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CAN THO UNIVERSITY
COLLEGE OF AQUACULTURE AND FISHERIES
NGUYEN QUANG TRUNG
EFFECTS OF QUINALPHOS INSECTICIDE ON
PHYSIOLOGY, BIOCHEMICAL CHANGES
AND GROWTH OF COMMMON CARP
(Cyprinus carpio Linnaeus, 1758)
Major: AQUACULTURE
Code: 62.62.03.01
ABSTRACT OF THESIS OF DOCTOR DEGREE OF
AQUACULTURE

Can Tho, November 2013
1
The study was conducted at Department of Nutrition and Fisheries
Processing, College of Aquaculture and Fisheries, Can Tho University
Supervisor: Asso.Dr. Đo Thi Thanh Huong – Can Tho University
Commentator 1:
Commentator 2:
Commentator 3:
Thesis will be defensed at final thesis commentary council at:
………………………………………………………………………
At: o’clock date
i
TABLE OF CONTENTS
AT: O’CLOCK DATE I
TABLE OF CONTENTS II
LIST OF FIGURE VI
VI
CHAPTER 1: INTRODUCTION 1
1.1 Introduction 1

1.2 Objective 1
1.3 Contents 2
1.4 Signification of thesis 2
1.5 New results of thesis 2
CHAPTER 2: LITERATURE 4
2.1 Chemical 4
2.2 Active of mechanism of AChE 4
Figure 2.3: Active mechanism of AChE (A) and mechanism 4
of inhibition of organophosphorus pesticide (B) 4
(Richard and David, 2008) 4
CHAPTER 3: METHODOLOGY 5
3.1 Experimental site 5
ii
3.2 Materials 5
3.3 Experimental fish 5
3.4 Experimental chemical 5
3.6 Feed 5
3.7 Methods 5
3.7.1 Survey on pesticide use in Can Tho city 5
3.7.2 Determination of LC50- 96 hrs of quinalphos on common carp 5
Experiment was conducted in glass tank of 50 L, 10 fish/tank, weight
from 8-10 g. During the experiment, no exchange of water, no aeration
and no feeding was applied 5
3.7.3 Effects of quinalphos on respiratory physiology of common carp 6
3.7.3.1 Oxygen comsumption 6
Experiment included in five treatments: control 10%, 20%, 50% and
75% value of LC50-96 hrs., six replicates, 2 fish/2 L. Oxygen
comsumption was determined by method of closed-vessel. There were
two cases: contamination design before 24 hours and direct design 6
Experiment included in five treatments: control 10%, 20%, 50% and

75% value of LC50-96 hrs., six replicates, 4 fish/2 L. Oxygen
comsumption was determined by method of closed-vessel. There were
two cases: contaminating design before 24 hours and direct design.
When observed mortality of 50% in vessel, sampling oxygen to
determine oxygen threshold 6
3.7.4 Effects of quinalphos on ChE sensitivity and inhibition threshold of
common carp 6
3.7.5 Effects of quinalphos on haematological parameters, ChE activity,
digestive enzymes and growth of common carp 7
3.7.5.1 Effects of quinalphos on haematological parameters, and enzyme
activities 7
3.7.6 Effects of quinalphos on cholinesterase activity and growth
performances of common carp cultured in rice-field 8
3.7.6.1 Implementary method 8
3.8 Methods of analysis 8
3.8.1 Measurable method of haematological parameters 8
3.8.2 Measurable method of oxygen consumption 8
3.8.3 Analytic method of biochemical parameters 8
3.8.4 Measurable methods of growth parameters 9
iii
3.8.5 Measurable method of environmental parameters 9
3.8.6 Measurable method of quinalphos residue 9
3.9 Method of data treatment 9
VALUE OF LC50 WAS DETERMINED BY METHOD OF
PROBIT. ONE-WAY ANOVA AND TWO-WAY ANOVA
WERE USED TO COMPARE THE DIFFERENCES
BETWEEN TREATMENTS AT A 5% SIGNIFICANT
LEVEL, USING SOFTWARE SPSS 11.5 9
CHAPTER 4: RESULTS AND DISCUSSIONS 10
4.1 Survey on pesticide use in Can Tho city 10

4.1.1 General information 10
4.1.2 Information on Autumn-Summer crop 10
OM 2514 was the most common variety (26,7%). Most of farmers began
Autumn-Summer crop from Lunar March to June (88,9%). Rice growth
duration was 95 days 10
4.1.3 Technical parameters on rice-fish system 10
4.1.4 Economic efficiency of rice and rice-fish 10
4.1.5 Present status of pesticide use 10
4.1.6 Information on agents of pesticide trade 10
4.2 Acute toxicity of quinalphos insecticide on common carp 11
4.2.1 Environment parameters during the experiment 11
4.2.2 Acute toxicity of quinalphos insecticide on common carp 11
4.3 Effects of quinalphos on respiratory physiology 11
4.3.1 Oxygen consumption 11
4.3.2 Oxygen threshold 11
Increased concentration of decamethrin increased oxygen consumption
of common carp (James et al., 1994). Murty (1988) and Đo Thi Thanh
Huong (1997) stated that oxygen consumption increased at the beginning
of exposing to insecticide because respiratory activity increased but
decreasing after 12
4.4 Effects of quinalphos on ChE sensitivity and inhibition threshold 12
4.4.1 Environment parameters during the experiment 12
iv
4.4.2 ChE sensitivity 12
4.4.3 ChE inhibition threshold 12
4.5 Effects of quinalphos on haematological parameters, ChE activity,
digestive enzymes and growth of common carp 12
4.5.1 Effects of quinalphos on haematological parameters and enzyme
activities of common carp 12
4.5.2 Effects of quinalphos on haematological parameters, cholinesterase

