1. QUINALPHOS INDUCED BIOCHEMICAL
AND PATHOPHYSIOLOGICAL CHANGES
IN FRESHWATER EXOTIC CARP,
CYPRINUS CARPIO (LINNAEUS)
By
Mr. Sameer Gopal Chebbi
Under the Guidance of
Lt. Dr. M. David
Professor
Department of Zoology
Karnatak University
Dharwad

2. Introduction



Water resources have been the most
exploited natural system since man stepped
the earth.
On the one hand rapid population growth,
increasing living standards, wide spheres of
human activities and industrialization have
resulted in greater demand of good quality
water while on the other, pollution of water
resources is increasing steadily.
Water pollution is one of the main concerns
of the world today.

3. 

The pollution of rivers, canals, and lakes with
chemical substances of anthropogenic origin
may have adverse consequences, the waters
become unsuitable for drinking and other
household purposes, irrigation, and fish
cultivation, and also the animal communities
living in them may suffer seriously (Koprucu
and Aydln, 2004; Ural and Saglam, 2005).
The increasing use of synthetic pesticides is
intensifying worldwide pollution risks.

4. 

More than 50% of India’s economy is
dependent upon agriculture and India’s
agriculture has to go a long way to match the
yield of the advanced countries.
Indiscriminate discharge of waste by
agriculture has variety of problems including
those owing to pesticide and other exotic
chemicals that are unfavorable to all life forms
(Nagaraju, et al., 2011).

5. 


PESTICIDE POLLUTION
Pesticides are toxic and designed to repel or
kill unwanted organisms, and when applied to
the land they may be washed into surface
waters and kill or, at least adversely influence,
the life of aquatic organisms (El-Sayed et al.,
2009).
Dependence on pesticides for pest control has
been increasing since the onset of the green
revolution.

6. Many of the more than 1000 pesticides
currently used in most of the countries of the
world inadvertently reach aquatic
ecosystems.
 All pesticides are potentially toxic to living
organisms.
 Pesticide use, therefore, is one of the many
factors contributing to the decline of fish
and other aquatic species in tropical areas
(Helfrich et al., 1996).


7. 

Many pesticides can be grouped into
chemical families. Prominent insecticide
families
include
organochlorines,
organophosphates, and carbamates.
Among different classes of pesticides,
organophosphates are more frequently used,
because of their high insecticidal property,
low mammalian toxicity, less persistence
and
rapid
biodegradability
in
the
environment (Singh et al., 2010).

8. 


Organochlorine toxicities vary greatly, but they
have been phased out because of their persistence
and potential to bioaccumulate. Organophosphate
and carbamates largely replaced organochlorines.
Both operate through inhibiting the enzyme
acetylcholinesterase, allowing acetylcholine to
transfer nerve impulses indefinitely and causing a
variety of symptoms such as weakness or paralysis.
Organophosphates are quite toxic to vertebrates,
and have in some cases been replaced by less toxic
carbamates (Kamrin, 1997).

9. 


Organophosphate pesticides are irreversibly
inactivate acetylcholinesterase, which is
essential to nerve function in insects,
humans, and many other animals.
Organophosphate pesticides affect this
enzyme in varied ways, and thus in their
potential for poisoning.
Although organophosphates degrade faster
than the organochlorides, they have greater
acute toxicity, posing risks to people who
may be exposed to large amounts.

10. 


It is estimated that only 0.1% of the insecticide
reaches the specific target (Aguiar, 2002) or often
less, leaving 99.9% as an unintended pollutant in the
environment, including in the soil, air, and water, or
on nearby vegetation (Pimentel, 1995).
The aquatic environment is the ultimate sink for all
anthropogenic chemicals and global pollutants. Any
compound that has been used in large quantities
ultimately reaches the aquatic ecosystem (Zitko,
1974).
The size and nature of the water body and the extent
of possible dilution influence the level of
accumulation of residue by organisms.

11. 


SCOPE FOR THE PRESENT STUDY:
Fish is a vital source of food for people. It is man’s
most important single source of high-quality
protein, providing ~16% of the animal protein
consumed by the world’s population, according to
the Food and Agriculture Organisation of the
United Nations (2000).
The FAO estimates that about one billion people
world-wide rely on fish as their primary source of
animal protein (FAO, 2000). Fish is a food of
excellent nutritional value, providing high quality
protein and a wide variety of vitamins and
minerals.

12. 

Fish species are most sensitive to aquatic
pollutants during their early life stages
(Jiraungkoorskul et al., 2002). The indiscriminate
use of pesticides, careless handling, accidental
spillage or discharge of untreated effluents into
natural waterways have harmful effects on fish
population and other forms of aquatic life and may
contribute long term effects in the environment.
Fish communities are often used as an indicator of
high water quality areas, as these individuals have
extremely specific water quality requirements
(Bauer and Ralph, 2001).

13. 

The evaluation and assessment of the
ecotoxicological risks caused by pesticides
are based on the toxicity and effects of
pesticide
preparations
to
non-target
organisms like fish, on which exert a wide
range of effects (Velisek, et al., 2008).
With increased interest from public and
scientific communities in man’s use, abuse
and misuse of the aquatic environment, a
diverse array of bioassay methods have
evolved..

14. 


Quinalphos
(O,
O-diethyl-O-quinoxalin-2-yl
phosphorothioate) is one of the most widely used
organophosphorus insecticides in agriculture, due
to its acaricidal and an insecticidal property is in
large scale use in this country. Quinalphos
formulations are widely used as contact and
systematic insecticide against broad range of
insects.
Quinalphos effectively controls caterpillars on fruit
trees, cotton, vegetables and peanuts; scale insect
on fruit trees and pest complex on rice.
Quinalphos also controls aphids, bollworms,
borers, leafhoppers, mites, thrips, etc. on vines,
ornamentals, potatoes, soya beans, rice, tea, coffee,
cocoa, and other crops.

15. 

Among exotic carps Cyprinus carpio, is
widely cultured in ponds and lakes of this
region. This fish is largely preferred for table
purpose by the people because of its low
cost proteins and vitamins. This fish being
specifically a bottom feeder in habit is
widely considered for composite fish culture.
As the water bodies are the ultimate
recipient of all the toxic chemicals and
wastes emitted from industries, agricultural
practices, household application, forest
spraying, etc aquatic organisms especially
the fishes becomes the target.

16. 

Hence, present study focus on the
toxicological impact of organophosphate
pesticide, quinalphos on the freshwater
exotic carp, Cyprinus carpio.
The objective of the present investigation is
to understand the toxic effects lethal and
sublethal concentration of quinalphos (EC
25%) on freshwater exotic carp Cyprinus
carpio with following objectives.

17. Chapter 1: Studies on Toxicity Evaluation
 Chapter 2: Studies on Behavioral changes
 Chapter 3: Studies on Respiratory distress
 Chapter 4: Studies on Quinalphos
Accumulation
 Chapter 5: Ions and associated ATPases
 Chapter 6: Studies on Histopathology
 Chapter 7: Study on Metabolic and
Enzymological aspects
 Chapter 8: Studies on Haematology


18. Aim of Study
QUINALPHOS INDUCED BIOCHEMICAL AND PATHOPHYSIOLOGICAL
CHANGES IN FRESHWATER EXOTIC CARP, CYPRINUS CARPIO (LINNAEUS)
Test Animal:
CYPRINUS CARPIO
Test Toxicant:
QUINALPHOS
(OP)
Dose: Toxicity test
LC50
Subacute Dose: 0.75 μl/l
Exposure Periods:
1, 5, 10 and 15 days
Acute Dose: 7.5 μl/l
Exposure Periods:
1, 2, 3 and 4 days
Study on
Biochemical
Parameters:
Proteins,
Oxidative stress
and AchE, Ions
and ATPase
Tissue selected:
Gill, Kidney and
Liver
Studies on Respiratory distress
Studies on Accumulation
Result
Discussion
Conclusion
Study on
Pathological
Parameters:
Histology and
Haematology

19. 

Toxicant:
The commercial grade organophosphate
insecticide, quinalphos (25% emulsified
concentration) is procured from local market as
Quinalphos 25% EC (VAZRA-25).

20. 


Quinalphos
(O,
O-diethyl-O-quinoxalin-2-yl
phosphorothioate) is one of the most widely used
organophosphorus insecticides in agriculture, due
to its acaricidal and an insecticidal property is in
large scale use in the country.
Quinalphos effectively controls caterpillars on fruit
trees, cotton, vegetables and peanuts; scale insect
on fruit trees and pest complex on rice.
Quinalphos also controls aphids, bollworms,
borers, leafhoppers, mites, thrips, etc. potatoes,
soya beans, rice, tea, coffee, cocoa, and other
crops.

21. 














Some chemical and physical properties of quinalphos.
Chemical Structure:
Molecular formula: C12H15N2O3PS
Molecular Weight: 298.3
Common Name: Quinalphos
Common trade names: Vazra
IUPAC name: O,O-diethyl O-quinoxalin-2-yl phosphorothioate
Formulation in solvent: 2-methoxy and 2-ethoxyethnol
CAS chemical name: O,O-diethyl O-2-quinoxalinyl phosphorothioate
Melting point: 31 – 32oC
Boiling point: 107 oC at 0.05mmHg, 86 oC at 0.01 mmHg
Solubility in water (21 oC) up to 17.8 gm/L
Half- Life in aqueous media, at pH 2-7, relatively stable.
Relative molecular mass: 298.25
Volatility: 1.235 mg/m3

22. The consumption of quinalphos during the year 2007 – 2010 in
Dharwad district (Karnataka) is presented below in terms of liters
used to control different pests.
Name of the
Pesticides
Year
Quantity (in
liters)
Quinalphos
2007
1,380
Quinalphos
2008
1,650
Quinalphos
2009
2,220
Quinalphos
2010
4,310
Source: Joint Director of Agriculture, Dharwad, Karnataka, India.

23. 
Test Toxicant: Quinalphos

24. 




