Basrah Journal of Veterinary Research,Vol.17, No.3,2018
Proceeding of 6th International Scientific Conference,College of Veterinary Medicine
University of Basrah,Iraq
621
EVALUATION OF HISTOPATHOLOGICAL CHANGES OF
HYDATID CYST ON KIDNEY OF EXPERIMENTALLY
INFECTED MICE
* Amal KH. Khalaf and ** Salih K. Majeed
*Department of microbiology , College of Medicine, University of Thi-Qar
Thi-Qar,Iraq.
** college of Veterinary Medicinem,University of Basrah,Basrah,Iraq
Keywords: Hydatid cyst,kidney,mice.
Corresponding Author: amalkhudair111@yahoo.com
ABSTRACT
Hydatidosis is an endemic disease in Iraq till now and it causes highly economic
looses . the aim of the following study was to investigate the histopathological effect
of hydatid cyst on kidney function .
11% total percentage were recorded among kidney of experimentally infected male
mice with hydatid cyst in the current study and many pathological changes were also
reported.
The present study has explained that the infection with hydatid disease affect on the
kidney function after examination of Blood urea nitrogen that related to kidney
function and reporting increase of its level .
INTRODUCTION
Hydatid disease, hydatidosis, cystic echinococcosis, unilocular hydatid disease , E.
granulosus Echinococcosis , and Al - akyas al-mai'yah' and 'al atash' all terms
describing infections which are caused by cestodes of the genus Echinococcus
particularly E. granulosus (1 ). The organ most commonly involved is the liver (50-
70%), followed by lungs (20-30%) and other organs (like the spleen, kidney, heart,
bones, central nervous system etc.) in less than 10% of the cases (16).
The hydatid cyst remains a significant public health hazard in endemic areas such
as Iraq , Turkey, the Middle East, South America, New Zealand , Africa , China,
northern Kenya, Australia, and other sheep-raising areas ( 1 ) . As an endemic disease,
it causes social and economic losses for countries. WHO reports stated that
approximately 100,000 people in the world are infected with this disease every year (
10) which is common in rural populations of underdeveloped countries because of
their close association with domestic and wild animals (11,12).
Basrah Journal of Veterinary Research,Vol.17, No.3,2018
Proceeding of 6th International Scientific Conference,College of Veterinary Medicine
University of Basrah,Iraq
622
MATERIALS AND METHODS
Parasite Materials and Protoscolices Preparation
Fresh hydatid cysts were obtained from livers and lungs of naturally-infected sheep,
which had been slaughtered at local abattoirs in Basrah city, human hydatid cysts
were obtained from Al- Sadir teaching hospital in Basrah city. They were wrapped
carefully in clean plastic bags, placed in an ice box, and transported to the Department
of Biology, College of Education, Basrah University, where protoscolices were
isolated according to (14) method . Protoscolices were counted according to method
cited by (2). The viable protoscolices were counted in 1ml based on the formula :
Viability in 1 ml = number of protoscolices in (10 μl) × 100
Eight male of Mus musculus mice Balb\C strain were injected with 0.2 ml 480/ ml
(2400/5ml rate of viability) of protoscolices intraperitoneally and consider as positive
group and left for six months. also, negative control group were included in the study
and involve eight of uninfected male mice.
The Study of the effect of infection on blood urea nitrogen (BUN):
Blood samples were obtained from the heart of each animal after anesthesia using
1ml volume syringe. The samples were collected in vials without any anticoagulant
for the determination blood urea nitrogen (BUN) . blood urea nitrogen (BUN) was
tested by the spectrophotometer using a suitable kit for each of BUN based on (13)
methods as follows:
Blood Urea Nitrogen Test (BUN)
BUN was used to determine the functional status of the kidney and it was
measured by using a special kit (Biomerieux \ France) as follow :
Hydatid cysts after removal from liver Germinal layer containing protoscolices
removed from hydatid cyst
Basrah Journal of Veterinary Research,Vol.17, No.3,2018
Proceeding of 6th International Scientific Conference,College of Veterinary Medicine
University of Basrah,Iraq
623
Test Principle :
Urea + H2O 2NH3 + CO2
NH4 + Salycilate + NaClO indophenol + NaCl
Test Procedure
Solution Blank Standard Sample
Working reagent 1000 μl 1000 μl 1000 μl
Sample ‾ - 10 μl
Standard ‾ 10 μl -
The tubes were mixed and incubated 5 minutes at 37.
