Download Comparative Estimation of Total Protein Content and Enzymatic

2nd International Conference on Ecological, Environmental and Biological Sciences (EEBS'2012) Oct. 13-14, 2012 Bali (Indonesia)
Comparative Estimation of Total Protein
Content and Enzymatic Activities of Hydatid
Cyst of Ecchinococcus granulosus Isolated from
Sheep and Goats in Duhok Province, Kurdistan
Region of Iraq
Wijdan M.S. Mero and Arshad Mohammad Abdullah
become infected accidentally by ingesting food, drink or other
materials contaminated with the egg of this parasite[2]. There
are 4 known species of Echinococcus, namely: Echinococcus
granulosus, Echinococcus multilocularis, Echinococcus vogeli
and Echinococcus oligarthrus. [9]. Three of these species are
of medical importance, Echinococcus granulosus, causing
cystic echinococcosis (CE) which is the most prevalent
species in all continents, causing a considerable public health
problems in many regions of the world [2]. Furthermore, it is
also common in Iraq [1]. E. multilocularis, causing alveolar
echinococcosis (AE) is more widely distributed in the
northern hemisphere, and represents a considerable public
health burden as it’s the most virulent species and infection
with it is lethal in most untreated patients. However,
biochemical studies are also useful in differentiating strain
variations of E.granulosus in different countries [11]. The
present study was designed to evaluate the Some biochemical
studies on the parasite and its intermediate hosts, which
included the activity of some key enzymes and total protein
contents. These findings will give us some information about
the strain identification and treatment of hydatid disease.
Abstract—These This study included comparative Enzymatic
and total protein contents of hydatid cyst of Ecchinococcus
granulosus isolated from liver and lungs of infected sheep and goats
in Duhok abattoirs. With respect to enzymes the activity of the
following enzymes were measured namely acid phosphatases (ACP),
alcaline phosphatase (ALP), lactate dehydrogenase (LDH), glutamate
oxaloacetate transminase (GOT) and glutamate Pyruvate transminase
(GPT) in cysts isolated from both liver and lungs of infected sheep
and goats. Regarding protein content of protoscolices was higher
than hydatid cyst fluid, but it was lower than infected host
tissues.The activities of all these enzymes were higher in prtoscolices
as compared with their activities in hydatid fluid, but in general
infected host tissue showed the highest enzymatic activities as
compared with hydatid cyst.
II. MATERIALS AND METHODS
For this study a total of 36 hydatid cysts (Fluid and tissue of
infected organs) of sheep and goats in addition to 8 samples of
organs (liver and lung) from non infected animals (sheep and
goat) were collected to be used as a control. In this study we
used total protein kit and some enzyme kits acid phosphatases
(ACP), alcaline phosphatase (ALP), lactate dehydrogenase
(LDH), glutamate oxaloacetate transminase (GOT) and
glutamate Pyruvate transminase (GPT) and some Buffer
systems (Citrate buffer, Carbonate-Bicarbonate buffer and
Tris-HCl buffer) for homogenization of tissue. Hydatid cyst
fluids were taken in a special test tubes (5-8 mL), separated by
centrifugation at 4000 rpm for 30 minutes and the
supernatants was separated and stored in deep freezer at 40˚C
until used. The protoscolices separated by
centrifugation of hydatid cyst fluid then homogenization of
protoscolices and centrifugation of extract at 4000 rpm for 30
Fig. 1 Hydatid cysts and infected organs
Keywords— ACP, ALP, Ecchinococcus granulosus, GOT, GPT.
I. INTRODUCTION
YDATID disease (hydatidosis or echinococcosis) is a
silent cyclozoonotic infection of man and domestic
animals, caused by the larval stage of Echinococcus
granulosus (small dog tapeworm). Man and domestic animals
H
Arshad Mohammad Abdullah, M.Sc. in Biology/ Parasitology, University
of Duhok, phone: 009647504572331, e-mail: [email protected].
Prof. Dr.Wijdan M.S. Mero, Professor of Parasitology, Head of Biology
School, Faculty of Science,University of Zakho,
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2nd International Conference on Ecological, Environmental and Biological Sciences (EEBS'2012) Oct. 13-14, 2012 Bali (Indonesia)
minutes and the supernatants were collected and stored in a
deep freezer at -40˚C till used. The liver and lung tissues of
infected and non infected animals
homogenized with a
desired buffer, the supernatant was collected and stored in
deep freezer at -40˚C till used [8]. Hydatid cyst fluid and
extracts of protoscolices and host tissues were used for
determination of total proteins and enzyme activities.
enzyme in high activity in both hydatid fluids and
protoscolices of E. granulosus as well as infected host tissue.
