JZS)Journal of Zankoy Sulaimani , 2005,vol 8(1) part A (

A )1 (8 2005

The Prevalence of Tuberculosis Among The

Cattle in Sulaimani Districts

Jalal Majeed Shareef, Rizgar Raheem Sulaiman and Ihsan Kadir Zangana

College of Veterinary Medicine, University of Sulaimani /Kurdistan Region Iraq

College of Veterinary Medicine, University of Duhok //Kurdistan Region Iraq

Abstract
In spite of the devastating effect of tuberculosis in man and animals, studies regarding bovine
tuberculosis have not been done, in the Sulaimani districts so far, The aim of the present study was to
investigate for the first time, the prevalence of tuberculosis in cattle in the Sulaimani districts. The study
was carried out among five groups of bovine (the pregnant cattle at last two months of gestation, recently
parturient cattle (two months after parturition), under six months age calves, calves of six months to two years
old heifers and cattle above two years old (but not belonging to the first and second groups), from June to
December 2000. The study includes the field observation, using the single intradermal comparative
tuberculin test (SIDT) which involved (4012) cattle from different Sulaimani districts (Sulaimani center,
Halabja, Penjwen, Rania, Chamchamal, Darbandikhan). The field observation has shown 207 (5.1%) positive
reactors to (SIDT). The results have revealed highest positive rate in Halabja district, where the rate was (9%)
and lowest rate in Darbandikhan, where the rate was (1.5%), while different prevalence were detected (3.5%,
4%, 5.5% and 6%) in Chamchamal, Penjwen, Sulaimani center and Rania respectively. The prevalence rate
was also high (6.1%) in a group of cattle that were classified at a stage of two months after delivery (two
months from parturition) and no incidence was detected in a group of calves under six months of age.
Out of 207 SIDT positive reactor cattle, (117) sputum samples were examined by direct smear with
Zihle –Neelsen stain, positive result was 37 (31.7%) only. The samples were also cultured on Modified
Lowenstein-Jensen media, which showed 45 (38.5%) positive growth. This study included also the abattoir
observation, by routine meat inspection of 7375 carcasses that have been slaughtered in Sulaimani abattoir,
the result revealed 65 (0.9%) positive carcasses that showed typical lesions of tuberculosis. The samples from
the lesions were submitted to the microbiological and histopathological examinations that confirmed the
preliminary diagnosis. The photos are also shown.
Keywords: Tuberculosis, zoonotic disease, bovine.
Introduction
Among many diseases, which transferred considered as enormous threats of the
from animals to man, precisely among those health of both human and animal. Since in
diseases which transferred from cattle to an uncontrolled environment the spread of
man, tuberculosis (TB) is one of the most TB from animal to man is ease and
devastating diseases. It is known as one of frequent, it considered as an important
the world wide distributed diseases, TB is of anthropozoonotic disease. Concerning
major importance particularly in dairy cattle the precautionary measures to protect the
[1]. It causes a great damage to the health of people and cattle in the area, no
country’s economy, moreover TB is serious attempt has been implemented so