NaoH 1000 μl 1000 μl 1000 μl
The tubes were mixed and incubated for 5 minutes at 37 . Within 60 minutes the
absorbance of sample was read against the reagent blank by the spectrophotometer at
a 600 nm wave length.
Calculation
ΔA Sample
× Standard concentration = urea concentration
ΔA Standard
Standard concentration: 50 mg/dl.
Histopathological study
It included pathological and histological Studies. After anesthesia of mice with
chloroform, kidney of the mice were visually inspected and dissected. Portions of
kidney tissue and cysts in them were prepared for the histopathological study. Tissues
were processed according to (2), briefly the tissue from the kidney collected and
placed in 10% formalin for histopathological studies and dehydrated by several
urease
Nitroprusside
Basrah Journal of Veterinary Research,Vol.17, No.3,2018
Proceeding of 6th International Scientific Conference,College of Veterinary Medicine
University of Basrah,Iraq
624
dilutions of ethanol alcohol dealcoholization with xylol, then embedded with paraffin
wax blocked (3-5) mm thickness sections were obtained by microtome. The sections
were put on glass slides deparaffinised with xylol, rehydrated by alcohol and stained
by hematoxyline and eosin.
RESULTS
The examination of experimentally infected males Balb/c mice with protoscolices at
6 months – post infection revealed 11% of experimentally animals have hydatid cyst
on kidney as well as presence of hydatid cyst in liver, spleen, mesenteries, and lungs .
Male of infected m ice with hydatid cysts after six month- post infection
A B
Kidney of infected mice with hydatid cysts after one (A) and six (B) month- post
infection
Blood urea test (BUN) for each of the infected and control group were investigated.
The results showed increase in the level of blood urea ( 40. 5μg\ml) in the infected
animals compared with 37.37 in the control 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
625
Groups BUN μg\ml
Control group 37.37
Infected group 40.5
Granulation tissue, congestion, edema, peri- vascular inflammatory cell , fat necrosis
in the adipose tissue and inflammation around tubules in the adjacent area were
recorded after the examination of the kidney tissue six months after infection . also
The results have showed increases in the focal inflammatory cell of the infected
organs of the infected group compared to control group .
- A- -BThe
kidney: A : before infection , B : After six month – post infection
The structure of the hydatid cyst was also examined in the present study and it
was found that the hydatid cyst of E. granulosus consisted of three layers starting with
the fibrous layer , the second layer of the cyst was the laminated layer and the
germinal is the third (figures ,C, D).
At Initial infection of kidney superficially with hydatid cyst and in about one month
, the protoscolex becomes with multilayer epithelial lining with prominent acidophilic
cytoplasm (epithelial layer between 5-6 layer enclosed directly by fibrous capsule
like, in the outside adipose tissue at the far exterior part. See the protoscolex was
found in the perirenal region. 4X macro kidney & protoscolex in perirenal region
(figure , A, B, E, F).
Infl.
Con.
Basrah Journal of Veterinary Research,Vol.17, No.3,2018
Proceeding of 6th International Scientific Conference,College of Veterinary Medicine
University of Basrah,Iraq
626
-A- -B-
-C- -D-
- E- - FE
F
Figure A, B, E, F.: The protoscolex was found in the perirenal region. kidney And
protoscolex in perirenal region. H&E,4X
Basrah Journal of Veterinary Research,Vol.17, No.3,2018
Proceeding of 6th International Scientific Conference,College of Veterinary Medicine
University of Basrah,Iraq
627
DISCUSSION
In Iraq , the studies about the infectio n with hydatid cyst are huge (2,3, 4,5, and 6 )
but it is rarely discussed or deals with effect of the infection with hydatid cyst on
kidneys structure and kidneys function .
The infection with hydatid cyst is primary from ingestion of egg after release with
stool of dog or definitive host or secondary hydatid cyst which is produced from
rupture of primary hydatid cyst ( 7,8) . the current study were designed from the
second route of infection , therefore, kidneys were recorded about 11 % of total
percent for infection with hydatid cyst but kidneys are rarely infected with primary
hydatid cyst because liver and lung are mainly involved with primary hydatid cyst,
further , if there is unsuitable condition in liver and lung the parasite will migrate to
the other organ to form hydatid cyst like kidneys or may be brain .
Involvement of the kidney is rare (3% of cases). Renal HCs are usually located in the
upper or lower pole. HCs are frequently solitary and located in the cortex, and they
may reach 10 cm in diameter before any clinical symptoms are noted (16).