activity, digestive enzymes and growth performances of common carp
cultured in tank 16
4.5.2.1 Environmental parameters and quinalphos residue 16
4.5.2.2 Effects of quinalphos on haematological parameters 16
4.5.2.3 Effects of quinalphos on ChE activity and digestive enzymes
activities of common carp 17
4.6 Effects of quinalphos on ChE activity and growth of common carp
cultured in rice-field 19
4.6.1 Environmental paramters during the experiment 19
4.6.2 Effects of quinalphos on ChE activity 19
4.6.3 Effects of quinalphos on growth, survival and yield of common
carp cultured in rice-field 19
4.7 General discussion 20
CHAPTER 4: CONCLUSIONS AND
RECOMMENDATIONS 22
4.1 Conclusions 22
4.2 Recommendations 22
REFERENCES 23
LIST OF RESEARCH WORK 27
LIST OF TABLE
Table 4.9: Survival of common carp exposing to quinalphos 10
Table 4.25: Growth of common carp …………………………. 17
v
LIST OF FIGURE
Figure 2.3: Active mechanism of AChE 3
Figure 4.1: Changes of brain ChE activity after 28 days ………. 12
Figure 4.2: Brain ChE inhibition after 28 days………………… 12
Figure 4.3: Changes of muscle ChE activity after 28 days ……. 12
Figure 4.4: Muscle ChE inhibition after 28 days ………………. 12
Figure 4.5: Changes of gill ChE activity after 28 days………… 13

Figure 4.6: Gill ChE inhibition after 28 days …….……………. 13

vi
CHAPTER 1: INTRODUCTION
1.1 Introduction
Quinalphos is among the most widely used organophosphorus
insecticides (OPs) in agriculture in India and some other countries
among them Viet Nam (Chebbi et al., 2009).
Haematological parameters such as hemoglobin, hematocrit,
erythrocyte,…can be used to find physiological reaction in fish when
environment was pollution (Dethloff et al, 2001). AChE inhibition
affected to respiration, swimming activity, feeding and aquatic animal
behaviours leading to imbalance, convulsion and even death (Peakall,
1992). Digestive enzymepatterns can reflect the feeding rate and
digestive capacity of fish; hence, digestive enzyme activity can be used
as bio-indicators of growth and health status of fish (Debnath et al.,
2007; Suarez et al., 1995; cited by Li et al., 2010).
Common carp Cyprinus carpio was the most common species in
rice-field in Mekong Delta and Can Tho (Nguyen Van Hảo et al.,
2001; Vromant et al., 2002). Quinalphos was widely used in
agriculture and residue of quinalphos can affect to fish (Das et al.,
2000). In order to study effects of insecticide on physiological
parameters, biochemical changes in tank and rice-field, this study was
conducted at College of Aquaculture and Fisheries – Can Tho
University.
1.2 Objective
- General objectives: Search scientific data of toxicity of insecticide
on aquatic animals in order to recommend farmers suitable pecticide use
in rice-field basically.
- Specific objectives: Search effects of quinalphos insecticide to

change physiological parameters (haematology and respiratory
physiology), biochemical changes (enzyme activities) and growth
performances of common carp. As a result, physiological parameters
or biochemical changes as a biomarker for organophosphorus
insecticides like quinalphos in rice-filed.
1
1.3 Contents
a) Survey on pesticide use in Can tho city.
b) Determination of LC
50
-96 hrs. of quinalphos on common carp.
c) Effects of quinalphos on respiratory physiology of common
carp Cyprinus carpio.
d) Effects of quinalphos on ChE sensitivity and inhibition
threshold of common carp.
e) Effects of quinalphos on haematological parameters,
cholinesterase activity, digestive enzymes and growth performances of
common carp cultured in tank.
f) ) Effects of quinalphos on cholinesterase activity, digestive
and growth performances of common carp cultured in rice-field.
1.4 Signification of thesis
The results of thesis are scientific basic to recommend farmers
suitable pecticide use in rice-field.
1.5 New results of thesis
Thesis determined quinalphos changed haematological parameters
and respiratory physiology (oxygen consumption and threshold).
Quinalphos considerably inhibited the brain, muscle, gill and liver
ChE activities of common carp exposed to sublethal concentrations in
96 hours of acute test and 28 days and 60 days of long test in tank and
44 days in rice-filed. ChE activity was completly recovered at least 21

days in tank and 14 days in rice-field.
Thesis also determined GST activity of common carp didn’t play
an important role for decontamination. Quinalphos considerably
inhibited digestive enzymes. Thesis determined brain was the most
sensitive organ of common carp. ChE inhibition affected to swimming
behaviour of fish. ChE inhibition threshold causing died fish was 95%.
Thesis determined quinalphos affected to growth performmances,
feed conversion rate and survival rate of common carp cultured in tank.
Recommended dose affected to growth performmances, survival rate and
productivity of common carp as compared to the control in rice-field.
Concentrations of quinalphos which gradually decreased during the
experiment were under the limit of determination (<LOD) at least 14
days.
2
3
CHAPTER 2: LITERATURE
2.1 Chemical
- Trade name: Kinalux 25EC
- Chemical name: 0,0-diethyl 0–2 quinoxalin
phosphorothioate
- Active ingredient: Quinalphos
- Source of chemical: organophosphosphat
- Molecular formula: C
12
H
15
N
2
O
3