Email:- drjalalshareef@yahoo.com

1
JZS)Journal of Zankoy Sulaimani , 2005,vol 8(1) part A (
A )1 (8
far, also there is no any national program or
strategy to point out the disease, which
transmitted to human through animals and
their products. In the neighboring countries
the above status of controlling zoonotic
diseases is common, particularly when it is
dealing with tuberculosis. According to
World Health Organization (WHO) report,
it is estimated that human tuberculosis
incidence and deaths for years 1990 and
1999 were 88 and 30 millions respectively,
with most cases in developing countries.
Anthropozoonotic TB (caused by
Mycobacterium bovis) is present in animals
of most developing countries, where
surveillance and control activities are often
inadequate or unavailable; therefore, many
epidemiological and public health aspects of
infection mainly remain unknown. From 36
Asian nations, 16 reported a sporadic/low
occurrence of bovine TB, and one (Bahrain)
described the disease as enzootic; ten did
not report bovine TB; and the remaining
nine did not have data. Within the Asian
region, seven countries apply disease
control measures as part of a test-and-
slaughter policy and consider bovine TB
noticeable. In the remaining 29 countries,
bovine TB is partly controlled or not
controlled at all. Of the total Asian cattle
and buffalo populations, 6% and less than
1%, respectively, were found in countries
where bovine TB is notifiable and a test-
and-slaughter policy was used; 94% of the
cattle and more than 99% of the buffalo
populations in Asia are either only partly
controlled for bovine TB or not controlled
at all. Thus, the human population in Asia
lives in countries where cattle and buffaloes
undergo no control or only limited control
for bovine TB.
TB is produced by Mycobacterium
species, the term is derived from the small
2
2005
granulomatous nodules or tubercles that are
seen in advanced infections by
Mycobacterium tuberculosis or
Mycobacterium bovis. The disease
affects particularly all species of
vertebrates, producing signs and lesions,
which are generally similar in the various
species. Three main types of TB and their
causative agents are well known. These
types are (human) Mycobacterium
tuberculosis, (bovine) Mycobacterium bovis
and (avian) Mycobacterium avium. Human
TB is the most species specific of the three
types, rarely being transmitted to non-
human species. Avian TB is typically
restricted to birds however pigs and a few
other animals are susceptible. Bovine TB is
the most infectious TB infecting humans
and most warm-blooded animals, [2, 3, 1].
There are many methods for diagnosis. The
single most important diagnostic test for TB
in vivo is the intradermal tuberculin test.
This test is applied by the intradermal
injection of 0.05 ml of tuberculin to anal
fold or into the skin of the neck. The
reaction is read between 48 and 96 hrs after
injection with preference for 48 to 72 hrs
for maximum sensitivity and at 96 hrs for
maximum specificity, and a positive
reaction constituted a diffuse swelling at the
injection site [4].
Intradermal tuberculin (4ml) is injected
subcutaneously into the neck of the cattle
with a rectal temperature of not more than
39˚C at the time of injection and for 2hours
later. If the temperature at 4, 6 and 8 hours
after injection rises above 40˚C, the animal
is classed as a positive reactor. This test is
succeeded to detect those cases of minimal
sensitivity, but it is not also sufficiently so
specific to differentiate between the
reactions due to infection with
JZS)Journal of Zankoy Sulaimani , 2005,vol 8(1) part A (
A )1 (8
Mycobacterium bovis and other avium
mycobacterium complex [4, 5, 1].
Where the presence of John’s disease or
avian tuberculosis is suspected, or skin
tuberculosis is apparent, not-specific
sensitization must be considered and a
comparative test should be used [2]. The
comparative test depends on the greater
sensitivity to (PPD) tuberculin. Avian and
mammalian tuberculin is injected
simultaneously into two separated sites on
the same side of the neck [6]. Three days
after injection, the thickness of the skin at
the two injection sites are compared.
Interpretations of the results are based on
differences in the increase in skin thickness
elicited by the two products and by the
disease history of the herd [1]. Diagnosis on
clinical signs alone is very difficult, even in
advanced cases. Radiography is useful in
non-human primates and small animals.
Microscopically examination of sputum and
other discharges is some times used.
Necropsy findings of classic TB granulomas
are often very suggestive of the disease.
Conformation of diagnosis required
isolation and identification of the organism,
with culture usually tacking 4-8 weeks [4, 7,
1]. Diagnostic samples include sputum,
tracheobronchial lavages, lymph nodes of
thoracic, abdominal, and other aspirates,
urine, feces, milk and biopsy specimens.
Necropsy material is obtained from the
lesions [8]. In laboratory the samples can be
used for diagnosis as following: Direct
microscopic examination, bacterial culture,
histopathology, animal inoculation, and
serology. TB is a public health hazards and
it is a zoonotic disease with the risk of
sharing the organism to the outside by
tuberculous animals, antituberculous
chemotherapy of animals is discourage and
controversial, so euthanasia is the most
3
2005
commonly accepted option [9]. The
complete control and eradication of bovine
TB are very difficult because
Mycobacterium bovis often does not
produce acute disease, persists in the carrier
stage, in human and non-human reservoirs,
and easily crosses species. No effective
vaccine or centralized global surveillance or
eradication programs currently exist. The
control measures result in significant
economic losses. The primary threat of
Mycobacterium bovis exists in wild life that
shares watering holes or pastureland with
domestic stock. In the developed world,
surveillance can minimize risk, but one-
third of the world’s population lacks
effective agricultural and food safety
programs, leaving them at substantial risk
for zoonotic infection by Mycobacterium
bovis [10]. Hence researches and
investigations for cattle TB are of great
importance, for this reason the present study
is designated in Sulaimani districts for the
first time.
Materials and Methods
1. Animals
A total of 4012 cattle were divided into
five groups, according to some facts, such
as; tuberculous cattle go through a period of
desensitization immediately before and after
calving, about 30% gives false negative
reactions, but returning to a positive status 4
to 6 weeks later, and calves drinking their
colostrum give positive reaction for up to 3
weeks after birth, even those are not
infected [1]. The groups were as following;
Group I: composed of (211) pregnant cows
at last two months of gestation.
JZS)Journal of Zankoy Sulaimani , 2005,vol 8(1) part A (
A )1 (8
Group II: composed of (151) recently
parturient cows (two months after
parturition).
Group III: composed of (360) calves under
six months of age.
Group IV : composed of (807) calves and
cattle six months to two years old.
Group V : composed of (2483) cattle above
two years old, but not belonging to the first
and second group.
T-test for the first two groups and Chi
square test for all groups of cattle were
applied for analysis statistically [11].
2. Area of the Study
The present study was conducted in
Sulaimani Governorate, north-east of Iraq,
including six districts, and involved the
following numbers of cattle: Sulaimani
center 2012 cattle, Halabja district 400
cattle, Penjwen district 400 cattle, Rania
district 400 cattle, Chamchamal district 400
cattle, and Darbandikhan district 400 cattle.
3. Tuberculin Test
All groups of cattle were tested by
single intradermal comparative tuberculin
test. The test carried out in June to end of
December 2000. The method and allergen
of single intradermal comparative
tuberculin test were used as described by [1,
4, and 6].
For performing single intradermal
comparative tuberculin test, caliper for
measuring of skin fold thickness, scissors
for clipping the site of injection and
multidose resetting intra-dermal syringes, at
least two sets of types Mucklintock
tuberculin syringe were used. Two types of
tuberculin have been used; Mycobacterium
bovis and Mycobacterium avium were
received from Razi Laboratory-Iran Islamic
4
2005
Republic. The vials contain 1mg/ml (25000
C.T.U/ml) straw color bovine antigen and
pink color avian tuberculin, contains
0.5mg/ml (2500 C.T.U/ml).
Two different types of tuberculin
were injected into the skin simultaneously,
fairly close together, so that any resulted
reaction may be more readily assessed for
interpretation purposes. For the test,
the intra-dermal injection (0.1 ml) in the
middle portion of the neck was used, and
the sites prepared by clipping away below
the crest and the lower site 12cm lower
down on a line roughly parallel with the
slope of the shoulder. The avian tuberculin
has being injected at the upper site and the
bovine tuberculin at the lower one. The
distance between two sites of injection is 10
cm. In calves the injection sites have being
located one on each side at the central point
of the neck. Prior to injection, the thickness
of the skin at each site was recorded, using
a caliper. The appearance of a small pea-
sized swelling, which forms a bulge on the
skin surface, was a good indicator for the
achievement of the process successfully [7].
The test was evaluated by repeated
skin measurement 72 hours later after the
injection using a caliper. The interpretation
of the single intradermal comparative
tuberculin test was classified as negative,
suspected or positive reactors. A negative
reactor was identified when there was a
negative reaction to bovine tuberculin, or a
positive or suspected reaction to bovine
tuberculin which was equal to or less than a
positive or suspected reaction to avian
tuberculin. A suspected reactor was
indicated by a positive or suspected reaction
to bovine tuberculin, which was 1-4
millimeter greater than the reaction to avian
tuberculin. A positive reactor was identified
when there was a positive reaction to bovine
JZS)Journal of Zankoy Sulaimani , 2005,vol 8(1) part A (
A )1 (8
tuberculin, which was more than 4mm