Biology of fish Cyprinus carpio:
Body elongated and somewhat compressed. Lips
thick. Two pairs of barbels at angle of mouth,
shorter ones on the upper lip.
Dorsal fin base long with 17-22 branched rays and
a strong, toothed spine in front; dorsal fin outline
concave anteriorly. Anal fin with 6-7 soft rays;
posterior edge of 3rd dorsal and anal fin spines
with sharp spinules.
Lateral line with 32 to 38 scales. Pharyngeal teeth
5:5, teeth with flattened crowns. Colour variable,
wild carp are brownish-green on the back and
upper sides, shading to golden yellow ventrally.
The fins are dusky, ventrally with a reddish tinge.
Golden carp are bred for ornamental purposes.

25. 
Test Animal : Cyprinus carpio

26. Classification: Systematic position of Cyprinus carpio:










Phylum
Sub-Phylum
Division
Super Class
Class
Sub Class
Super order
Order
Genus
Species
:
:
:
:
:
:
:
:
:
:
Chordata
Vertebrata
Gnathostomata
Pisces
Osteichthyes
Actinopterygii
Teleostei
Cypriniformes
Cyprinus
carpio

27. 



Procurement and maintenance of fish:
Fish, Cyprinus carpio weighing 4 ± 2 g and
measuring an average length of 5 ± 2 cm were
collected from the State Fisheries Department,
Dharwad, and maintained in large cement tank
previously washed with potassium permanganate
solution.
The water was aerated twice a day so as to provide
sufficient oxygen. The fish were fed daily with
commercial fish pellets (40% protein content)
procured from market.
They were acclimated to laboratory conditions for
fifteen days. The tanks were cleaned periodically to
avoid infection to fish. The temperature of the
water in the aquaria was 29 ± 1oC and the same was
maintained throughout the course of investigation.

28. 
Experimental Design:

Five groups of ten fish each were exposed to
lethal and sub lethal concentrations (1/10th
of the LC50) of quinalphos.

Similarly one such group of thirty fish was
taken as control.

29. 


Fixation of Exposure Periods:
The effect of the lethal and sub lethal
concentrations of quinalphos on fishes were
studied at different periods of exposure in
order to understand the influence of time
over toxicity.
Thus in lethal 1, 2, 3 and 4 days and in the
sub lethal 1, 5, 10 and 15 days were chosen to
observe the short term and long term acute
and chronic effects of quinalphos on fish,
Cyprinus carpio, respectively.

30. 

Fixation of Lethal and Sub Lethal
Concentrations:
The LC50 96 h of quinalphos concentration
was taken is fixed as lethal concentration
(7.5 μl/l) and sub lethal concentration
(1/10th of the LC50 i.e., 0.75 μl/l) to study
the
physiological,
biochemical,
haematological and histological responses of
fish.

31. 
Tissue selected:

Biochemical Parameters: Gill, kidney and
liver

Histopathology: Gill, kidney and liver

32. 


















Methods followed
Parameters
Methods
Toxicity evaluation : Finney, 1971 and Carpenter, 1975
Behavioural studies : David et al., 2008
Estimation of AChE activity: Ellman et al., 1961
Studies on Whole animal oxygen consumption: Welsh and Smith, 1953
Quinalphos bioaccumulation : Rao et al., 2003
Catalase activity: Luck et al., 1974
Protease activity: Davis and Smith, 1955
Malondialdehyde (MDA) Placer et al., 1966,
Free amino acids Moore and Stein, 1954
Protein Lowry et al., 1951
Estimation of Ca2+, K+, Mg2+ and Na+ ions Dall, 1967
Ca2+, Mg2+ and Na+/K+ ATPases activities Watson and Beamish,
1981
RBC count Donald and Henry, 1969
WBC count Donald Hunter and Bomford, 1963
Haemoglobin Dacie and Lewis, 1961
Histopathology Humason, 1972
Statistical analysis of data Duncan, 1955

33. Chapter 1: Studies on Toxicity Evaluation



The toxicity tests have been historically played an
important role in assessing the effect of human
activities on animals.
These tests have wide applicability in evaluating
the acute toxicity of pesticides to fish.
Teleost fish have proved to be good models to
evaluate the toxicity and effects of contaminants
on animals, since their biochemical responses are
similar to those of mammals and of other
vertebrates (Sancho et al., 2000).

34. 


The LC50 96 hr value for the fish, Cyprinus carpio
was determined after conducting static renewal
bioassay test.
In the present test, acute toxicity is expressed as
the median lethal concentration (LC50) that is the
concentration in water which kills 50% of a test
batch of fish within a continuous period of
exposure (96 h).
One tenth of the LC50 was selected as sub lethal
concentration for sub acute studies (1, 5, 10 and 15
days) to find out quinalphos induced metabolic
changes.

35. 




Toxicity evaluation (RESULT)
Hence the present study is initiated with the determination
of 96 h LC50 value on exposure on Cyprinus carpio to
different concentrations of quinalphos as described by
Finney, 1971 and Carpenter (1975).
96 h exposure is preferred with a view that the effects of the
toxicant on these animals become consistent within this
period (Eisler, 1977).
The dosage response studies conducted for 96 h reveled
that the LC50 value was found to be 7.4 μl/l and 7.6 μl/l
(sigmoid curve/linear curve, Finney method) and 7.7 μl/l
according Dragstedt-Behrens’s method.
Thus, the average 96 h LC50 was found to be 7.5 μl/l.

36. Table 2. Mortality of Cyprinus carpio in different concentrations of
Quinalphos at 96-hour exposure period.
Concentration
of Quinalphos
25 % EC
(µl/L)
5.0
5.5
6.4
6.5
7.0
7.1
7.3
7.5
8.5
Log
concentrati No. of fish No. of
on of
exposed fish alive
quinalphos
0.6989
0.7403
0.8016
0.8129
0.8450
0.8526
0.8678
0.8772
0.9294
10
10
10
10
10
10
10
10
10
10
9
8
7
5
4
2
1
0
No.
Fish
dead
0
1
2
3
5
6
8
9
10
Perce Probit
nt kill
kill
0
10
20
30
50
60
80
90
100
----3.72
4.16
4.48
5.00
5.52
5.84
6.28
8.09

37. 

Toxicity evaluation
LC50 studies are believed to provide
information on the relative lethality of a
toxicant to an organism and establishing
the tolerance limits and safe levels of toxic
agents for the biota of aquatic environment
and also for the evaluation of lethal and
sub lethal concentrations.

38. 



LC50 studies are highly useful in determining the
sublethal concentrations of a particular compound.
Most of the information on the effects of pesticides
in aquatic animals is on short-term experiments
carried out at the lethal concentrations (Tilak and
Swarna Kumari, 2009).
Hence, there is need for the sublethal toxicity
studies, which prove to be of great worth in
evaluating the sequence of events involving in
response of the test animal to the sublethal
concentrations (Sprague, 1971).
So, to derive such sublethal concentrations and to
compare responses of animal at the sublethal
concentration with those of the lethal, LC50s are
prime requisites (Das and Mukherjee, 2000).

39. 

Toxicity of quinalphos to C. carpio is
relatively lower when compared with other
species of fishes.
The 96 hour LC50 value (7.5µ l/L) obtained
in the present study is lower than the values
reported in literature for other species of fish
(Das and Mukherjee, 2000; Tilak and
Swarna Kumari, 2009; Nikam, et al., 2011).

40. 
The results obtained form the present
toxicity study shows that the quinalphos EC
25% was more toxic to the aquatic organism
because of its liquid formulation of technical
quinalphos in the solvent, 2-methoxy and 2ethoxyethnol (Das and Mukherjee, 2000).

41. 
Chapter 2: Studies on Behavioral changes

The pesticide brings tremendous changes in
the organism. Changes in regular
behavioural pattern of the organism link it
with the disturbed physiology.
In the laboratory fish behaviour can be a
sensitive marker of toxicant induced stress.
Hence an attempt has been made to study
physical, morphological and behavioural
changes in quinalphos exposed fish.


42. 


Behavioural study (RESULT)
The behavioural tests are useful in
evaluating the toxicant induced effects on
whole population, small change in
learning, dominance, parental behaviour,
food selection, migration etc.
Further they are excellent tool for quick
screening and for understanding the toxic
effects of the organophosphate pesticide
such as quinalphos.

43. 
The fishes were exposed to the lethal
concentration of quinalphos exhibited the
behavioural changes like excitability, erratic
swimming, jumping, respiratory disruptions
like discomfort movement (S jerks, partial
jerk, fin flicker, burst swimming), mucus
secretion all over the body, precipitation of
mucus on the gills and caudal bending etc.
were observed.

44. 

Whereas in sub lethal concentration such
behavioural changes were negligible.
Suffocation caused by the mucus film on
gills could be one of the reasons for the
death of fish in the lethal concentration of
quinalphos.
Hence an approximate concentration of
pesticide in the ambient medium can be
predicted based on the behavioural response
of the inhabitatiting animals.

45. 



Behavioural characteristics are obviously sensitive
indicators of toxicant effect.
It is necessary, however, to select behavioural indices
for monitoring that relation to the organism’s
behaviour in the field in order to derive a more
accurate assessment of the hazards.
In the present study as evidenced by the results the
abnormal changes in the fish exposed to lethal
concentration of quinalphos are time dependent.
However, the normal behaviour of the fish at 10 and
15 days on exposed to sub lethal concentrations
indicates its adaptability to the sub lethal
concentration due to long term exposure of
quinalphos.

47. 


Chapter 3: Studies on Respiratory distress
A change in respiration rate is one of the
common
physiological
responses
to
toxicants and is easily detectable through
changes in oxygen consumption rate, which
is frequently used to evaluate the changes in
metabolism
under
environmental
deterioration.
Respiration may be defined as an oxidative
process during which food materials
undergo oxidation and get converted into
carbon dioxide, water and energy.

48. 



A reduction in oxygen consumption is observed
when the fish is exposed to the toxicant and the
mortality is due to effect of metabolism of energy
synthesis (Tilak and Swarna Kumari, 2009).
Determination of oxygen consumption of aquatic
animals will undoubtedly provide information on
the effects of interactions of toxicants on the
physiology of aquatic life (Sarkar, 1999).
Oxygen consumption is an important parameter to
assess the toxicological stress, since it serves as
index of energy expanded and speaks of
physiological and metabolic state of an organism.
Generally when toxicants gain entry through food
chain or respiratory surfaces, the physiological
function to be affected is oxygen consumption.

