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			<p class="title- ParaOverride-1">Epidemiological Study of Sheep Anaplasmosis (<i>Anaplasma ovis</i> Infection) in Kairouan, Central Tunisia</p>
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			<p class="Authors ParaOverride-1">&nbsp;</p>
			<p class="Authors ParaOverride-1"><span class="CharOverride-3">Mohamed Gharbi</span><span class="CharOverride-4">1*</span><span class="CharOverride-3">, Houda Omri</span><span class="CharOverride-4">1</span><span class="CharOverride-3">, Mohamed Jedidi</span><span class="CharOverride-4">1</span><span class="CharOverride-3">, Samir Zorii</span><span class="CharOverride-4">2</span><span class="CharOverride-3">, Mohamed Aziz Darghouth</span><span class="CharOverride-4">1</span></p>
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			<p class="Affiliations ParaOverride-1"><span class="CharOverride-5">1</span>Laboratoire de Parasitologie, École Nationale de Médecine Vétérinaire, Université de Manouba, 2020 Sidi Thabet, Tunisia; <span class="CharOverride-5">2</span>Office des Terres Domaniales, Agro-Combinat El Alem, Tunisia.</p>
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			<p class="Abstract">&nbsp;</p>
			<p class="Abstract"><span class="CharOverride-6">Abstract</span> | Specific diagnosis of infection with <span class="CharOverride-7">Anaplasma ovis</span> is difficult, if based only on clinical signs. A cross-sectional survey was carried out in 26 flocks totalling 8,049 ear tagged Barbarin sheep. All the animals were clinically examined and Giemsa stained blood smears were performed. A total number of 282 <span class="CharOverride-7">Anaplasma ovis</span> infected sheep were examined for clinical symptoms. The cumulative incidence of sheep anaplasmosis was 11.6%, rickettsiaemia was significantly higher in symptomatic (median=16/50 field microscope) compared to asymptomatic animals (median=6/50 field microscope) (p&lt;0.001). Symptoms of anaplasmosis were pale mucosa (84.4%), anorexia (25%), prostration (51.6%), nasal discharge (18.3%), respiratory distress (22.6%) and fever (100%). Clinical cases with no mortality, and positive blood smears, only occurred in lambs, and adults were apparently immune. The situation may therefore be considered as enzootic stable. We recommend for field veterinarians to suspect erythrocytic anaplasmosis in regions where <span class="CharOverride-7">Rhipicephalus turanicus</span> is present in lambs with prostration, pale mucosa, and fever.</p>
		  <p class="Abstract">&nbsp;</p>
			<p class="Abstract ParaOverride-1"><span class="CharOverride-6">Keywords </span>| <span class="CharOverride-7">Anaplasma ovis</span>, Sheep, Erythrocytic anaplasmosis, Clinical symptoms, Tunisia</p>
		  <p class="Abstract ParaOverride-1">&nbsp;</p>
		</div>
		<div class="Basic-Text-Frame">
			<p class="Editor----Citation"><span class="CharOverride-6">Editor</span> | Muhammad Imran Rashid, Department of Parasitology, University of Veterinary and Animal Sciences, Lahore, Pakistan.</p>
			<p class="Editor----Citation"><span class="CharOverride-6">Received</span> | May 31, 2015; <span class="CharOverride-6">Revised</span> | June 11, 2015; <span class="CharOverride-6">Accepted </span>| June 12, 2015; <span class="CharOverride-6">Published</span> | June 16, 2015&#9;&#9;</p>
			<p class="Editor----Citation"><span class="CharOverride-6">*Correspondence</span> | Mohamed Gharbi, Laboratoire de Parasitologie, École Nationale de Médecine Vétérinaire, Univ. Manouba, 2020 Sidi Thabet, Tunisia; <span class="CharOverride-6">Email: </span>gharbim2000@yahoo.fr</p>
			<p class="Editor----Citation"><span class="CharOverride-6">Citation</span> | Gharbi M, Omri H, Jedidi M, Zorii S, Darghouth MA (2015). Epidemiological study of sheep anaplasmosis (<span class="CharOverride-21">Anaplasma ovis</span> infection) in Kairouan, Central Tunisia. J. Adv. Parasitol. 2(2): 30-34.  </p>
			<p class="Editor----Citation"><span class="CharOverride-6">DOI </span>| <a href="http://dx.doi.org/10.14737/journal.jap/2015/2.2.30.34"><span class="Hyperlink">http://dx.doi.org/10.14737/journal.jap/2015/2.2.30.34</span></a></p>
			<p class="Editor----Citation"><span class="CharOverride-6">ISSN</span> | 2311-4096 </p>
