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		<div class="Basic-Text-Frame">
			<p class="Type-of-Article">&nbsp;</p>
			<p class="Type-of-Article"><span class="CharOverride-1">Research Article</span></p>
	</div>
		<div class="Basic-Text-Frame">
			<p class="title- ParaOverride-1">&nbsp;</p>
			<p class="title- ParaOverride-1">Prevalence Survey of Bovine Brucellosis in Apparently Healthy Dairy Animals in Karachi, Pakistan</p>
		</div>
		<div class="Basic-Text-Frame">
			<p class="Authors ParaOverride-1">&nbsp;</p>
			<p class="Authors ParaOverride-1"><span class="CharOverride-2">NaeemUllah Durrani</span><span class="CharOverride-3">1*</span><span class="CharOverride-2">, RahmatUllah Rind</span><span class="CharOverride-3">1</span><span class="CharOverride-2">, Asghar Ali Kamboh</span><span class="CharOverride-3">1</span><span class="CharOverride-2">, Faiza Habib</span><span class="CharOverride-3">1</span><span class="CharOverride-2">, Saba Parveen Samo</span><span class="CharOverride-3">2</span><span class="CharOverride-2">, Shujat Ali khan</span><span class="CharOverride-3">3</span><span class="CharOverride-2">, ZiaUllah</span><span class="CharOverride-3">4</span><span class="CharOverride-2">, Muhammad Shahid</span><span class="CharOverride-3">5</span></p>
		</div>
		<div class="Basic-Text-Frame">
			<p class="Affiliations ParaOverride-1"><span class="CharOverride-5">1</span>Department of Veterinary Microbiology; <span class="CharOverride-5">2</span>Department of Animal Nutrition; <span class="CharOverride-5">3</span>Department of Veterinary Anatomy and Histology; <span class="CharOverride-5">4</span>Department of Animal Reproduction, Faculty of Animal Husbandry and Veterinary Sciences, Sindh Agriculture University, Tandojam, Pakistan; <span class="CharOverride-5">5</span>Veterinary Research Institute, Bacha Khan Chowk, Peshawar, KPK, Pakistan.</p>
		</div>
		<div>
			<p class="Abstract ParaOverride-1">&nbsp;</p>
			<p class="Abstract ParaOverride-1"><span class="CharOverride-6">Abstract</span> | Brucellosis is one of the major zoonosis diseases that are still prevalent throughout the world. The present modified cross-sectional study aims to determine the prevalence of brucellosis in apparently healthy dairy cattle/buffaloes in Karachi, Pakistan. A total of 100 farms at different localities of Karachi were visited and milk and blood samples (n=200 each) were collected. These were analysed by Rose Bengal Plate Test (RBPT), complement fixation test (CFT) and milk ring test (MRT). Results indicated a 17.5%, 12.5% seropositive samples by RBPT and CFT, respectively. While, 14% were recorded in the milk samples analysed by MRT. In different areas (towns) prevalence percentage varied highly, ranging from 5.88% to 25%. From these results it could be concluded that brucellosis is present in the apparently healthy dairy cattle and buffaloes of Karachi, however, its prevalence is low comparing with small cities of Pakistan. Furthermore, disease prevalence is diverse in different areas of Karachi. These results emphasize the need for a better monitoring of the disease by government policies or local veterinarians to overcome the disease prevalence.  </p>
		  <p class="Abstract ParaOverride-1">&nbsp;</p>
			<p class="Abstract ParaOverride-1"><span class="CharOverride-6">Keywords </span>| Brucellosis, Cattle, Buffalo, Prevalence, Karachi</p>
		  <p class="Abstract ParaOverride-1">&nbsp;</p>
		</div>
		<div class="Basic-Text-Frame">