greater than the reaction to avian tuberculin.
Cattle which had been classified as
suspected reactors were being re-tested after
interval of 60 days, If they were again
classified as suspected they would be
considered as positive [1, 12, 4, 6].
4. Abattoir observation
From the periods of March 2000 to
December 2000, the carcasses of 7375 cattle
of different ages were examined at interval
of four visits per a week by the routine meat
inspection. The samples of different organs,
which contain tuberculosis like lesions (Fig
1), were submitted to laboratory for testing
by Ziehl-Neelsen direct smear for detection
of acid-fast bacilli, and the positive samples
were sent for histopathological examination.
5. Sputum specimen collection
Out of the (117) sputum samples collected
by sputum cup from the animals that were
positive to tuberculin
test at the early morning, the cattle were
being fasted to about 12 hrs, before the
sputum was collected in a plastic container.
The sputum volume was about (10-20) ml.
The samples were reserved in a refrigerator
under (4˚C) until processing.
The samples were homogenized by
adding sodium hydroxide then neutralized
with acid [7]. The sample was centrifuged at
(3000rpm) for 30 minutes. The supernatant
was excluded; the sediment was used for
direct smear and inoculation in the specific
media for culturing. The smear was
taken from the above sediment, using
microscopical slide and stained by Ziehl-
Neelsen stain for detection of acid-fast
bacilli under the light microscope (400X)
[8]. Modified Lowenstein-Jensen media
5
2005
were used for culturing of sputum
samples. The direct swab was also
prepared from the abattoir lesions, which
were opened by scalpel on microscopical
slides and stained by Zeihl-Neelsen as
described above. Histopathological
specimens obtained from suggestive
tuberculosis lesions were processed;
sections were stained with haematoxylin,
eosin (H&E) and Zeihl-Neelsen, and then
examined for typical tuberculous lesions
and acid-fast bacilli [13], see Fig 2.
Results and Discussion
:Intradermal Tests .1
The preliminary results of single
intradermal comparative tuberculin test are
shown in Table 1.
A total number of 157 (3.9%) out of 4012
animals were positive reactors with the
single intradermal comparative tuberculin
test, the remaining 3664 (91.3%) animals
were negative and 191 (4.8%) animals were
inconclusive (suspected). The highest rate
was in Halabja district 26 (6.5%) and non
positive reactors were detected in
Darbandikhan while in Sulaimani center,
Chamchamal, Penjwen and Rania were 98
(4.9%), 8 (2%), 7 (1.7%) and 18 (4.5%)
respectively. Each inconclusive reactor
was subjected to a retest after an interval of
60 days from the preliminary test, as shown
in Table 1. In Sulaimani center from 77
suspected reactors 6 (7.8%) were positive,
in Halabja district from 42 suspected 8
(19.1%) were positive, in Chamchamal
from 25 suspected 5 (20%) were positive, in
Penjwen from 14 suspected 6 (42.9%) were
positive, in Rania from 22 suspected 3
(13.7%) were positive and in Darbandikhan
from 11 suspected 4 (36.7%) were positive.
The heighest positive rate was in Penjwen 6
JZS)Journal of Zankoy Sulaimani , 2005,vol 8(1) part A (
A )1 (8
(42.9%) and low positive rate was in
Sulaimani center 6 (7.8%) while in Rania,
Halabja, Chamchamal and Darbandikhan
were 3 (13.7%), 8 (19.1%), 5 (20%) and 4
(36.7%) respectively. The final result of
prevalence rate after retest is also shown in
Table 1. The prevalence rate increased from
157 (3.9%) to 207 (5.1%) and the highest
prevalence rate was in Halabja
district 36 (9%) and the lowest rate was
Darbandikhan district, which was 6 (1.5%)
while in Sulaimani center, Chamchamal,
Penjwen and Rania were 111 (5.6%), 14
(3.5%), 16 (4%) and 24 (6%) respectively.
The preliminary results of single
intradermal comparative tuberculin test
regarding the five different age groups are
shown in Table 2. The highest rate 11
(5.2%) was in group of cattle of two months
before delivery (in last two months of
gestation), and non-positive reactors were
detected in group of calves under six
months of age, while groups of cattle that
were two months after delivery, 6 months to
2 years age and above 2 years age were 6
(4%), 22 (2.7%) and 118 (4.8%)
respectively. All suspected reactors were
subjected to retest after 60 days from
preliminary test, also according to [1, 6] the
negative reactor cattle of two months before
and after delivery were subjected to retest
because these cattle were going on to
desensitization stage and must be tested
after 60 days.
In group of cows, which were
pregnant at last two months before delivery,
from 38 suspected cases, 7 (18.4%) were
positive, among 34 cows, which were two
months after delivery, 8 (23.5%) were
positive, in 32 cattle which were 6 months
to two years old, 4 (12.5%) were positive
and in 75 cattle which were above 2 years
old, 13 (17.3%) were positive, while no
6
2005
positive reactor was detected among calves
which were under six months.
From 162 negative or desensitized cattle
which were 2 months before delivery, 11
(6.8%) were positive, after retest and of 111
cattle which were two months after delivery,
7 (6.3%) were positive.
The final results of the five different
age groups are also shown in Table (2). The
highest prevalence rate, 21 (13.9%) was in
group of cows which were two months after
delivery, and no prevalence rate was
detected in group of calves which were
under six months of age, while in cattle
which were two months before delivery, 6
months to 2 years age or above 2 years age,
the results were 29 (13.7%), 26 (3.3%) and
131 (5.6%) respectively.
The single intradermal comparative
tuberculin test had a great performance in
identifying cattle with tuberculosis. The test
had sensitivity range from 90.9% to 95%
and specificity is estimated to be 96% to
99% [14, 15]. On the other hand, when
John’s disease or avian tuberculosis is
suspected or skin tuberculosis is apparent,
non-specific sensitization must be
considered and a comparative test must be
used [16, 14, 12]. In Sulaimani region
there are large populations of wild and
domestic birds. Moreover, the presence of
stagnant water as a source of animal
drinking water, so it is very logical to use
single intradermal comparative tuberculin
test to detect the incidence of this disease.
The bovine (PPD) was selected since the
comparative tuberculin test, using avian
(PPD) and bovine (PPD) had a much more
efficient than human (PPD), for separation
between tuberculous and non-tuberculous
cattle, in comparative tuberculin test [17,
18, 19]. In this study the result of
single intradermal comparative tuberculin
JZS)Journal of Zankoy Sulaimani , 2005,vol 8(1) part A (
A )1 (8
test was in the same range (5.3%) as that
reported in Dar Salaam region of Tanzania
[20], and in a lower rate comparatively with
that obtained in Ninevah province (11.9%)
[21]. However, various rates were recorded
from different parts of the world. In Ghana
the rate was 13.8% (24), in Britain 20.8%
[23] and in Czech Republic 21.03% [24].
In the present study the prevalence
rate was higher than that obtained from
other countries. In Republic of South Africa
it was 0.062% [25], in Britain it was 0.4%
[26] and in Latin American and the
Caribbean it was 1% [27].
Regarding bovine tuberculosis in
Sulaimani region and according to F.A.O.
[28] information no work has been done or
published so far. Comparing results
obtained by single intradermal comparative
tuberculin test in the present study with
results from other studies in other areas,
higher rate in Sulaimani districts was
observed. This may be possibly explained
by the absence of irradication campaign in
the past and the present time in this region.
In UK (1900 - 1930) Mycobacterium bovis
was isolated in 6-30% of human population,
and from each 20 cows 5 cows (25%) had
tuberculosis, but when the policy of test and
slaughter had been conducted, the rate
reduced to about (0.4%) in 1996 [26].
The construction of old fashioned
animal house, bad ventilation, over crowded
and non-hygienic places in rural areas may
contribute in high prevalence of TB among
cattle of Sulaimani region. Other workers
also considered these as important attributed
factors [29]. In Sulaimani region there are
inadequate regulations for stamping out or
immediate quarantining or strict prevention
system for animal movement from and to
the region. This may favored the
distribution of the disease among the
7
2005
animals. On the other hand, a poor
quality of feed, malnutrition, poverty and
low educated owners, concerning hygiene
and animal management, may cause
improper conditions for animals. Protein
and energy deficiency and subsequently
inhibition of cell mediated immunity, which
is the main route of immunity against
tuberculosis, accentuated the propagation of
the disease among the animals [30, 31, 32,
and 33]. Among different districts of
Sulaimani region the rate of prevalence was
highest in Halabja than others. This is
probably due to maintaining the animals in
overcrowded and closely housing, which
might provide more chances for
transmission. Similar observation was
previously reported by [34, 1]. The limited
pasture and closely grazing cattle may
enhance further transmission of the
tuberculosis among cattle [35, 36]. A high
significant effect of age on the disease
incidence among cattle is well known [37,
38, 1, and 39]. The pregnancy and calving
has been implicated in allergy to tuberculin
test [40, 41, 1]. There was no any statistical
difference (P< 0.01) between groups of two
months before and after calving, both have
a high incidence rate (13.9%) and (13.7%)
respectively, but statistical difference was
observed (P< 0.01) among the first two
groups and other groups of age. The mean
age of cattle of two months before and after
delivery were between 7 –8 years old while
the mean age in group of animals over two
years old were between 2.5-3.5 years. The
prevalence of the disease is elevated with
the age of the animals. There was
statistically significant relationship between
the age and prevalence of the disease (P
<0.01). This agrees with the finding of [37,
38, 1, 39]. In UK it was found [42] that the
risk of TB in cows over eight years of age
 JZS)Journal of Zankoy Sulaimani , 2005,vol 8(1) part A (
A )1 (8 2005