			<p class="Editor----Citation"><span class="CharOverride-6">Copyright</span> © 2015 Gharbi 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">S</span><span>heep </span><span class="CharOverride-8">production in Tunisia (North Africa) is an important sector, involving 4,044,000 females; the density of sheep in Tunisia is the highest in Africa (40 sheep/km²) (</span><a href="#Ministere-De-L-agriculture-Et-Des-Ressources-Hydrauliques--Tunisie--2006"><span class="Hyperlink">Ministère de l’Agriculture et des Ressources Hydrauliques, 2006</span></a><span class="CharOverride-8">). This species contributes to 50.3% of national red meat production (</span><a href="#Ministere-De-L-agriculture-Et-Des-Ressources-Hydrauliques--Tunisie--2006"><span class="Hyperlink">Ministère de l’Agriculture et des Ressources Hydrauliques, 2006</span></a><span class="CharOverride-8">). Sheep are facing several health problems with high prevalence such as cystic echinococcosis, brucellosis, toxoplasmosis, gastrointestinal and respiratory nematodes, fasciolosis, etc. (</span><a href="#Akkari-H--Gharbi-M--Darghouth-MA--2011"><span class="Hyperlink">Akkari et al., 2011</span></a><span class="CharOverride-8">; </span><a href="#Akkari-H--Gharbi-M--Darghouth-MA--2012"><span class="Hyperlink">Akkari et al., 2012</span></a><span class="CharOverride-8">; </span><a href="#Darghouth-MA--Gharbi-M--2011-"><span class="Hyperlink">Darghouth and Gharbi, 2011</span></a><span class="CharOverride-8">; </span><a href="#Lahmar-S--Trifi-M--Naceur-SB--Bouchhima-T--Lahouar-N--Lamouchi-I--Ma-mouri-N--Selmi-R--Dhibi-M--Torg"><span class="Hyperlink">Lahmar et al., 2012</span></a><span class="CharOverride-8">). The sheep population is mainly owned by small farmers with poor financial resources, since farmers having a land surface comprised between 0 and 10 ha own 70% of the sheep (</span><a href="#Ministere-De-L-agriculture-Et-Des-Ressources-Hydrauliques--Tunisie--2006"><span class="Hyperlink">Ministère de l’Agriculture et des Ressources Hydrauliques, 2006</span></a><span class="CharOverride-8">).</span></p>
		  <p class="Caps-on-First-Para ParaOverride-1">&nbsp;</p>
			<p class="Body-Text ParaOverride-1">According to <a href="#Friedhoff-KT--1997-.-Tick-borne"><span class="Hyperlink">Friedhoff (1997)</span></a> sheep anaplasmosis due to <span class="CharOverride-7">Anaplasma ovis </span>infection is a subclinical infection with little economic importance; <a href="#Camus-E--Uilenberg-G--2010"><span class="Hyperlink">Camus and Uilenberg (2010) </span></a>also consider it to be usually subclinical or moderately virulent. It is frequently misdiagnosed due to the absence of specific symptoms and consequently its confusion with other diseases. According to <a href="#Klabi-I--2011"><span class="Hyperlink">Klabi (2011)</span></a> this long-lasting infection may nevertheless cause an important decrease of weight gain, estimated in carrier lambs as over 14%. Ovine erythrocytic anaplasmosis seems to be an emergent disease in some Tunisian regions (North and Centre). Recently, <a href="#Chochlakis-D--Ioannou-I--Tselentis-Y--Psaroulaki-A--2010"><span class="Hyperlink">Chochlakis et al. (2010)</span></a> reported on what may be a human case of <span class="CharOverride-7">A. ovis </span>infection.</p>
		  <p class="Body-Text ParaOverride-1">&nbsp;</p>
			<p class="Body-Text ParaOverride-1">We report herein a cross-sectional epidemioclinical survey carried out in farms where clinical cases of sheep anaplasmosis were reported for the first time in 2003.</p>
		  <p class="Body-Text ParaOverride-1">&nbsp;</p>
			<p class="Heading-1--Introduction----">Material and methods</p>
		  <p class="Heading-1--Introduction----">&nbsp;</p>
			<p class="Body-Text ParaOverride-1">This cross-sectional survey was carried out during the summer season (from June to late August) concerned 26 flocks totalling 8,049 ear tagged Barbarin sheep (kept within a distance of about 3 km) in the Governorate of Kairouan (Central Tunisia, 35-40N; 010-06E). This locality is characterized by a semi-arid climate with a mean annual rainfall of 290 mm and mean monthly minimum and maximum temperatures of 13 and 36°C during December and July respectively.</p>
		  <p class="Body-Text ParaOverride-1">&nbsp;</p>
