Basrah Journal of Veterinary Research,Vol.17, No.3,2018
Proceeding of 6th International Scientific Conference,College of Veterinary Medicine
University of Basrah,Iraq
738
STUDY THE ROLE OF TITANIUM DIOXIDE (TIO2) NANOPARTICLES
AGAINST THE TOXICITY OF Echinococcus granulosus IN ADULT
ALBINO MALE RATS
Abeer A. Ali*, Shahad Abbas Alattar**
*Technical college of Kirkuk,University North Technical ,Kirkuk, Iraq
**College of Veterinary Medicine university of Kirkuk , Kirkuk, Iraq
Keywords: Tio2 nanoparticles; MDA (malonedialdehyied); glutathione (GSH).
Corresponding Author: ahmedeagle72@gmail.com
ABSTRACT
The present study was designed to show the role of green Tio2 nanoparticles against toxicity
of Echinococcus granulosus. The present study used 16 adult albino male rats that distributed
randomly to following groups (each group consist 5 rats); control group received ad libidium,
second group injected with 2,5 X 103 of Echinococcus granulosus protoscolices third group
injected with protoscolices and treated with 50mg/kg green Tio2 nanoparticles, fourth group
injected with protoscolices and treated with 100mg/kg green Tio2 nanoparticles. The results
show high significant increased (P < 0.05) in levels of Alanine Aminotransferase (ALT),
Aspartate Aminotransferase (AST), Alkaline Phosphatase (ALP) in group injected with
protoscolices compared with control group. Oxidative stress factor in group injected with
protoscolices show significant increased (P < 0.05) in levels of MDA (malonedialdehyied) and
significant decreased (P < 0.05) in levels of glutathione (GSH) and catalase compared with
control group. While, after used green Tio2 nanoparticles with Echinococcus granulosus, the
results showed non-significant changes (P < 0.05) in liver functions and MDA, GSH and catalase
also showed non-significant changes (P < 0.05) compared with control group. It was concluded
that green Tio2 nanoparticles has been potential role against toxicity of Echinococcus granulosus
in male rats.
Basrah Journal of Veterinary Research,Vol.17, No.3,2018
Proceeding of 6th International Scientific Conference,College of Veterinary Medicine
University of Basrah,Iraq
739
INTRODUCTION
Titanium dioxide (TiO2) Nanoparticles possess interest in a different applications such as gas
sensor, nanomedicine, photocatalyst, dye-sensitized solar cells [1-4] and antibacterial agents [5].
Titanium dioxide (TiO2) Nanoparticles are synthesis and produced by a different techniques
ranging from simple methods including mechanical, chemical and vacuum methods, including
variants of chemical and physical vapor deposition methods [6], or also by biological synthesis
[7]. Eruca sativa (Rocket) is important a medicinal plant with many different properties,
including strong aphrodisiac effects [8]. Several studies found that the presence of vitamin C,
carotenoids., flavonoids including luteolin, appiin, and glucosinolates the precursors of
sulfaraphene and isothiocyanates [9-10] and volatile oils like phellandrene , apiole and myristicin
[11-12] Echinococcus parasite are responsible of hydatid cyst (HC). It is a
parasite which causes a lot of economic and social problems [13]. Theinfection is made by
eating parasite eggs through the mouth.
Potoscoleces phaseare the infective stage of the Echinococcus spp. in definitive host, but they
are also able to differentiate asexually into secondary cysts are released in host
by accidental rupture of the primary cyst [14].
MATERIALS AND METHODS
Animal model
In this study twenty adult male albino rats, (wt 200-250 gm with age5-8 month) obtained
from Technical college/ North Technical University, and kept on standard pellet diet for two
weeks to insure its normal and there aren’t any infection.
Plant Extraction
leafs of Eruca sativa were obtain from local market. Then, fine powder was obtain and then
extracted with ethanol using sahxulate apparatus. The extracts of the leaf was evaporated to
dryness in a rotary evaporator at 40ºC. T. The filtrate autoclaved at 121ºC and 15 lb pressure for
20 min.[15].
Basrah Journal of Veterinary Research,Vol.17, No.3,2018
Proceeding of 6th International Scientific Conference,College of Veterinary Medicine
University of Basrah,Iraq
740
TiO2 nanoparticles synthesis
To prepared and synthesis TiO2 nanoparticles, dissolve 1.0 N of Titanium Chloride (TiCl4)
in 100 ml of distal water. Then, added leaves extract . The mixture was leaved to stirring for 4
hours. In this process TiO2 nanoparticles were formed, then separate these nanoparticles using
filter paper and washed the materials with water to remove the by-products. The nanoparticles
were dried at 100°Cfor overnight and storage until using [16].