49. 


Oxygen consumption (RESULT)
Generally when a toxicant gains entry
through food chain or respiratory surfaces,
the first physiological function to be
affected is whole animal oxygen
consumption.
In view of the vital role ascribed to the
enzymes involved in the energy pathways
through respiratory process by utilizing
oxygen.

50. 

The study of respiration at whole animal
level needs emphasis to ascertain the oxygen
requirements
of
fish
under
toxic
environmental condition.
In the present investigation the whole
animal oxygen consumption was decreased
under lethal and sub lethal concentrations of
quinalphos
indicating
prevalence
of
respiratory distress due to interference of
quinalphos in oxidative metabolism.

51. 


In the present study, the oxygen consumption was
gradually decreasing with increasing exposure periods as
observed by Mathivanan (2004) in Oreochromis
mossambicus exposed to sublethal concentrations of
quinalphos.
Oxygen consumption is widely considered to be a critical
factor for evaluating the physiological response and a useful
variable for an early warning for monitoring aquatic
organisms (Chinni et al., 2000). Like most fish, common
carp (Cyprinus carpio) are oxygen regulators, i.e., they
maintain their oxygen consumption at a constant level
along a gradient of environmental oxygen concentrations,
until critical oxygen concentration is reached, and below
which oxygen consumption begins to fall.
Under conditions of stress, this critical oxygen
concentration is likely to increase, reflecting the decreased
capacity of the fish to cope with environmental
perturbations.

52. Table 3: Whole animal oxygen consumption (ml/gm wet wt/h) of the fish, Cyprinus carpio
on exposure to the lethal and sub lethal concentrations of quinalphos.
Exposure period in days
Estim
ations
Cont
rol
Mean 0.83 A
Lethal
1
2
Sub lethal
3
4
1
0.0823 0. 0333 0.1646 0.2597 0.1391
E
H
C
B
D
0.001
0.011
0.002
0.003
SD ±
0.004
0.002
%
Chang
e
-----
-90.08 -95.98
5
10
15
0.075
0.0590
0.0288 I
G
8F
0.013
0.012
0.003
-80.16 -68.71 -83.24 -90.86
-96.53
-92.89
Values are Means ± SD (n=6) for oxygen consumption in a column followed by the same letters
are not significantly different (P ≤ 0.05) from each other according to Duncan’s multiple range
(DMR) test.

54. 


Chapter 4: Studies on Quinalphos
Accumulation (residue analysis)
In environmental studies, certain organisms
provide valuable information about chemical
states of their environment not through their
absence or presence but ability to concentrate
environmental toxins within their tissues.
Since fish spend their entire lives in the
aquatic environment and are often found at
higher feeding levels of the aquatic food
chain, they incorporate chemicals from the
environment into their body tissues through
feeding relationships.

55. 

Persistent hydrophobic chemicals may
accumulate in aquatic organisms through
different mechanisms: via the direct uptake
from
water
by
gills
or
skin
(bioconcentration), via uptake of suspended
particles
(ingestion)
and
via
the
consumption
of
contaminated
food
(biomagnification).
Parameters affecting these processes include
lipid and water solubility, degree of
ionization, chemical stability and molecular
size.

56. 
In the present study an attempt has been
made to determine the residue levels of
quinalphos in different tissues viz., gill,
muscle, and liver of fish Cyprinus carpio at
different exposure periods under median
lethal
and
sublethal
concentrations
employing High Performance Liquid
Chromatographic (HPLC) technique.

57. 


Bio-accumulation (RESULT)
A very important biological property of
pesticide
is
their
tendency
to
bioaccumulate. A significant amount of
quinalphos is accumulated in the organs of
the fish in both lethal and sub lethal
concentrations of exposure.
Relative to control a significant increase in
the concentration of quinalphos is
observed in the organs of the fish.

58. 

However this increase is predominantly
more in lethal concentration and sub lethal
concentration. Further, it appeared that the
amount of quinalphos accumulated in the
fish is time dependent, as it increased with
the increase in the exposure period in both
lethal and sub lethal concentrations.
The rate of accumulation of quinalphos in
the fish exposed to sub lethal concentration
could be due to the activation of the toxicant
bioconcentration and biomagnification
processes.

59. 

Several authors have reported that, pesticide
residue can cause cellular damage to the gill tissue,
as it is the first organ to face pesticide medium
(Kalavathy
et
al.,
2001;
Kanabur
and
Sannadurgappa, 2001, Chanchal et al., 1990
Bashamohideen et al., 1989), which offers support
to the present findings.
Simultaneously, the residues are absorbed by the
aquatic organisms, bioaccumulated in the tropic
chain and deposit in the tissues. Earlier studies
revealed that, the accumulation of pesticides in
fish tissues (David and Philip, 2005; .Tilak. et al.,
2001; Gupta et al., 2001; Sharma, 1994) shows
similar result of present study.

60. Table 4: Accumulation of quinalphos (µg/g wet wt.) in the organs of fish, Cyprinus carpio on exposure to the lethal
and sublethal concentrations of quinalphos.
Exposure period in days
Organ
Lethal
1
Gill
98.21 H
SD ±
0.013
Kidney
78.54 H
SD ±
0.002
Liver
65.91 H
SD ±
0.001
Sub lethal
2
3
4
155.34
312.54
F
D
443.78 C
0.001
0.002
0.011
133.67
198.23
G
E
218.99 C
0.001
0.002
112.23
163.91
F
0.013
1
5
10
15
148.23
257.73
G
E
489.87 B
502.33 A
0.012
0.001
0.003
0.012
136.85
215.33
F
D
281.34 B
376.54 A
0.001
0.001
0.003
0.001
0.011
D
197.92 C
98.79 G
E
203.73 B
261.81 A
0.003
0.004
0.002
0.001
0.001
0.003
156.82
Values are Means ± SD (n=6) for a tissue in a column followed by the same letters are not significantly
different (P ≤ 0.05) from each other according to Duncan’s multiple range (DMR) test.

61. 


Chapter 5: Ions and associated ATPases
Ions play a vital role in several body functions, viz.
the monovalent ions sodium, potassium and
chloride are involved in neuromuscular excitability,
acid base balance and osmotic pressure (Verma et
al., 1981), whereas divalent cations, calcium and
magnesium facilitate neuromuscular excitability,
enzymatic reactions and retention of membrane
permeability.
Alteration in osmotic regulatory mechanism under
toxic conditions may cause severe imbalance in
biochemical composition of the tissue fluids
followed by undesirable metabolic consequences

62. 

In freshwater fishes, blood and electrolyte
concentration are regulated by interacting
processes, such as absorption of electrolytes
from surrounding medium through active
mechanisms predominantly at the gill,
control of water permeability and selective
re-absorption of electrolytes from urine.
Any alteration in one or more of these
processes results in a change in the plasma
electrolyte composition.

63. 


Sodium (Na+) ion plays an important role in the
osmotic regulation of body fluids and also serves
as an essential activating ion for specific enzyme
system.
Potassium ion (K+) is the prominent intracellular
cation of animals. It is an important co-factor in
the regulation of osmotic pressure and acid-base
balance (Sexena, 1957).
Calcium ion (Ca2+) is also important osmotic
effectors and plays significant role in the
regulation of cellular metabolism and is involved
in conferring stability to the cell membrane.

64. 




Adenosine triphosphatase (ATPase) is a membrane bound
enzyme group for regulating oxidative phosphorylation,
ionic transport, muscle function and several other
membrane transport dependent phenomena.
ATPase have the central role in physiological function of
cells as energy transducers by coupling the chemical
reactions of ATP hydrolysis (Laugher, 1987).
Membrane bound Na+- K+ ATPase is the enzymatic
machinery for the active transport of sodium and potassium
across the cell membrane.
Magnesium translocation is dependent on membrane
bound Mg2+ ATPase (Beeler, 1983).
The implication of ATP inhibition as the mechanism of
toxic action by insecticides has been proposed from studies
on the inhibition of ATPases activities by the pesticides
(Henery et al., 1996).

65. 




Results
In this investigation, it is evident that Na+ loss is higher in
the case of gill indicating the derangement in Na+
transport and rupture in the respiratory epithelium of gill
tissue (David et al., 2003).
The decreases in K+ ion content in the tissues of Cyprinus
carpio exposed to quinalphos might attribute to the
derangement in whole animal oxygen consumption and
ionic content at tissue levels as observed in the present
investigation.
The decrease in Ca2+ ion level indicates increased
decalcification. Ca2+ is concerned with neuromuscular
excitability, cell membrane permeability and regulation of
protein binding capacity (Walser, 1960).
In the present study, the restlessness in fish during
organophosphate stress corresponds to structural change in
mitochondrial integrity.

66. 

In the present study, the decrease in the levels of
Na+- K+, Ca2+ ions in the gill, kidney and liver
exposed to lethal and sublethal concentrations of
quinalphos indicates changes in the permeable
properties of the cell membrane of these organs
and of deranged Na+-K+ and Ca2+ ionic pumps
due to the probable consequences of tissue
damage.
The imperative reason for the diminish of sodium,
potassium and calcium ion levels in the organs of
fish, exposed to quinalphos could be attributed to
the suppressed activities of Na+- K+, Mg2+ and
Ca2+ ATPase (Renfro et al., 1974), since ATPases
have been described as prominent energy linked
enzymes in fishes (Desaiah et al., 1975).

67. 

However this change increased over time of
exposure in lethal but decrease in sub lethal
concentration. The results indicted the
changes in the permeable properties of the
cell membrane on exposure to quinalphos.
These decreases in the ionic levels by high
quinalphos concentration could be due to
the consequences of tissue damage and
severe disruption in cellular ionic regulation
of the fish leading the osmo-regulatory
failure.