			<p class="Body-Text">The animals pasture from late March till early September in stubbles of wheat, maize, and barley. The rest of the year, they graze in natural poor pastures. During the lambing period (from September to November), ewes receive complementation with concentrate and oaten hay. Acaricide treatments (diazinon) are implemented on animals once a year in early summer. The sheep are treated against gastro-intestinal and respiratory nematodes with a subcutaneous injection of ivermectin at the conventional dose of 0.2 mg/kg.</p>
		  <p class="Body-Text">&nbsp;</p>
			<p class="Body-Text">Weekly, the sheep were examined to evaluate their general state, presence of pale mucosa (classified as present or absent), rectal temperature, respiratory and cardiac frequencies and any other symptoms. A clinical case of anaplasmosis was defined as an animal presenting positive blood smears and at least anorexia with prostration. Animals with positive blood smears but no symptoms were classified as carriers. </p>
			<p class="Body-Text ParaOverride-1">From each animal, 5 ml of EDTA blood were collected in order to make a Giemsa-stained blood smear. The slides were examined for <span class="CharOverride-7">Anaplasma ovis</span> and the presence of any other haemopathogens (<a href="#Gharbi-M--Uilenberg-G--2004-."><span class="Hyperlink">Gharbi and Uilenberg, 2004</span></a>). The infection intensity was estimated by calculating the number of <span class="CharOverride-7">Anaplasma</span> in 50 microscope fields (each containing approximately 300 erythrocytes).</p>
		  <p class="Body-Text ParaOverride-1">&nbsp;</p>
			<p class="Body-Text ParaOverride-1">Weekly prevalence was calculated as follows: </p>
		  <p class="Body-Text ParaOverride-1">&nbsp;</p>
			<p class="Body-Text ParaOverride-2"><span class="CharOverride-7">Weekly prevalence (%) = 100 x number of new cases per week/number of examined animals</span></p>
			<p class="Body-Text ParaOverride-1">&nbsp;</p>
			<p class="Body-Text ParaOverride-1">Comparison of percentages was made by using chi square test, rickettsiaemia was compared with student t test with a significance threshold of 0.05 (<a href="#Schwartz-D--1993"><span class="Hyperlink">Schwartz, 1993</span></a>).</p>
		  <p class="Body-Text ParaOverride-1">&nbsp;</p>
			<p class="Heading-1--Introduction----">Results</p>
		  <p class="Heading-1--Introduction----">&nbsp;</p>
			<p class="Body-Text ParaOverride-1">The only haemopathogen found in blood smears was <span class="CharOverride-7">A. ovis</span> with a peak incidence during late June-early July, a second peak was observed during mid-July. No clinical cases were observed after late August (<a href="#Figure-1-"><span class="Hyperlink">Figure</span><span class="Hyperlink"> 1</span></a>). Clinical cases were observed exclusively in lambs of both sexes with a cumulative incidence of 11.6%, no clinical cases were detected in the other age categories. The disease was more frequent in females than in males (p&lt;0.001) (<a href="#Table-1-"><span class="Hyperlink">Table 1</span></a>); diseased animals presented various symptoms (<a href="#Table-2-"><span class="Hyperlink">Table 2</span></a>). No mortality was observed or reported.</p>
		  <p class="Body-Text ParaOverride-1">&nbsp;</p>
			<p class="Body-Text ParaOverride-1">All clinically ill animals had positive Giemsa-stained smears except six, rickettsiaemia varied between 2 rickettsia to 267 per 50 microscope fields. The median rickettsiaemia was estimated to be 16 rickettsia/50 microscope fields (<a href="#Table-3-"><span class="Hyperlink">Table 3</span></a>). They were all lambs; none of the older animals had positive blood smears. Animals without symptoms but with positive blood-smears were considered as healthy carriers of the infection. The rickettsiaemia in these animals was significantly lower than clinically ill animals (p&lt;0.001), it varied between 2 and 300 rickettsia/50 field microscope in one animal (<a href="#Figure-2-"><span class="Hyperlink">Figure 2</span></a>). These animals were not in incubation, since these farms were visited periodically (data not shown). No statistically significant relation was observed between rickettsiaemia and the other symptoms.</p>
		  <p class="Body-Text ParaOverride-1">&nbsp;</p>
			
            <div class="pt" > <a href="http://nexusacademicpublishers.com/uploads/figures/20150617150056.png" target="new"><img class="img_display" src="http://nexusacademicpublishers.com/uploads/figures/20150617150056.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/20150617150056.png">Weekly incidence of clinical cases of ovine anaplasmosis</a></p>
