Sero – Prevalence of Brucellosis in Cattle in Southern Area of Khyber Pakhtunkhwa , Pakistan

Department of Zoology, Hazara University Mansehra, Khyber PakhtunKhwa, Pakistan; Microbiology and Biotechnology Center, Veterinary Research Institute, Khyber PakhtunKhwa, Peshawar, 25000, Pakistan; Department of Microbiology, Kohat University of Science and Technology, Khyber PakhtunKhwa, 26000, Pakistan ;Department of Microbiology, Quid –e–Azam University Islamabad, Pakistan; Department of Animal Health, The University of Agriculture Peshawar, Pakistan *Corresponding author: aabdul.9090@gmail.com

Brucellosis is the contagious and zoonotic disease of humans and animals worldwide especially in the Middle East, Africa, Asia, South and Central America (Yohannes et al., 2012 andMunir et al., 2010).It is an infectious bacterial disease caused by member of genus Brucella, Brucella melitensis and Brucella abortus are the principal cause of brucellosis in bovines (Karaca et al., 2007 andRadostits et al., 2000).Brucellosis is primary reproductive disease with potential of great economic loss in bovines resulting abortion, death of young stock, birth of weak calves, stillbirth, delayed calving and infertility in males and reduced milk production in females (Abubakar et al., 2011 andMaadi et al., 2011).This disease occurs by means of contaminated water or feed, inhalation and via conjunctiva, and it can be transmitted through direct or indirect contact with infected animals or animal secretions.Moreover ingestion of the excretion in uterine discharge and milk of infected animals also the source of transmission (Yohannes et al., 2012).
Signs and symptoms of this disease are nonspecific or may be atypical.Laboratory confirmation is therefore essential for the diagnosis of brucellosis.Moreover, combinations of serological, cultural and molecular techniques are essential for diagnosis.Serological blood tests and Milk ring test (MRT) are mostly used for diagnosis of brucellosis (Al-Mariri and Haj-Mahmoud, 2010).Detection of brucella antibodies in milk considered the principal technique for screening and monitoring the infected dairy herds or an individual animal for diagnosing brucellosis at regular intervals (Godfroid and Kasbohrer, 2002).Serological tests are comparatively easy to carry out and provide a practical benefit in estimating the prevalence of Brucella infection in bovines (Abubakar et al., 2011).
Brucellosis is one of the major disease problems in Pakistan.Many studies documented the prevalence of brucellosis in livestock of different districts and provinces of Pakistan (Rabab et al., 2000;Iftikhar et al., 2008;Mukhtar and Kokab, 2008;Abubakar et al., 2010;Shafee et al., 2011;Ullah et al., 2013).Moreover, livestock populations of Pakistan are large and well adapted to the local environmental conditions.Therefore this disease has a