PS
- Molar mass: 298,3 g/mol
- Chemical formula:
2.2 Active of mechanism of AChE
AChE serves as a regulating agent of nervous transmission. AChE
catalyzed hydrolysis of ACh into choline (Ch) and acetic acid (A). When
AChE is inactivated by an organophosphorus, the enzyme is no longer
able to hydrolyze ACh; the concentration of ACh in the junction remains
high, and continuous stimulation of the muscle or nerve fiber occurs,
resulting eventually in muscle twitching, tremors or exhaustion
Figure 2.3: Active mechanism of AChE (A) and mechanism
of inhibition of organophosphorus pesticide (B)
(Richard and David, 2008)
4
CHAPTER 3: METHODOLOGY
3.1 Experimental site
The study was carried out at College of Aquaculture and Fisheries,
Cantho University and Thoi Hung commune, Co Do district, Can Tho
city from May 2009 to July 2012.
3.2 Materials
The study used main materials such as:
- Composite tank (capacity of 500 L, 2 m
3
, 4 m
3
), glass tank 84 L.
- Spectrophotometer, centrifugal machine and so on.
3.3 Experimental fish
Common carp (8-12 grams/individual) were selected from
hatcheries in Can Thome. Fish were acclimated in composite tank at

least 7 days before experiment.
3.4 Experimental chemical
Kinalux 25 EC insecticide which had acitve ingredient of quinalphos
(organophosphate) bought in Agent of Pesticide Trade of Can Tho.
3.5 Experimental water
Water was supplied from tap water. Tap water was aerated 48
hours before starting experiment.
3.6 Feed
Fish were fed with commercial feed (30% crude protein) and
feeding on satiation
3.7 Methods
3.7.1 Survey on pesticide use in Can Tho city
- Secondary data collection: The secondary data was collected
from local management office.
- Primary data collection: Total of samples was 105 including
three groups: 45 rice cultivation farmers, 45 rice-fish farmers and 15
agencies of pesticides in Co Do, Thoi Lai and Vinh Thanh district.
3.7.2 Determination of LC
50
- 96 hrs of quinalphos on common carp
Experiment was conducted in glass tank of 50 L, 10 fish/tank,
weight from 8-10 g. During the experiment, no exchange of water, no
aeration and no feeding was applied.
Range-finding toxicity test
Experiment was designed 10 concentrations (one replicate for
each) from 0-4 mg/L. The mortality was recorded in 96 hours. Range-
finding toxicity was calculated to find value of LC
50
-96 hrs.
5

LC
50
-96 hrs. definitive test
Experiment was designed 7 concentrations (three replicates for
each) from 0-1,8 mg/L. The mortality was recorded in 96 hours.
Temperature, pH and oxygen were measured twice daily.
3.7.3 Effects of quinalphos on respiratory physiology of common carp
3.7.3.1 Oxygen comsumption
Experiment included in five treatments: control 10%, 20%, 50%
and 75% value of LC
50
-96 hrs., six replicates, 2 fish/2 L. Oxygen
comsumption was determined by method of closed-vessel. There were
two cases: contamination design before 24 hours and direct design.
3.7.3.2 Oxygen threshold
Experiment included in five treatments: control 10%, 20%, 50%
and 75% value of LC
50
-96 hrs., six replicates, 4 fish/2 L. Oxygen
comsumption was determined by method of closed-vessel. There were
two cases: contaminating design before 24 hours and direct design.
When observed mortality of 50% in vessel, sampling oxygen to
determine oxygen threshold.
3.7.4 Effects of quinalphos on ChE sensitivity and inhibition
threshold of common carp.
3.7.4.1 ChE sensitivity
a) Experimetal design
Experiment was conducted based on two way anova. Two factors
were sampling time and concentrations including in control, 1%, 10%,
20%, 50% and 75% value of LC

50
-96 hrs., three replicates, 30 fish/tank
100 L. Experimental time was 96 hours.
b) Method of sampling
Three fish per tank were collected in 96 hours. Brain was collected
to determine sensitivity of ChE. Temperature, pH and oxygen was
measured twice daily.
3.7.4.2 ChE inhibition threshold
a) Experimetal design
Treaments consisted of control, 1%, 10%, 20%, 50%, 75% and
100% value of LC
50
-96 hrs., three replicates, 10 fish/tank 50 L.
Experimental time was 96 hours.
b) Method of sampling
Brain was collected when obversed abmormal swimming activities
such as imbalance, dull swimming and died fish to determine ChE
inhibition threshhold.
6
3.7.5 Effects of quinalphos on haematological parameters, ChE
activity, digestive enzymes and growth of common carp
3.7.5.1 Effects of quinalphos on haematological parameters, and
enzyme activities
a) Experimetal design
Experiment was conducted based on two way anova. Two factors
were sampling time and concentrations including in control, 10%,
20%, 50% and 75% value of LC
50
-96 hrs., six replicates, 15 fish/tank
60 L. Experimental time was 28 days.