68. Table 5:
Sodium ion content (µg/g wet wt.) in the tissues of fish, Cyprinus carpio on exposure to the lethal
and sublethal concentrations of quinalphos.
Exposure periods in days
Tissue
Gill
± SD
% Change
Kidney
± SD
% Change
Liver
± SD
% Change
Contro
l
0.5443
Lethal
1
Sublethal
2
3
4
1
5
10
15
0.3324
0.3112
0.3064
0.4343
0.3422
0.3321
0.3395
F
H
I
B
D
G
E
A
0.4211 C
0.002
0.001
0.012
0.013
0.003
0.004
0.012
0.011
0.013
----
-22.63
-38.93
-42.82
-43.70
-20.20
-37.13
-38.98
-37.62
0.4227
0.3835
0.3338
0.2748
0.2443
0.3974
0.3468
0.2948
0.2887
A
C
E
H
I
B
D
F
G
0.003
0.011
0.002
0.003
0.013
0.004
0.011
0.003
0.013
----
-9.27
-21.03
-34.98
-42.21
-5.98
-17.95
-30.25
-31.70
0.5033
0.4960
0.4128
0.3765
0.3284
0.4995
0.4674
0.3877
0.3855
A
C
E
H
I
B
D
G
F
0.002
0.001
0.013
0.004
0.011
0.004
0.012
0.004
0.012
----
-1.45
-17.98
-25.19
-34.75
-3.63
-7.13
-22.96
-23.41
Values are Means ± SD (n=6) for a tissue in a column followed by the same letters are not significantly
different (P ≤ 0.05) from each other according to Duncan’s multiple range (DMR) test.

69. Table 6: Potassium ion content (µg/g wet wt.) in the tissues of fish, Cyprinus carpio on exposure to the
lethal and sublethal concentrations of quinalphos.
Exposure periods in days
Tissue
Contro
l
Lethal
Sublethal
1
2
3
4
1
5
10
15
0.9937
0.9025
0.8737
0.8152
0.7430
0.9832
0.9453
0.8743
0.7998
A
D
F
G
I
B
C
E
H
± SD
0.011
0.001
0.002
0.003
0.013
0.011
0.012
0.001
0.011
% Change
----
-15.12
-23.46
-41.58
-45.71
-7.85
-13.75
-20.55
-11.95
1.0743
0.9833
0.9023
0.8764
0.7289
0.9673
0.9567
0.8346
0.7321
A
B
E
F
I
C
D
G
H
± SD
0.002
0.011
0.002
0.001
0.004
0.001
0.002
0.004
0.012
% Change
----
-8.47
-16.01
-18.42
-32.15
-9.95
-10.94
-22.31
-31.85
0.8837
0.8027
0.7655
0.7121
0.6525
0.8665
0.7964
0.7343
0.6678
A
C
E
G
I
B
D
F
H
± SD
0.003
0.012
0.002
0.004
0.003
0.011
0.003
0.004
0.013
% Change
----
-9.16
-13.37
-19.41
-26.16
-1.94
-9.87
-16.91
-24.43
Gill
Kidney
Liver
Values are Means ± SD (n=6) for a tissue in a column followed by the same letters are not
significantly different (P ≤ 0.05) from each other according to Duncan’s multiple range (DMR) test.

70. Table 7:
Calcium ion content (µg/g wet wt.) in the tissues of fish, Cyprinus carpio on exposure to the lethal
and sublethal concentrations of quinalphos.
Exposure periods in days
Tissue
Gill
± SD
% Change
Kidney
± SD
% Change
Liver
± SD
% Change
Contro
l
Lethal
Sublethal
1
2
3
4
1
5
10
15
0.8627
0.7932
0.7457
0.7217
A
D
E
H
0.7035 I
0.8112 B
0.7976 C
0.7523
0.7398
F
G
0.001
0.003
0.011
0.002
0.001
0.012
0.004
0.003
0.013
----
-8.05
-13.56
-16.34
-18.45
-5.96
-7.54
-12.79
-14.24
1.2232
1.0538
0.9425
0.9228
1.1123 B
0.9758 D
0.9566
0.8985
G
0.8847 I
A
C
F
E
H
0.011
0.001
0.003
0.002
0.013
0.004
0.002
0.003
0.012
----
-13.84
-22.94
-24.55
-27.67
-9.06
-20.22
-21.79
-26.54
0.9128
0.8835
0.8663
0.7968
E
G
0.8967 B
0.8856 C
0.7769
D
0.7062 I
0.8563
A
F
H
0.004
0.003
0.011
0.002
0.003
0.004
0.013
0.012
0.001
----
-3.21
-5.09
-12.71
-22.63
-1.76
-2.97
-6.18
-14.88
Values are Means ± SD (n=6) for a tissue in a column followed by the same letters are not significantly
different (P ≤ 0.05) from each other according to Duncan’s multiple range (DMR) test.

71. Table 8: Na+-K+ ATPase activity (µM of Pi formed / mg protein / h) in the organs of fish, Cyprinus carpio on
exposure to the lethal and sub lethal concentrations of quinalphos.
Exposure period in days
Organ
Gill
SD ±
% Change
Kidney
SD ±
% Change
Liver
SD ±
% Change
Contro
l
Lethal
1
3.7512 A 3.3112 C
Sub lethal
2
3
4
1
5
10
15
3.0245
2.9137
2.0621
3.4185
3.2233
2.7823
3.0579
D
G
I
B
E
H
F
0.003
0.012
0.004
0.003
0.001
0.002
0.004
0.011
0.003
------
-11.64
-19.34
-22.52
-44.88
-9.11
-19.43
-25.69
-18.63
3.2234
2.8544
2.0653
1.1647
0.9853
2.9785
2.6355
A
C
G
H
I
B
E
2.1866 F
0.002
0.001
0.003
0.002
0.004
0.011
0.012
0.004
0.002
-----
-11.44
-35.92
-63.86
-69.43
-7.59
-18.23
-32.16
-16.65
3.5495
3.0344
2.5755
2.2785
2.6507
2.1756
2.7864
B
F
G
1.5893 I
2.9709
A
C
E
H
D
0.001
0.002
0.003
0.004
0.011
0.013
0.012
0.002
0.004
------
-14.51
-27.44
-35.81
-55.22
-16.30
-25.32
-38.71
-21.49
Values are Means ± SD (n=6) for a tissue in a column followed by the same letters are
not significantly different (P ≤ 0.05) from each other according to Duncan’s multiple
range (DMR) test.
2.6866
D

72. Table 9: Mg2+ ATPase activity (µM of Pi formed / mg protein / h) in the organs of fish, Cyprinus carpio on
exposure to the lethal and sublethal concentrations of quinalphos.
Exposure period in days
Organ
Gill
SD ±
% Change
Kidney
SD ±
% Change
Liver
Contro
l
Lethal
Sub lethal
1
2
3
1
5
10
15
3.2233
2.6553
2.0656
1.7785
A
D
G
H
0.9365 I
2.8577
2.3754
2.4743
2.7873
B
F
E
C
0.004
0.002
0.003
0.011
0.002
0.012
0.004
0.011
0.001
-------
-17.62
-35.91
-44.82
-70.94
-11.34
-26.31
-23.23
-13.52
3.5434
3.1112
D
2.5754
1.4865
1.2668
3.3843
2.6987
2.2378
3.3874
F
G
H
C
D
F
B
0.011
0.002
0.004
0.012
0.004
0.001
0.002
0.013
0.003
------
-12.19
-27.31
-58.04
-64.24
-4.49
-23.83
-36.84
-4.41
3.8745
2.6873
2.4836
1.4884
3.6775
3.2862
1.8833
A
D
E
H
B
C
G
A
4
0.1896 I
2.3885 F
0.003
0.002
0.004
0.011
0.012
0.003
0.002
0.001
0.003
SD ±
% are Means ------30.64
-35.89
-61.58
-95.11
-5.08
-15.18
-51.39
Values Change ± SD (n=6) for a tissue in a column followed by the same letters are not significantly -38.35
different (P ≤ 0.05) from each other according to Duncan’s multiple range (DMR) test.

73. Table 10: Ca2+ ATPase activity (µM of Pi formed / mg protein / h) in the organs of fish, Cyprinus carpio on exposure
to the lethal and sublethal concentrations of quinalphos.
Exposure period in days
Organ
Gill
SD ±
% Change
Kidney
SD ±
% Change
Liver
SD ±
% Change
Contro
l
Lethal
Sub lethal
1
2
3
4
1
5
10
15
3.9687
3.5646
3.3881
2.0965
1.2793
3.6945
3.2899
2.0455
3.6235
A
D
E
G
I
B
F
H
C
0.012
0.013
0.001
0.003
0.014
0.013
0.014
0.013
0.011
------
-10.18
-14.62
-47.17
-67.76
-6.91
-17.11
-48.45
-8.69
3.2676
2.2443
1.9677
1.7894
1.1673
2.8673
2.4879
2.2853
2.9878
A
F
G
H
I
C
D
E
B
0.022
0.011
0.012
0.015
0.023
0.014
0.013
0.014
0.012
------
-31.31
-39.78
-45.23
-64.27
-12.25
-23.86
-30.06
-8.56
2.4784
2.2894
2.1984
1.7783
1.5835
2.2569
1.9963
1.8343
2.3785
A
C
E
H
I
D
F
G
B
0.001
0.013
0.011
0.014
0.012
0.014
0.011
0.002
0.014
-----
-7.62
-11.29
-28.24
-36.10
-8.93
-19.45
-25.98
-4.03
Values are Means ± SD (n=6) for a tissue in a column followed by the same letters are not significantly
different (P ≤ 0.05) from each other according to Duncan’s multiple range (DMR) test.

75. 




Chapter 6: Studies on Histopathology
Histopathology is mainly directed to study the effect
of chemicals and pesticides on the structural
components of the living system and the ways in
which cells and tissues respond to injury.
As an indicator of exposure to pollutants, histology
represents a useful tool to assess the degree of
pollution, particularly for lethal and chronic effects.
The severity of histological damages in any
particular aquatic organism is directly proportional
to the concentration of a pollutant in the medium.
Gill, kidney and liver (Bucher and Hofer, 1993) are
suitable organs for histological examination to
determine the effect of pollution.

76. 



Histopathological study (RESULT)
Gill
Gills are the vital organs for respiration in
fish which establish a direct contact with
the medium through which a pollutant
largely enters into the body.
The gills of fish are the main target organs
for toxic action of chemical pollutants, as
well as for detoxification process.

77. 