was twelve times more than the risk for the visible tubercles. The total number of
cows of one to two years old. Petukhove carcasses have shown typical lesions of
[37] revealed that the onset of the diseases tuberculosis was 65 (0.9%) as shown in
was among the six years age in Latvia, Table (3).
while some workers [39] mentioned that the The rate in animals under one year old, 1-3
most infected cattle in Mexico were adult year old and above 3 years of age were 5
females. The pregnancy acts as immune (0.6%), 33 (0.9%) and 27 (0.9%)
suppressor in dairy cattle due to nutritional respectively. The lesions were located
deficiency. The productions of the mainly in the parenchyma of the lungs,
corticosteriods initiated by the fetus also pleura, mediastinal, bronchial and
enhance peri-parturient immuno- retropharyngeal lymph nodes. The lesions
suppression [40, 41]. These factors that are varied in size from 2 mm to about 5 cm in
mentioned above may explain the highest diameter and characterized by hard gray to
prevalence rate in the groups of two months yellowish deeply embedded and others have
before and after calving cattle. been showing dry yellowish, caseous,
In the present study, tuberculosis necrotic center and solid abscess.
was not detected among calves less than six Calcification was common in many
months old. The evidence of the disease in tubercles as well as a gritty sensation and a
calves even if contacted with infected stock gritty sound when sectioned Fig. 1. In
is remaining low until they enter into the addition to that, samples have been
cowshed [43, 44, and 45]. The lymphocyte submitted to the laboratory for
count in bovine colostrums is up to histopathological section and showed
1Х106cell/ml, about half of which are T- typical feature of tuberculosis lesion. The
cell and the CD4/CD8 ratio are section of lung tissue showed caseous
approximately 0.57, which is somewhat necrotic area surrounded by a wide zone of
higher than the ratio in the blood. Through epithelioid cells. Several lymphocytes are
this procedure the tuberculin sensitivity, and also present within the zone of epithelioid
T-cell mediated hypersensitivity reaction, cells. Also other section showed several
may transfer to the calves from infected or acid-fast bacilli within the cytoplasm of the
sensitive dam. This may give the base of epithelioid cells. Several lymphocytes,
immunity to the offspring and protection neutrophils and erythrocytes were also seen
from the infection, since the cell-mediated in the section. Prevalence of tuberculosis
immunity is the major defense mechanisms lesion on carcasses in abattoir was (0.9%),
against the tuberculosis infection and this
may explain why Total
no any incidence of calve
Negative Positive
Ages tuberculosis has been detected [46, 1].
number No. (%) No. (%)
Table (3): Abattoir 840 5
Observation2. Abattoir observation:
845 )99.4( )0.6(
Under one years ages
A total number of 7375 carcasses of animals
3484 33
One year to three years
3517 were examined )by
in the Sulaimani abattoir 99.1( )0.9(
ages
routine meat inspection in order to detect
2986 27
Above three years ages 3013 )99.1( )0.9(
7310 65
Total 7375 )99.1( )0.9(
8
Table (3): Abattoir Observation
 JZS)Journal of Zankoy Sulaimani , 2005,vol 8(1) part A (
A )1 (8
Which is low if compared with the result of
single intradermal comparative tuberculin
test (5.1%) in the present work, this may be
due to inefficient routine meat inspection in
abattoir. In a sample of cattle that were
positive reactors to the tuberculin test,
abattoir inspection failed to detect an
estimated 47% of the cattle with lesions.
The details necropsy examination of
positive tuberculin test cattle was identified
in 21 sites of infection as compared with the
15 sites of infections that were identified by
a roughly routine meat examination [47,
48]. On the other hand, in the U.S.A. it was
found that not all the tuberculosis lesions in
cattle have been visible by the routine
inspection [49].
In this work, most lesions were located on
the retropharyngeal, mediastinal, tracheo-
bronchial and parenchyma of the lungs.
This was in agreement with that seen by
[47, 50, 49, and 51].
3. Laboratory Finding:
From (207) positive reactors to the single
intradermal comparative tuberculin test
(117) sputum samples were stained with
Table (4): Direct Ziehl-Neelsen
Smear Examination byexamined
stain and Zhiel- Neelsen Stains and Culturing Sputum
directly.
Samples. A total number of 37 (31.7%) were positive
for Mycobacterium bacilli, and in addition
to that, these samples have been submitted
for culturing on a modified Lowenstien –
Technique Total Number
Jensen media. A total number of 45 (38.5%) )%( .No
had shown a typical colonies of
Mycobacterium bovis. The colonies were a
rather rough. Their centers were raised and
Direct smears examination
117
Culturing
117
2005
surrounded by a fairly large, spreading
growth with irregular borders. The colonies
grow very slowly at 37˚C in a period of
about 67 days and the color of colonies was
off-white to yellowish. Direct smears were
also prepared from the colonies and stained
by Ziehl-Neelsen stain. The smears showed
a large number of acid-fast bacilli; see Table
4 and Fig. 2. Since both of the direct smear
staining by Zhile-Neelsen stain and culture
on Modified Lowenstein-Jensen media were
diagnostic methods.
The latter was significantly (P<0.01) more
accurate and sensitive than direct smear.
This was in agreement with the reports of
[22, 9, 52], and may be attributed to a
number of the microorganisms
in the inoculum. To find positive results in a
direct smear it should contain a large
number of mycobacterium bacilli while
very few bacilli even one bacilli may be
sufficient for culture method (in favorite
cultivated media) to give a positive result
[53, 54, 55, 22].
Samples for culture purpose were not
collected from all positive reactors by single
intradermal comparative tuberculin test,
since some owners generally do not like
cattle fasting and keeping them at home for
24 hours.
Out of (117) samples (sputum) which were
collected from (207) positive
Positi
Negative ve
No.
(%)
37
80
(31.7)
(68.3)
9 72 45
(61.5) (38.5)
 JZS)Journal of Zankoy Sulaimani , 2005,vol 8(1) part A (
A )1 (8 2005