b) Management of experimental fish
Renewal water was periodically carried out. No aeration and
siphon daily were applied. Fish were feeding on satiation.
c) Method of sampling
- Fish were sampled before exposure (day 0), day 1, 7, 14, 21 and 28
after exposure. Each sampling time, six fish per treatment. Blood was
collected to measure haematological parameters such as erythrocyte,
hemoglobin, hematocrit, MCV, MCH and MCHC. Brain, muscle, gill and
liver was collected to measure ChE, GST, CAT and LPO activities.
- Temperature, pH and oxygen was measured daily.
3.7.5.2 Effects of quinalphos on haematological parameters, ChE,
activity, digestive enzymes and growth performances of comon carp
a) Experimetal design
Experiment was conducted based on two way anova (biochemical
experiment). Two factors were sampling time and concentrations
(control, 10%, 20%, 50% and 75% value of LC
50
-96 hrs.), three
replicates, 80 fish/tank 380 L. Experimental time was 90 days.
Quinalphos insecticide was applied twice.
b) Method of sampling
Fish were collected at day 0 (before exposure), 6 hours, day 3, day 30
after one exposing time; day 31 (6 hours), day 33 and day 60 after two
exposing time. Blood was collected to measure haematological parameters
(erythrocyte, hemoglobin, hematocrit, MCV, MCH and MCHC).
Brain, muscle, gill and liver was collected to measure ChE activity.
Gut was collected to determine digestive enzymes (trypsin,
chymotrypsin, alpha-amylase). Each time collected 5 fish/tank.
c) Management of experimental fish
Renewal water was periodically carried out. Tanks were slightly

aerated. Fish were fed on commercial feed (30% crude protein) based on
satiation. Feed were weighted before and after feeding to determine FCR.
7
d) Recorded parameters
Fish growth were measured at day 30, 60 and 90, sampling 30
fish/tank. Data was collected to determine growth parameters such as
weight gain, DWG, SGR, FCR and survival rate. Temperature, pH and
oxygen was measured daily.
3.7.6 Effects of quinalphos on cholinesterase activity and growth
performances of common carp cultured in rice-field
3.7.6.1 Implementary method
Experiment was conducted based on two way anova (enzyme
activity). Two factors were sampling time and concentrations (control and
recommended dose), three replicates, 2 fish/m
2
(each plot of 1.000 m
2
approximately. Experimental time was 132 days. Quinalphos insecticide
was applied twice: OM 4218 variety was used in this experiment.
3.7.6.2 Method of sampling
- Fish were collected at day 0 (before exposure), 1, 7 and 14 after
one exposing time. Sampling was similarity in two exposing time.
- Brain, muscle, gill and liver was collected to measure ChE
activity. Each time collected 5 fish/plot. Fish were collected by net.
3.7.6.3 Management of plot
Fish were fed on commercial feed (30% crude protein) based on
satiation. Renewal water was periodically carried out twice a month by
pump.
3.7.6.4 Recorded parameters
Fish growth were measured at day 50, 100 and 132. Sampling 20-

30 fish/plot. Recorded parameters were weight gain, DWG, SGR,
survival rate and productivity. Temperature, pH and oxygen was
measured once a week. Ammonia was measured twice a month.
3.8 Methods of analysis
3.8.1 Measurable method of haematological parameters
- Erythrocyte was counted by Neubauer method.
- Blood parameters related to erythrocyte (MCV, MCH, MCHC)
were measured by Svobodova et al .(1991).
- Hemoglobin was measured by spectrophotometer 540 nm.
- Hematocrit was measured by percent of plasma as compared to
total of volume
3.8.2 Measurable method of oxygen consumption
Oxygen consumption was measured by Đo Thi Thanh Huong, 1997:
3.8.3 Analytic method of biochemical parameters
- Cholinesterase (ChE) was determined by Ellman et al. (1961).
8
- Catalase (CAT) was determined by Baudhuin et al. (1964).
- Glutathione S-transferase (GST) was determined by Habig et al. (1974).
- Lipid peroxidation (LPO) was determined by Fatima et al. (2000).
- Digestive enzymes: Trypsin activity was determined by Tseng et
al. (1982). Chymotrypsin activity was determined by Worthington
(1982). Alpha-amylase was determined by Bernfeld (1951).
- Protein was determined by Lowry et al. (1951) and Bradford (1976).
3.8.4 Measurable methods of growth parameters
Weight gain – WG
WG(g) = W
final
– W
initial
Specific growth rate –SGR

SGR (%/day)=100x(LnW
final
- LnW
initial
)/T
Daily weight gain – DWG
DWG (g/day)= (W
final
– W
initial
)/T
Among them:: W
initial
: Fish weight of starting experiment
W
final
: Fish weight of ending experiment
T : Experimental time (day)
Feed conversion ratio - FCR
FCR= Dry food fed/ Weight gain of fish
Survival rate – SR
SR (%)=100x(number of fish at harvest/number of fish at stock)
Fish yield
Yield (kg/ha)= [Total fish weight at harvest x10.000] x
Experimental area (m
2
).
3.8.5 Measurable method of environmental parameters
- Temperature, pH and oxygen were measured by oxygen meter.
- Ammonia was measured by method of Indophenolblue.

