Download Sudan - ARPN Journal of Science and Technology

VOL. 5, NO. 4, April 2015
ISSN 2225-7217
ARPN Journal of Science and Technology
©2011-2015. All rights reserved.
http://www.ejournalofscience.org
Seroprevelance of Brucellosis in Different Animals Species in Northern
State (Sudan)
1
1 Department
Zein A.M, 2 Adris M.A
of Microbiology and Parasitology, Faculty of Medicine, University of Dongola, Sudan
2 Department of Biochemistry, Faculty of Medicine and Health Sciences, University of Dongola, (SUDAN)
1
[email protected]
ABSTRACT
Brucellosis is considered to be one of the most important zoonotic infectious diseases because of its impact on public
health and the economy. In Northern State, Sudan, brucellosis is prevalent in Dongola, Daba and Marawe localities and
new cases are reported annually (Anon, 2001), but the exact situation of the disease is not known in human and animals in
the Northern State. Objectives of the study were to: Investigate the prevalence of the disease in different animal species in
the Northern State. Prospective study was conducted in Northern State, Sudan. Blood Samples collected over 6 Months in
the period January ( 201 to June 2011) form (7607) domesticated animals (2009 cattle, 2170 Sheep, 2303 goats and 1225
camels) were tested using. RBPT and CELISA. Amongst live stocks 23.8% of cattle, 11.2% of sheep 16.3% of goats and
23.6%. Of camels were seropostive. The overall prevalence of brucellosis in the 7607 domesticated animals was found to
be 17.9% in Northern State. In conclusion, there is an urgent need to perform a control strategy at national level on animal
brucellosis.
Keywords: Brucellosis, domesticated animals
1. INTCTIONRODU
Brucellosis is a contagious disease which affects
different animal species including cattle, camels, small
ruminants, swine, dogs and man (Nicoletti, 1989). The
disease is caused by Brucella abortus, Br. melitensis, Br.
ovis, Br. suis, Br. canis and Br. neotomae. The disease
cause different clinical manifestation, abortion and to less
extent orchitis and infertility in males (Solera, 1997). It is
essentially a disease of animals caused by bacteria of the
genus Brucella, with humans as accidental host (Taylor et
al. 1989). The disease is usually transmitted to human by
consumption of unpasteurized milk and dairy products or
by direct contact with infected animals or both through
surveys and countries reports, it was confirmed that
animal brucellosis is wide spread in Africa (Pandey, et al.
1999). Several published reports indicate that, animal
brucellosis is a common disease in Sudan. Control
measures were adopted, but still the disease is widely
spread and new cases were reported annually (Anon
2001). This Study was carried out to investigate the
prevalence of the disease in the Northern State, Sudan, in
different animal species.
study area are delivered through district health net-work,
consisting of a local hospital and a few health centers in
the cities.
2.2 Localities of the Northern State
The state was divided into seven localities for the
purpose of the study, and the localities were divided into
three geographical areas:
a.
b.
c.
Northern area: Consisted of Wadi Halfa and
Dangola localities which were divided into five
units (Wadi Halfa, Abre, Dalgo, Berka, and
Farage).
Middle area: Consisted of Alburgage, Dongola,
and Goled Localities which were divided into
seven units (Hafeer, Karma, Argo, Dongola city,
Goled, Kaddar and shark Elneel).
Southern area: Consisted of Dabba and Marawe
localities which were divided into seven units
(Gaba, Dabba City, Tadamon, Karema City,
Marawe city, Gorear and Alshohada)
2. MATERIALS AND METHODS
2.1 Area of the study
The study was carried in Northern State which is
one of the biggest States in the Sudan. It is about 348,796
Km2. It is bordered by Egypt in the North and Libya and
North Darfur in the West, Nahr Elneel in the East,
Khartoum and Northern Kordofan State in the South. The
population of the Northern State is about 699,065while
the population of animals is about 3,150,000 distributed in
seven localities, Wadi Halfa, Dalgo, Al burgage, Dongla,
Goled, Dabba, and Marawe. The Northern State is mainly
an agricultural State (dates, wheat and beans). The
veterinary services facilities depend on veterinary
hospitals in all parts of the State as well as dispensaries
and primary veterinary care units. Health services in the
210 VOL. 5, NO. 4, April 2015
ISSN 2225-7217
ARPN Journal of Science and Technology
©2011-2015. All rights reserved.