Echinococcus granulosus
hydatid cysts were collected and obtained from infected sheep livers. They were put in
plastic bags, and transported to the Department of Microbiology , College Technical , North
Technical University, where protoscolices were isolated from livers according to [17] method .
Protoscolices indicates the fertility of hydatid cyst and it’s were counted according to method
cited by [18]. The viable protoscolices for parasite were counted in 1ml from supernatal based on
the formula : Viability in 1 ml = number of protoscolices in (10 μl) × 100.
Experimental design
sixteen adult male albino rats were used and divided as follow (each group consist four rats):
I. Rats were received standard pellet diet only for seven days and then killed.
II. Rats injected with protoscolices, and then killed.
III. Rats injected with protoscolices and treated with 50mg/kg TiO2 NPs for month, and then
killed.
IV. Rats injected with protoscolices and treated with 100mg/kg TiO2 NPs for month, and then
killed.
Prepare of blood solution
The blood collecting from rats by cardiac puncher, under anesthesia, and put in test tubs.
After clotting, the tubes were centrifugation for 10 min to obtain sera. The serum was taken and
stored by deep freezing until used.
Basrah Journal of Veterinary Research,Vol.17, No.3,2018
Proceeding of 6th International Scientific Conference,College of Veterinary Medicine
University of Basrah,Iraq
741
Homogenization
liver samples were removed immediately and the put in glass dish contents 0.9% NaCl buffer
for washing and removed the blood. To oxidative stress factors determination, 10% from organ
weight was dissolved with buffer (PH 7.4) and the organ tissue was crashed by use ceramic
mortar. Then mixture was centerfigation for 10 min. Supernated was taken and stored by deep
freezing until used [19].
Measurements
ALT, AST & ALP
Serum ALT, AST & ALP were measured by technique according to the instructions of
manufacturer company kit (Randox).
Plasma Peroxidation levels (MDA)
MDA (malonedialdehyied), was measured based on the colorimetric reaction with
thiobarbituric acid (TBA) using spectrophotometer [20].
Glutathione (GSH) and Catalase
GSH level estimated bymixed 2.3 ml buffer with 0.2ml of the sample and then added 0.5ml of
5,5-dithio-bis-(2-nitrobenzoic acid) (DTNB). The mixture was analyzed by spectrophotometer
[21]. Catalase was measured by using the procedure of Biovision-USA kits.
Statistical analysis
The Data were analyzed using a statistical Minitab program. A statistical difference between
the means of the experimental groups was analyzed using one way analysis of variance
(ANOVA).
Basrah Journal of Veterinary Research,Vol.17, No.3,2018
Proceeding of 6th International Scientific Conference,College of Veterinary Medicine
University of Basrah,Iraq
742
RESULTS
Liver function tests
ALT levels (72.46 ± 5.18), AST (81.24 ± 6.29) and ALP (95.3 ± 6.12) in rats injected with
protoscolices show high significant increased (P < 0.05) compared with control rats (17.4 ± 2.15,
19.45 ± 2.29 and 57.8 ± 3.32 respectively). The levels of ALT (29.18 ± 1.33), AST (28.21 ±
3.45) and ALP (68 ± 4.52) in rats injected with protoscolices and received 50mg/kg TiO2 NPs
show high significant increased (P < 0.05) compared with control rats. While, the levels of ALT
(15.34 ± 4.23), AST (20.2 ± 1.73) and ALP (68.75 ± 5.3) in rats injected with protoscolices and
received 100mg/kg TiO2 NPs show no significant decreased (P < 0.05) compared with control
rats as shown in table (1).
Table (1): The levels of ALT, AST and ALP in serum
Parameters
Groups
ALT (IU/L) AST (IU/L) ALP (IU/L)
I 17.4 ± 2.15 c 19.45 ± 2.29 c 57.8 ± 3.32 c
II 72.46 ± 5.18 a 81.24 ± 6.29 a 95.3 ± 6.12 a
III 29.18 ± 1.33 b 28.21 ± 3.45 b 68 ± 4.52 b
IV 15.34 ± 4.23 c 20.2 ± 1.73 c 68.75 ± 5.3 c
Note: same letters mean non-significant changes and different letters mean significant changes.