The tissues and cysts from kidney of each uninfected and infected animals were
examined in the present study, The results have showed increases in the focal
inflammatory cell of the infected organs of the infected group compared to control
group . the number of lymphocytes and macrophages were increased in the tissues of
organs infected with hydatid cysts (9).
The vascular congestion had been seen in infected organs and this corresponded with
the study of 2, 12, 15.
The structure of the hydatid cyst was also examined in the present study and it was
found that the hydatid cyst of E. granulosus consisted of three layers starting with the
fibrous layer which were resulted from the stimulation of host tissues by hydatid cysts
as a foreign body (12) . The second layer of the cyst was the laminated layer and the
germinal is the third from which the protoscolex proliferated as recognized in the
present study
REFERENCES
1. Akhan , O. ; Ensari , S. and Ozmen, M. (2002) . Precutaneous treatment of
a period gland hydatid cyst . Eur. Radiol. , 12: 597-599.
2. Al – Humairy , A. KH. (2010). Evaluation of the Activity Datura
stramonium Seeds Extracts on Growth and Development of Hydatid Cysts for
Echinococcus granulosus in White Mice Balb/c (Therapeutic , Histologic and
Immunologic Study ).Ph. D. Thesis . College of Science , Kufa University.
3. Al – Mobarak, Z.A. (2006). Effect of peel extracts of Punica fruits on on
echinococcosis of Echinococcus granulosus . M. Sc. Thesis, College 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
628
Education . , Kufa University Dar, F.K. and Alkarmi ,T. (1997). Public
health aspects of cystic echinococcosis in the Arab countries. Acta. Tropica.
67 : 125-132.
4. Al-Saimary, I.E. and Zeki, A. (1999). In vitro chemotherapeutic effects of
antibiotics and plant extract on protoscolices viability of human hydatid cysts.
J. Basrah Res., 20: 79-98.
5. Al-Saimary,I.E., Al-Shemari, M.N. and Al-Fayadh, M.M.A.(2010).
Epidemiological and immunological findings on human Hydatidosis . Medical
Practice and Review ., 1: 26-34.
6. Al-Sanafi, A. E. (1996). Possible new treatments for hepatic hydatid disease:
Methotrexate and praziquantel. Ph. D. Thesis, College of Medicine ,
University of Baghdad: 107 pp.
7. Georgopoulos , S. ; Korres, S. ; Riga, M. and Ferekidis, E. (2007). Hydatid
cyst in the bile duct of the submandibular gland. Int. J. Oral. Surg. , 36: 177 –
179.
8. John , D.T. and Petri , W.A. (2006). Markell and Voge’s Medical
parasitology . 9th ed. , Elsevier , United states of America : 468 pp.
9. Mendez-Sanchez, N. ; Vega, R. ; Cortes, R. ;Corona, R. ; Bahena, R. and
Uribe, M. (2003). Hepatic Echinococcus granulosus. A case report. Annals ,
hepatol. , 2(2) : 99-100.
10. Parija, S.C. and Sheela, D. C. (1999). Current concepts in the diagnosis of
cystic echinococcosis in humans and Livestocks and intestinal echinococcosis
in canine hosts. J. vet. Parasit., 13: 93-102.
11. Pedrosa , Saiz , Arrazola , Ferreiros and Pedrosa . (2000) . HD: radiologic
and pathologic features and complications. RadioGraphics . 20:795–817.
12. Rigano, R. ; Profumo, E. ; Bruschi, F. ; Carulli, G. ; Azzara, A. ; Ioppolo,
S. ; Buttari, B. ; Ortona, E. ; Margutti, P. ; Teggi, A. and Siracusano, A.
(2001). Modulation of human immune response by Echinococcus granulosus
antigen B and its possible role in evading host defence. Infec. Immunol. , 69
(1): 288-269.
13. Schalm, O.W.; Jain, N.C. and Carrol, E.J. (1975). Veterinary hematology.
Leo and Febiger, ed. Philadelphia. Pp: 152-140.
14. Smyth, J. D. (1964). The biology of hydatid organisms. Adv. Parasitol. , 2:
169-219.
15. Sida , L. A. (2005). Trails in treatment of secondary hydatid disease in
laboratory mice . Ph.D. Thesis , College of Education , University of Baghdad
.(In Arabic)
16. Tuzun and Hekimoglu . (2001). Various locations of cystic and alveolar HD:
CT appearances. J Comput Assist Tomogr . 25:81–87.
