CAMEL'S MILK AS AN IMMUNE IMPROVER AGAINST THE HYDATICYSTOSIS IN LABORATORY MICE

Basrah Journal of Veterinary Research,Vol.17, No.3,2018
Proceeding of 6th International Scientific Conference,College of Veterinary Medicine
University of Basrah,Iraq
346
CAMEL'S MILK AS AN IMMUNE IMPROVER AGAINST THE
HYDATICYSTOSIS IN LABORATORY MICE
*Bassad A. AL-Aboody; Enas Abdul Kareem Jabbar ; Shaymaa Z. Al-
Rumaidh, Sabah H. Enayah
*College of Science, University of Thi-Qar, Thiqar ,Iraq
Keyword :, Echinococcus granulosus, immune response , camel's milk
.
Corresponding Author:[email protected]
ABSTRACT
Hydaticystosis is one of the most common diseases in the world, and had a
zoonotic pattern ,surgical intervention is one of the most important treatment attempt,
despite the serious problems the patient experiences during surgery. The current study
were aimed to use camel's milk as an immune improver against this disease in
laboratory mice .Experimental study was conducted in department of biology ,
College of Science University of Thi-Qar, during the period from November 2015
till April 2016 so four groups of mice(Mus musculus) of Balb /c strain age (6- 8)
weeks were used as following .The first group was orally treated with 0.2 ml of
physiological solution for one week (Negative control ) .The second group was orally
treated with 0.2 ml of physiological solution for a week and then injected
intraperitoneally with 0.2 ml of protoscolices of Echinococcus granulosus (3000
protoscolex / ml ) ( Positive control ).The third group was orally treated with 0.2 ml
of camel's milk for a week and then injected intraperitoneally with 0.2 ml of
protoscolices (3000 protoscolex / ml ). The fourth group was orally treated with 0.2
ml of camel's milk for two week and then injected intraperitoneally with 0.2 ml of
protoscolices (3000 protoscolex / ml).
The results of the current study showed that the groups of animals treated with
camel's milk for one week and two weeks had a decrease in the number and weights
of secondary hydatid cysts and the group treated with camel's milk for two weeks
showed a significant decrease in the number and weights of cysts compared to the
Basrah Journal of Veterinary Research,Vol.17, No.3,2018
Proceeding of 6th International Scientific Conference,College of Veterinary Medicine
University of Basrah,Iraq
347
positive control group. A significant reduction in weights and size ratio of liver and
spleen was observed in the group treated with camel's milk for two weeks compared
with control group .
It has been concluded that camel's milk has the ability to reduce the incidence of
secondary hydatid cysts in infected mice and treated with camel's milk and the
increased duration of treated with camel's milk significantly reduces the incidence of
infection
INTRODUCTION
Human cystic echinococcosis or hydatidosisis is a zoonotic infection caused by
the larval stage of cestode Echinococcus granulosus [1].The disease is one of the most
common diseases in various parts of the world and is endemic in the countries of the
Middle East and parts of America, Australia, the Mediterranean, Central Asia and
Central and Eastern Europe [2].
The final host of these parasites is Canine(Canidae) which infected by the feeding of
the hydatid cyst from the intermediate host as a domesticated animal [3], while the
human is accidental or incidental host do not play a role in the continuation of the life
cycle [4].
Children are more susceptible to infection with this disease especial those whom close
association with dogs [5].In addition to the human health effects, this disease is still an
endemic disease in Iraq and the economic impact which prompted many studies to
investigate the means of treatment was surgical intervention is the most important
methods of treatment despite the serious problems experienced by the patient during
surgery, the patient is not surgically qualified due to age, anesthesia or cyst in places
where it is difficult for the surgeon to deal with it as in the brain cyst or in certain
cases such as age or in the case of pregnancy or the patient has severe diseases such as
heart disease, diabetes and hypertension [6]
The medicinal importance of camel's milk in the treatment of different diseases caused
by fungi, parasitic worms and intestinal , bacteria has been studied by many
researchers and its chemical properties have found that camel's milk contains a good
quality of proteins that are effective against bacteria and viruses such as Lactoferrin,
Lactoperoxidase, Lysozyme which makes it far superior to cow's milk in terms of
Basrah Journal of Veterinary Research,Vol.17, No.3,2018
Proceeding of 6th International Scientific Conference,College of Veterinary Medicine
University of Basrah,Iraq
348
nutrients [7; 8; 9 ]. Several studies conducted in different areas in Iraq were used
different materials such as medical plant to reduce the infectivity of infection and
enhance the immune response , [ 10 ] studied the effect of volatile oil to
Cymbopogon citrates with or without usage protoscolisces antigen against infection in
hydated cysts of Echinococcus granulosus. [11] they studied the effect of the aqueous
extract of the Allium sativum plant on the vitality of the protoscolisces of
Echinococcus granulosus .No one trialed to tested camel's milk therefore this study
was aimed to use camel's milk as immune improver against the infection of hydatid
disease in laboratory mice
MATERIAL AND METHODS
study design
Experimental study was conducted in department of biology , college of Science
,University of Thi-Qar, during the period from November 2015 till February 2016
Source of hydatid cysts
The liver hydatid cysts of sheep were obtained from the Nasiriyah massacre and
placed in a refrigerated case and transferred directly to the laboratory of the college of
Science at Thi- Qar University within 10-30 minutes to conduct the isolation
According to [12 ].