Sero-Prevalence of Brucellosis in Cattle in Southern Area of Khyber Pakhtunkhwa, Pakistan
ISSN 2308-2798 significant impact on human, animal health and socioeconomic aspects; where the people of rural areas of Pakistan mainly dependent on livestock, land cultivation and their domestic animals (Shafee et al., 2011 andMaadi et al., 2011).Due to the lack of diagnostic facilities and financial limitations in Pakistan, veterinarians generally prefer conventional tests for screening of brucellosis.Serum Agglutination Test, Rose Bengal Test and Milk Ring Test are usually performed at both private and government livestock laboratories (Asif et al., 2009;Gul and Khan, 2007).
This study was carried to estimate the prevalence of brucellosis in milk and serum samples collected from of different breed of cattle located in Bannu and Laki Marwat area of Khyber PakhtunKhwa region of Pakistan.
The study was conducted on the lactating dairy cattle in District Bannu and Lakki Marwat, Khyber PakhtunKhwa Pakistan, from May to July, 2011(Figure 1).A total (n = 302) blood sera and milk samples of Cattle were randomly collected.Approximately 5 mL of milk was collected from four quarters of each cattle into sterile glass bottle.All the blood serum and milk samples were then kept in an ice box and transported directly to the VRI, Peshawar.
The serum samples were subjected to SPAT for screening brucella antibodies as described by Alton et al., (1975).The results of agglutination in SPAT were recorded.A titer of 1:80 or above was considered positive for brucellosis according to the instruction of the antigen manufacturer (Global invitro.LLP Sudbury Hill, London UK).
Milk ring test was conducted on milk as describe by Alton et al., (1988) with slight modification.The antigen hematoxylin used in this test was supplied by Veterinary Research Institute (VRI) Peshawar.The positive samples were differentiated on the basis of blue ring present on the top of milk after overnight reaction.
Statistical analysis was executed by using SPSS (version 16.0) software.By using chi square test significance of difference was determined.Value of p < 0.05 considered as statistically significant.
In the present study, from 302 samples total 170 and 132 blood and milk samples were collected from District Bannu and District Lakki Marwat respectively.Among 170 samples from District Bannu, 13 (7.64 %) and 5 (2.94%) cattle were found positive for brucellosis on the basis of SPAT and MRT respectively (Table 1).Similarly from 132 samples from District Lakki Marwat, (11 8.33%) and 7 (5.30%)samples were positive on the bases of SPAT and MRT respectively (Table 1).With respect to the districts, incidence rate of brucellosis was found higher in Lakki Marwat (10.8%) compared to Bannu district (5.64%) (Table 1).2).
The rest of 23 cattle were with unknown source of breeding pattern, showed 3 (13.04%)and 2 (8.69%) samples positive on SPAT and MRT, respectively.The incidence of brucellosis was found higher in cattle mated with unknown source followed by community bull and then artificially inseminated (Figure 1).Moreover no significant association of brucella infection in cattle with respect to breeding practices were found (P > 0.05).
Brucellosis is considered most important zoonotic disease.As this disease make significant economic losses to livestock industry, therefore common diagnostic techniques are essential for its efficient diagnosis.Diagnostic laboratories mostly utilize serological test and MRT for identification of Brucella species in the host.
In present study, sero-prevalence of brucellosis was found higher on SPAT (7.94%) as compared to MRT (3.97%).These findings are in agreement of Saleha et al., (2014), they also revealed in their study that SPAT test showed more positive results compared to MRT.This might be due to fact that milk proteins hinder the brucella antibodies isolation (Akhtar et al., 2010).
In present study overall prevalence of brucellosis in District of Bannu and Lakki Marwat was found 5.29% and 10.9% respectively on the bases of SPAT and MRT.Shafee et al., (2011), in their study reported the prevalence of 8.5% in Quetta Pakistan.Similar study was also conducted by Song et al., (2009) who estimated the sero-prevalence brucellosis 5.0% and 0.5% in cattle located in Bangladesh Agriculture University, by using antigens of Brucella abortus and Brucella melitensis.The same patterns of results were observed by Wali et al., (2005), who assessed the sero-prevalence of brucellosis in livestock at different districts of Khyber PakhtunKhwa.The variation in results might be due to sample size and different habitat of animals.
In the present study the prevalence of brucellosis in age group of > 5 showed higher prevalence compared to the other age group of Cattle (Table 3).These findings are in agreement with Ahmad et al., (2009), who reported the high prevalence of brucellosis in age group of more than five compared to the younger animals.The reason might be due to fact that with the passage of time animals are likely to be exposed to the Brucella infection.Radostits et al., (2000) also revealed that this disease is chronic and increases with age.

Figure 1 :
Figure 1: Map of Khyber PakhtunKhwa province showing the study area (Highlighted Red)

Figure 1 :
Figure 1: Prevalence of brucella infection in the cattle with respect to breeding practice in Bannu and Lakki Marwat distracts, Khyber PakhtunKhwa Pakistan

Table 1 :
Prevalence of brucellosis in the cattle located in Bannu and Lakki Marwat Districts,

Table 2 :
Prevalence of brucellosis in locally available breed of the cattle, in Bannu and Lakki Marwat distracts, Khyber

Table 3 :
Age wise distribution of brucella infection in the cattle in Bannu and Lakki Marwat distracts, Khyber PakhtunKhwa Pakistan

Table 4 :
Prevalence of brucella infection in the cattle on the basis of feeding practices in Bannu and Lakki Marwat distracts, Khyber PakhtunKhwa Pakistan