Similar high activities of ACP enzyme was observed by
McManus and Bryant (1995) [6] in sheep hydatid cysts of E.
granulosus, Sherif et al., (1984) [17], they reported it in
hydatid fluid.
TABLE2
ACTIVITY OF ACP ENZYME
III. RESULTS AND DISCUSSION
A. Total protein content of hydatid cyst isolated from sheep
and goats liver and lung: Table (1).
Similarly Smyth (1979) [18], reported lower protein content
in infected host tissue, and this low protein content was due to
its use by the parasite, since the protoscolices of E. granulosus
can digest host proteins. Meduri et al., (1990) [7], also
reported low protein content in hepatic and pulmonary hydatid
cysts of infected cattle which were 0.2 mg/100ml,
0.09mg/100ml respectively, these values were much lower
than the values reported in the present study. On the other
hand, the results of the present study disagree with those
reported by Frayha and Haddad (1980) [3]; Ul-Haq (1994)
[21] and Radfar and Iranyar (2004) [11], who stated that
hydatid fluid and protoscolices of sheep liver hydatid cysts
contain high concentration of protein.
TABLE 1
TOTAL PROTEIN
Specimens
Hydatid
Fluids
( g/dL )
Protoscolices
(IU/100g)
Infected
Organ
(IU/100g)
Control
Organ
(IU/100g)
Sheep Liver
M ± SEM
Sheep Lung
M ± SEM
Goats Liver
M ± SEM
Goats Lung
M ± SEM
1.600 ±
0.084
N=7
2.086 ±
0.265
N=7
1.840 ±
0.13
N=5
2.386 ±
0.143
N=7
2.186 ±
0.135
N=7
2.229±0.18
7
N=7
1.925 ±
0.2810
N=4
±
2.557 ±
0.146
N=7
2.420 ±
0.1463
N=5
2.050 ±
0.1708
N=4
2.904 ±
0.029
N=2
2.705 ±
0.033
N=2
2.503 ±
0.43
N=2
2.206 ±
0.128
N=2
1.557 ±
0.036
N=7
2.757
0.187
N=7
TABLE 3
ACTIVITY OF ALP ENZYME
B. ACP activity in hydatid cyst isolated from sheep and
goats liver and lung: Table (2).
ACP is an important enzyme and is found in Liver and
other organs [19]. McManus and Barrett (1985) [5], reported
the presence of ACP enzyme in the sheep liver protoscolices.
The results of the present study revealed the presence of this
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2nd International Conference on Ecological, Environmental and Biological Sciences (EEBS'2012) Oct. 13-14, 2012 Bali (Indonesia)
TABLE 4
ACTIVITY OF LDH ENZYME
TABLE 5
ACTIVITY OF GPT ENZYME
Specimens
Sheep Liver
M ± SEM
Sheep Lung
M ± SEM
Goats Liver
M ± SEM
Goats Lung
M ± SEM
Specimens
Sheep Liver
M ± SEM
Sheep Lung
M ± SEM
Goats Liver
M ± SEM
Goats Lung
M ± SEM
Hydatid
Fluids
(IU/L)
583.7 ±
21.70
N=6
705.6
±20.25
N=7
722.8 ±
4.176
N=5
749.3 ±
41.29
N=6
Hydatid
Fluids
(IU/L)
93.17 ±
2.393
N=5
108.0 ±
5.007
N=5
103.5 ±
3.08
N=7
107.0 ±
6.484
N=6
Protoscolices
(IU/100g)
645.5 ±
33.70
N=4
707.3
±30.06
N=7
740.5 ±
34.38
N=6
751.1
±20.61
N=7
Protoscolices
(IU/100g)
100.5 ±
4.786
N=6
112.2 ±
8.801
N=5
107.7 ±
6.53
N=7
109.9 ±
5.870
N=6
Infected
Organ
(IU/100g)
735.5 ±
16.61
N=6
732.1±
10.67
N=7
889.6 ±
37.82
N=5
896.8 ±
48.13
N=4
Infected
Organ
(IU/100g)
106.9 ±
7.951
N=7
114.0 ±
12.73
N=7
109.8 ±
8.68
N=7
113.2 ±
10.94
N=7
442.4 ±
2.568
N=2
453.2 ±
1.843
N=2
430.7 ±
0.391
N=2
427.1 ±
0.948
N=2
84.59 ±
1.359
N=2
79.74 ±
0.172
N=2
76.92 ±
0.931
N=2
67.65 ±
1.508
N=2
Control
Organ
(IU/100g)
Control
Organ
(IU/100g)
increases dramatically in acute liver damage and in infected
host tissue with echinococcusis [24, 10]. In this study high
GPT activity was observed in hydatid cyst and infected tissue.
