
)
          
 
         
Res. J. Animal, Veterinary 
and
 
Fishery
 
Sci
.
 
  
 
International Science Congress Association
 
 
3
 
the RBC, Hb and PCV of fishes exposed to heavy metals was 
also noticed
20,21
. A specific toxic effect on fish blood and tissues 
occurs due to various heavy metals and toxins which enters the 
aquatic environment
. Prolonged reduction in haemoglobin 
content is deleterious to oxygen transport and any blood 
dyscrasia and degeneration of the erythrocytes could be ascribed 
as pathological conditions in fishes exposed to toxicants
22
. Our 
results are in good concurrence 
with the earlier works of
 
Buckley 
et al
., and Palanisamy 
et al
.
23,
24
. The alterations in 
these hematological indices may be due to a defence reaction 
against toxicity through the stimulation of erythropoises. 
 
 
The anemic conditions in fish may be detected
 
using 
haematocrit 
25
. The PCV values always decrease when a fish 
loses appetite or become diseased or stressed. At present, the 
distinct decrease in the level of Haemoglobin and PCV after 
exposure to heavy metal chromium clearly suggests a 
haemodilution m
echanism possibly due to gill damage or 
impaired osmoregulation. The haemodilution has been 
interpreted as a mechanism that reduces the concentration of the 
irritating factor in the circulatory system
26
. Our result are in line 
with 
Smit 
et al.
27
 
that heavy
 
metal exposure results in the 
decrease in  RBC count, Hb and PCV values are due to the 
impaired intestinal absorption of iron. The increase in WBC in 
the present study has been attributed to several factors like 
increase in thrombocytes, lymphocytes or sq
ueezing of WBC’s 
in peripheral blood. Increase in WBC count can be correlated 
with an increase in antibody production which helps in survival 
and  recovery of fishes exposed to toxicant. High WBC counts 
indicate damage due to infection of body tissues, sev
ere 
physical stress as well as Leukemia. Similar increase was 
reported by
 
Banerjee and Banerjee
28
 
in Channa punctatus due to 
Copper sulphate and Potassium dichromate induced toxicity
29,30
 
in Channa punctatus exposed to Copper.
 
 
 
The erythrocyte constants M
CV, MCH, and MCHC offer 
relationship on size, form and Hb constants of erythrocytes. 
They allow the determination of morphological anaemia that 
whether Normocyte, Macrocyte or Microcytic anaemia. The 
alterations in the haematological indices i.e. increase 
in MCV 
and decrease of MCH and MCHC in the present study may be 
due to a defense against the toxic effect of chromium and in turn 
due to decrease in RBC’s, Hb and PCV and the disturbances 
occurred both in metabolic and haemopoitic activities in fish. 
Incre
ase in MCV and WBC count suggests that the anemia is of 
macrocytic type
31
. The MCV gives an indication of the status of 
size of Red blood cell and 
reflects an
 
abnormal or normal cell 
division during erythropoises. The increase in MCV may be 
attributed to t
he swelling of erythrocytes as a result of Hypoxic 
condition or Osmotic stress or Macrocytic anaemia in fishes 
exposed to metal pollution
32. 
Our results are in line with the 
findings of
 
Larsson 
et al.
33
. The decrease in MCH and MCHC 
in the present study cl
early indicates that the concentration of 
hemoglobin in RBC is reduced. The MCH is a good indicatior 
of Red Blood Cell swelling
 
34
. The significant decrease in the 
MCHC values in the present study may be due to swelling of 
RBC or decrease in hemoglobin syn
thesis. The present results 
are in line with the previous findings of
 
Ovie Kori
-
Siakpere
 
35
.
 
 
 
 
Table
-
2
 
Haematological parameters of control and experimental fresh water fish,  
Labeo rohita
 
exposed to Tannery effluent, 
Chromium
 
 
Haematological parameters
 
 
Control
 
 
Experimental
 
 
7 days
 
 
30 days
 
RBC (cumm)
 
1.27±0.012
 
1.22±0.009*
 
(
-
3.937)
 
1.08±0.008*
 
(
-
14.961)
 
WBC (cumm)
 
6.18±0.011
 
7.63±0.007*
 
(+23.463)
 
9.47±0.008*
 
(+53.236)
 
Hb (%)
 
4.2±0.063
 
3.9±0.105*
 
(
-
7.143)
 
3.0±0.052*
 
(
-
28.571)
 
PCV (%)
 
10.7±0.075
 
1
0.3±0.089*
 
(
-
3.738)
 
9.2±0.105*
 
(
-
14.019)
 
MCV (cuµ)
 
84.38±0.010
 
85.08±0.073*
 
(+0.829)
 
86.11±0.008*
 
(+2.050)
 
MCH (pg)
 
32.81±0.008
 
32.23±0.009*
 
(
-
1.768)
 
27.78±0.012*
 
(
-
15.331)
 
MCHC (%)
 
38.88±0.011
 
37.87±0.008*
 
(
-
2.598)
 
32.25±0.009*
 
(
-
17.052)
 
Values are ex
pressed as Mean ± SD (n=6), * = Significant at p.05 level
,
 
Values in Parenthesis indicates percent change over 
control. 
 
 
Research Journal of 
Animal, Veterinary and Fishery Sciences
 
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_
_
_______________________________
___
_
 
ISSN 2320
 
–
 
6535
 
 
Vol. 
1
(
6
), 
1
-
5
, 
July
 
(201
3
)
          
 
         
Res. J. Animal, Veterinary 
and
 
Fishery
 
Sci
.
 
  
 
International Science Congress Association
 
 
4
 
Conclusion
 
 
Thus it is concluded that the present study clearly indicates that 
chromium, a toxic heavy metal discharge via effluent
s into 
aquatic environments caused severe anemia and alterations in 
hematological indices in the fresh water fish, 
Labeo rohita
.
 
 
Acknowledgement
 
One of the authors (MP) 
is
 
grateful to the UGC, New Delhi for 
providing financial assistance through RGNF.
 
 
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Research Journal of 
Animal, Veterinary and Fishery Sciences
 
___
_
_
_______________________________
___
_
 
ISSN 2320
 
–
 
6535
 
 
Vol. 
1
(
6
), 
1
-
5
, 
July
 
(201
3
)
          
 
         
Res. J. Animal, Veterinary 
and
 
Fishery
 
Sci
.
 
  
 
International Science Congress Association
 
 
5
 
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