http://www.ejournalofscience.org
2.3 Mapif the Northern State and localitcs
N
Naher Alneel State
Northern Darfur State
Libya
Egypt
Khartoum State
211 VOL. 5, NO. 4, April 2015
ISSN 2225-7217
ARPN Journal of Science and Technology
©2011-2015. All rights reserved.
http://www.ejournalofscience.org
2.4 Population of the study and sampling
The study was focused on: adult sexually mature
non vaccinated cattle, sheep, goats, and camels in the
study area.
2.5 Rose Bengal Plate Test
A standardized RBPT was provided by the
Central Veterinary Research laboratory, Soba, Sudan, was
used to screen the samples as by Alton et al. (1975):
a.
b.
c.
d.
e.
f.
The serum sample and antigen were brought to
room temperature (22±C).
A 30µl of each serum sample was placed on a
white enamel or plastic plate.
After shaking gently the antigen bottle, an equal
volume of the antigen was placed near each
serum spot.
Immediately after the last drop of antigen has
been added to the plate, the serum and antigen
were mixed thoroughly, using a clean glass or
plastic rod for each test, to produce a circular or
oval zone approximately 2 cm in diameter.
The mixture was agitated gently for 4 minutes at
ambient temperature on a rocker.
The result of agglutination was read at the end of
the 4 minutes period and reported as positive or
negative according to the OIE (2008).
2.6 Competitive Enzyme-linked Immuno-Sorbent
Assay (c-ELISA)
The RBPT positive serum samples were
confirmed by the c-ELISA (OIE, 2008). The test was
performed using commercial kits (Sabnova- Sweden).
Material and reagents were provided by the University of
Dongola, Faculty of Medicine. The test was performed
according to the method described by the producing
company as follows:
2.7 Test Procedure
a. The PBS-Tween Solution 20x concentrate 1/20
was diluted in distilled water. 500 ml were
prepared by adding 25ml PBS-Tween Solution to
475 ml distilled water and mixed thoroughly.
b. The freeze dried mAb was reconstituted
immediately before use with 6ml sample dilution
buffer by adding the buffer carefully into the
bottle.
c. All reagents were equilibrated to the room
temperature before use.
d. The samples and controls were diluted by adding
them to the wells of the plates prefilled with
buffer.
e. A50µl of each serum controls (positive, weak
positive and negative) were added into each of
the appropriate wells, respectively. Each control
was run in duplicate.
f. A5µl of each sample dilution buffer were added
into two appropriate wells (designated as
Conjugate control, Cc).
g.
A5µl of each test sample was added to each of
appropriate well. For confirmation each samples
was run in duplicates.
h. A50µl of mAb-solution were added to all wells
of the samples and controls used for controls
within 10 minutes.
i. The test plate was sealed and the reagents were
mixed thoroughly by tipping the sides of the
plate.
j. The plate was incubated at room temperature for
30 minutes.
k. The plate was rinsed 4 times with PBS-Tween
using Sanofi Pasteur washer.
l. A100 µl substrate solutions were added to each
well and incubated for 10 minutes at room
temperature timing began after the first well was
filled.
m. The reaction was stopped by adding 50µl of stop
solution in the same order as the substrate
solution was added to each well and mixed
thoroughly.
n. The optical densities (OD) of the controls and
samples were measured using a micro plate
photometer at 450 nm after 15 minutes from the
addition of the stop solution.