Fish on exposure to the lethal concentration on day 1, the
enlargement of the base of primary gill lamellae was
observed.
On day second lamellar oedema and secondary gill lamellae
clubbing at the distal end was seen leading towards
telangiectatic secondary lamellae.
On day 3rd lamellar telangiectasis was observed, which
continued up to day 4. Lamellar hypertrophy and lamellar
hyperplasia was observed. Fusion of these lamellae was
noticed all along their length on day 4.
The cells seemed to have undergone a clear increased
hyperplasia further desquamation of the hyperplasic
epithelium and capillaries with loss of the lamellar structure
of the gill were seen all along the gill filaments.

78. H&E:X 100 and
X200
24 hr exposed gill to lethal
dose of quinalphos
Control Fish Gill
48 hr exposed gill to lethal
dose of quinalphos

79. H&E:X 100 and
X200
72 hr exposed gill to lethal
dose of quinalphos
Control Fish Gill
96 hr exposed gill to lethal
dose of quinalphos

80. 


In the sublethal concentration of quinalphos a mild
degree of degenerative changes and sign of shrinkage in
the primary gill lamellae was observed at day 1
exposure. The slight damage to the base of secondary
lamellae and inter lamellar tissue was observed.
On day 5 the changes were more marked when
compared to day 1. The degeneration of epithelial cells
encapsulating primary and secondary gill lamellae with
necrosis. Fusion of secondary lamellar, bubbling of
primary gill lamellae, atrophy is also observed. The
excess secretion of mucus and necrosis of basal
filament was observed.
But on further exposures, for 10 and 15 days the gill
structure was just similar to that of control fish except
mild degree of precipitation of mucus over the gill
lamellae.

81. H&E:X 100 and
X200
Day 1st exposed gill to
lethal dose of quinalphos
Control Fish Gill
Day 5th exposed gill to
lethal dose of quinalphos

82. H&E:X 100 and
X200
Day 10 exposed gill to
lethal dose of quinalphos
th
Control Fish Gill
Day 15th exposed gill to
lethal dose of quinalphos

83. 


Liver
Fish liver is regarded as a major site of
storage, biotransformation and excretion of
pesticides. These organs have been proven
to be indicative of pollution.
The liver has the ability to degrade toxic
compounds, but its regulating mechanisms
can
be
overwhelmed
by
elevated
concentrations of these compounds, and
could subsequently result in structural
damage.

84. 



On day 1 of exposure to the lethal concentration of
quinalphos, the liver of fish exhibited enlarged nuclei and
vacuolization in hepatic cells. Liver cords were seen
disarrayed.
On day 2 of exposure, the parenchymatous nature of the
liver was greatly disrupted with congested blood vessels.
The hepatocyte cell membranes were ruptured and
granular degeneration was evident in most of the
hepatocytes. Nuclei became slightly hypertrophic.
Further on day 3 severe degrees of atrophic changes were
noticed in the liver cords. Hemorrhagic condition was
prominent with heavy vacuolization in the liver tissue. At
some regions exfoliation and congregation of hepatocytic
nuclei and focal necrosis were seen.
This was followed by the severe degree of vacuolization,
shrinkage
of
hepatocytes,
atrophy,
cytoplasmic
degeneration, rupture of blood vessels, diffused necrosis,
dissolution of laminar structure and cytoplasmic
disintegration in hepatocytes on day 4 of exposure.

85. H&E:X 200
Control Fish
and X400
Liver
24 hr exposed fish liver to
lethal dose of quinalphos
48 hr exposed fish liver l to
lethal dose of quinalphos

86. H&E:X 200
Control Fish
and X400
Liver
72 hr exposed fish liver to
lethal dose of quinalphos
96 hr exposed fish liver l to
lethal dose of quinalphos

87. 


Compared to the structure of the liver of control fish,
exposed to sublethal concentration of quinalphos
initially exhibited few changes like slight disarray of
liver lobes, mild degree of degeneration of cytoplasm,
occasional blood clots and congregation of nuclei at day
1 and cloudy swelling of hepatocytes, granulization of
cytoplasm, hypertrophic and pyknotic nuclei on day 5.
However, on further exposure to day 10 certain degree
of reorganization in the structure of liver cords was
observed. The nuclei appeared normal, with a very little
degree of cytoplasmic vacuolization.
At 15 days of exposure, no significant changes were
seen different from controls, except a slight degree of
hyperchromatic condition of the nuclei.

88. H&E:X 200
Control Fish
and X400
Liver
Day 1st exposed fish liver to
lethal dose of quinalphos
Day 5th exposed fish liver to
lethal dose of quinalphos

89. H&E:X 200
Control Fish
and X400
Liver
Day 10th exposed fish liver to lethal
dose of quinalphos
Day 15th exposed fish liver to
lethal dose of quinalphos

90. 


Kidney
In fish, the kidney performs an important
function related to electrolyte and water
balance and the maintenance of a stable
internal environment.
The kidney of fish receives much the largest
proportion of post-branchial blood, and
therefore renal lesions might be expected to
be good indicators of environmental
pollution.

91. 



In lethal concentration of quinalphos the kidney showed
reduction in renal cell number in the proximal and distal
collecting tubules, which have resulted in narrowness of
lumen. On day 1 the tubular cells have undergone
hypertrophy and some of the renal tubules have lost their
normal shape.
On day 2 Vacuolization due to degeneration of cytoplasm
is quite obvious. The nuclei of epithelial cells have become
quite dominant and are found infiltrating into the
surrounding tissue. The perforation of kidney tubules is
commonly observed.
On day 3 the kidney demonstrated hyperplasia,
vacuolization, degeneration and necrosis leading to the
complete necrosis. Cubiodal epithelial cells lining the
tubules showed complete vacuolization with degenerating
cytoplasm and more nuclear division and their disorderly
scattering nature.
The hemopoietic tissue was fully studded with lymphatic
cells at the highest rate of nuclear division. The lumen of
the tubules was found to be dilated. Kidney tubules were
also found to be perforated on day 4.

92. H&E:X 200
Control Fish
and X400
Kidney
24 hr exposed kidney to lethal
dose of quinalphos
48 hr exposed kidney to lethal
dose of quinalphos

93. H&E:X 200
Control Fish
and X400
Kidney
72 hr exposed kidney to lethal
dose of quinalphos
96 hr exposed kidney to lethal
dose of quinalphos

94. 




In sublethal concentration of quinalphos the kidney of the fish
exhibited a mild degree of changes.
1st, 5th and 10th day of shows more changes i.e., epithelial cells
of the tubules which showed desquamation, irregular orientation
of the nuclei in the cells, lumen of the tubules became wider as a
result of flattening of epithelial cells.
Ruptures of the tubules were quite prominent, cell fragments
could be seen inside the lumen of some tubules.
Vacuolar degeneration was seen in the few tubules. Hemopoeitic
tissue was degenerated. But on 15th day kidney showed recovery
tendency. Glomerular cells attained normalcy in structure.
Cytoplasm appeared clear and vacuolization and karyolysis of cell
was completely reduced. Necrotic changes in uriniferous tubules
were reduced. Clumping of damaged blood cells was seen.

95. H&E:X 200
Control Fish
and X400
Kidney
Day 1 exposed kidney to lethal dose
of quinalphos
st
Day 5th exposed kidney to lethal
dose of quinalphos

96. H&E:X 200
Control Fish
and X400
Kidney
Day 10th exposed kidney to lethal
dose of quinalphos
Day 15th exposed kidney to lethal
dose of quinalphos

97. Ultrastructure
Control Fish liver
In control fish the hepatocytes are characterized by a system of
parallelly stacked cisternae of rough ER in the vicinity of the
centrally located round nucleus, associated by a few
mitochondria and a few other cell organelles such as
lysosomes and peroxisomes. The cellular compartmentation is
clearly separated into a central, organelle-rich area and a
peripheral cell area with storage material, consisting mainly of
glycogen.

98. Ultrastructure of
Fish liver exposed
to sublethal dose
of quinalphos
In comparison to the controls, hepatocytes of Cyprinus carpio exposed to
quinalphos, demonstrated a loss of cellular compartmentation. The organelles were
distributed through the entire cytoplasm and the amount of glycogen was decreased
compared to controls. The rough ER showed numerous structural alterations including
proliferation, fragmentation and vesiculation of cisternae and little dilation and
degranulation. Lysosomes, peroxisomes, macrophages and myelin-like bodies increased
in number in quinalphos exposed fish compared to controls. Generally, ultrastructural
responses in liver of carp were significantly different between the control.

99. Ultrastructure of
Fish liver exposed
to sublethal dose
of quinalphos
Hepatocytes of common carp exposed to the
sublethal concentration of quinalphos. Cells with a
disturbed cellular compartmentation. The RER is
dilated and vesiculated, the amount of glycogen is
reduced. ga: Golgi apparatus, RER: rough
endoplasmic reticulum.

100. 


Chapter 7: Study on Metabolic and
Enzymological aspects
To
understand
quinalphos
induced
oxidative damage on changes in the protein,
malondialdehyde and protein carbonyls
levels in the tissue of control and
experimental fish tissues.
Enzymes associated also studied i.e., AChE
activity, and catalase activity in the control
and exposed fish.

101. 




Biochemical Studies (RESULT)
Organophosphates are neuro toxins that disrupt the
central nervous system of animals by inhibiting the
enzyme acetylcholinesterase (Rangarsdottir, 2000).
Inhibition of acetylcholinesterase activity results in
changes in behaviour and morphological alterations seen
in the test animal.
The enzyme AChE and its closely related ones are
responsible for the toxicity of organophosphates (OP)
compounds to vertebrates.
In the present study, the level of AChE activity in gill,
kidney and liver tissues of fish, C. carpio exposed to
quinalphos were decreased suggesting the increase in the
level of ACh

102. 

Depression of acetyl cholinesterase activity
suggests decreased cholinergic transmission
and
consequent
accumulation
of
acetylcholine in the tissues leading to
cessation of nerve impulses.
This has lead to behavioral and
morphological changes due to impaired
neurophysiology of the fish.