was in the Darbandikhan district. When

Single intradermal comparative tuberculin
test, only (37) samples were positive by
direct smear staining with Zhile-Neelsen
stain and (45) samples were positive by
culturing method. The culturing method and
direct smear were useful for the detection of
only active tuberculosis with open lesions.
This result has been supported by many
studies done by [55, 47], regarding 55
lungs from positive tuberculin test reactors,
Mycobacterium bovis was only isolated in
(19%) of the confirmed cases, So not all
positive tuberculin test reactors show
positive result in direct smear or culture
method.
Conclusion
In view of the results obtained from
the presented study, the following points
have been concluded:
The highest incidence of tuberculosis was in
the adultcattle that their age was above
seven years and noincidence of the disease
was detected in the calves that are under six
months old. The highest incidence rate was
in the Halabja district and the lowest rate
result of single intradermal comparative
tuberculin test compared with the abattoir
observation, the former was higher than the
latter. The examination of sputum revealed
that not all-positive reactors to (SID)
comparative tuberculin test give positive
result to direct smear and culturing
examination, examinationof sputum by
culturing was more sensitive than direct
smear examination. It is recommended to
determine the distribution of the disease in
the remaining districts of the governorate,
set up legislation for test and slaughter with
compensation for the owners, educating the
farming community and livestock owners to
be apprized of the economic and public
health significance of the disease, also to
improve husbandry, nutritional status and
housing of the animals, isolation and
identification of the mycobacterium among
the livestock owners, which may be useful
for finding infection focal among the cattle
herds, and monitoring the incidence of the
disease in wild life, because the wild life is
an important source of the disease for
livestock.

Fig. 1: Yellowish gray tubercles Fig. 2: Section through the lung of

and infected Cow, show acid-fast
10
The cut section of the tubercles tubercle bacilli (darts).
show caseous necrotic centere
(darts) .
References