2.8 Calculations
a. The mean OD-values for each of the controls and
samples were calculated.
b. The percentage inhibition (PI) values for the
controls and samples were calculated using the
following formula:
PI = 100 – (mean OD samples / control ×100)
Mean OD conjugate control Cc
2.9 Test Validity
To ensure validity of the test, the values of the
controls must fall within the following limits:
OD Cc = 0.75 – 2.0
PI Positive Control =90 – 100
PI Weak Positive Control =35–65
PI Negative Control = (-10) – 15
2.10 Interpretation of the test
The status of the test was determined as follows:
PI
< 30%
≥ 30%
Status
Negative
Positive
3. RESULTS AND DISCUSSION
3.1 Field Observations
A surveillance was carried out during (2011) in
Northern State in Northern Sudan. Indigenous
domesticated animals includes sheep, goats, cattle, camels
and equine and foreign breeds were kept in the area of the
study to supply milk and meat for private consumption
and also to support the farmer’s income. The animals
were kept in backyards of the houses and villagers raised
211 VOL. 5, NO. 4, April 2015
ISSN 2225-7217
ARPN Journal of Science and Technology
©2011-2015. All rights reserved.
http://www.ejournalofscience.org
small flocks ranging from 2-12 animals which are usually
managed by women or children. No housing was
available except night shelters (small holdings). Nomadic
and semi-nomadic systems of breeding were adopted in
southern area of the State. Local green fodders were given
in the morning and evening to the animals kept in small
holdings and concentrates were not fed except to animals
in governmental farms and those kept in confined system.
Sources of water for the livestock are wells and
the River Nile. Natural breeding is used in the area of the
study and the farmers exchange breeding bulls. Animal
diseases were reported in the area included mainly
pneumonia, external and internal parasites and nutritional
deficiency diseases. There are several small abattoirs,
which were poorly equipped or even lacked facilities.
Sometimes they are next to meat shops in the
vicinity of houses. Home slaughter and consumption of
sheep, goats, cattle and camel meat was wide spread and
their offal’s are usually given to dogs. Dead animals are
often abandoned and or are usually thrown into the River
Nile. Fresh milk and dairy products, meat and other
animal products are often consumed raw or insufficiently
cooked and constituted a special risk. In some areas, dried
animal feces are kept near human dwellings and used for
cooking .There are houses which lack running water and
toilets. Small markets are scattered all over the State and
large- permanent markets were only in big towns.
3.2 Prevalence of brucellosis in different animal’s
species
The (7607) animals examined, 1368 (17.9%)
were positive for brucellosis (Table 1).
Accordingly, sero-prevalence of brucellosis in sheep,
goats, cattle and camels in the study area was found to be
11.2%, 16.3, 23.8% and 23.6%, respectively.
Table 1: The prevalence of brucellosis in the different
animal according to species
Animal
Total of samples
Number
(%)
species
examined
positive positive
1-Sheep
2170
242
11.2%
2-Goat
2203
359
16.3%
3-Cattle
2009
478
23.8%
4-Camel
1225
289
23.6%
Total
7607
1368
17.9%
3.3 Prevalence of brucellosis in different areas
The serological survey showed that 13.6% of the
animals examined in the Northern area were positive for
brucellosis. The prevalence rate of Brucella antibodies in
Wadi Halfa locality was 13.2% while in Dongola locality
was 14%. The overall prevalence of brucellosis in 386
sheep, 360 goats, 311 cattle and 158 camels in the
Northern area was found to be 9%, 9.7%, 13.8% and 34.1,
respectively. The serological survey showed that 13.3% of
the animals examined in the middle area were positive for
brucellosis. The prevalence rate of Brucella antibodies in
Dongola locality was 10.6%, Alburgage locality was
11.7% and Al Goled locality was 17.1%.The overall
prevalence of brucellosis in the 954 sheep, 949 goats, 995
and 573 camels examined was found to be 7.7%, 17.4%,
17.1%, 16.7 respectively. The serological survey showed
that 15% of the animals examined in the southern area
were positive for brucellosis. The prevalence rate of
brucella antibodies in Aldaba locality was 10.4% while in
Marawe locality was 19.9%. The overall prevalence of
brucellosis in the 830 sheep, 894 goats, 703 cattle and 494
camels equine examined in this area was found to be
14.3%, 14.9%, 23.4% and 22% respectively. In the
present study the overall prevalence of brucellosis in
the7607 domesticated animals was found to be 17.9%.