       </div>

			
		  <p class="Figure--and-Table-Heading ParaOverride-1">&nbsp;</p>
		  <p class="Figure--and-Table-Heading ParaOverride-1">&nbsp;</p>
		  <p class="Figure--and-Table-Heading ParaOverride-1"><span class="CharOverride-6"><a id="Table-1-"></a>Table 1: </span><span class="CharOverride-13">Prevalence of clinical cases in different sheep categories with erythrocytic anaplasmosis</span></p>
			<table width="658" height="203" class="Table-Style-1" id="table-1">
				<colgroup>
					<col />
					<col />
					<col />
					<col />
					<col />
					<col />
					<col />
				</colgroup>
				<tbody>
					<tr class="_idGenTableRowColumn-1">
						<td width="136" height="59">
							<p class="Basic-Paragraph"><span class="CharOverride-14">Age category</span></p>
						</td>
						<td width="77">
							<p class="Basic-Paragraph"><span class="CharOverride-14">No. of sheep examined</span></p>
						</td>
						<td width="89">
							<p class="Basic-Paragraph"><span class="CharOverride-14">No. of positive blood smears</span></p>
						</td>
						<td width="56">
							<p class="Basic-Paragraph"><span class="CharOverride-14">No. of diseased animals</span></p>
						</td>
						<td width="89">
							<p class="Basic-Paragraph"><span class="CharOverride-14">Infection prevalence (S.E.)</span></p>
						</td>
						<td width="90">
							<p class="Basic-Paragraph"><span class="CharOverride-14">Carrier state prevalence (S.E.)</span></p>
						</td>
						<td width="89">
							<p class="Basic-Paragraph"><span class="CharOverride-14" xml:lang="en-US">Disease prevalence (S.E.)</span></p>
						</td>
					</tr>
					<tr class="_idGenTableRowColumn-2">
						<td height="21">
							<p class="Basic-Paragraph"><span class="CharOverride-15">Female lamb</span></p>
						</td>
						<td>
							<p class="Basic-Paragraph"><span class="CharOverride-15">1,756</span></p>
						</td>
						<td>
							<p class="Basic-Paragraph"><span class="CharOverride-15">280</span></p>
						</td>
						<td>
							<p class="Basic-Paragraph"><span class="CharOverride-15">224</span></p>
						</td>
						<td>
							<p class="Basic-Paragraph"><span class="CharOverride-15">19.14 (0.002)</span></p>
						</td>
						<td>
							<p class="Basic-Paragraph"><span class="CharOverride-15">3.19 (0.004)</span></p>
						</td>
						<td>
							<p class="Basic-Paragraph"><span class="CharOverride-15">12.76 (0.008)</span></p>
						</td>
					</tr>
					<tr class="_idGenTableRowColumn-1">
						<td height="21">
							<p class="Basic-Paragraph"><span class="CharOverride-15" xml:lang="en-US">Male lamb</span></p>
						</td>
						<td>
							<p class="Basic-Paragraph"><span class="CharOverride-15">675</span></p>
						</td>
						<td>
							<p class="Basic-Paragraph"><span class="CharOverride-15">65</span></p>
						</td>
						<td>
							<p class="Basic-Paragraph"><span class="CharOverride-15">58</span></p>
						</td>
						<td>
							<p class="Basic-Paragraph"><span class="CharOverride-15">10.65 (0.004)</span></p>
						</td>
						<td>
							<p class="Basic-Paragraph"><span class="CharOverride-15">1.04 (0.0039)</span></p>
						</td>
						<td>
							<p class="Basic-Paragraph"><span class="CharOverride-15">8.59 (0.010)</span></p>
						</td>
					</tr>
					<tr class="_idGenTableRowColumn-2">
						<td height="21">
							<p class="Basic-Paragraph"><span class="CharOverride-15" xml:lang="en-US">Yearling female lamb</span></p>
						</td>
						<td>
							<p class="Basic-Paragraph"><span class="CharOverride-15">1,133</span></p>
						</td>
						<td>
							<p class="Basic-Paragraph"><span class="CharOverride-15">0</span></p>
						</td>
						<td>
							<p class="Basic-Paragraph"><span class="CharOverride-15">0</span></p>
						</td>
						<td>
							<p class="Basic-Paragraph"><span class="CharOverride-15">0</span></p>
						</td>
						<td>
							<p class="Basic-Paragraph"><span class="CharOverride-15">0</span></p>
						</td>
						<td>
							<p class="Basic-Paragraph"><span class="CharOverride-15">0</span></p>
						</td>
					</tr>
					<tr class="_idGenTableRowColumn-1">
						<td height="21">
							<p class="Basic-Paragraph"><span class="CharOverride-15" xml:lang="en-US">Ewe</span></p>
						</td>
						<td>