- Oxygen content were measured by method of Winkler.
3.8.6 Measurable method of quinalphos residue
Quinalphos insecticide residue was measured by method of GC-
ECD (Nguyen Quoc Thinh et al., 2012).
3.9 Method of data treatment
Value of LC
50

was determined by method of probit. One-way Anova
and Two-way Anova were used to compare the differences between
treatments at a 5% significant level, using software SPSS 11.5.
9
CHAPTER 4: RESULTS AND DISCUSSIONS
4.1 Survey on pesticide use in Can Tho city
4.1.1 General information
Area of rice cultivation was 1.93 ha (70% of total area), area of
rice-fish was 2.02 ha (75%).
4.1.2 Information on Autumn-Summer crop
OM 2514 was the most common variety (26,7%). Most of farmers
began Autumn-Summer crop from Lunar March to June (88,9%). Rice
growth duration was 95 days.
4.1.3 Technical parameters on rice-fish system
Common carp was the most common species in rice-fish system
(100% household). Density of fish was 0.42±0.25 con/m
2
. Culture
duration was 149 days. Fish productivity was 577±322 kg/ha.
4.1.4 Economic efficiency of rice and rice-fish
Productivity of mono-rice was 4.5 tons/ha and rice crop in rice-fish
model was 4.4 tấn/ha. Profit of rice crop in rice-fish model was 7.7

millions VND/ha and significant difference as compared to mono-rice
(4,3 millions VND/ha).
4.1.5 Present status of pesticide use
The results showed that number of spray ranged from 2.7 from 2.9
times/crop. The most extensive spray time was from day 31 to day 60
of mono-rice (89.7%) and rice-fish (73.7%). Chess was the most
widely used organophosphorus insecticides in mono-rice (48.9%
household), next to Basa (26,7%) and Kinalux (active ingredient of
quinalphos) (20%). For rice-fish system, Basa was the most widely
used insecticides (37.8% household), Kinalux 6.7%. In agents of
pesticide trade, Dragon was the most widely consumed insecticide
(80%), next to Kinalux (66.7% household).
4.1.6 Information on agents of pesticide trade
There were about 133 products of pesticides which bought at 15
agents in Can Tho city. The most consuming volume of pesticide was
Basa 3,041 liters/year, next to Kinalux 1,497 liters/year.
Ngo Van Ngoc et al. (2001), Nguyen Van Hao et al. (2001) indicated
that common carp were among the most common species in rice-field in
Mekong Delta. According to survey on pesticide use in Can Tho city in
2007 (Tam, 2008) showed that fenobucarb (Basa),
Chlorpyriphos+Cypermethrin (Dragon) and quinalphos (Methink) were
10
among the most widely used insecticides. The present study recorded that
quinalphos was one of the most extensively used insecticides.
4.2 Acute toxicity of quinalphos insecticide on common carp
4.2.1 Environment parameters during the experiment
Temperature in the morning was 26.9
o
C and in the afternoon was
27.6

o
C; fluctuation of temperature was under 1
o
C. pH ranged 7.8-7.9.
Oxygen dissolve ranged 3.6

mg/L.
4.2.2 Acute toxicity of quinalphos insecticide on common carp
Mortality and value of LC
50
of quinalphos on common carp were
presented in Table 4.9. After 1 hour, died fish (13,3%) appeared at
concentration of 1.8 mg/L; 80% after 3 hours and 100% after 72 hours.
Value of LC
50
of quinalphos on common carp (size of 8.6 g) was determined
1.16, 0.76 and 0.76 mg/L after 24, 72 and 96 hours, respectively.
Table 4.9: Mortality of common carp exposed to quinalphos in 96 hours
Time
(hour)
Concentrations of quinalphos (mg/L)
LC
50
(mg/L)
0
0.2 0.5 0.8 1.1 1.5 1,8
1 0 0 0 0 0 0 13.3
3 0 0 0 0 0 23.3 83.3
24 0 0 13.3 26.7 36.7 56.7 90 1.16 (0.96-1.44)
72 0 0 23.3 53.3 70 93.3 100 0.76 (0.66-0.85)

96 0 0 23.3 53.3 70 93.3 100 0.76 (0.66-0.85)
Value of LC
50
-96 hrs. of quinalphos on tilapia (Oreochromis
niloticus) was 0.84 mg/L (Đo Van Buoc, 2010) and silver barb
(Barbonymus gonionotus) was 0.856 mg/L (Tran Thien Anh. 2011).
4.3 Effects of quinalphos on respiratory physiology
4.3.1 Oxygen consumption
In case of direct exposing, oxygen consumption significantly
increased with increased concentrations, ranged 382-518 mgO
2
/kg
fish/hour (p<0.05). In case of contamination 24 hours, oxygen
consumption tended to decrease but there was no significant
difference, ranged 356-371 mgO
2
/kg fish/hours (p>0.05).
4.3.2 Oxygen threshold
Oxygen threshold significantly increased with increased
concentrations in case of direct exposing, ranged 0.13-0.45 mg/L
(p<0.05). In case of contamination 24 hours, oxygen threshold
considerably increased with increased concentrations, ranged 0.13-0.65
11
mg/L (p<0.05).
Increased concentration of decamethrin increased oxygen
consumption of common carp (James et al., 1994). Murty (1988) and Đo
Thi Thanh Huong (1997) stated that oxygen consumption increased at the
beginning of exposing to insecticide because respiratory activity increased
but decreasing after.
4.4 Effects of quinalphos on ChE sensitivity and inhibition threshold