103. Table 11: Ach level (µM/g wet wt.) in the organs of fish, Cyprinus carpio on exposure to the lethal and sublethal
concentrations of quinalphos.
Exposure period in days
Organ
Contr
ol
Lethal
1
2
133.15 G 147.73 D
Sub lethal
3
156.22
4
1
5
10
154.55
C
Gill
86.54 I
SD ±
% Change
0.001
0.001
0.002
0.012
0.003
0.002
0.013
0.011
0.002
------
53.86
70.71
80.52
88.21
41.97
58.48
66.55
78.59
Kidney
59.58 I
94.89 G
119.11 A
84.34 H
97.23 F
108.51 D
113.63
B
SD ±
% Change
0.003
0.012
0.013
0.011
0.003
0.001
0.012
0.003
0.002
------
59.26
71.75
89.06
99.92
41.56
63.19
82.12
90.72
Liver
66.98 I
118.72 F
123.91 C
132.24 A
95.45 H
119.93 E
121.23 D
B
102.33 E 112.64 C
128.44
B
162.88 A 122.86 H 137.15 F 144.13 E
15
105.34
G
0.002
0.003
0.001
0.002
0.011
0.001
0.013
0.003
0.002
SD ±
% Change
77.25
91.76
40.51
57.27
79.05
80.99
Values are Means ± -----SD (n=6) for a tissue 85.00
in a column followed97.43 same letters are not significantly different (P
by the
≤ 0.05) from each other according to Duncan’s multiple range (DMR) test.

104. Table 12: AChE activity (nM of acetylthiocholine iodide hydrolyzed/mg protein/min) in the organs of fish, Cyprinus
carpio on exposure to the lethal and sublethal concentrations of quinalphos.
Exposure period in days
Organ
Contro
l
Lethal
Sub lethal
1
2
3
135.34
123.29
D
E
110.87 G
4
102.67
1
5
10
166.47
Gill
197.21 A
SD ±
% Change
0.001
0.003
0.002
0.012
0.011
0.001
0.021
0.003
0.011
------
-31.37
-54.61
-70.03
-85.27
-20.31
-48.94
-79.69
-30.33
112.23
152.86
133.51
169.99
H
C
147.15 D
F
B
Kidney
178.28 A 138.21 E 121.87 G 103.22 I
H
157.15 C 120.29 F 101.34 I
15
B
SD ±
% Change
0.003
0.002
0.012
0.002
0.011
0.001
0.003
0.011
0.003
------
-22.47
-40.81
-61.59
-63.98
-14.25
-20.36
-30.42
-6.20
Liver
186.44 A
SD ±
% Change
0.003
0.001
0.002
0.003
-----
-33.90
-54.81
-71.48
123.23
E
118.89 G 101.45 I 111.67 H
155.97
C
131.71 D
0.012
0.001
-73.70
-16.34
119.53
F
161.23 B
0.013
0.003
0.011
-35.09
-58.394
-35.88
Values are Means ± SD (n=6) for a tissue in a column followed by the same letters are not significantly different
(P ≤ 0.05) from each other according to Duncan’s multiple range (DMR) test.

105. 


The assessment of the protein content can be
considered as a diagnostic tool to determine the
physiological process of the cell (David et al.,
2004). Proteins are involved in major physiological
events.
Therefore, the estimation of protein under toxic
stress is useful to determine the physiological
phases of organisms (Kapila and Ragothaman,
1999).
The pesticides are found to alter the structural and
soluble proteins by causing histopathological and
biochemical changes in the cell (Shakoori et al.,
1976).

106. 



Among the exposure periods, in lethal
concentrations, the levels of soluble, structural and
total proteins significantly decreased in the gill,
kidney and liver relative to its controls.
Protein synthesis is an energetically expensive
process.
It appears that protein degradation is in active
phase over synthesis in the gill, kidney and liver of
fish, at day 1 and 5 of exposure to the sublethal
concentration of quinalphos as evidenced from the
decrease in soluble, structural and total proteins
with the significant increase in amino acid levels.
An increase in free amino acid content, which
might have come as a result of tissue damage, is
also suggestive finding of the present study.

107. Table 13: Soluble protein contents (mg of protein/g wet wt. of tissue) in the organs of fish, Cyprinus carpio on
exposure to the lethal and sublethal concentrations of quinalphos.
Exposure period in days
Organ
Control
Lethal
Sub lethal
1
2
3
4
1
5
10
15
Gill
16.54 A
15.52 B
14.44 D
13.21 G
10.71 I
15.01 C
12.96 H
13.63 F
14.12 E
SD ±
0.001
0.002
0.011
0.004
0.003
0.013
0.002
0.003
0.011
% Change
------
-6.16
-12.69
-20.13
-35.24
-9.25
-21.64
-17.59
-14.63
Kidney
15.92 A
14.97 B
13.88 C
13.72 D
9.64 I
13.65 E
11.12 H
12.91 G
13.36 F
SD ±
0.011
0.001
0.003
0.013
0.014
0.002
0.003
0.004
0.012
% Change
------
-5.96
-12.81
-13.81
-39.44
-14.25
-30.15
-18.91
-16.08
Liver
22.92 A
20.86 B
19.05 C
17.25 F
10.88 I
18.81 D
16.87 H
17.18 G
17.97 E
SD ±
0.002
0.001
0.003
0.002
0.011
0.004
0.013
0.012
0.004
% Change
-----
-8.98
-16.88
-24.73
-52.53
-17.93
-26.39
-25.04
-21.59
Values are Means ± SD (n=6) for a tissue in a column followed by the same letters are not significantly different
(P ≤ 0.05) from each other according to Duncan’s multiple range (DMR) test.

108. Table 14: Structural protein contents (mg of protein/g wet wt. of tissue) in the organs of fish, Cyprinus carpio
on exposure to the lethal and sublethal concentrations of quinalphos.
Exposure period in days
Organ
Control
Lethal
Sub lethal
1
2
3
4
1
5
10
15
Gill
22.39 A
20.11 B
17.65 E
15.56 H
11.82 I
19.21 C
15.91 G
17.34 F
18.02 D
SD ±
% Change
0.001
0.003
0.012
0.001
0.021
0.003
0.004
0.003
0.013
------
-10.18
-21.17
-30.51
-47.21
-14.20
-28.94
-22.55
-19.51
Kidney
19.88 A
17.45 B
15.66 E
12.52 H
10.61 I
16.88 C
14.22 G
15.35 F
16.54 D
SD ±
% Change
0.012
0.002
0.004
0.011
0.014
0.021
0.013
0.001
0.013
------
-12.22
-21.22
-37.02
-46.62
-15.09
-28.47
-22.78
-16.81
Liver
29.77 A
25.92 B
22.13 D
19.04 G
15.64 I
22.73 C
18.23 H
19.22 F
20.71 E
SD ±
0.003
0.001
0.021
0.004
0.011
0.023
0.013
0.004
0.002
% Change
-----
-12.93
-25.66
-36.04
-47.46
-23.64
-38.76
-35.43
-30.43
Values are Means ± SD (n=6) for a tissue in a column followed by the same letters are not significantly
different (P ≤ 0.05) from each other according to Duncan’s multiple range (DMR) test.

109. Table 15: Total protein contents (mg of protein/g wet wt. of tissue) in the organs of fish, Cyprinus carpio on
exposure to the lethal and sublethal concentrations of quinalphos.
Exposure period in days
Organ
Control
Lethal
Sub lethal
1
2
3
4
1
5
10
15
Gill
55.87 A
50.12 B
47.44 D
39.84 G
35.35 I
48.65 C
39.77 H
42.07 F
45.78 E
SD ±
0.012
0.001
0.002
0.013
0.004
0.014
0.004
0.003
0.013
% Change
------
-10.29
-15.08
-28.69
-36.72
-12.92
-28.81
-24.71
-18.05
Kidney
47.93 A
42.67 B
39.03 E
33.67 H
28.32 I
40.65 C
35.99 G
37.45 F
39.11 D
SD ±
0.003
0.002
0.013
0.021
0.003
0.013
0.002
0.001
0.002
% Change
------
-10.97
-18.56
-29.75
-40.91
-15.18
-24.91
-21.86
-18.41
Liver
95.32 A
91.34 B
88.11 D
82.78 G
75.99 I
89.98 C
82.06 H
85.27 F
87.66 E
SD ±
0.013
0.003
0.001
0.012
0.021
0.013
0.004
0.002
0.001
% Change
-----
-4.17
-7.56
-13.15
-20.27
-5.61
-13.91
-10.54
-8.03
Values are Means ± SD (n=6) for a tissue in a column followed by the same letters are not significantly
different (P ≤ 0.05) from each other according to Duncan’s multiple range (DMR) test.

110. Table 16: Free amino acid levels (µmol of tyrosine equivalents/g wet wt.) in the organs of fish, Cyprinus
carpio on exposure to the lethal and sublethal concentrations of quinalphos.
Exposure period in days
Organ
Contro
l
Lethal
Sub lethal
1
2
3
4
1
5
10
15
Gill
7.04 I
7.55 H
8.22 F
9.33 D
9.88 C
7.87 G
8.99 E
10.51 B
10.92 A
SD ±
0.011
0.002
0.003
0.004
0.001
0.013
0.012
0.014
0.003
% Change
------
2.17
13.06
26.28
29.54
6.84
17.57
34.21
39.51
Kidney
5.33 I
5.82 H
6.68 F
7.76 D
7.92 C
6.05 G
6.96 E
7.97 B
8.29 A
SD ±
0.003
0.013
0.021
0.002
0.001
0.004
0.023
0.002
0.003
% Change
------
9.26
25.45
45.61
48.66
13.61
30.71
49.60
55.51
Liver
12.65 I
13.42 H
SD ±
% Change
0.021
0.002
0.011
0.013
0.004
0.003
0.001
0.023
0.014
-----
6.08
21.50
32.96
38.65
13.75
32.56
49.17
57.86
15.37
F
16.82 D 17.54 C 14.39 G
16.77
E
18.87 B 19.97 A
Values are Means ± SD (n=6) for a tissue in a column followed by the same letters are not significantly
different (P ≤ 0.05) from each other according to Duncan’s multiple range (DMR) test.

111. 



Oxidative stress
Among the most commonly used biomarkers,
those related to oxidative stress assume an
important position, being frequently used both in
environmental monitoring and laboratory assays
(Pandey, et al., 2003).
Rates or amounts of reactive oxygen species (ROS)
production can be increased by the presence of a
wide range of natural and man-made xenobiotics
(Livingstone, 2001).
The stimulation of free radical production,
induction of lipid peroxidation, and disturbance of
the total antioxidant capability of the body are
mechanisms of toxicity for most pesticides
(Abdollahi, et al., 2004).