							<p class="Basic-Paragraph"><span class="CharOverride-15">4,169</span></p>
						</td>
						<td>
							<p class="Basic-Paragraph"><span class="CharOverride-15">0</span></p>
						</td>
						<td>
							<p class="Basic-Paragraph"><span class="CharOverride-15">0</span></p>
						</td>
						<td>
							<p class="Basic-Paragraph"><span class="CharOverride-15">0</span></p>
						</td>
						<td>
							<p class="Basic-Paragraph"><span class="CharOverride-15">0</span></p>
						</td>
						<td>
							<p class="Basic-Paragraph"><span class="CharOverride-15">0</span></p>
						</td>
					</tr>
					<tr class="_idGenTableRowColumn-2">
						<td height="21">
							<p class="Basic-Paragraph"><span class="CharOverride-15" xml:lang="en-US">Yearling male lamb</span></p>
						</td>
						<td>
							<p class="Basic-Paragraph"><span class="CharOverride-15">59</span></p>
						</td>
						<td>
							<p class="Basic-Paragraph"><span class="CharOverride-15">0</span></p>
						</td>
						<td>
							<p class="Basic-Paragraph"><span class="CharOverride-15">0</span></p>
						</td>
						<td>
							<p class="Basic-Paragraph"><span class="CharOverride-15">0</span></p>
						</td>
						<td>
							<p class="Basic-Paragraph"><span class="CharOverride-15">0</span></p>
						</td>
						<td>
							<p class="Basic-Paragraph"><span class="CharOverride-15">0</span></p>
						</td>
					</tr>
					<tr class="_idGenTableRowColumn-1">
						<td height="21">
							<p class="Basic-Paragraph"><span class="CharOverride-15" xml:lang="en-US">Ram</span></p>
						</td>
						<td>
							<p class="Basic-Paragraph"><span class="CharOverride-15">257</span></p>
						</td>
						<td>
							<p class="Basic-Paragraph"><span class="CharOverride-15">0</span></p>
						</td>
						<td>
							<p class="Basic-Paragraph"><span class="CharOverride-15">0</span></p>
						</td>
						<td>
							<p class="Basic-Paragraph"><span class="CharOverride-15">0</span></p>
						</td>
						<td>
							<p class="Basic-Paragraph"><span class="CharOverride-15">0</span></p>
						</td>
						<td>
							<p class="Basic-Paragraph"><span class="CharOverride-15" xml:lang="en-US">0</span></p>
						</td>
					</tr>
				</tbody>
			</table>
			<p class="Figure--and-Table-Heading ParaOverride-1">&nbsp;</p>
			<p class="Figure--and-Table-Heading ParaOverride-1"><span class="CharOverride-16">S.E.:</span><span class="CharOverride-17"> Standard error</span></p>
			<p class="Figure--and-Table-Heading ParaOverride-1">&nbsp;</p>
		  <p class="Figure--and-Table-Heading ParaOverride-1">&nbsp;</p>
			
		  <p class="Figure--and-Table-Heading ParaOverride-1"><span class="CharOverride-6"><a id="Table-2-"></a>Table 2: </span><span class="CharOverride-13">Symptoms in sheep with erythrocytic anaplasmosis</span></p>
			<table width="657" height="255" class="Table-Style-1" id="table-2">
				<colgroup>
					<col />
					<col />
				</colgroup>
				<tbody>
					<tr class="_idGenTableRowColumn-1">
						<td colspan="2">
							<p class="Basic-Paragraph"><span class="CharOverride-14">Range</span></p>
						</td>
					</tr>
					<tr class="_idGenTableRowColumn-2">
						<td>
							<p class="Basic-Paragraph"><span class="CharOverride-15">Rectal temperature (°C)</span></p>
						</td>
						<td>
							<p class="Basic-Paragraph ParaOverride-1"><span class="CharOverride-15">38.9 -42.5</span></p>
						</td>
					</tr>
					<tr class="_idGenTableRowColumn-1">
						<td>
							<p class="Basic-Paragraph"><span class="CharOverride-15" xml:lang="en-US">Respiratory frequency (cycles/ min.)</span></p>
						</td>
						<td>
							<p class="Basic-Paragraph ParaOverride-1"><span class="CharOverride-15">14 - 42</span></p>
						</td>
					</tr>
					<tr class="_idGenTableRowColumn-2">
						<td>
							<p class="Basic-Paragraph"><span class="CharOverride-15" xml:lang="en-US">Cardiac frequency (pulsations/min.)</span></p>
						</td>
						<td>
							<p class="Basic-Paragraph ParaOverride-1"><span class="CharOverride-15">96 - 138</span></p>
						</td>
					</tr>
					<tr class="_idGenTableRowColumn-1">
						<td colspan="2">
							<p class="Basic-Paragraph"><span class="CharOverride-14">Number of sheep (percentage)</span></p>
						</td>
					</tr>
					<tr class="_idGenTableRowColumn-2">
						<td>
							<p class="Basic-Paragraph"><span class="CharOverride-15">Pale mucosa</span></p>
						</td>
						<td>