4.4.1 Environment parameters during the experiment
pH in the morning and afternoon was 6.8±0.1 and 6,9±0,1,
respectively. Temperature in the morning and afternoon was 25.4±0.1
o
C
and 26.9±0.1
o
C, respectively. Oxygen dissolve ranged 5.9-6.3 mg/L.
4.4.2 ChE sensitivity
The study showed that brain ChE activity decreased with increased
concentrations and time. At 0.0076 mg/L, ChE activity was significantly
inhibited (25%) as compared to the control (p<0.05). The most inhibited
ChE activity was recorded at 0.57 mg/L (91,6%). During the experiment,
ChE inhibition was 74.8% and there was no sign of complet recovery as
compared to the control after 96 hours (p<0.05).
4.4.3 ChE inhibition threshold
The present study reported that ChE inhibition affected swimming
behaviours such as imbalance (inhibited 82.8%), dull swimming
(91.8%) and died fish (95.1%).
Fleming and Grue (1981) reported that brain AChE activity
decreasing 20% as compared to the control considered as fish exposing
to organophosphorus insecticide. The relation between ChE inhibition
and dead fish was unknown because some species could live when brain
ChE inhibition was 90-95% (Fulton and Key, 2001; Ferrari et al., 2004).
ChE inhibition causing change of behaviours might affect to fish
survival in experimetal condition (Ferrari et al., 2007).
4.5 Effects of quinalphos on haematological parameters, ChE
activity, digestive enzymes and growth of common carp
4.5.1 Effects of quinalphos on haematological parameters and
enzyme activities of common carp

4.5.1.1 Environment parameters during the experiment
Temperature fluctuated from 25.6 to 26.1
o
C. Oxygen dissolve
ranged from 3.2 to 3.4 and pH was around 7.9.
4.5.1.2 Effects of quinalphos on haematological parameters
Erythrocyte (1.6-2 triệu/mm
3
), hematocrit (30.7-33,5%) at
concentration of 0,57 mg/L significantly decreased as compared to the
12
control (p<0.05), whereas MCV (148-200 fl), MCH (48-64 pg) and MCHC
(28-32%) significantly increased as compared to the control (p>0.05).
Hemoglobin (9.3-10 g/dL) didn’t change (p>0.05).
Erythrocyte decreased in common carp exposed to diazinon (Svobova
et al., 2001). Ali and Rani (2009) showed that MCH (39-59 pg) and MCHC
(29-50%) was increased after 45 days when Oreochromis niloticus
exposing to phosalone tended to increase based on concentration and time.
4.5.1.3 Effects of quinalphos on enzyme activities of common carp
a) Cholinesterase acticity (ChE)
The study presented that there was interation between concentration
and time on changes of brain, muscle and gill ChE activity.
- Brain: Brain ChE activity was the most inhibited organ after 4
days, level of inhibition at four concentrations ranged 89.0-94.1%.
After 21 days, ChE activity at 0.076 mg/L completely recovered. At
higher concentrations, ChE actitvity recovered so slowly.

Figure 4.1: Changes of brain ChE activity Figure 4.2: Brain ChE inhibition
- Muscle: After 1 day, ChE activity was reamarkably inhibited at
four concentrations, ranged 83-93% (p<0,05). After 28 days, muscle

ChE activity had a sign of incompletly recovery at all concentratons


Figure 4.3: Changes of muscle ChE activity Figure 4.4: Muscle ChE inhibition
a
a
a
a
a
a
e
e
a
a
a
e
e
cd
ab
a
e
e
cde
bc
ab
e
e
de
de
0

40
80
120
160
200
240
280
0 1 4 21 28
Experimental time (day)
nmol/min/mg protein
0 mg/L
0,076 mg/L
0,152 mg/L
0,38 mg/L
0,57 mg/L
0
20
40
60
80
100
1 4 21 28
Experimental time (day)
Brain ChE Inhibition (%)
0,076 mg/L
0,152 mg/L
0,38 mg/L
0,57 mg/L
ab
bc

abc
a
a
fgh
h
de
abc
gh
h
de
bc
h
h
defg
bc
h
h
def
0
20
40
60
80
100
120
140
0 1 4 28
Experimental time (day)
nmol/min/mg protein
0 mg/L

0,076 mg/L
0,152 mg/L
0,38 mg/L
0,57 mg/L
0
20
40
60
80
100
1 4 28
Experimental time (day)
Muscle ChE Inhibition (%)
0,076 mg/L
0,152 mg/L
0,38 mg/L
0,57 mg/L
13
- Gill: Gill ChE activity was inhibited at all concentrations and
ranged 84-92% after 4 days (p<0,05). After 21 days, ChE activity at
0.076 and 0.152 mg/L had a sign of complet recovery. After 28 days,
ChE activity at 0.38 and 0.57 mg/L incomletly recovered as compared to
the control (p<0.05).
Figure 4.5: Changes of gill ChE activity Figure 4.6: Gill ChE inhibition
- Liver: ChE inhibition increased with increased concentrations and
ranged 45-78%. After 1 day, liver ChE activity was inhibited 61,3% as
compared to the control. After 28 days, liver ChE activity had a sign of
recovery incompletly (p<0,05).
The study showed that after 1 day, at the lowest concentration of
0.076 mg/L, brain, muscle, gill and liver ChE inhibition were 88.1, 79.6;