112. 
Results

The increase in catalase activity and H2O2
level observed in all the organs of fish at all
the exposure periods studied in the lethal
concentration of quinalphos.
The increase in MDA level in all the organs
of fish at all the exposure periods studied.
The steep increase in protein carbonyl
observed in all the organs of fish at all the
exposure periods studied.



113. 


In the present study, lethal and sublethal
concentrations of quinalphos were resulted in the
significant decrease of antioxidant enzymes with
concomitant increase in the lipid peroxidation in
time-dependent manner when compared with
corresponding control groups.
The cellular defense machinery against H2O2 is
very efficient and involves both low molecular
weight antioxidants and enzymes such as catalase
and glutathione peroxidase.
Catalase activity increased during experimental
periods and is probably a response to toxicant
stress and serves to neutralize the impact of
increased ROS generation (John et al., 2001; Zaidi
and Slotani, 2010)

114. 


The most widely used assay for lipid peroxidation is
the malondialdehyde (MDA) formation, which
represents the secondary lipid peroxidation product
with the thiobarbituric acid reactive substances test
(Draper et al. 1993; Janero 1990).
Malondialdehyde (MDA) is the final product of lipid
peroxidation. The concentration of MDA is the direct
evidence of toxic processes caused by free radicals
(Sieja and Talerczyk 2004).
The formation of carbonyl proteins is non-reversible,
causing conformational changes, decreased catalytic
activity in enzymes and ultimately resulting, owing to
increased susceptibility to protease action, in
breakdown of proteins by proteases (Zhang et al.,
2008).

115. 




Increase in the protease activity as evidenced from the
present study suggests that damage to proteins thus
releasing their monomers due to oxidative damage and
chopping by protease.
Protein degradation is in active phase over synthesis in the
kidney, gill and liver of fish during experimental periods in
both the lethal and sublethal concentration of quinalphos.
Moreover our results recommended that oxidative stress
may, in part, be contributing to quinalphos induced
hepatic, renal and gill damage.
It may provide an indication of quinalphos is the affected
carp C. carpio.
Also our results indicated that the adverse effects of
quinalphos on most of biochemical parameters, lipid
peroxidation and enzymatic activities if used in low
concentration. Also, produced oxidative stress in fish gill
more than liver and kidney both at catalase activity and
MDA levels.

116. Table 17: Catalase activity (mmol of hydrogen peroxide decomposed/mg protein/min) in the organs of fish,
Cyprinus carpio on exposure to the lethal and sublethal concentrations of quinalphos.
Exposure period in days
Organ
Contro
l
Lethal
1
2
Sub lethal
3
4
1
5
10
15
Gill
6.56 I
7.12 H
7.99 E 9.23 B 7.33 G
7.38 F
8.16 D 9.47 A 8.56 C
SD ±
0.014
0.002
0.004
0.013
0.011
0.023
0.001
0.002
0.012
% Change
------
8.53
21.79
40.71
11.73
12.51
24.39
44.35
30.48
Kidney
3.44 I
3.96 H
4.57 E 4.96 B 4.11 G
4.23 F
4.78 D 5.23 A 4.83 C
SD ±
0.011
0.001
0.002
0.003
0.004
0.002
0.021
0.013
0.001
% Change
------
15.11
32.84
44.18
19.47
22.96
38.95
52.03
40.41
Liver
4.96 I
5.42 H
6.11 E 7.12 B 5.72 G
5.88 F
6.54 D 7.32 A 6.77 C
SD ±
0.001
0.003
0.002
0.004
0.011
0.013
0.021
0.012
0.002
% Change
-----
9.27
23.18
43.54
15.32
18.54
31.85
47.58
36.49
Values are Means ± SD (n=6) for a tissue in a column followed by the same letters are not significantly different (P ≤ 0.05)
from each other according to Duncan’s multiple range (DMR) test.

117. Table 18: Hydrogen peroxide levels (nmol of hydrogen peroxide/mg protein) in the organs of fish, Cyprinus carpio
on exposure to the lethal and sublethal concentrations of quinalphos.
Exposure period in days
Organ
Control
Lethal
Sub lethal
1
2
3
4
1
5
10
15
Gill
6.45 I
6.56 H
6.67 F
6.92 C
6.76 E
6.72 G
6.83 D
7.24 B
7.37 A
SD ±
0.002
0.011
0.021
0.004
0.013
0.003
0.003
0.014
0.003
% Change
------
1.71
3.41
7.28
4.81
4.18
5.89
12.24
14.26
Kidney
2.83 I
2.96 H
3.12 F
3.22 C
3.15 E
3.07 G
3.18 D
3.31 A
3.27 B
SD ±
0.001
0.004
0.001
0.023
0.024
0.013
0.004
0.012
0.004
% Change
------
4.59
10.24
13.78
11.30
8.48
12.36
16.96
15.54
Liver
4.53 I
4.66 G
4.81 E 5.02 C
4.64 H
4.78 F
4.96 D
5.44 B
5.54 A
SD ±
0.004
0.011
0.023
0.001
0.004
0.003
0.002
0.021
0.012
% Change
-----
2.86
6.18
10.81
2.42
5.51
9.49
20.08
22.29
Values are Means ± SD (n=6) for a tissue in a column followed by the same letters are not significantly different (P ≤ 0.05)
from each other according to Duncan’s multiple range (DMR) test.

118. Table 19: MDA levels (nmol of TBARS formed/mg of protein) in the organs of fish, Cyprinus carpio on exposure to
the lethal and sublethal concentrations of quinalphos.
Exposure period in days
Organ
Control
Lethal
1
2
Sub lethal
3
4
1
5
10
15
F
3.031 D
2.656 E
2.198 G
3.075 C
3.262 A
3.11 B
2.522
Gill
1.423 I
1.835 H
SD ±
0.001
0.011
0.013
0.003
0.021
0.014
0.002
0.001
0.004
% Change
------
28.87
77.46
113.38
86.61
54.22
116.19
129.57
119.01
Kidney
0.298 I
0.323 H
F
0.407 B
0.393 D
0.335 G
0.383 E
0.425 A
SD ±
0.003
0.003
0.011
0.013
0.001
0.031
0.013
0.004
0.002
% Change
------
8.38
21.81
36.57
31.87
12.41
28.52
42.61
36.24
Liver
0.402 I
0.468 H
F
0.633 C
0.597 E
0.501 G
0.612 D
0.696 A
0.636 B
SD ±
0.011
0.002
0.004
0.0013
0.021
0.015
0.001
0.014
0.013
% Change
-----
16.41
35.57
57.46
48.51
24.62
52.23
73.13
58.20
0.363
0.545
0.406
C
Values are Means ± SD (n=6) for a tissue in a column followed by the same letters are not significantly different (P ≤ 0.05)
from each other according to Duncan’s multiple range (DMR) test.

119. Table 20: Protein carbonyls (nmol of DNPH incorporated/mg protein) in the organs of fish, Cyprinus carpio
on exposure to the lethal and sublethal concentrations of quinalphos.
Exposure period in days
Organ
Lethal
Contr
ol
Sub lethal
1
2
3
4
1
5
10
15
0.331
0.348
0.402
0.422
0.345
0.395
0.442
0.468
H
F
D
C
G
E
B
A
Gill
0.323 I
SD ±
0.013
0.001
0.012
0.011
0.003
0.002
0.021
0.022
0.004
% Change
------
2.47
7.73
24.45
30.65
6.81
22.29
36.84
44.89
Kidney
0.847 I
0.896
0.977
1.112
1.218
0.932
1.011
1.233
1.341
H
F
D
C
G
E
B
A
SD ±
0.004
0.002
0.001
0.003
0.004
0.001
0.021
0.013
0.022
% Change
------
5.78
15.34
31.05
42.85
10.03
19.36
45.21
58.20
Liver
0.511 I
0.553
0.591
0.644
0.674
0.577
0.624
0.702
0.737
H
F
D
C
G
E
B
A
SD ±
0.021
0.002
0.001
0.004
0.014
0.011
0.003
0.021
0.001
% Change
-----
8.21
15.65
26.02
31.89
12.91
22.11
37.37
44.22
Values are Means ± SD (n=6) for a tissue in a column followed by the same letters are not significantly different (P ≤ 0.05)
from each other according to Duncan’s multiple range (DMR) test.

120. Table 21: Protease activity (µmol of tyrosine equivalents/mg protein/h) in the organs of fish, Cyprinus
carpio on exposure to the lethal and sublethal concentrations of quinalphos.
Exposure period in days
Organ
Control
Lethal
1
Gill
0.634 I
SD ±
0.665
2
Sub lethal
3
4
1
5
0.822
0.847
0.696
D
C
G
0.764 E
H
0.735 F
0.002
0.001
0.021
0.013
0.021
0.004
% Change
------
4.88
15.93
29.65
33.59
9.77
Kidney
0.523 I
0.628 F
0.654 E
0.675
0.595
H
C
G
0.657 D
SD ±
0.011
0.033
0.003
0.001
0.014
0.003
% Change
------
10.32
20.07
25.04
29.06
Liver
0.753 I
0.783
0.837
0.948
0.958
H
G
D
SD ±
0.003
0.002
0.001
% Change
-----
3.98
11.15
0.577
10
0.874
15
B
0.902 A
0.003
0.011
0.001
20.50
37.85
42.27
0.711
B
0.733 A
0.021
0.013
0.022
13.76
25.62
35.94
40.15
C
0.838 F
0.933 E
0.014
0.004
0.002
25.89
27.22
11.28
1.112
B
1.231 A
0.011
0.001
0.021
23.91
47.67
63.47
Values are Means ± SD (n=6) for a tissue in a column followed by the same letters are not significantly different (P ≤ 0.05)
from each other according to Duncan’s multiple range (DMR) test.

123. 