							<p class="Basic-Paragraph ParaOverride-1"><span class="CharOverride-15">291 (84.3)</span></p>
						</td>
					</tr>
					<tr class="_idGenTableRowColumn-1">
						<td>
							<p class="Basic-Paragraph"><span class="CharOverride-15" xml:lang="en-US">Prostration</span></p>
						</td>
						<td>
							<p class="Basic-Paragraph ParaOverride-1"><span class="CharOverride-15">178 (51.6)</span></p>
						</td>
					</tr>
					<tr class="_idGenTableRowColumn-2">
						<td>
							<p class="Basic-Paragraph"><span class="CharOverride-15" xml:lang="en-US">Anorexia</span></p>
						</td>
						<td>
							<p class="Basic-Paragraph ParaOverride-1"><span class="CharOverride-15">86 (25)</span></p>
						</td>
					</tr>
					<tr class="_idGenTableRowColumn-1">
						<td>
							<p class="Basic-Paragraph"><span class="CharOverride-15" xml:lang="en-US">Respiratory discordance</span></p>
						</td>
						<td>
							<p class="Basic-Paragraph ParaOverride-1"><span class="CharOverride-15">78 (22.61)</span></p>
						</td>
					</tr>
					<tr class="_idGenTableRowColumn-2">
						<td>
							<p class="Basic-Paragraph"><span class="CharOverride-15" xml:lang="en-US">Nasal discharge</span></p>
						</td>
						<td>
							<p class="Basic-Paragraph ParaOverride-1"><span class="CharOverride-15">63 (18.3)</span></p>
						</td>
					</tr>
					<tr class="_idGenTableRowColumn-1">
						<td>
							<p class="Basic-Paragraph"><span class="CharOverride-15" xml:lang="en-US">Diarrhea</span></p>
						</td>
						<td>
							<p class="Basic-Paragraph ParaOverride-1"><span class="CharOverride-15"> 1 (0.29)</span></p>
						</td>
					</tr>
				</tbody>
			</table>
			<p class="Figure--and-Table-Heading ParaOverride-1">&nbsp;</p>
			<p class="Figure--and-Table-Heading ParaOverride-1">&nbsp;</p>
		  <p class="Figure--and-Table-Heading ParaOverride-1"><span class="CharOverride-6"><a id="Table-3-"></a>Table&#160;3: </span><span class="CharOverride-13">Comparative indicators of </span>rickettsiaemia <span class="CharOverride-13">in the two sheep groups</span></p>
			<table width="657" height="163" class="Table-Style-1" id="table-3">
				<colgroup>
					<col />
					<col />
					<col />
				</colgroup>
				<tbody>
					<tr class="_idGenTableRowColumn-1">
						<td />
						<td colspan="2">
							<p class="Basic-Paragraph"><span class="CharOverride-14">Rickettsiaemia /50 microscope fields</span></p>
						</td>
					</tr>
					<tr class="_idGenTableRowColumn-2">
						<td>
							<p class="Basic-Paragraph"><span class="CharOverride-15">Indicator</span></p>
						</td>
						<td>
							<p class="Basic-Paragraph"><span class="CharOverride-15">Asymptomatic animals</span></p>
						</td>
						<td>
							<p class="Basic-Paragraph"><span class="CharOverride-15" xml:lang="en-US">Diseased animals</span></p>
						</td>
					</tr>
					<tr class="_idGenTableRowColumn-1">
						<td>
							<p class="Basic-Paragraph"><span class="CharOverride-15">Minimum</span></p>
						</td>
						<td>
							<p class="Basic-Paragraph"><span class="CharOverride-15">2</span></p>
						</td>
						<td>
							<p class="Basic-Paragraph"><span class="CharOverride-15" xml:lang="en-US">0</span></p>
						</td>
					</tr>
					<tr class="_idGenTableRowColumn-2">
						<td>
							<p class="Basic-Paragraph"><span class="CharOverride-15">1st quartile</span></p>
						</td>
						<td>
							<p class="Basic-Paragraph"><span class="CharOverride-15">6</span></p>
						</td>
						<td>
							<p class="Basic-Paragraph"><span class="CharOverride-15" xml:lang="en-US">8</span></p>
						</td>
					</tr>
					<tr class="_idGenTableRowColumn-1">
						<td>
							<p class="Basic-Paragraph"><span class="CharOverride-15">Median</span></p>
						</td>
						<td>
							<p class="Basic-Paragraph"><span class="CharOverride-15">8</span></p>
						</td>
						<td>
							<p class="Basic-Paragraph"><span class="CharOverride-15" xml:lang="en-US">16</span></p>
						</td>
					</tr>
					<tr class="_idGenTableRowColumn-2">
						<td>
							<p class="Basic-Paragraph"><span class="CharOverride-15">3 rd quartile</span></p>
						</td>
						<td>
							<p class="Basic-Paragraph"><span class="CharOverride-15">12</span></p>
						</td>
						<td>
							<p class="Basic-Paragraph"><span class="CharOverride-15" xml:lang="en-US">26</span></p>
						</td>
					</tr>