			<p class="Editor----Citation"><span class="CharOverride-8">Editor</span><span class="CharOverride-9"> | </span>Sanjay Kumar Singh (MVSc., Ph.D. (Veterinary Gynaecology &amp; Reproduction)), Senior Scientist, In-Charge Animal Gynecology Lab, Animal Reproduction Division, Indian Veterinary Research Institute, Izatnagar 243122, Bareilly (UP), India.</p>
			<p class="Editor----Citation"><span class="CharOverride-6">Received</span> | March 10, 2015; <span class="CharOverride-6">Revised</span> | April 16, 2015; <span class="CharOverride-6">Accepted</span> | April  17, 2015; <span class="CharOverride-6">Published</span> | April  29, 2015&#9;&#9;</p>
			<p class="Editor----Citation"><span class="CharOverride-6">*Correspondence</span> | NaeemUllah Durrani, Sindh Agriculture University, Tandojam, Pakistan; <span class="CharOverride-6">Email: </span>drnaeemdurrani12@gmail.com</p>
			<p class="Editor----Citation"><span class="CharOverride-6">Citation</span> | Durrani N, Rind R, Kamboh AA, Habib F, Samo SP, khan SA, ZiaUllah, Shahid M (2015). Prevalence survey of bovine brucellosis in apparently healthy dairy animals in Karachi, Pakistan. J. Anim. Health Prod. 3(2): 35-38.</p>
			<p class="Editor----Citation"><span class="CharOverride-8">DOI</span><span class="CharOverride-9"> </span>| <a href="http://dx.doi.org/10.14737/journal.jahp/2015/3.2.35.38"><span class="Hyperlink">http://dx.doi.org/10.14737/journal.jahp/2015/3.2.35.38</span></a></p>
			<p class="Editor----Citation"><span class="Editor---Citation CharOverride-6">ISSN </span>| 2308–2801</p>
			<p class="Editor----Citation"><span class="CharOverride-8">Copyright </span>>© 2015 Durrani et al. This is an open access article distributed under the Creative Commons Attribution License, which permits unrestricted use, distribution, and reproduction in any medium, provided the original work is properly cited.</p>
		</div>
		<div class="Basic-Text-Frame">
			<p class="Heading-1--Introduction----">&nbsp; </p>
		  <p class="Heading-1--Introduction----">INTRODUCTION </p>
			<p class="Caps-on-First-Para ParaOverride-1">&nbsp;</p>
			<p class="Caps-on-First-Para ParaOverride-1"><span class="_idGenDropcap-1">T</span><span>he </span>economic importance of livestock goes beyond direct food production. Skins, fibers, manure (fertilizer or fuel), draught power, and capital are also livestock benefits. Livestock provides a lifeline for a large proportion of 95% of the world’s rural population that lives in the developing world and cultivates 64% of the world’s arable land. The major products of livestock are draught power, meat, milk, eggs, manure, feathers, fiber, hides, and horn (<a href="#Hoffmann-D--1999"><span class="Hyperlink">Hoffmann, 1999</span></a>).</p>
		  <p class="Caps-on-First-Para ParaOverride-1">&nbsp;</p>
			<p class="Body-Text">Livestock is facing several health issues, however, the most important livestock diseases endemic in rural areas include brucellosis and calf scour (<a href="#Mailk-S--Kumar-A--Verma-AK--Gupta-MK--Sharma-SD--Sharma-AK--Rahal-A--2013"><span class="Hyperlink">Mailk </span><span class="Hyperlink">et al., 2013</span></a>). Brucellosis is one of the major zoonotic diseases. It is still prevalent throughout the world. Although many developed countries have eradicated <span class="CharOverride-11">Brucella abortus </span>from cattle, but it continues to pose a major public and animal health problem. Even though, <span class="CharOverride-11">Brucella </span>has definite host preferences, bovine brucellosis caused mainly by <span class="CharOverride-11">B. abortus</span>.<span class="CharOverride-11"> </span>The main factors of concern for the farmers to brucellosis are the low fertility and the high mortality in young animals (<a href="#Soomro-AH--Kamboh-AA--Rind-R--Dawani-P--Sarwar-M--Abro-SH--Awais-M-2014."><span class="Hyperlink">Soomro et al., 2014</span></a>).</p>