77.5 and 61.3%, respectively. As a result, brain ChE inhibition was the
highest or brain was the most sensivtive organ of common carp exposed
to quinalphos.
b) Glutathione-S-transferase activity (GST)
- Brain: GST activity (140-212 nmol/min/mg protein) at 0.57
mg/L was significant difference as compared to other concentrations.
After 1 day, GST activity significantly increased as compared to the
control. GST activity completely recoverd after 14 days.
- Muscle: GST activity was no significant difference at all
concentrations and ranged 32-41 nmol/min/mg protein. After 28 days,
GST activity didn’t significantly changed (32-40 nmol/min/mg protein).
- Gill: GST activity was no change at all concentrations, ranged
101-122 nmol/min/mg protein. After 28 days, GST activity had no
significant difference (p>0.05).
ab
bcdef
bcdef
abc
abc
k
cdefg
abcd
abc
bcdef
k
hijk
k
bcdef
defgh
efghij

hijk
k
bcdef
0
5
10
15
20
25
30
0 4 21 28
Experimental time (day)
nmol/min/mg protein
0 mg/L
0,076 mg/L
0,152 mg/L
0,38 mg/L
0,57 mg/L
efghi
0,0
20,0
40,0
60,0
80,0
100,0
4 21 28
Experimental time (day)
Gill ChE Inhibition (%)
0,076 mg/L
0,152 mg/L

0,38 mg/L
0,57 mg/L
14
- Liver: GST activity was the highest and ranged 559-630
nmol/min/mg protein. There was no significant difference of GST
activity during the experiment.
c) Catalase activity (CAT)
The present study presented that brain and gill CAT activity had a
tendency to increase significantly; liver CAT was decreased at 0.57
mg/L as compared to the control (p<0.05). Meanwhile, quinalphos
didn’t affect to muscle CAT activity (p>0.05).
d) Lipid peroxidation activity (LPO)
- Brain: LPO activity at concentrations fluctuated 162-179 nmol/g
protein (p>0.05). After 4 days, brain LPO activity was significantly
decreased (p<0.05). From day 7 to ending the experiment, brain LPO
completely recoverd as compared to the control (p>0.05).
- Muscle: Muscle LPO activity at 0.57 mg/L significantly differed as
compared to other concentrations (p<0.05). After 21 days, muscle LPO
activity was sinificant differrence (p<0.05) and after 28 days, muscle LPO
had a sign of complet recovery as compared to the control (p>0.05).
- Gill: Gill LPO activity was significantly increased at 0.57 mg/L
as compared to other concentrations (p<0.05). After 4 days, gill LPO
was significantly increased and completely recoverd as compared to
the control after 28 days (p>0.05).
- Liver: After 7 days, liver LPO was significanly increased as
compared to the control (p<0.05). After 28 days, liver LPO had a sign
of complet recovery as compared to the control (p>0.05).
Tran Thien Anh (2012) reported that quinalphos considerably
decreased brain, muscle and gill of silver barb Barbonymus gonionotus. Đo
Van Buoc (2010) stated brain GST of tilapia Oreochromis niloticus

exposing to quinalphos, brain GST was no change at lower concentrationsin
in 96 hours. Pangasianodon hypophthalmus exposing to quinalphos
showed that brain and liver CAT activity increaed 71.5% and 11.9%,
respectively; muscle and gill CAT decreased 36.4% and 28%, respectively
(Nguyen Thi Que Tran, 2010). Durmaz et al. (2006) indicated that
Oreochromis niloticus exposing to diazinon (0.1, 1 and 2 ppm), gill LPO
was no significant change, whereas muscle LPO increased at day 15 and
30.
15
4.5.2 Effects of quinalphos on haematological parameters,
cholinesterase activity, digestive enzymes and growth
performances of common carp cultured in tank.
4.5.2.1 Environmental parameters and quinalphos residue
a) Environmental parameters during the experiment
Temperature in the morning and afternoon ranged 27.1-27.6
o
C and
28.1-28.4
o
C, respectively. Fluctuation of temperature daily ranged about
1
o
C. pH was from 6.7 to 7.1. Oxygen dissolve ranged 4.7-5.6 mg/L in
the morning and 4.8-5.6 mg/L in the afternoon.
b) Residue of quinalphos
After 30 days at one exposing time and 44 days at two exposing time,
quinalphos concentrations were under limit of determination (<LOD).
4.5.2.2 Effects of quinalphos on haematological parameters
- Erythrocyte: Number of erythrocyte tended to decrease with
increased concentrations but there was no significant change (p>0.05),

ranging 1.7-2 million cells/mm
3
. After 3 days, erythrocyte significantly
decreased as compared to the control (p>0,05). After 30 days, erythrocyte
had a tendency of complete recovery (p>0.05). At the second exposing
time, erythrocyte tended to decrease but there was no significant
difference as compared to the control (p>0.05).
- Hemoglobin: Hemoglobin content was no change at concentrations
(p>0.05). After 3 days, hemoglobin was decreased at both of two exposing to
insecticide but no significant change was recorded (p>0.05).
- Hematocrit: Hematocrit was significantly decreased at most of
concentrations as compared to the control (p<0.05). After 3 days,
hematocrit was significantly decreased as compared to the control
(p<0.05). Hematocrit completely recoverd as compared to the control
after 30 days (p>0.05).
- MCV: MCV (ranging 177-198 fl) tended to increase with
increased concentrations but there was no significant change as
compared to the control (p>0.05). During the expriment, MCV didn’t
significantly change (p>0.05), ranging 159-210 fl.
- MCH: MCH (41-56 pg) significantly increased with increased
concentrations as compared to the control (p<0.05). After 3 days,
MCH (62 pg) significantly increased and complete recovery as
compared to the control after 30 days (p>0.05).
16
- MCHC: MCHC (26-30 g/dL) was significantly increased at all
concentrations as compared to the control (p<0.05). After 3 days,
MCHC was no significant difference (p>0.05). After 6 hours at the second
exposing time, MCHC was significantly decreased (p<0.05). MCHC was
no complet recovery as compared to the control after 60 days (p<0.05).
4.5.2.3 Effects of quinalphos on ChE activity and digestive enzymes

