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			<p class="Type-of-Article">Research Article</p>
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			<p class="title- ParaOverride-1"><span class="CharOverride-1">Clinical Association of Chicken Anaemia Virus with Other Infectious Poultry Diseases in North India and Nepal: Its Pathological Studies, Molecular Epidemiology and RFLP Pattern of PCR Amplified Full Length Viral Genome </span></p>
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			<p class="Authors ParaOverride-1">&nbsp;</p>
			<p class="Authors ParaOverride-1"><span class="CharOverride-2">Gopal Krishan</span><span class="CharOverride-3">1</span><span class="CharOverride-2">, Santosh Kumar Shukla</span><span class="CharOverride-3">2</span><span class="CharOverride-2">, Prakash Bhatt</span><span class="CharOverride-3">3</span><span class="CharOverride-2">, Maqbool Yaqoob Wani</span><span class="CharOverride-3">4</span><span class="CharOverride-2">, Kuldeep Dhama</span><span class="CharOverride-3">5</span><span class="CharOverride-2">, Yash Pal Singh Malik</span><span class="CharOverride-3">6</span><span class="CharOverride-2">, Rajesh Kumar</span><span class="CharOverride-3">7*</span></p>
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		<div class="Basic-Text-Frame">
			<p class="Affiliations ParaOverride-1"><span class="CharOverride-4">1</span>Department of Animal Husbandry, Himachal Pradesh, India; <span class="CharOverride-4">2</span>Department of Veterinary Medicine;<span class="CharOverride-4"> 3</span>Veterinary Clinics, <span class="CharOverride-4">7</span>Department of Veterinary Microbiology, College of Veterinary and Animal Sciences, Govind Ballabh Pant University of Agriculture and Technology, Pantnagar, Udham Singh Nagar, Uttarakhand - 263 145, India; <span class="CharOverride-4">4</span>Immunology Section, <span class="CharOverride-4">5</span>Avian Diseases Section, Division of Pathology, <span class="CharOverride-4">6</span>Division of Biological Standardization, Indian Veterinary Research Institute (IVRI), Izatnagar (U.P.) – 243 122, India.  </p>
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			<p class="Abstract ParaOverride-1">&nbsp;</p>
			<p class="Abstract ParaOverride-1"><span class="CharOverride-5">Abstract</span> | The aim of the study was to assess the role of chicken anaemia virus (CAV) in precipitating important poultry diseases in commercial poultry flocks and its molecular surveillance in poultry flocks of North India (Uttarakhand and Uttar Pradesh states) and Nepal (Mahendra Nagar).<span class="CharOverride-5" xml:lang="en-US"> </span>Study was conducted during September-2012 to April-2013 on 13 randomly selected poultry flocks. Necropsy was performed for making tentative diagnosis of other concomitant diseases and samples were collected and epidemiological data was recorded. Flocks were screened for CAV by VP2 gene based PCR followed by PCR-RFLP analysis of viral genome. Haemato-biochemical parameters of flocks were also studied. Out of 185 tissue samples<span class="CharOverride-5" xml:lang="en-US">, </span>PCR screening confirmed the presence of CAV genome in 95 samples, while 50 showed other viruses like infectious bursal disease (IBD), Inclusion Body Hepatitis (IBH) and Marek’s Disease (MD). PCR-RFLP analysis of the full genome of different CAV isolates with <span class="CharOverride-6" xml:lang="en-US">Hin</span>d III, <span class="CharOverride-6" xml:lang="en-US">Pst </span>I, <span class="CharOverride-6" xml:lang="en-US">Nhe </span>I and <span class="CharOverride-6" xml:lang="en-US">Mbo </span>II restriction enzymes showed indistinguishable restriction patterns. A significant decline in erythrocytic and leukocytic cell counts, and increase in the activities of degenerating enzymes was observed in CAV