1. Radostits, O. M., Gay, C. C. Blood, D. C. and Hinchcliff, K. W., 2000 Veterinary

edicine, A Textbook of the Diseases of Cattle, Sheep, Pigs, Goats and Horses. pp.
909-934. 9th.W. B. Saunders -Harcourt Publishers Ltd.London.
William, C. R., 1995. Disease of Dairy Cattle. Chapter 14, pp.474-6. First edition .2
.Lippincott Williams and Wilkins Company. U. S. A
3. Bradford P. Smith, 1996. Large Animal Internal Medicine. pp. 672-674. 2nd. Mosby
Company. U. S. A.
4. Hungerford, T. G., 1990. Diseases of Livestock. pp.354-9. 9th ed., Lindsay Costelloe
Publisher., Australia.
5. John, F. T., James, H. G., Fredric, W. S. and Jeffery, E. B., 1992. Hagan and Bruner’s
Microbiology and Infection Diseases of Domestic Animals. 8th. pp270-89.
Comstock Publishing Associates. London.
6. Kelly, W. R. 1984. Veterinary Clinical Diagnosis. pp. 417-423. 3rd ed. Bailliere
Tindall. London.
7. OFFICE INTERNATIONAL DES EPIZOOTIES (O.I.E), 1996.
8. Ernst, L. B., and Dwight, C. H., 1999. Mycobacterium Species: The Agents of
Animal Tuberculosis. Chapter
30, pp. 158-164. In Veterinary Microbiology by Dwight, C. H., and Tuan, C. Z., editor.
First edition. Blackwell Science, Inc. U.S.A.
.Bourne, J., Donnelly, C. A., Cox, D. R., Gettinby, G., Mclnerney, J. P., Morrison, I .9
and Woodruff, R., Bovine tuberculosis: towards a future control strategy.Vet
.Rec2000.Feb.19;146 (8):207-10
10. Nelson, A. M., The cost of disease eradication of small pox and bovine
tuberculosis. Annual of the NewYork Academy of Science, 1999. 894,83-91.
Aviva, P., and Paul, W., 1999. Statistics for Veterinary and Animal Science. First .11
.edition. Blackwell Science Ltd. London
12. Doherty, M, L., Bissett, H. F, Quinn, P. J., Davis, W. C., Kelly, A. P. and Mongahan,
M. L., 1996. Asequential study of the bovine tuberculin reaction. Immunology Jan ;
87 (1):9-14
13. Hernandez, A. J., Renteria, E. T., Lopez, V. G., and Montano, H. M., 1997. An
abattoir monitoring system fordiagnosis of tuberculosis in Baja California, Mexico.
J. Am. Vet. Med. Assoc. Sep.211;(6): 709-11.
14. Wood, P. R., and Rothel, J., 1994. In vitro immuno diagnostic assay for bovine
tuberculosis. Vet. Microbial.May;40(1-2):125-35.
15. Monaghan, M. L., Doherty, M. L., Collins, J. D., Kazda, J. F. and Quinni, P. J.,
1994. The tuberculin test.Veterinary Microbiology, 40: 111-124.
16. Fifis, T.,Corner, L. A., Rothel, J. S. and Wood, P. R., 1994. Cellular and humoral
immune responses of cattleto purified Mycobacterium bovis antigens. Scandinavian
Journal of Immunology, 39, 267-274.
17. Lesslie, I. W. and Herbert, C. N., 1975. Comparsion of the specificity of humman
and bovine tuberculin PPD for testing cattle.3. National trial in Great Britain. Vet.
Rec. 1976. Apr 12 ; 46 (15) : 338-41.
18. Lesslie, I. W., Hebert C. N. and Frerichs, G. N., 1976. Practical application of
bovine tuberculin PPD intesting cattle in Great Britain. Vet. Rec. Feb 28 ; 98(9):
170-2.
19. Radunz, B. L. and Lepper, A. W., 1985. Suppression of skin reactivity to bovine
tuberculin in repeat test.Aust. Vet. J. Jun;62 (6):191-4.
20. Weinhauple, I., Schopf, K. C., Khaschabi, D., Khaschabi, D., Kapaga, A. M. and
Msami, H. M., 2000 Investigation on the prevalence of bovine tuberculosis and
brucellosis in dairy cattle in Dar es Salaam region and inZebu cattle in Lugoba area,
Tanzania. Trop. Anim. Health Prod., Jun 32 (3): 147-54.
21. Ihsan, M. A., 1996. Isolation and identification of Mycobacteria from the human
and cow in NinevahProvince. M.Sc. thesis, college of veterinary medicine.,
University of Mosul.
Bonsu, O. A., Laing, E. and Akanmori, B. D., 2000. Prevalence of tuberculosis in .22
attle in the Dangme-West distrit of Ghana, public health implications. Acta.
.Trop. Jul 21; 76 (1):9-14
Wielesmith, J. W., 1983. Epidemiological feature of bovine tuberculosis in cattle .23
.herds in Great Britain. J. Hyg.(London) 90,159-76
24. Kauba, V., 1999.Prevalence of TB in the Czech Republic. Cas Lek Cesk Aug2; 138
(15):460-3.
25. Bakunzi f. R., Zambo, G. C. and Morris, S., 1995. Bovine tuberculosis survey in
the Molopo district of theNorth West Province. J. S. Afr. Vet. Assoc., Mar; 66 (1):
28-9.
26. Vincent, A. J., 1996. TB testing. Vet. Rec. Jan 13 ;138(2):47.
27. De kantor, I. N. and Ritacco, V., 1994. Bovine tuberculosis in Latin America and
the Caribbean current status, control and eradication programs. Vet Microbial 40:5-
14.
28. Food and Agriculture Organization (FAO), Sulaimani sub-office (personel
communication).
29. Hejlicek, K. and Chloupek, B. 1999. Hisrtory of elimination of the bovine
tuberculosis in the Czech Republic.Cas Lek Cesk; Aug.2;138 (15); 456-9.
30. Pritchard, D. G. 1988. A century of bovine tuberculosis 1888-1988; conquest and
controversy. J. Comp.Pathol. 99,357-99.
31. Young. D., Garbe, T., Lathigra, R. and Aboud-Zeid, C. 1991.Proteins antigens:
structure, function andregulation. In Molecular Biology of the Mycobacterium
(ed.J.Mcfadden), pp.1-35,Surrey University Press.(Internet)www.maff.gov.uk/animalh/tb.
32. McCoy, H. and Kenny, M. A., 1992. Magnesium and immune function, recent
finding. Magnesium Research 5, 281-293.33. Phillips S. J. C., Chiy, P. C., Arney, D.
R. aand Kart, O., 2000. Effects of sodium fertilizers and supplements on milk
production and mammary gland health. Journal of Dairy Reserch 67, 1-12.
34. Neill, S. D., Hanna, J., O’Brien, J. J. and McCraken, R. M. 1989. Transmissin of
tuberculosis fromexperimentally infected cattle to in-contact calves. Vet. Rec. 124,
269-71.
35. Schellner, H. 1956. Risk of infection in cattle grazing pastures contaminated with
tubercle bacilli. dieRindertuberkulose 5,179-188.
36. Jackson, R., deLisle, G. W. and Morris, R. S. 1995. A study of the environmental
survival of Mycobacteriumbovis on a farm in NewZealand. Newzeland Veterinary
Journal 43, 346-352.
37. Petukhove, V. L. 1981. Genetics of resistance in cattle to tuberculosis age at illness
milk productivity andinfluence of mothers on frequncy of disease in progony.
Genetika 17 ,729-733.
38. Benham, P. F. J. 1985. "Astudy of cattle and badger behaviour and farm husbandry
practices relevant to thetransmission of the bovine tuberculosis (Mycobacterium
bovis). In the report to the Ministry of Agriculture,Fisheries and Food. (Internet)
www.maff.gov.uk/animalh/tb.
39. Milian-Suazo, F.,Salman, M. D., Ramirez, C., Payear, J. B., Rhyan, J. C. and
Santilloan, M. 2000.Identification of tuberculosis in cattle slaughtered in Mexico.
Am. J. Vet. Res. 61, 86-9.
40. Kerri, W. R., Lamont, H. G. and McGirr, J. L., 1949. Farther studies on tuberculin
sensitivity in the bovine.Veterinary Record 61, 466-75.
41. Buddle, B. M., Aldwell, F. E., Pfeffer, A., Delisle, G, W. and Corner, L. A. 1994.
Experimental Mycobacterium bovis infection of cattle-effect of dose of M. bovis and
pregnancy on immune- responses anddistribution of lesions. New Zealand
Veterinary Journal 42, 167-172.
42. Morris, R. S., Pfieffer, D, U. and Jackson, R., 1994. The epidemilogy of
Mycobacterium bovis infection. Veterinary Microbiology, 40, 153-177.
.Francis, J. 1972. Rout of infection in tuberculosis. Aus. Vet. J. 48, 578-580 .43
Fallon, R. J., 1994. “Cattle of cattle enterprise type on the rate, of disclosure of .44
tuberculin reactors.” Tuberculosis Investigation Unit, University College Dublin. In
the report to the Ministry of Agriculture, Fisheries and Food.
Internet).www.maff.gov.uk/animalh/tb
45. Martin, D.O. 1994.“The effect of enterprise type on the prevalence of tuberculin
reactors in the East Offaly badger research project,” Selected papers,18-19.
Tuberculosis Investigation Unit, University College Dublin. In the report to the
Ministry of Agriculture, Fisheries and Food. (Internet)
www.maff.gov.uk/animalh/tb.
46. Ian, R. Tizard. 1996. Veterinary Immunology An Introduction. 5th ed. W. B.
Saunders Company.Philadelphia. pp. 381-390.
47. Mcllroy, S. G., Neill, S. D. and McCraken, R. M. Pulmonary lesions and
Mycobacterium bovis excretion from respiratory tract of the tuberculin reacting
cattle. Vet. Rec. 1986. 118, 718-21.
48. Corner, L., Wood, P. R. and Rothel, J. S., 1990. Efficincy of inspection procedures
for the detection of tuberculous lesion in cattle. Aust. Vet. J. 67, 389-92.
49. Whipple, D. L., Bolin, C.A. and Miller, J. M.,1996. Distribution of lesions in cattle
infected with Mycobacterium bovis. J. Vet. Diagn. Inverst., July,8(3); 351-4.
50. Neill, S. D., O’Brien, J. J., and McCracken, R. M. 1988. Mycobacterium bovis in
the anterior respiratory tracts in the heads of tuberculin-reacting cattle. Vet. Rec.,
122,184-6.
51. Palmer, M. V., whipple, D. l., Rhyan, J. C., Bolin, C. A. ans Saari, D. A., 1999
Granuloma development incattle after intratonsilar inoculation with Mycobacterium
bovis. Am. J. Vet. Res. March; 60(3):310-5.
52. O’Brien, R.,Danilowicz, B. S., Bailey, L., Flynn, O., Costello, E., O’grady, D. and
Rogers, M., 2000.Characterization of the Mycobacterium bovis restriction fragment
length polymorphism DNA probe PUCD and performance comparison with
standard methods. J. Clin. Microbiol.,September:38(9):3362-9.
Grosset, J. 1976. The role of the bacteriology laboratory in the identification of the .53
sources of tuberculous infection. Cited by Isolation and Identification of
Mycobacteria from Human and Cows in Ninevah Province, M.Sc. thesis, college of
.Veterinary Medicine, University of Mosul by Ihsan M. A. 1996
54. Collines, C. H. and Grange, J. M., 1985. Isolation and identification of
microorganisms of medical and veterinary importance. The society for applied
bacteriology Technical series. No 21 Academic press, INC. U. S. A.
55. Corner, L. A., and Nicolacopoulos, C., 1988. Comparsion of media used for the
primary isolation of Mycobacterium bovis by vetrinary and medical diagnostic
laboratories. Aust. Vet. Jour. 65 (7): 202-5.
56. De Kantor, I. N., and Roswurm, J. D., 1978. Mycobacteria isolated from asal secretion
of tuberculin test reactor cattle. Am. J. Vet. Res. July: 39 (7); 1233-4.
‫رةشََََة وول َََََخى‬
‫بلَوبونةوةى نةخؤشََََى سََََيل لة ِ‬
‫ناوضةكانى سةر بة سليَمانى‬
‫جلل مجيد شريف و ِرزطار رحيم سليمان‬
‫كؤليجى ثزيشكيى ظيَتيَرنةرى ‪ /‬زانكؤى سليَمانى‪ /‬هةريمى كوردستان ‪-‬عيراق‬
‫أحسان زةنطةنة‬
‫كؤليجى ثزيشكيى ظيَتيَرنةرى ‪ /‬زانكؤى دهؤك ‪ /‬هةريمى كوردستان ‪-‬عيَراق‬