This was higher than those reported by Suliman (1987) in
El Gazera and Khartoum States 11.6% and 15.8%,
respectively and lower, than that reported by Musa (1995)
in Darfur States 20%. The results indicated that, there was
a marked increase in brucellosis in animals in the
Northern State due to the absence of brucellosis control
programmer. The prevalence reported here was also
higher than those reported in some Arabian and African
countries; 12% in Moroco, 4.8% in Egypt, 5.2% in
Kuwait, 8% in Jordan and 3.8% in Libya (Refai , 2002)
and 3.8% in Chad, 6% in Ivory Coast, 7.5% in Cameron,
8% in Upper Volta and 10% in South Africa (Kubuafor et
al. 2000) Several reports indicated that, brucellosis in
animals is common in the Sudan. El Nasri (1960) reported
14% to 18% sero-prevalence of brucellosis among cattle,
6.6% in goats and 35% in sheep in Southern Sudan.
Abdalla (1964) reported prevalence of 3% in
cattle and 1.7% and 1.5% in sheep and goats in Northern
Sudan, respectively. In Darfur States Western Sudan,
Musa (1995) found 13.9%, 3.5%, 5.9% and 7.7%
prevalence in cattle, sheep, goats and camels,
respectively. In Gezera State Central Sudan, Dafalla
(1962) reported 8.7% to 10.7% sero prevalence of the
disease in cattle, 4.2% to 50% in sheep and 2.5% to 30%
in goats. In Khartoum State, Suliman (1987) found a
prevalence of 15.8% among cattle and more recently Ali
(2007) reported a prevalence of 27.4% in cattle. The
results confirm the finding of Omer, et al. (2007) who
reported that the prevalence of brucellosis increased
during the last years among sheep, goats, cattle and
camels in Kassala area, Eastern Sudan. Therefore, an
urgent organized program for monitoring and control of
animal brucellosis should be adopted current study
revealed a prevalence of 23% in cattle, which is high,
according to the criteria of Thimm and Wundit (1976)
who considered that a percent range between16-25 was
high. The prevalence was higher in cattle compared with
that in other animals in the study area 11.2% in sheep,
15.6% in goats. Only camels showed a similar prevalence
rate of 23.6%. This might be due to the system of
breeding adopted in the study area that large ruminants
were kept in small holdings. The results are in agreement
with these, obtained in Togo (22.5%) (Akakpo et al.
1981), Rwanda (25.7%) (Kabagmbe et al. 1988), India
(Punjab) (22.6%) (Aulakh et al., 2008) and Iran (22.8%)
(Moshelani et al. 2011). However, were lower than, those
found in Kenya (77.5%) ( waghela, et al. 1978),
Cameroon (40%) (Kubuafor et al. 2000) and Turkey
212 VOL. 5, NO. 4, April 2015
ISSN 2225-7217
ARPN Journal of Science and Technology
©2011-2015. All rights reserved.
http://www.ejournalofscience.org
(68.1%) (Gen et al. 2005). The prevalence of bovine
brucellosis is increasing in the Northern State when it is
compared with the previous study by Abdalla (1966) who
found 3% prevalence in cattle. The overall prevalence of
brucellosis in camels, in Northern, middle and southern
areas of the Northern State was found to be 34.1%, 16.7%
and 22%, respectively. The prevalence was higher in the
Northern area which might be due to the expansion of
large-scale trading of camels transported from Western
Sudan through the area to Egypt without proper attention
being payed to the possibility of disease transmission.
Some of these camels are kept in feed-lots before being
exported across the Nubian lake to Egypt. The 23.6%
prevalence of the disease in camels in the area is in
agreement with that reported by Musa (1995) who found
23.8% prevalence in Darfur, Western Sudan, but is
considerably higher than those recorded by several
investigators from different parts of the country. Yagoub
et al. 1990 and Bitter, 1986) and lower than those found
by Majid et al., 1999; Fayza et al. 1990 and Omer, 2006.