		  <p class="Body-Text">&nbsp;</p>
			<p class="Body-Text ParaOverride-1">Brucellosis is a major reproduction problem, and unfortunately in developing countries farmers not taken keen interest to regularly tests their animals for brucellosis. Some small scale farms cannot afford to practice the system of culling for positive animals. These animals become carriers of the <span class="CharOverride-11">Brucella </span>organism and thus a potential source of infection. Even though many countries have extensive eradication programs, some even having eradicated the disease, brucellosis is still a serious disease challenging the veterinary and medical professions. The reported incidences and the prevalence of the disease vary from country to country and region to region (<a href="#Radostits-OM--Gay-CC--Blood-DC--Hinchcliff-KW--2000"><span class="Hyperlink">Radostits et al., 2000</span></a>).</p>
		  <p class="Body-Text ParaOverride-1">&nbsp;</p>
			<p class="Body-Text ParaOverride-1"><span class="HTML-Cite CharOverride-12">The present study was therefore, carried out to record the prevalence of brucellosis in apparently healthy dairy cattle and buffaloes of Karachi, Pakistan.</span></p>
		  <p class="Body-Text ParaOverride-1">&nbsp;</p>
			<p class="Heading-1--Introduction----">MATERIALS AND METHODS</p>
		  <p class="Heading-1--Introduction----">&nbsp;</p>
			<p class="Heading-2--History-in-MM-">Study Design</p>
			<p class="Body-Text ParaOverride-1">A modified cross-sectional study design was used to find out the prevalence of brucellosis in cattle and buffaloes in Karachi. A total of 100 farms at different localities of Karachi were visited during February-August 2011 and samples were collected from lactating animals. Visits were made to all 4 districts of Karachi and a total of 200 dairy buffaloes and cattle were examined and samples including blood and milk samples were collected. A total of 5 ml milk or blood sample were collected ascetically in sterilized sample bottles and transported to laboratory under refrigerated conditions for analysis (<a href="#Soomro-AH--Kamboh-AA--Rind-R--Dawani-P--Sarwar-M--Abro-SH--Awais-M-2014."><span class="Hyperlink">Soomro et al., 2014</span></a>). Upon coagulation, serum were separated from blood samples and it was further centrifuged at 3000 rpm for 10 min to remove any cellular material. </p>
		  <p class="Body-Text ParaOverride-1">&nbsp;</p>
			<p class="Heading-2--History-in-MM-">Laboratory Procedures</p>
		  <p class="Heading-2--History-in-MM-">&nbsp;</p>
			<p class="Heading-3">Milk Ring Test (MRT)</p>
			<p class="Body-Text ParaOverride-1">Milk Ring Test (MRT) was performed according to the standard  procedure (<a href="#OIE--2008-"><span class="Hyperlink">OIE, 2008</span></a>). Hematoxylin stained <span class="CharOverride-11">Brucella abortus</span> (Strain-99) antigen was purchased from Veterinary Research Institute (VRI, Lahore, Pakistan) and used for MRT as recommended by manufacturer. </p>
		  <p class="Body-Text ParaOverride-1">&nbsp;</p>
			<p class="Heading-3">Rose Bengal Plate Test (RBPT)</p>