Chapter 8: Studies on Haematology
The study of blood parameters in fishes has been
widely used for the detection of physiopathological
alterations in different conditions of stress (Nussey
et al., 1995).
Moreover, haematological parameters are closely
associated to the response of the fish to the
environment (Tiwari and Sing, 2006).
Accordingly, haematology can be used as clinical
tool for the investigations of physiological and
metabolic alterations in fish caused by pollution of
the aquatic environment (Anand Kumar, 1994).

124. 

Blood is a vehicle for quickly mobilizing defense
against trauma and diseases. Since, fishes differ
considerably in their activity patterns and respond
to the pollutant.
The blood parameters like Red blood corpuscle
(RBC),
white
blood
corpuscle
(WBC),
Haemoglobin (Hb), Packed cell volume (PCV),
Mean corpuscular volume (MCV), Mean
corpuscular haemoglobin (MCH) and Mean
corpuscular haemoglobin concentration (MCHC)
are commonly studied in fishes to assess the
impact of pesticides in aquatic biota.

125. 


Hematological Study (RESULT)
There was decrease in RBC, Hb, and
values of MCH, MCHC, PCV and MCV on
the fish exposed to both lethal and sub
lethal concentrations of quinalphos.
Whereas WBC recorded an elevation.
The effect is conspicuous under
quinalphos
toxicity
suggesting
augmentation of additive effect on RBC
and
haemoglobin
synthesis
and
leucocytosis.

126. 


Measurement of haematological parameters are important
in diagnosing the structural and functional status of
animals exposed to the toxicant because blood parameters
are highly sensitive to environmental or physiological
changes and health conditions.
Pesticides are known to alter the blood parameters of
fishes. A significant decrease in RBC, Hb content and PCV
has been observed earlier in fishes exposed to different
pesticides.
The findings of the present investigation also reveal a
similar decreasing trend in all the parameters such as RBC,
Hb
content
and
PCV
suggesting
that
the
Organiphosphorous pesticides also induce changes which
give evidence for decrease haematopoiesis followed by
anemia induction in test fishes.

127. Table 22: RBC count (x106/mm3) in the blood of the fish, Cyprinus carpio on exposure to lethal and sublethal
concentrations of quinalphos.
Exposure periods in days
Parame
ter
Lethal
Control
Sublethal
1
2
3
4
1
5
10
15
1.53 A
1.42 B
1.36 D
1.11 H
0.95 I
1.29 E
1.17 G
1.21 F
1.39 C
SD ±
0.003
0.025
0.013
0.012
0.015
0.014
0.012
0.016
0.012
%
Change
----
-7.18
-11.11
-27.45
-37.90
-15.68
-23.52
-20.91
-9.15
RBC
Values are Means ± SD (n=6) for a parameter in a row followed by the same letters are not significantly
different (P ≤ 0.05) from each other according to Duncan’s multiple range (DMR) test.

128. Table 23: WBC count (x103/mm3) in the blood of the fish, Cyprinus carpio on exposure to lethal and
sublethal concentrations of quinalphos.
Exposure periods in days
Paramete Contro
r
l
Lethal
Sublethal
1
2
3
4
10.45 F
10.67 E
9.78 G
8.59 H
6.49 I
SD ±
0.022
0.011
0.013
0.011
%
Change
----
2.11
-6.41
-17.79
WBC
1
5
10
D
12.31 B
12.87 A
0.0123
0.013
0.012
0.013
0.012
-37.89
2.67
17.79
23.15
14.16
10.73
15
11.93
C
Values are Means ± SD (n=6) for a parameter in a row followed by the same letters are not significantly
different (P ≤ 0.05) from each other according to Duncan’s multiple range (DMR) test.

129. Table 24: Haemoglobin level (g/100 ml) in the blood of the fish, Cyprinus carpio on exposure to lethal
and sublethal concentrations of quinalphos.
Exposure periods in days
Paramet
er
Haemogl
obin
Lethal
Control
Sublethal
1
2
3
4
1
5
10
15
7.64 A
6.55 D
5.94 G
3.87 H
3.53 I
6.88 B
6.12 F
6.34 E
6.62 C
SD ±
0.011
0.012
0.013
0.012
0.014
0.003
0.015
0.012
0.011
%
Change
----
-14.26
-22.25
-49.34
-53.79
-9.94
-19.89
-17.01
-13.35
Values are Means ± SD (n=6) for a parameter in a row followed by the same letters are not significantly
different (P ≤ 0.05) from each other according to Duncan’s multiple range (DMR) test.

130. Table 25: PCV level (%) in the blood of the fish, Cyprinus carpio on exposure to lethal and
sublethal concentrations of quinalphos.
Exposure periods in days
Param
Control
eter
25.22 A
PCV
Lethal
Sublethal
1
2
3
4
23.21
20.55
16.73
C
D
H
10.49 I
1
23.58
B
5
10
15
19.96 E 18.64 G 19.66 F
SD ±
0.012
0.001
0.013
0.012
0.014
0.015
0.013
0.002
0.021
%
Chang
e
----
-7.96
-18.51
-33.66
-58.41
-6.50
-20.85
-26.09
-22.04
Values are Means ± SD (n=6) for a parameter in a row followed by the same letters are not
significantly different
(P ≤ 0.05) from each other according to Duncan’s multiple range
(DMR) test.

131. Table 26: MCV level (cu mm) in the blood of the fish, Cyprinus carpio on exposure to lethal
and sublethal concentrations of quinalphos.
Exposure periods in days
Param
eter
Contr
ol
Lethal
Sublethal
1
2
3
70.32
75.33
82.21
92.83
H
G
C
A
87.37 B
SD ±
0.013
0.001
0.014
0.032
%
Chang
e
----
7.12
16.91
32.01
MCV
4
1
5
78.73 79.26
10
15
75.65 F 72.81 I
E
D
0.002
0.021
0.003
0.013
0.012
24.24
11.95
12.71
7.57
3.52
Values are Means ± SD (n=6) for a parameter in a row followed by the same letters are not
significantly different
(P ≤ 0.05) from each other according to Duncan’s multiple range (DMR)
test.

132. Table 27: MCH level (pg) in the blood of the fish, Cyprinus carpio on exposure to
lethal and sublethal concentrations of quinalphos.
Exposure periods in days
Parame
ter
MCH
SD ±
%
Change
Cont
rol
Lethal
1
2
3
Sublethal
4
1
5
10
15
22.38 23.84 24.92 26.79 28.66 24.87 27.23 27.48 25.12
I
H
F
D
0.022 0.003 0.023 0.012
----
6.52
11.34
A
0.011
19.71 28.06
G
C
B
E
0.014 0.001 0.015 0.013
11.12
21.67 22.78 12.24
Values are Means ± SD (n=6) for a parameter in a row followed by the same letters are not
significantly different
(P ≤ 0.05) from each other according to Duncan’s multiple range
(DMR) test.

133. Table 28: MCHC level (%) in the blood of the fish, Cyprinus carpio on
exposure to lethal and sublethal concentrations of quinalphos.
Exposure periods in days
Paramet Contr
er
ol
Lethal
Sublethal
1
2
3
4
1
22.67
20.54
20.94
18.84
18.55
19.88
A
E
D
G
H
F
SD ±
0.011
0.001
0.013
0.021
0.013
0.002
0.012
0.014
0.023
%
Change
----
-9.39
-7.63
-16.89 -18.17 -12.31
-18.79
-7.41
-5.51
MCHC
5
10
15
18.41 I 20.99 C 21.42 B
Values are Means ± SD (n=6) for a parameter in a row followed by the same letters are
not significantly different
(P ≤ 0.05) from each other according to Duncan’s
multiple range (DMR) test.

134. 


In the present investigation, fish Cyprinus carpio
treated to the lethal and sublethal concentrations of
quinalphos, showed considerable alteration in the
level of different blood parameters.
Hematological parameters in fish can significantly
change in response towards chemical stressors;
however, these alterations are non-specific to a wide
range of substances. Some of these changes may be
the result of the activation of protective mechanisms
(Cazenave et al., 2005) such as the results of the
blood parameters observe in the present work.
The anaemia produced due to quinalphos treatment
might be the reasons for the decrease in the RBC
count, haemoglobin percentage and haematocrit
value at lethal concentration, which therefore clearly
suggests the species response to pesticide and other
toxicants.

135. 


Conclusion
As food source, fish interferes on man’s life quality
and so more detailed analysis of the action of what
may lethal and sub lethal concentrations of
pesticide and insecticide substances provoke in
these organisms is necessary.
The over all study inferred that, the death of fish
under lethal concentration might be due to the
collective interference of the pesticide with all the
systems at physiological, biochemical and
histopathological levels.

136. 

From the above assessment pertinent to
physiological, behavioural and biochemical
response of freshwater fish, C. carpio to
quinalphos (EC 25%), the conclusion could
be drawn that the changes arrived at are
dependent on concentration of pesticide and
the duration of exposure.
Irreparable damage was caused to the
physiological, histological, biochemical and
behavioural activities of the fish at higher
concentration. The damage increased and
prevailed over time of exposure.

137. 

Under low concentration, i.e., sublethal
concentration stress in fish was observed
only for short period (1 to 5 days) and on
later days of exposure the stress appeared to
lessen and the fish seemed to adapt the toxic
environment.
The recovery tendency shown by the fish,
perhaps, could be due to physiological
resistance developed by the animal, which
also be reasoned as possible enhancement of
detoxification mechanism and quinalphos
elimination processes.

138. 


Therefore, the above statement suggests that the
fish can adapt to low concentration of quinalphos
toxicity during long-term exposure periods.
The over all study inferred that, the death of fish
under lethal concentration might be due to the
collective interference of the pesticide with all the
systems at physiological, biochemical and
histopathological levels.
But in sub lethal concentration the fish can survive
to a grater extent with significant metabolic
compensation to overcome the chronic stress. The
results are processed with statistical treatment and
discussed in the light of available literature.

139. 
It is hoped that based on some of the significant
differences observed in all the aspects of the
present study, useful in determining the safe
concentration of quinalphos to ensure protection of
worthy fishery resources and provide base line
information for future monitoring of pesticides in
the aquatic environment.

140. Thank you

141. 







Acknowledgement
Dr. M. David sir and family
My Parents and In-laws'
My Family, Wife and Daughter
University authorities
My College Karnatak Science College,
Dharwad
All Teaching and Non-teaching staff
Friends