results in were reported by Sanchez and Sanchez, (1971)
[13]; Frayha and Haddad (1980) [3]; McManus and Bryant
(1995) [6] ; Zeheer (1997) [24]. Furthermore Zeheer (1997)
[24], reported high GPT activity in hydatid cysts wall also.
C. ALP activity in hydatid cyst isolated from sheep and
goats liver and lung: Table (3).
ALP is an enzyme which plays an important role in the
metabolism, physiology, immunology and nutrient absorption
of the cystic echinococcosis [4, 20, 15]. Frayha and Haddad
(1980) [3], reported a higher activity of ALP enzyme in
protoscolices as compared with hydatid cyst fluid.
The higher activity of ALP in hydatid cysts (both
protoscolices and hydatid fluid) than that of uninfected host
tissue, is in agreement with the finding of Sarciron et al.,
(1991) [14], who also reported higher activity of this enzyme
in hydatid cyst as compared to control host liver.
F. GOT activity in hydatid cyst isolated from sheep and
goats liver and lung: Table (6).
The enzyme glutamate oxaloacetate transminase (GOT) is
distributed in all body tissue with a high activity in liver
parenchymal cells, and its level increases during liver damage
[24, 19, 22]. In the present study, high GOT activity was
observed in hydatid cysts(hydatid cyst fluid and protoscolices)
isolated from sheep and goats liver and lung as well as
infected sheep and goats liver and lung tissue. These results
are similar to what have been reported by Sanchez and
Sanchez (1971) [13]; Frayha and Haddad (1980) [3];
McManus and Bryant (1995) [6] ; Zeheer (1997) [24], who
also detected high GOT activity in sheep hydatid cyst fluids,
protoscolices, liver and hydatid cysts wall.
D. LDH activity in hydatid cyst isolated from sheep and
goats liver and lung: Table (4).
LDH is an enzyme that catalyzes the conversion of lactate to
pyruvate, it is present in large amounts in heart, kidney, liver,
and other body organs [16]. The higher activity in sheep
protoscolices as compared with the hydatid fluid LDH in this
study is in agreement with the finding of Frayha and Haddad
(1980) [3] and Zeheer (1997) [24], while McManus and
Bryant, (1995) [6], Rouhani and Vatankhah (2008) [12],
reported high LDH in hydatid fluid of sheep.
E. GPT activity in hydatid cyst isolated from sheep and
goats liver and lung: Table (5).
Glutamate Pyruvate transminase (GPT) is an enzyme present
in hepatocytes (liver cells), kidney and other body organs, it is
important for metabolism of amino acid in krebs cycle and
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2nd International Conference on Ecological, Environmental and Biological Sciences (EEBS'2012) Oct. 13-14, 2012 Bali (Indonesia)
TABLE 6
ACTIVITY OF GOT ENZYME
Specimens
Sheep Liver
M ± SEM
Sheep Lung
M ± SEM
Goats Liver
M ± SEM
Goats Lung
M ± SEM
Hydatid
Fluids
(IU/L)
90.03 ±
2.78
N=7
98.69 ±
3.91
N=7
73.93 ±
5.316
N=4
100.9 ±
4.04
N=5
Protoscolices
(IU/100g)
94.05 ±
7.16
N=7
114.5 ±
12.87
N=7
81.78 ±
4.566
N=6
103.3 ±
4.98
N=6
Infected
Organ
(IU/100g)
105.2 ±
6.12
N=7
125.4 ±
12.71
N=7
94.55 ±
4.203
N=6
115.5 ±
4.796
N=7
63.46 ±
1.151
N=2
71.41 ±
1.04
N=2
54.47 ±
0.359
N=2
57.48 ±
0.982
N=2
Control
Organ
(IU/100g)
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