			<p class="Body-Text">Rose Bengal Plate Test (RBPT) was performed according to the method as described by <a href="#Khalil-MA--Gabbar-A--1992"><span class="Hyperlink">Khalil and Gabbar (1992)</span></a>. The Rose Bengal stained antigen containing <span class="CharOverride-11">Brucella abortus</span> cells (strain-99) suspended in buffer at pH 3.6 were used, as per the recommendation of supplier (VRI, Lahore, Pakistan). </p>
		  <p class="Body-Text">&nbsp;</p>
			<p class="Heading-3">Complement Fixation Test (CFT)</p>
			<p class="Body-Text">Serum samples which were reacted positive to RBPT were reanalysed by CFT to confirm and to eliminate any cross reaction. All test protocols and interpretations of fixation ratios were followed as per standard procedure (<a href="#OIE--2008-"><span class="Hyperlink">OIE, 2008</span></a>). </p>
			<p class="Heading-1--Introduction----">&nbsp;</p>
			<p class="Heading-1--Introduction----">RESULTS</p>
		  <p class="Heading-1--Introduction----">&nbsp;</p>
			<p class="Body-Text ParaOverride-1">During present survey, a total of 200 blood and milk samples were collected and examined for bovine brucellosis and results have been summarized in <a href="#Figure-1-"><span class="Hyperlink">Figure 1</span></a>. During sera analysis through RBPT, 35 (17.5%) were found sero-positive for bovine brucellosis (<a href="#Figure-1-"><span class="Hyperlink">Figure 1A</span></a>). However, when same sera were analyzed by CFT only 25 (12.5%) were shown reactivity (<a href="#Figure-1-"><span class="Hyperlink">Figure 1B</span></a>). </p>
		  <p class="Body-Text ParaOverride-1">&nbsp;</p>
			
			<div class="pt" > <a href="http://nexusacademicpublishers.com/uploads/figures/20150501172645.png" target="new"><img class="img_display" src="http://nexusacademicpublishers.com/uploads/figures/20150501172645.png" width="80" height="80"></a>
            
            <p class="Figure--and-Table-Heading ParaOverride-1"><span class="CharOverride-6"><a id="Figure-1-"></a>Figure 1: </span><a href="http://nexusacademicpublishers.com/uploads/figures/20150501172645.png">Prevalence of bovine brucellosis in Karachi:<span class="CharOverride-6"> A. </span>Analysed by RBPT;<span class="CharOverride-6"> B. </span>Analysed by CFT;<span class="CharOverride-6"> C. </span>Analysed by MRT</a></p>
       </div>

			
		  <p class="Body-Text ParaOverride-1">
		  <p class="Body-Text">For milk samples a total of 100 farms were visited and 200 milk samples were collected from cattle and buffalos of different towns of Karachi. These were analyzed by MRT and results have been summarized in <a href="#Table-1-"><span class="Hyperlink">Table 1</span></a> and <a href="#Figure-1-"><span class="Hyperlink">Figure 1C</span></a>. An overall, prevalence of <span class="CharOverride-11">Brucella abortus</span> in milk samples was recorded as 14% (28/200). Highest prevalence of bovine brucellosis was recorded in dairy farms of Landhi Cattle Colony (25%), followed by Hospital Chorangi (22.2%), Filter Plant (20.0%) and Karadar (16.67%), while lowest one was found at farms of Nagori dairies (5.88%). </p>
		  </p>
		  <p class="Body-Text">&nbsp; </p>
		  <p class="Figure--and-Table-Heading ParaOverride-1"><span class="CharOverride-6"><a id="Table-1-"></a>Table 1: </span>Prevalence of bovine brucellosis in milk samples in different towns of Karachi</p>
			<table width="658" height="201" class="Table-Style-1" id="table-1">
				<colgroup>
					<col />
					<col />
					<col />
					<col />
				</colgroup>
				<tbody>
					<tr class="_idGenTableRowColumn-1">