					<tr class="_idGenTableRowColumn-1">
						<td>
							<p class="Basic-Paragraph"><span class="CharOverride-15">Maximum</span></p>
						</td>
						<td>
							<p class="Basic-Paragraph"><span class="CharOverride-15">300</span></p>
						</td>
						<td>
							<p class="Basic-Paragraph"><span class="CharOverride-15" xml:lang="en-US">267</span></p>
						</td>
					</tr>
				</tbody>
			</table>
		  <p>&nbsp;</p>
			<p><br>
			  
		  </p>
			<div class="pt" > <a href="http://nexusacademicpublishers.com/uploads/figures/20150617152756.png" target="new"><img class="img_display" src="http://nexusacademicpublishers.com/uploads/figures/20150617152756.png" width="80" height="80"></a>
            
           <p class="Figure--and-Table-Heading ParaOverride-1"><span class="CharOverride-6"><a id="Figure-2-"></a>Figure 2: </span><a href="http://nexusacademicpublishers.com/uploads/figures/20150617152756.png">Rickettsiaemia in asymtomatic and diseased animals</a></p>
       </div>
			
		  <p class="Figure--and-Table-Heading ParaOverride-1"><span class="CharOverride-12" xml:lang="en-US">*: not statistically significant&#160;; **&#160;: statistically significant</span></p>
			<p class="Heading-1--Introduction----">&nbsp;</p>
			<p class="Heading-1--Introduction----">Discussion and conclusions</p>
		  <p class="Heading-1--Introduction----">&nbsp;</p>
			<p class="Body-Text ParaOverride-1">Ovine anaplasmosis due to <span class="CharOverride-7">A. ovis</span> infection is an under-diagnosed disease; it seems to be an emergent disease in some regions of Tunisia (mainly the North and Centre of Tunisia) (<a href="#Klabi-I--2011"><span class="Hyperlink">Klabi, 2011</span></a>).</p>
		  <p class="Body-Text ParaOverride-1">&nbsp;</p>
			<p class="Body-Text ParaOverride-1">To our knowledge, there is no gold standard for ovine erythrocytic blood smears examination associated with clinical signs should represent the best way to establish a diagnosis of sheep erythrocytic anaplasmosis. The overall prevalence of clinical cases was estimated as 3.5%, affecting exclusively lambs in their first tick season (less than 10 months of age) with a global incidence of 11.6% in the lambs. These prevalences are by far lower than those estimated by PCR in Silica (south-eastern Slovakia) which reached 70.2% (<a href="#Derdakova-M--Stefancikova-A--Spitalska"><span class="Hyperlink">Derdáková et al., 2011</span></a>); this is logical as prevalence in our study was based solely on Giemsa blood smear examination and clinical observation.</p>
		  <p class="Body-Text ParaOverride-1">&nbsp;</p>
			<p class="Body-Text ParaOverride-1">Since the farmers implement oestrus synchronization, lambing occurs exclusively during early autumn. As for several other haemopathogens, the infection intensity depends on the equilibrium between the immune system of the host and the pathogen leading to a continuous fluctuation of infection prevalence and intensity, and they were certainly underestimated in the present survey. The establishment of protective immunity can be explained by the infection of animals during the first months of the life, and consecutive infections boost their immunity. These data can explain the state of enzootic stability since neither positive blood smears nor clinical cases were observed in adult animals despite the fact that both of them were borne on the farms and exposed to the same risk factors (<a href="#Table-1-"><span class="Hyperlink">Table 1</span></a>). The lambs become immunized; the negative blood smears in adults should be explained by the very low rickettsiaemia. Infection was more frequent in female lambs (12.8%) than in male lambs (8.6%), which can be explained by the fact that only female lambs graze, and males are kept in the barns for fattening.</p>
		  <p class="Body-Text ParaOverride-1">&nbsp;</p>
			<p class="Body-Text ParaOverride-1">Maximum incidence was observed during late June-early July with a second smaller peak in mid-July and no clinical cases during late August. This period is characterized by high temperatures (peaks of 42°C), favorable for tick activity, a weakness of the immune system of lambs and colostral antibodies dramatically decreasing after the third month. Moreover, during this period, several veterinary actions were performed (vaccinations and injections of ivermectin) which are important risk factors since the veterinarian uses the same syringe for many of the animals.</p>