Source of camel's milk
Milk was obtained from camel group in SiddhKhel area east of Thi-Qar up to 25 km.
Milk samples were obtained by hand milking method. The milk was transferred
directly to the laboratory and kept in the refrigerator under 2-5 ° C until use.
Experimental design
Twenty eight of male mice (Mus musculus) of Balb /c strain age (6- 8) weeks and
weighted (25-27)gm by divided into four groups (7 males for each group ) as
following.:-
1-The first group was orally treated with 0.2 ml of physiological solution for one
Basrah Journal of Veterinary Research,Vol.17, No.3,2018
Proceeding of 6th International Scientific Conference,College of Veterinary Medicine
University of Basrah,Iraq
349
week (negative control )
2-The second group was orally treated with 0.2 ml of physiological solution for a
week and then injected intraperitoneally with 0.2 ml of protoscolices (3000
protoscolex / ml ) ( positive control ) (
3- The third group was orally treated with 0.2 ml of camel milk for a week and then
injected intraperitoneally by 0.2 ml of protoscolices (3000 protoscolex / ml ) (
4- The fourth group was orally treated with 0.2 ml of camel milk for two week and
then injected intraperitoneally by 0.2 ml of protoscolices (3000 protoscolex / ml).
On the day 90th of the treated with camel's milk and injections with protoscolices the
animals were dissected. The growth or non-growth of the hydatid cyst was
determined and their weights measured and determined changes in liver and spleen
weights and determination of their enlarged ratio.
Statistical analysis
SPSS program applied to find the Mean and SD with statistic significant by determine
p value>0.05 using 2-way ANOVA[13]
RESULT
Examination of secondary hydatid cyst
The results of the dissection of the groups of mice which treated with camel's milk
and infected with the protoscolices of the Echinococcus granulosus and 90 days after
the added It was noted that the second group( Positive control group) and the presence
of secondary hydatid cyst in the peritoneal cavity and distributed irregular and found
hydatid cyst on the liver is not implanted in it has been the average number of 7 cyst
of different sizes and a weight of 0.31 g the third group treated with camel milk for
one week decreased the number of secondary hydatid cyst compared to the positive
control group, the number of cysts in it to 4 and weight of 0.15 g. The fourth group
treated with camel milk for two weeks the number of hydatid cyst decreased
Basrah Journal of Veterinary Research,Vol.17, No.3,2018
Proceeding of 6th International Scientific Conference,College of Veterinary Medicine
University of Basrah,Iraq
350
significantly compared with the positive control group, where the number of cysts in
it to 0.4 and a weight of 0.036 g . As shown in Table (1)
Table (1) Changes in the numbers and weights of the hydatid cyst in study
groups
Weight of cyst / gm(meanSD)
P (value)
Number of cyst (meanSD)
P (value)
Group
Negative control 00 00
0.310.59
0.000*
71.58
0.000*
Positive control
0.150.051
0.002*
1.40.54
0.096
Milk 1 week
0.0360.049
0.62
0.40.54
0.88
Milk 2 week
* meaning statically significant comparing to the control P<0.05
Changes in liver weight and its size ratio in treated mice and compared with
positive and negative control
According to result of Table (2) it is show that the fourth group treated with camel's
milk for two weeks had the lowest weight in the liver and size ratio where the liver
weight in this group was 1.64 gm and the size ratio 55.18 and the differences were
significant between this group and the positive control group that recorded liver
weight and size ratio3.21gm and 90.99 Respectively ,the fourth group did not record
any significant differences from the negative control group, while the third group
treated with camel milk for one week showed a decrease in weight and size ratio liver
but less than in the fourth group.
Basrah Journal of Veterinary Research,Vol.17, No.3,2018
Proceeding of 6th International Scientific Conference,College of Veterinary Medicine
University of Basrah,Iraq
351
Table (2) Changes in liver weight and its size ratio in study groups
Liver size ratio( meanSD)
P (value)
Liver weight /gm( meanSD)
P (value)
Group
1.580.083 54.082.62
Negative control
90.9911.35
0.001*
3.210.318
0.003*
Positive control
62.913.22
0.252
1.940.114
0.04*
Milk 1 week
55.187.83
0.99
1.640.273
0.009*
Milk 2 week
* meaning statically significant comparing to the control P<0.05
Changes in spleen weight and its size ratio in treated mice and compared with
positive and negative control
The result of present study found that the groups treated with camel milk for one
week and two weeks showed less weight and enlarged ratio in the spleen where the
fourth group treated with camel milk for two weeks showed less weight of the spleen
0.11 g and the enlarged ratio 3.96 and the third group treated with camel milk for
one week was the weight of the spleen has 0.17 g and the enlarged ratio 5.63 .The
differences were significant between camel milk treatment groups and positive control
group, which recorded weight of spleen 0.25 g and enlarged ratio of 7.3,while no
significant differences noticed between fourth group and negative control group as shown in
Table (3).