						<td>
							<p class="Basic-Paragraph"><span class="CharOverride-15">Town name</span></p>
						</td>
						<td>
							<p class="Basic-Paragraph"><span class="CharOverride-15">No. of individual farm’s milk examined</span></p>
						</td>
						<td>
							<p class="Basic-Paragraph"><span class="CharOverride-15">No. of positive</span></p>
							<p class="Basic-Paragraph"><span class="CharOverride-15">farms</span></p>
						</td>
						<td>
							<p class="Basic-Paragraph"><span class="CharOverride-15">Sample prevalence </span></p>
							<p class="Basic-Paragraph"><span class="CharOverride-15">(%)</span></p>
						</td>
					</tr>
					<tr class="_idGenTableRowColumn-2">
						<td>
							<p class="Basic-Paragraph"><span class="CharOverride-16">Landhi Cattle Colony</span></p>
						</td>
						<td>
							<p class="Basic-Paragraph"><span class="CharOverride-16">4</span></p>
						</td>
						<td>
							<p class="Basic-Paragraph"><span class="CharOverride-16">1</span></p>
						</td>
						<td>
							<p class="Basic-Paragraph"><span class="CharOverride-16">25.00</span></p>
						</td>
					</tr>
					<tr class="_idGenTableRowColumn-1">
						<td>
							<p class="Basic-Paragraph"><span class="CharOverride-16">Nagori dairies</span></p>
						</td>
						<td>
							<p class="Basic-Paragraph"><span class="CharOverride-16">34</span></p>
						</td>
						<td>
							<p class="Basic-Paragraph"><span class="CharOverride-16">2</span></p>
						</td>
						<td>
							<p class="Basic-Paragraph"><span class="CharOverride-16">5.88</span></p>
						</td>
					</tr>
					<tr class="_idGenTableRowColumn-2">
						<td>
							<p class="Basic-Paragraph"><span class="CharOverride-16">Hospital Chorangi</span></p>
						</td>
						<td>
							<p class="Basic-Paragraph"><span class="CharOverride-16">9</span></p>
						</td>
						<td>
							<p class="Basic-Paragraph"><span class="CharOverride-16">2</span></p>
						</td>
						<td>
							<p class="Basic-Paragraph"><span class="CharOverride-16">22.22</span></p>
						</td>
					</tr>
					<tr class="_idGenTableRowColumn-1">
						<td>
							<p class="Basic-Paragraph"><span class="CharOverride-16">Karadar </span></p>
						</td>
						<td>
							<p class="Basic-Paragraph"><span class="CharOverride-16">48</span></p>
						</td>
						<td>
							<p class="Basic-Paragraph"><span class="CharOverride-16">8</span></p>
						</td>
						<td>
							<p class="Basic-Paragraph"><span class="CharOverride-16">16.67</span></p>
						</td>
					</tr>
					<tr class="_idGenTableRowColumn-2">
						<td>
							<p class="Basic-Paragraph"><span class="CharOverride-16">Filter Plant</span></p>
						</td>
						<td>
							<p class="Basic-Paragraph"><span class="CharOverride-16">5</span></p>
						</td>
						<td>
							<p class="Basic-Paragraph"><span class="CharOverride-16">1</span></p>
						</td>
						<td>
							<p class="Basic-Paragraph"><span class="CharOverride-16">20.00</span></p>
						</td>
					</tr>
					<tr class="_idGenTableRowColumn-1">
						<td>
							<p class="Basic-Paragraph"><span class="CharOverride-16">Total</span></p>
						</td>
						<td>
							<p class="Basic-Paragraph"><span class="CharOverride-16">100</span></p>
						</td>
						<td>
							<p class="Basic-Paragraph"><span class="CharOverride-16">14</span></p>
						</td>
						<td>
							<p class="Basic-Paragraph"><span class="CharOverride-16">14.00</span></p>
						</td>
					</tr>
				</tbody>
			</table>