activities of common carp
a) Cholinesterase activity (ChE)
The study indicated that there was interaction between concentration
and time on changes of brain, muscle, gill and liver ChE activity.
- Brain: After 6 hours, brain ChE activity was signicantly inhibited at
all concentrations as compared to the control (p<0.05), ranging 86.2-
91.4%. Brain ChE activity had a sign of complete recovery after 30 days
and 60 days (p>0.05).
- Muscle: Muscle ChE activity was signicantly inhibited at all
concentrations as compared to the control after 6 hours, ranging 83.5-
93%. After 30 days, muscle ChE activity completely recovered
(p>0.05). However, muscle ChE activity incompletely recovered as
compared to the control after 60 days (p<0.05).
- Gill: ChE activity was signicantly inhibited at all concentrations as
compared to the control after 6 hours, ranging 84.6-91.4%. Gill ChE
activity completely recovered after 30 days (p>0.05). Similarity in
muscle ChE, gill ChE activity incompletely recovered as compared to
the control after 60 days (p<0.05).
- Liver: Liver ChE activity was considerably inhibited at all
concentrations as compared to the control after 6 hours (p<0.05), ranging
81.6-85.4%. Similarity in brain ChE, liver ChE activity had a sign of
complete recovery at all concentrations after 30 and 60 days.
Banaee et al. (2008) reported common carp exposing to diazinon,
number of erythrocyte, hemoglobin content and hematocrit were
decreased. Boone and Chambers (1996) stated recovery of ChE
activity was different from a week to four weeks more.
Similarity in section 4.5.1.3, this experiment showed that after 6
hours, brain, muscle, gill and liver ChE inhibition at the lowest
concentration (0.076 mg/L) was 86.2, 83.5, 84.6 and 81.6%. Therefore, it
might be reported that brain ChE inhibition was the highest or brain was

the most sensivtive organ of common carp exposing quinalphos.
b) Digestive enzymes activities
17
- Trypsin: Trypsin activity (12-22 mU/min/mg protein) was
significatly inhibited 20-45% at all concentrations as compared to the
control (p<0.05). After 30 days, trypsine activity (21-28 mU/min/mg
protein) completely recovered as compared to the control (p>0.05).
However, trypsin activity (17-28 mU/min/mg protein) had a sign of
incomplete recovery after 60 days.
- Chymotrypsin: Chymotrypsin activity (881-1.058 mU/min/mg
protein) insignificantly decrease at all concentrations as compared to
the control (p>0.05). After 60 days, chymotrypsin activity was no
change, ranging 897-1.190 mU/min/mg protein (p>0.05).
- Alpha-amylase: Alpha-amylase activity (108-148 mU/phút/mg
protein) was significantly inhibited 21-27% at all concentrations as
compared to the control (p<0.05). After 6 hours at both of two exposing
time, alpha-amylase activity was significantly inhibited 48-50%.
Alpha-amylase activity incompletely recovered after 30 days and 60
days (p<0.05).
Nguyen Thi Kim Ha et al. (2012) realized that trypsin, chymotrypsin
and alpha-amylase activity of silver barb Barbonymus gonionotus
exposing to quinalphos was significantly inhibited after 6 hours. Tran
Thien Anh (2012) reported that quinalphos considerably affected to
growth parameters of silver barb Barbonymus gonionotus.
4.5.1.4 Growth, survival and FCR
Fish weight at the end of experiment (18.9 g/con), day weight gain
(DWG) (0.12 g/ngày) and specific growth rate (SGR) (1,26%/ngày) at
0.57 mg/L were the lowest and significant difference as compared to
the control (p<0.05).
Table 4.25: Growth of common carp

Nồng độ
(mg/L)
Wđ Wc
DWG
(g/ngày)
SGR
(%/ngày)
0 11.04±0.12
a
29.42±1.91
a
0.22±0.02
a
1.68±0.14
a
0.076 11.14±0.52
a
30.43±4.68
a
0.23±0.06
a
1.83±0.43
a
0.152 11.30±0.41
a
24.22±2.47
b
0.16±0.02
b
1.47±0.07

a
0.38 10.51±0.56
a
20.98±0.79
bc
0.15±0.02
b
1.55±0.16
a
0.57 10.52±0.36
a
18.89±1.60
c
0.12±0.01
b
1.26±0.06
a
Values represent mean ±standard deviation
Vaues in a column with the same letter (a, b, c) are not different to each other (p>0.05)
During 90 days, the highest survival rate was at the control (98.8%) and
significant difference as compared to other treatments (p<0.05), next to at
0.076 mg/L (89.2%), 0.152 mg/L (73.3%), 0.38 mg/L (67.5%) and the
18