			<p class="Body-Text ParaOverride-1">&nbsp;</p>
			<p class="Body-Text ParaOverride-1">The symptoms were prostration, anorexia, fever, pale mucosa and less frequently cases of respiratory distress and nasal discharge. The anemia can be due to several other causes such as haemonchosis, fasciolosis or eperythrozoonosis (<span class="CharOverride-7">Mycoplasma ovis</span> infection). All the animals receive periodically ivermectin, the region is arid with no foci of fasciolosis and all the blood samples were negative for <span class="CharOverride-7">Mycoplasma ovis</span>. Hemoglobinuria was not observed in any of the clinical cases, an unusual symptom reported in Hungary (<a href="#Hornok-S--Elek-2007"><span class="Hyperlink">Hornok et al., 2007</span></a>). No relation was observed between rickettsiaemia and the other symptoms. This is an important finding leading to conclude that there is no threshold of rickettsiaemia to distinguish between carrier and diseased sheep. Hence, we recommend to the field veterinarian to usually associate any of the symptoms cited above with the presence of any level of rickettsiaemia.</p>
		  <p class="Body-Text ParaOverride-1">&nbsp;</p>
			<p class="Body-Text ParaOverride-1"><span class="CharOverride-7">Anaplasma ovis</span> has been reported to be transmitted by soft ticks (<span class="CharOverride-7">Ornithodoros lahorensis</span>), hard ticks (<span class="CharOverride-7">Rhipicephalus bursa</span>, <span class="CharOverride-7">R. turanicus</span> and <span class="CharOverride-7">Dermacentor andersoni</span>) and biting Diptera (which act as mechanical vectors), and transmission by sheep keds has been suspected (<a href="#Friedhoff-KT--1997-.-Tick-borne"><span class="Hyperlink">Friedhoff, 1997</span></a>; <a href="#Camus-E--Uilenberg-G--2010"><span class="Hyperlink">Camus and Uilenberg, 2010</span></a>; <a href="#Hornok-S--De-La-Fuente-J--Biro-N--Fernandez-D"><span class="Hyperlink">Hornok et al., 2011</span></a>).</p>
		  <p class="Body-Text ParaOverride-1">&nbsp;</p>
			<p class="Body-Text ParaOverride-1">The present study is one of the scarce studies about clinical symptoms of ovine anaplasmosis. Positive blood smears concerned exclusively lambs that become protected thereafter. It would appear from our study that practitioners should take the possibility of erythrocytic anaplasmosis as the cause of disease symptoms in lambs into account.</p>
		  <p class="Body-Text ParaOverride-1">&nbsp;</p>
			<p class="Heading-1--Introduction----">Acknowledgements</p>
		  <p class="Heading-1--Introduction----">&nbsp;</p>
			<p class="Body-Text ParaOverride-1">This study was supported by Laboratoire d’épidémiologie des infections enzootiques des herbivores en Tunisie (Ministry of Higher Education, Scientific Research and Information, Technology and Communication, Tunisia) and the Deutsche Forschungsgemeinschaft project ‘‘Molecular epidemiology network for promotion and support of delivery of live vaccines against <span class="CharOverride-7">Theileria</span> <span class="CharOverride-7">parva</span> and <span class="CharOverride-7">Theileria</span> <span class="CharOverride-7">annulata</span> infection in Eastern and Northern Africa’’ (AH 41/7-1). The authors are grateful to Dr Kamel Haouala (Office des Terres Domaniales, Tunisia) for supporting this work.</p>
		  <p class="Body-Text ParaOverride-1">&nbsp;</p>
			<p class="Heading-1--Introduction----">Conflict of Interest</p>
		  <p class="Heading-1--Introduction----">&nbsp;</p>
			<p class="Body-Text ParaOverride-1">We declare that we have no conflict of interest.</p>
		  <p class="Body-Text ParaOverride-1">&nbsp;</p>
			<p class="Heading-1--Introduction----">Author’s Contribution</p>
		  <p class="Heading-1--Introduction----">&nbsp;</p>
			<p class="Body-Text ParaOverride-1">Mohamed Gharbi performed the analyses and wrote the paper and Houda Omri collected the samples, examined the animals, performed the analyses. Mohamed Jedidi performed the analyses; Samir Zorii collected the samples and Mohamed Aziz Darghouth conceived the study.</p>
		  <p class="Body-Text ParaOverride-1">&nbsp;</p>
			<p class="Heading-1--Introduction----">References</p>
		  <p class="Heading-1--Introduction----">&nbsp;</p>
		
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