			<p class="Heading-1--Introduction----">&nbsp;</p>
		  <p class="Heading-1--Introduction----">DISCUSSION</p>
			<p class="Body-Text ParaOverride-1">&nbsp;</p>
			<p class="Body-Text ParaOverride-1">The prevention of human brucellosis depends on the control of the disease in animals. The successful eradication of the bovine disease has been achieved mainly in industrialized countries (Europe and Americas). However, most countries in Asia do have control programs as well, but they do not exist in Pakistan (<a href="#Corbel-MJ--1997-.-B"><span class="Hyperlink">Corbel, 1997</span></a>). The people of this country are frequently infected with <span class="CharOverride-11">B. abortus</span>, but it generally produces a less serious disease than virulent strains of <span class="CharOverride-11">B. melitensis</span> or <span class="CharOverride-11">B. suis</span> (<a href="#Alton-GG--Nielsen-K--Duncan-JR--1990"><span class="Hyperlink">Alton et al., 1990</span></a>). The findings of this study clearly underline the necessity to improve the legislation on animal disease controls. As in present investigation, among 200 blood samples examined, a proportion of 17.5% to 12.5% were found sero-positive by RBPT and CFT respectively. These values were quite higher (i.e., 1.84% and 1.38% by RBPT and CFT respectively) than recorded in a recent study in eastern Ethiopia (<a href="#Degefu-H--Mohamud-M--Hailemelekot-M--Yohannes-M--2011"><span class="Hyperlink">Degefu et al., 2011</span></a>). However, in an Irish study, 51% prevalence of bovine brucellosis was recorded in CFT (<a href="#Hayes-M--Ashe-S--Collins-DM--Power-S--Kenny-K--Sheahan-M--O-Hagan-G--More-SJ--2009"><span class="Hyperlink">Hayes et al., 2009</span></a>). These differences might be due to different geographical locations as prevalence of bacterial diseases having been found varied from region to region (<a href="#Ansari-ARMIH--Rahman-MM--Islam-MZ--Das-BC--Habib-A--Belal-SMSH--Islam-K--2014"><span class="Hyperlink">Ansari et al., 2014</span></a>; <a href="#Sachan-N--Nautiyal-B--Agarwal-RK--Singh-VP--Verma-AK--2013"><span class="Hyperlink">Sachan et al., 2013</span></a>). Similar type of findings has been observed in our results of samplings from different farms/towns. However, difference of testing techniques could also be factor for variation of prevalence recorded, because it is well known that different techniques have numerous level of sensitivity (<a href="#Gall-D--Nielsen-K--2004-"><span class="Hyperlink">Gall and Nielsen, 2004</span></a>). In general, the ELISA is preferable to the complement fixation test (CFT) because it is more sensitive and it is not affected by pro- or anti-complementary factors. If ELISA reagents are not available, or if subtyping is pursued, then CFT could be performed as a most sensitive test (<a href="#Ferris-NP--Dawson-M--1988"><span class="Hyperlink">Ferris and Dawson, 1988</span></a>).</p>
		  <p class="Body-Text ParaOverride-1">&nbsp;</p>
			<p class="Body-Text ParaOverride-1">Milk ring test (MRT) is known to be an essential screening test for brucellosis in milk. We have used this test in our study and found 14% cases as positive. This prevalence is quite low as compared to our study in Hyderabad (<a href="#Soomro-AH--Kamboh-AA--Rind-R--Dawani-P--Sarwar-M--Abro-SH--Awais-M-2014."