The results were also higher than Kenya (8%) (Paling et
al. 1998), than that from Iran (10.5%) (Ahmed et al,
1995), from Libya (4.1%) (Gameel et al. 1993), from
Nigeria (11.42%) (Junaidu et al., 2006) and from Somalia
(3.9%) (Gen, et al. 2005). In countries where Br.
melitensis is prevalent in sheep and goats, other farm
animals such as pigs, cattle and camels, may become
infected from them (Altons, 1985). In the present study
the prevalence of brucellosis in sheep and goats in
Northern, middle and southern areas of the Northern State
was found to be 9%, 7.7%, and 14.3% in sheep and 9.7%,
17.4% and 15% in goats, respectively. The prevalence of
the disease was higher in the middle area as large numbers
of small ruminants were distributed in the farmers in this
area in the years 2007 and 2008, without testing for
brucellosis. Furthermore, some of these animals when
showed signs of the disease were not excluded and
attempts were made to treat them with a local therapeutic
(Om-regala). Treated animals might have constituted foci
of infection to the herds in the area. Results showed that
the prevalence of ovi-caprine brucellosis was increasing
when compared with the 1.7% in sheep and 1.5% in goats
which reported by Abdalla (1966). This increase might be
due to the spread of the disease due to absence of control
measures and to improvement of monitoring and
surveillance procedures applied. Although the prevalence
recorded in this study was 15.6%, it is lower than those
reported in Turkey (31%) (Gen et al. 2005), in Iran
(24.6%) (Yahya et al. 2009) in Jordan (27%) (Al-majali,
2004) and in Egypt (31%) (Mayada et al.2010). From the
results it appears that brucellosis is more prevalent in
cattle and camels than in sheep and goats and this is in
agreement with the finding of Omer et al. (2007). Sheep
and goat flocks in the study area were mobile. Free
movement of infected sheep and goats and mixing them
with other animals can spread brucellosis. Control of
movement of sheep and goats and control or elimination
of infection in them can reduce spread of the disease to
other animals.
ACKNOWLEDGEMENTS
I would like to thank to the staff of Veterinary
Research Institute, Dept. of Brucella, Dr. Enaam Elsanosi
and Mr.Salsh who provided the Rose Bengul antigen and
assets of the Department on hand to accomplish this work.
My thanks extend to Ministry of Health, Northern State,
Sudan for supporting this work.
REFERENCES
[1]
Abdalla,A.(1966).Incidence of brucellosis in Wadi
half a District.Sudan.J.Vet.Sci.Anim.Husb.,7:28-31
[2]
Ahmed, R, M.A, Munir (1995) Epidemiological
investigations of brucellosis in Pakistan.
Pakistan.Vet.J.,15:169-172
[3]
Akakapa, A.J,Bornarel, P, Sarradin, P. (1981).
Bovine brucellosis in Togo. Bull. Anim. Hlth.Prod
.,132:133-137
[5]
Ali, A ,A,I. (2007).Prevalence of brucellosis in
Kuku dairy, Khartoum State and the susceptibility
of the isolates to some chemotherapeutic agents.
MSc. Thesis, Faculty of Pharmacy, University of
Khartoum, Sudan.
[6]
Almajali, A.M,Talafha, A.Q, Ababenh, M.M. (
2004). Sero-prevalence and risk factors for bovine
brucellosis in Jordan .J. Vet. Sci .,10 : 6-15
[7]
Alton GG, Jones LM, Pietz DE. (1975). laboratory
techniques in brucellosis, World Health
Organization, Geneva.11-64
[8]
Alton GG, Jones LM, Pietz DE. (1975). laboratory
techniques in brucellosis, World Health
Organization, Geneva.11-64
[9]
Anczkowski F (1972). Further Studies on bovine
brucellosis .Vet. Bull., 42: 25-27.