positive flocks. In conclusion, CAV was detected at different poultry farms of India and Nepal and its association with precipitation of other poultry diseases through its marked immunosuppressive potential is evident from findings of present study. Thus, the role of CAV in precipitating other infectious diseases in birds along with molecular epidemiology and immunosuppressive effects needs to be studied in detail so that a comprehensive strategy may be formulated for timely adoption of effective prevention and control measures, subsequently alleviating economic losses in poultry rearing countries.</p>
		  <p class="Abstract ParaOverride-1">&nbsp;</p>
			<p class="Abstract ParaOverride-1"><span class="CharOverride-5">Keywords</span> | Chicken anaemia virus, Poultry, Immunosuppression, Haemato-biochemical parameters, PCR- RFLP, Molecular epidemiology </p>
		  <p class="Abstract ParaOverride-1">&nbsp;</p>
		</div>
		<div class="Basic-Text-Frame">
			<p class="Editor----Citation"><span class="CharOverride-5">Editor</span> | Muhammad Munir (PhD), Avian Viral Diseases Programme, Compton Laboratory, Newbury, Berkshire, RG20 7NN, UK.</p>
			<p class="Editor----Citation"><span class="CharOverride-5">Received</span> | May 20, 2015; <span class="CharOverride-5">Revised</span> | June 12, 2015; <span class="CharOverride-5">Accepted</span> | June 13, 2015; <span class="CharOverride-5">Published</span> | June 17, 2015&#9;&#9;</p>
			<p class="Editor----Citation"><span class="CharOverride-5">*Correspondence</span> | Rajesh Kumar, Govind Ballabh Pant University of Agriculture and Technology, Pantnagar, Udham Singh Nagar, Uttarakhand- 263 145, India; <span class="CharOverride-5">Email:</span> rajeshvet@rediffmail.com</p>
			<p class="Editor----Citation"><span class="CharOverride-5">Citation</span> | Krishan G, Shukla SK, Bhatt P, Wani MY, Dhama K, Malik YPS, Kumar R (2015). Clinical association of chicken anaemia virus with other infectious poultry diseases in North India and Nepal: Its pathological studies, molecular epidemiology and RFLP pattern of PCR amplified full length viral genome. Adv. Anim. Vet. Sci. 3(7): 395-405.  </p>
			<p class="Editor----Citation"><span class="CharOverride-5">DOI</span> | <a href="http://dx.doi.org/10.14737/journal.aavs/2015/3.7.395.405"><span class="Hyperlink">http://dx.doi.org/10.14737/journal.aavs/2015/3.7.395.405</span></a></p>
			<p class="Editor----Citation"><span class="Editor---Citation CharOverride-5">ISSN (Online)</span> | 2307-8316; <span class="Editor---Citation CharOverride-5">ISSN (Print)</span> | 2309-3331</p>
			<p class="Editor----Citation"><span class="CharOverride-5">Copyright</span> © 2015 Krishan 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="Heading-1--Introduction----">&nbsp;</p>
			<p class="Caps-on-First-Para ParaOverride-1"><span class="_idGenDropcap-1">C</span><span>hicken </span>anaemia virus (CAV), the causative agent of chicken infectious anaemia (CIA), belongs to the <span class="Emphasis">Gyrovirus</span> genus of the <span class="CharOverride-6">Circoviridae</span> family (<a href="#Pringle-CR--1999"><span class="Hyperlink">Pringle, 1999</span></a>; <a href="#Todd-D--McNulty-2000"><span class="Hyperlink">Todd et al., 2000</span></a>), and is an economically important and emerging pathogen affecting poultry industry worldwide (<a href="#McNulty-MS--1991-"><span class="Hyperlink">McNulty, 1991</span></a>; <a href="#Jiang-SJ--Meng-SS--Cui-ZZ--Tian-FL--Wang-ZF--2005"><span class="Hyperlink">Jiang et al., 2005</span></a>; <a href="#Dhama-K--Mahe-2008"><span class="Hyperlink">Dhama et al., 2008</span></a>; <a href="#Schat-KA--2009"><span class="Hyperlink">Schat, 2009</span></a>; <a href="#Kim-HR--Kwon-YK--Bae-YC--Oem-JK--Lee-OS--2010-."