‫ثوختة‬
‫َ‬
‫ل دذوارى نةخؤشببي سببيل بببؤ مرؤظ و ئاذةل‪ ،‬هةتببا ئي َببستا تويَذينةوة لةسببةر‬ ‫لة طة َ‬
‫نةخؤشى سيلي رِةشة وولَخ (طا ‪ ،‬مانطا ‪ ،‬طويَرةكة) لة ناوضةكةدا بةطشتىء ناوضةى‬
‫سببليَمانى بببة تايبةتببى نةكراوة‪ ،‬بؤيببة ئامانجببى ئةم تويَذينةوةيببة دياريكردنببى رِيِذةى‬
‫بلوبونةوةى نةخؤشببى سببيلة بببؤ يةكةم جار لةناو مانطاى ناوضةى سببليَمانى دا‪ ،‬كببة‬
‫بيَطومان سودى ئةبيَت لة كؤنترؤلَكردنى نةخؤشى سيل لة مرؤظ و ئاذةلَدا‪.‬‬
‫ى ليةنى لةخؤ طرتووة‪ ،‬ليةنى كيَلَطةيي بةهؤى بةكارهيَنانى (‬ ‫ئةم تويَذينةوةية س َ‬
‫‪ )Single Intradermal Comparative Tuberculin Test‬كببة لةسببةر ‪ 4012‬مانطببا لةضةنببد‬
‫ناوضةيةكبى جياجيادا ئةنجام درا و ‪ )%5,1( 207‬ئاذةلَى توش بوو دةركةوتبن‪ .‬بةرزتريبن‬
‫ريَذةى توش بوون (‪ )%9‬لةناوضةى هةلَةبجبببببة ‪ ،‬نزمتريبببببن ريَذة (‪ )%1,5‬لةناوضةى‬
‫دةربةنديخان بوو‪ .‬بةرِةضاوكردنبى تةمةن ء كاتبى زاووزي َبى ئاذةلَةكان بةرزتريبن ئاسبتى‬
‫تووش بوون (‪ )%13.9‬لة دوو مانبط دواى زاييبن دا بوو‪ ،‬لةخوار شةش مانبط تةمةنةوة‬
‫نةخؤشى نةبوو‪.‬‬
‫ليةنببببى دووةمببببى تويَذينةوةكببببة ثشكنينببببى رِاسببببتةوخؤى بةلَغةمببببى ئاذةلةَ‬
‫تووشبووةكان بوو ء بةهؤى سبليد و رِةنطبى زيبل – نيلسبن‪ ،‬هةروةهبا طةشةثيَكردنيان‬
‫لةسبةر (‪ .)Modified Lowenstein-Jensen Media‬لة كؤى ‪ 207‬ئاذةلَى تووش بوودا توانرا‬
‫‪ 117‬نموونببة وةربطيري َ بت و ئةو ثشكنينانةى سببةروةى بببؤ بكري َ بت كببة ئةنجامةكةى ‪37‬‬
‫ثؤزةتيفى رِةنطكراو بة زيل ‪ -‬نيلسن و ‪ 45‬ثؤزةتيف بةرِيَطةى طةشةثيَكردن بوون ‪.‬‬
‫ليةنبى سبىَيةمى تويَذينةوةكبة‪ ،‬ليةنبى كوشتارطبة بوو‪ ،‬بةمةبةسبتى دياريكردنبى‬
‫لشةى مانطاى سبيلوى‪ .‬لةئةنجامبى ثشكنينبى (‪ )7375‬لشبة بةرِيَطةى ثشكنينبى رؤتينبى‬
‫ئاسايي كة لة كوشتارطةدا ثيادة دةكريَت ‪ )%0,9( 65‬لشةى سيلوى دياريكرا كة هةموو‬
‫نيشانةكانبى ثاش مردن (‪ )Gross Lesion‬تايبةت ببة نةخؤشبى سبيليان لةسبةر دياربوو‪،‬‬
‫هةروةهببا بببؤ زياتببر دلَنيابوون نمونةشيان لىَوةرطيرا بببؤ ثشكنينببى مايكرؤبايؤلؤجببى و‬
‫‪ Histopathology‬ء لة ئةنجامدا هةموو نموونةكان ثؤزيَتيببف بوون بببؤ نةخؤشببى سببيل و‬
‫ويَنةكانيان ثيشاندراون ‪.‬‬