Basrah Journal of Veterinary Research,Vol.17, No.3,2018
Proceeding of 6th International Scientific Conference,College of Veterinary Medicine
University of Basrah,Iraq
352
Table (3) Changes in spleen weight and its size ratio in treated mice and
compared with positive and negative control.
Spleen size ratio (meanSD)
P (value)
spleen weight/gm
(meanSD)
P (value)
Group
3.580.402
0.1050.011
Negative control
7.31.589
0.002*
0.250.40
0.001*
Positive control
5.630.577
0.39
0.170.151
0.006*
Milk 1 week
3.961.31
0.94
0.110.36
0.898
Milk 2 week
* meaning statically significant comparing to the control P<0.05
DISCUSSION
The results of the current study showed that the groups of animals treated with camel
milk for one week and two weeks have decreased the weights and number of hydatid
cysts compared with positive control ,this confirms the efficacy of camel's milk in
reducing the numbers and weights of the hydatid cysts. The study agreed with the
findings of [12 ] , noting the low numbers and weights of hydatid cysts in animals
infected with E. granulosus parasites and treated with milk and urine camel ,
compared to the positive control group ,also this study agreed with other studies on
the effect of camel's milk on other parasites. [14 ] concluded to prevent the infected of
Leishmania donovaniin animals treated with milk and urine camel, [15] concluded
that camel's milk with or without leishmania antigen could be used as immune
improver against infection of Leishmania donovani in male mice. This can be
attributed to a high immune response that has an effect on controlling the growth and
development of protoscolices to secondary hydatid cysts and eliminate them this
effect may be attributed to the properties possessed by camel's milk as it has the
inhibitory ability of different types of bacteria, viruses and parasites. This is because
the immune molecules of camel's milk are smaller than in humans and other animals,
so they can easily penetrate the protein layer around the parasite and weaken the vital
effectiveness [16] .
Basrah Journal of Veterinary Research,Vol.17, No.3,2018
Proceeding of 6th International Scientific Conference,College of Veterinary Medicine
University of Basrah,Iraq
353
In this study, there was a significant decrease in liver weights and its size ratio in
groups treated with camel's milk compared with positive control group , the decrease
in liver weights and its size ratio is due to the immunological efficacy of camel's milk
proteins of medical importance in the treatment of many diseases such as ascites, liver
disease and liver function improvement [17].The results of the reduction in liver
weight and its enlarged ratio agree with the findings of [18] where she noted the
apparent decrease in liver weights and its enlarged ratio in infected mice with
Irradiated protoscolices compared to the positive control mice, which is due to
reduced liver volume and hence the number of parasites. Thus reducing the granular
tumor .This study agree with the results of the study of [10], which showed a clear
decrease in liver weights and its enlarged ratio of infected animals and treatment with
volatile oil of Cymbopogon citrates plant by different concentrations of oil compared
to positive control animals, which observed a large weight of the liver and high
enlarged ratio. The increase in liver weight may be attributed to the intensity of the
inflammatory reaction[12] , as well as the increase in the number of lymphocytes as
a result of their division and secretion of the cellular motor factors. The colony
stimulating factor (csf ) that secrete the macrophages stimulates their reproduction
[19]
The current study shows a decrease in the rates of spleen weight and its size ratio
among the groups treated with camel milk compared to the positive control group,
which observed large enlarged of the spleen ,changes in the weight of the spleen of
infected mice are caused by the increase in the production of T lymphocytes and B
lymphocytes [20] the study agrees with the findings of [21] that the weight of the
spleen increases and amplifies its size in the group of mice infected with E.
multilcularis due to the response of the spleen cells to the parasite and to the
associated bacterial infections and lymphocytic infiltration as an immunological
necessity .The decrease in spleen weights may be due to the chemical properties of
camel milk for its contents of immune proteins such as lyzosome , lactoferrin and
others [22].
It has been concluded that camel's milk has the ability to reduce the incidence of
secondary hydatid cysts in infected mice and treatment with camel's milk and the
increased duration of treatment with camel's milk significantly reduces the incidence
of infection . Therefore, the current study recommends the completion of research and
Basrah Journal of Veterinary Research,Vol.17, No.3,2018
Proceeding of 6th International Scientific Conference,College of Veterinary Medicine
University of Basrah,Iraq
354
research studies on the use of camel's milk as immune improver against other parasitic
infections and study the effect of camel's milk in the treatment of various parasitic
infections
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