><span class="Hyperlink">Soomro et al., 2014</span></a>) that showed 31-47% prevalence in cattle and buffaloes respectively. Likewise, in another study in Faisalabad, 20.1% was recorded in horses (<a href="#Gul-ST--Khan-A--Ahmad-M--Hussain-I--2013"><span class="Hyperlink">Gul et al., 2013</span></a>). These results clearly indicated that Karachi has a low prevalence of brucellosis as compared to other small cities of Pakistan i.e., Hyderabad and Faisalabad. This reflects the better managemental and sanitary conditions in dairy farms of Karachi as compared to other small cities of Pakistan. Because, it is well recognized that sanitary conditions like disposal of aborted material, contact with stray dogs and other wild animals etc., are the major risk factors for brucellosis (<a href="#Soomro-AH--Kamboh-AA--Rind-R--Dawani-P--Sarwar-M--Abro-SH--Awais-M-2014."><span class="Hyperlink">Soomro et al., 2014</span></a>). </p>
		  <p class="Body-Text ParaOverride-1">&nbsp;</p>
			<p class="Body-Text ParaOverride-1">From these results it could be concluded that brucellosis is present in the apparently healthy dairy cattle and buffaloes of Karachi, however, its prevalence is low comparing with other small cities of Pakistan. These results emphasize the need for a better monitoring of the disease by government policies or local veterinarians. Above all, it is essential to raise the awareness of the disease among all the dairy cattle/buffalo farmers.</p>
		  <p class="Body-Text ParaOverride-1">&nbsp;</p>
			<p class="Heading-1--Introduction----">References</p>
		  <p class="Heading-1--Introduction----">&nbsp;</p>
			
			  <li class="References ParaOverride-2" xml:lang="en-US"><a id="Alton-GG--Nielsen-K--Duncan-JR--1990"></a>Alton GG, Nielsen K, Duncan JR (1990). <span class="CharOverride-11">Brucella melitensis</span>, in Animal Brucellosis. CRC Press, Boca Raton. FL,<span class="CharOverride-11"> </span>383-422.</li>
				<li class="References ParaOverride-2" xml:lang="en-US"><a id="Corbel-MJ--1997-.-B"></a>Corbel MJ (1997). Brucellosis an Overview. Emerg. Infect. Dis.<span class="CharOverride-11"> </span>3(2): 213-21. <a href="http://dx.doi.org/10.3201/eid0302.970219"><span class="Hyperlink">http://dx.doi.org/10.3201/eid0302.970219</span></a></li>
				<li class="References ParaOverride-2" xml:lang="en-US"><a id="Ansari-ARMIH--Rahman-MM--Islam-MZ--Das-BC--Habib-A--Belal-SMSH--Islam-K--2014"></a>Ansari ARMIH, Rahman MM, Islam MZ, Das BC, Habib A, Belal SMSH, Islam K (2014). Prevalence and antimicrobial resistance profile of <span class="CharOverride-11">Escherichia coli</span> and <span class="CharOverride-11">salmonella</span> isolated from diarrheic calves. J. Anim. Health Prod. 2(1): 12 – 15. <a href="http://dx.doi.org/10.14737/journal.jahp/2014/2.1.12.15"><span class="Hyperlink">http://dx.doi.org/10.14737/journal.jahp/2014/2.1.12.15</span></a></li>
				<li class="References ParaOverride-2" xml:lang="en-US"><a id="Degefu-H--Mohamud-M--Hailemelekot-M--Yohannes-M--2011"></a>Degefu H, Mohamud M, Hailemelekot M, Yohannes M (2011). Seroprevalence of bovine brucellosis in agro pastoral areas of Jijjiga zone of Somali National Regional State, Eastern Ethiopia. Ethiop. Vet. J. 15(1): 37-47. <a href="http://dx.doi.org/10.4314/evj.v15i1.67683"><span class="Hyperlink">http://dx.doi.org/10.4314/evj.v15i1.67683</span></a></li>
				<li class="References ParaOverride-2" xml:lang="en-US"><a id="Ferris-NP--Dawson-M--1988"></a>Ferris NP, Dawson M (1988). Routine application of enzyme-linked immunosorbent assay in comparison with complement fixation for the diagnosis of foot-and-mouth and swine vesicular disease. Vet. Microbiol. 16: 201–209. <a href="http://dx.doi.org/10.1016/0378-1135(88)90024-7"><span class="Hyperlink">http://dx.doi.org/10.1016/0378-1135(88)90024-7</span></a></li>