Basrah Journal of Veterinary Research,Vol.17, No.3,2018
Proceeding of 6th International Scientific Conference,College of Veterinary Medicine
University of Basrah,Iraq
743
Oxidative stress parameters (MDA, GSH and catalase)
The levels of MDA (2.34 ± 0.13), GSH (0.307 ± 0.016) and catalase (1.12 ± 0.1) in rats
injected with protoscolices show high significant changes (P < 0.05) compared with control rats
(1.65 ± 0.21, 0.454 ± 0.031 and 1.12 ± 0.1 respectively). The levels of MDA (1.76 ± 0.21), GSH
(0.423 ± 0.034) and catalase (0.83 ± 0.09) in rats injected with protoscolices and received
50mg/kg TiO2 NPs show high significant changes (P < 0.05) compared with control rats. While,
the levels of MDA (1.51 ± 0.09), GSH (0.432 ± 0.056) and catalase (1.08 ± 0.07) in rats injected
with protoscolices and received 100mg/kg TiO2 NPs show no significant changes (P < 0.05)
compared with control rats as shown in table (3).
Table (3): The levels of MDA, GSH and catalase in serum
Parameters
Groups
MDA (mmol/l) GSH (mol/l) Cata (mmol/l)
I 1.65 ± 0.21 b 0.454 ± 0.031 a 1.12 ± 0.1 a
II 2.34 ± 0.13 a 0.307 ± 0.016 b 0.61 ± 0.04 c
III 1.76 ± 0.21 b 0.423 ± 0.034 a 0.83 ± 0.09 b
IV 1.51 ± 0.09 b 0.432 ± 0.056 a 1.08 ± 0.07 a
In liver
The levels of MDA (1.33 ± 0.12), GSH (0.13 ± 0.03) and catalase (0.37 ± 0.01) in rats
injected with protoscolices show high significant changes (P < 0.05) compared with control rats
(0.87 ± 0.09, 0.23 ± 0.05 and 0.67 ± 0.03 respectively). The levels of MDA (1.09 ± 0.02), GSH
(0.18 ± 0.02) and catalase (0.54 ± 0.08) in rats injected with protoscolices and received 50mg/kg
TiO2 NPs show high significant changes (P < 0.05) compared with control rats. While, the levels
of MDA (0.91 ± 0.04), GSH (0.24 ± 0.08) and catalase (0.63 ± 0.07) in rats injected with
Basrah Journal of Veterinary Research,Vol.17, No.3,2018
Proceeding of 6th International Scientific Conference,College of Veterinary Medicine
University of Basrah,Iraq
744
protoscolices and received 100mg/kg TiO2 NPs show no significant changes (P < 0.05)
compared with control rats as shown in table (4).
Table (4): The levels of MDA, GSH and catalase in liver
Parameters
Groups
MDA (mmol/l) GSH (mol/l) Cata (mmol/l)
I 0.87 ± 0.09 c 0.23 ± 0.05 a 0.67 ± 0.03 a
II 1.33 ± 0.12 a 0.13 ± 0.03 c 0.37 ± 0.01 c
III 1.09 ± 0.02 b 0.18 ± 0.02 b
0
.54 ± 0.08 b
IV 0.91 ± 0.04 c 0.24 ± 0.08 a 0.63 ± 0.07 a
DISCUSSION
The result of present study as previously showed increased in the levels of liver functions
when rats administrated with Echinococcus granulosus protoscolices, same finding was reported
by Khudhair et al (2014) who referred that Echinococcus granulosus lead to elevated liver
function parameters [22]. Also the results of present study may back to the toxic effects of
Echinococcus granulosus protoscolices on liver tissue, where its lead to lymphocytes infiltration,
degenerative changes, necrosis of hepatocytes and congestions [23]. About oxidative stress and
antioxidant factors, the results showed decreased in levels of antioxidant factors and increased in
MDA levels in rats administrated with Echinococcus granulosus protoscolices, these results is in
agreement with who found that the glutathione levels in patients with Echinococcus granulosus
show significant decreased compare with healthy subjects [24]. Also, Koltas et al. (2006)
referred that the infection with Echinococcus granulosus lead to eleveated the levels of MDA,
Basrah Journal of Veterinary Research,Vol.17, No.3,2018
Proceeding of 6th International Scientific Conference,College of Veterinary Medicine
University of Basrah,Iraq
745
where found the mean +/- SD of MDA levels of patients with Echinococcus granulosus [25] that
is in agreement with results of present study. About the green nanoparticles, Biosynthesis of
nanoparticles by a green method has more advantages over physical and chemical methods as
well as it is cost effective and eco-friendly and does not require use of pressure, temperature,
toxic chemicals and energy.. The nanoparticles synthesis process has diverse applications in the
field of healthcare, medicine, electronics [26]. the results of present study show the ability of
AgNPs as antimicrobial agent.
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