‫انتشار مرض السل فى ابقار المناطق السليمانية‬

‫جلل مجيد شريف و رزكار رحيم سليمان‬
 ‫كلية الطب البيطري ‪ /‬جامعة السليمانية ‪/‬اقليم كوردستان ‪-‬العراق‬

‫احسان زنكنة‬
‫جامعة دهوك ‪ /‬اقليم كوردستان ‪-‬العراق‬ ‫كلية الطب البيطري‪/‬‬
‫الخلصة‬
‫يعتبر مرض السل من المراض المنتشرة و الخطيرة و لكن لم تجرى لحد الن بحوث‬
‫كافيبة حول هذا المرض فبي المنطقبة بشكبل عام و بالخبص فبي المناطبق السبليمانية‪،‬‬
‫حيببببث أجري هذا البحببببث لول مرة‪ .‬ان أجراء تلك البحوث بات أمرا ضروريببببا لغرض‬
‫توضيببح نسبببة النتشار و خاصببة فببي البقار لنهببا تعتبببر مصببدرا خطيرا لهذا المرض‪.‬‬
‫أشتملت هذه الدراسبة على ثلثبة جوانبب‪ ،‬الدراسبة الحقليبة و ذلك باسبتعمال (‪Single‬‬
‫‪ )Intradermal Comparative Tuberculin Test‬و الذي اشتمبببل على ‪ 4012‬بقرة فبببي‬
‫مختلف مناطق السليمانية ‪ ،‬و تبين أن أعلى نسبة للصابة كانت في منطقة حلبجة حيث‬
‫بلغت (‪ )%9‬و اقل نسبة كانت في منطقة دربنديخان حيث لم تتجاوز( ‪.)%1,5‬‬
‫و مبع اخبذ العمبر و فترة الحمبل بعيبن العتبار فان أعلى نسببة للصبابة كانبت عنبد‬
‫البقار التبي مضبى على وضبع حملهبا (النجاب) فترة شهريبن‪ ،‬حيبث بلغبت إلى ‪ %13.9‬و‬
‫لم تسجل أية نسبة لدى العجول التي كانت أعمارها اقل من ستة اشهر‪.‬‬
‫أما الجانب الثاني من البحث فقد تم بأخذ عينات القشع من البقار المصابة و فحص‬
‫المسبحة المباشرة بعبد صببغها بصببغة زيبل ‪ -‬نيبل سبن‪ ،‬كذلك زرع العينات على الوسبط‬
‫الزرعي من نوع (‪ .)Modified Lowenstein-Jensen Media‬شمل هذا الجانب فحص ‪117‬‬
‫عينبة مأخوذة مبن مجموع ‪ 207‬بقرة مصبابة‪ ،‬و نتيجبة لهذه الفحوصبات ظهرت ‪ 37‬عينبة‬
‫موجببة عبن طريبق الشرائح المصببوغة بينمبا ‪ 45‬عينبة كانبت موجببة عبن طريبق الزرع‬
‫المذكور أعله‪.‬‬
‫أما الجانب الثالث من البحث فكان عن طريق المجزرة وذلك لتشخيص الذبائح‬
‫ذبيحة عن طريق الفحص العياني (‬ ‫المصابة بمرض السل‪ .‬و نتيجة لفحص ‪7375‬‬
‫‪ ) Gross Lesion‬الذي يتببم عادة فببي المجزرة تبببين أن ‪ 65‬ذبيحببة أي نسبببة (‪)%0,9‬‬
‫مصبابة بالمرض‪ .‬وقبد تبم أخبذ عينات منهبا لجراء الفحوصبات المايكروبايولوجيبة و‬
‫التقطيبع النسبيجي و ذلك للتأكبد مبن السبتنتاجات السبابقة‪ ،‬و أظهرت النتائج بان‬
‫كافبة العينات كانبت موجببة لمرض السبل‪ ،‬وقبد قمنبا بتصبوير نماذج منهبا و عرضهبا‬
‫ضمن نتائج هذا البحث‪.‬‬

‫‪.‬‬ ‫‪. 2 005 /21/2Resaved on 24/8/2004‬‬ ‫‪8/2004 /24‬‬

‫‪Accepted on 21/2/2005‬‬