				<li class="References ParaOverride-2" xml:lang="en-US"><a id="Gall-D--Nielsen-K--2004-"></a>Gall D, Nielsen K (2004). Serological diagnosis of bovine brucellosis: a review of test performance and cost comparison. Rev. Sci. Tech. 23(3): 989-1002.</li>
				<li class="References ParaOverride-2" xml:lang="en-US"><a id="Gul-ST--Khan-A--Ahmad-M--Hussain-I--2013"></a>Gul ST, Khan A, Ahmad M, Hussain I (2013). Seroprevalence of brucellosis and associated hemato-Biochemical changes in Pakistani horses. Pak. J. Agri. Sci. 50(4): 745-750. </li>
				<li class="References ParaOverride-2" xml:lang="en-US"><a id="Hayes-M--Ashe-S--Collins-DM--Power-S--Kenny-K--Sheahan-M--O-Hagan-G--More-SJ--2009"></a>Hayes M, Ashe S, Collins DM, Power S, Kenny K, Sheahan M, O’Hagan G, More SJ (2009). An evaluation of Irish cattle herds with inconclusive serological evidence of bovine brucellosis. Irish Vet. J. 62(3): 182-190. <a href="http://dx.doi.org/10.1186/2046-0481-62-3-182"><span class="Hyperlink">http://dx.doi.org/10.1186/2046-0481-62-3-182</span></a></li>
				<li class="References ParaOverride-2" xml:lang="en-US"><a id="Hoffmann-D--1999"></a>Hoffmann D (1999). Asian Livestock to the Year 2000 and beyond. Working Paper series<span class="CharOverride-11"> </span>1/2: 1-44. </li>
				<li class="References ParaOverride-2" xml:lang="en-US"><a id="Khalil-MA--Gabbar-A--1992"></a>Khalil MA, Gabbar A (1992). Procedure in Veterinary Microbiology. Field Documents No. 6.2<span class="CharOverride-17">nd</span> Ed. C.V.D.L. Tandojam, Sindh, Pakistan. 6-93.</li>
				<li class="References ParaOverride-2" xml:lang="en-US"><a id="OIE--2008-"></a>OIE (2008). Bovine brucellosis. In: Manual of diagnostic tests and vaccines for terrestrial animal. 6 th  Ed, Pp. 409-438.12, rue de Prony, Paris, France.</li>
				<li class="References ParaOverride-2" xml:lang="en-US"><a id="Radostits-OM--Gay-CC--Blood-DC--Hinchcliff-KW--2000"></a>Radostits OM, Gay CC, Blood DC, Hinchcliff KW (2000). Veterinary Medicine, A textbook of the Diseases of Cattle, Sheep, Pigs, Goats and Horses. Harcourt Publishers Limited, London. Pp. 867-882.</li>
				<li class="References ParaOverride-2" xml:lang="en-US"><a id="Mailk-S--Kumar-A--Verma-AK--Gupta-MK--Sharma-SD--Sharma-AK--Rahal-A--2013"></a>Mailk S, Kumar A, Verma AK, Gupta MK, Sharma SD, Sharma AK, Rahal A (2013). Incidence and drug resistance pattern of collibacillosis in cattle and buffalo calves in Western Utter Pradesh in India. J Anim. Health Prod. 1(2): 15-19</li>
				<li class="References ParaOverride-2" xml:lang="en-US"><a id="Sachan-N--Nautiyal-B--Agarwal-RK--Singh-VP--Verma-AK--2013"></a>Sachan N, Nautiyal B, Agarwal RK, Singh VP, Verma AK (2013). Sero-epidemiological Pattern of Salmonellosis in Animals of Rohilkhand Region, Uttar Pradesh, India. J. Anim. Health Prod. 1(1): 6-9.</li>
		  <li class="References ParaOverride-2" xml:lang="en-US"><a id="Soomro-AH--Kamboh-AA--Rind-R--Dawani-P--Sarwar-M--Abro-SH--Awais-M-2014."></a>Soomro AH, Kamboh AA, Rind R, Dawani P, Sarwar M, Abro SH, Awais M (2014). A study on prevalence and risk factors of brucellosis in cattle and buffaloes in district Hyderabad, Pakistan. J. Anim. Health Prod. 2 (3): 33 – 37. <a href="http://dx.doi.org/10.14737/journal.jahp/2014/2.3.33.37"><span class="Hyperlink">http://dx.doi.org/10.14737/journal.jahp/2014/2.3.33.37</span></a></li>
			
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