[10]
Anon (2001) Annual Report of Sudan Veterinary
Surices.
[11]
Aulakh, H.K,Patil, P.K, Sharma, S. ( 2008).Study
on the epidemiology of bovine brucellosis in
Punjab(India) using milk –ELISA. Acta. Vet.
Brono., 77:393-399
[13] Dafalla ,E. N and Khan, A.(1962) The occurrence,
epidemiology and control of animal Brucellosis in
Sudan. Bull.Epiz.Dis.Afr.,6:243:247
[14]
Elnasri, M.(1960) Brucellosis in the Southern
Sudan.Vet.Rec.72:1200-1201
[15]
Gameel, S. M, Mohamed, S. O, Mustafa, A. A,
Azwai, S. M.(1993). Prevelanc of camel brucellosis
in Libya. Trop .Anim .Hlth .Prod.,25:91-93
213 VOL. 5, NO. 4, April 2015
ISSN 2225-7217
ARPN Journal of Science and Technology
©2011-2015. All rights reserved.
http://www.ejournalofscience.org
[16]
[17]
[18]
Gen, O,Otlu, S, Pahun, J.P. (2005). Seroprevalence of brucellosis and Leptospirosis in
aborted dairy cows. Turk. J. Vet. Anim. Sci .,29:
359-366
Junaidu ,A.U, Oboegbulem, S.I. Salihu, M.D
(2006) Serological survey of Brucella antibodies in
breeding herds . J. Microbiol. Biotech. Res., 1: 6065
Kabagmbe, J.P,Elzer, P.H, Opuda, J, Smith, S.H.
(1988). Risk factors for Brucella seropositivity in
goats herds in eastern and western Uganda
.Prev.Vet.Med .,52:91-108
[19]
Kubuafor, D.K, Awumbia, B, Akanmoi, B.D.
(2000). Seroprevelance of brucellosis in cattle and
human in the Akawapin-South district of Ghana.
Acta.Trop .,76:45-47
[20]
Moshelani, S, Jaraheri-Koupae, M, Rabiee, S.
(2011). Detection of Brucella spp.and Liptospiras
spp. By multiplex polymerase chain reaction from
aborted bovine ,ovine and caprine fetuses in Iran.
Afr. J. Microbiol. Res .,5:627-630
[21]
Musa, M.T. (1995). The magnitude and the
problem of brucellosis in Darfur States
.Ph.D.Thesis ,University of Khartoum
[22]
Nicoletti, P. (1986). Diagnosis and Control of
Brucellosis in the Near East. FAO, Rome.
[23]
Omer, M.M, Abdelaziz, A.A, Abusalab, S.M.A and
Ahmed, A.M (2007). Survey of brucellosis among
sheep, goats, Camels and Caltle in Kassala Estern
Sudan. J. Anim. Vet. Ad., 6(3): 612-618.
[24]
Pandey, G.S, Kobayashi, K, Nomura, Y, Namota,
A, Mwima, H.K, Suzuki, A.K. (1999). Studies on
sero- prevalence of brucellosis Kafue lechwe in
Zambia. Ind. Vet. J., 76: 275-278
[25]
Refai, M. (2002). Incidence and control of
brucellosis in Near East region .Vet.Microbiol,
90:81-110
[26]
Solera, J, Martinez- Alfaro, E, Espinosa, A. (1997).
Recognition and optimum treatment of brucellosis.
Drugs, 53:245-256
[27]
Suliman, M.A. (1987). The prevalence of bovine
brucellosis in Khartoum and Gezira regions .Msc.
Thesis, Fculty of Veterinary Science,University of
Khartoum.
[28]
Waghela, S,Fazil, M.A, Gathuma, J.M. (1978). A
serological survey of brucellosis in camels in
northeastern
province
in
Kenya
.Trop.Anim.Hlth.Prod., 10:28-29.
[29]
Taylor JP, Perdue JN (1989). The changing
epidemiology of human brucellosis in Texas, 19771986. Am. J. Epidemiol., 130: 160-165
214