><span class="Hyperlink">Kim et al., 2010</span></a>; <a href="#Oluwayelu-DO--2010"><span class="Hyperlink">Oluwayelu, 2010</span></a>; <a href="#Bhatt-P--Shukla-2013"><span class="Hyperlink">Bhatt et al., 2013</span></a>; <a href="#Snoeck-CJ--Komoyo-GF--Mbee-BP--Nakoun--E--Le-Faou-A--Okwen-MP---Muller-CP--2012-"><span class="Hyperlink">Snoeck et al., 2012</span></a>; <a href="#Nayabian-H--Mardani-K--2013"><span class="Hyperlink">Nayabian et al., 2013</span></a>; <a href="#Wani-MY--Dhama-2013"><span class="Hyperlink">Wani et al., 2013</span></a>). CAV is a non-enveloped virus with a circular, covalently linked, negative-sense, ss DNA genome of about 2,298 to 2,319 nucleotides, and is one of the smallest avian viruses (23-25 nm size). The major transcript from the CAV genome is an unspliced polycistronic mRNA of about 2100 nucleotides encoding VP1 (51.6 kDa), the viral capsid protein; VP2 (24.0 kDa), a scaffold protein; and VP3 or apoptin (13.6 kDa) that is responsible for causing apoptosis in chicken thymocytes and chicken lymphoblastoid cells (<a href="#Todd-D--Mawhinney-KA--McNulty-MS--1992"><span class="Hyperlink">Todd et al., 1992</span></a>; <a href="#Natesan-S--Kataria-JM--Dhama-K--Rahul-S--Baradhwaj-N--2006"><span class="Hyperlink">Natesan et al., 2006</span></a>). </p>
		  <p class="Caps-on-First-Para ParaOverride-1">&nbsp;</p>
			<p class="Body-Text">Chicken anaemia virus infection is characterized by lymphoid atrophy, immunosuppression, increased mortality, aplasia of the bone marrow and anaemia, reduced body weight gain, and the development of subcutaneous and intramuscular haemorrhages in chickens (<a href="#Dhama-K--Mahe-2008"><span class="Hyperlink">Dhama et al., 2008</span></a>; <a href="#Schat-KA--2009"><span class="Hyperlink">Schat, 2009</span></a>; <a href="#Pope-CR--1991"><span class="Hyperlink">Pope, 1991</span></a>; <a href="#Miller-MM--Schat-KA--2004-."><span class="Hyperlink">Miller and Schat, 2004</span></a>). No specific treatment exists for CIA, supplementation with haematinics and immunostimulants can ameliorate anaemic condition and depressed immunity (<a href="#Bhatt-P--Shukla-2013"><span class="Hyperlink">Bhatt et al., 2013</span></a>; <a href="#Latheef-SK--Dhama-K--Wani-MY--Samad-HA--Tiwari-R--Singh-SD--2013"><span class="Hyperlink">Latheef et al., 2013</span></a>). Vaccines including of live-attenuated and killed vaccines are being used but have some limitations, and therefore DNA vaccine and other advanced vaccine options are being explored (<a href="#McNulty-MS--1991-"><span class="Hyperlink">McNulty, 1991</span></a>; <a href="#Schat-KA--2003"><span class="Hyperlink">Schat, 2003</span></a>; <a href="#Dhama-K--Mahe-2008"><span class="Hyperlink">Dhama et al., 2008</span></a>, <a href="#Dhama-K--Tiwari-2014"><span class="Hyperlink">2014</span></a>, <a href="#Dhama-K--Sam-2015"><span class="Hyperlink">2015</span></a>; <a href="#Sawant-PM--Dhama-K--Rawool-DB--Wani-MY--Tiwari-R--Singh-SD--Singh-RK--2015-."><span class="Hyperlink">Sawant et al., 2015</span></a>; <a href="#Zhang-X--Wu-B--Liu-Y--Chen-W--Dai-Z--Bi-Y--Xie-Q--2015-."><span class="Hyperlink">Zhang et al., 2015</span></a>). CAV is considered to play an important role in precipitation of number of poultry diseases (<a href="#Dhama-K--Mahe-2008"><span class="Hyperlink">Dhama et al., 2008</span></a>). Experimental co-infection of CAV with Marek’s disease virus (MDV) (<a href="#Zanella-A--Caro-R--Dall-AP--Grilli-G--Lavazza-A--Lombardi-G--1999"><span class="Hyperlink">Zanella et al., 1999</span></a>; <a href="#Miles-M--Reddy-M--Morgan-W--2001-."><span class="Hyperlink">Miles et al., 2001</span></a>), dual infection of CAV with infectious bursal disease virus (IBDV) (<a href="#Yuasa-N--Tani-1980"><span class="Hyperlink">Yuasa et al., 1980</span></a>; <a href="#Rosenberger-JK--Cloud-SS--1989"><span class="Hyperlink">Rosenberger et al., 1989</span></a>; <a href="#Imai-K--Mase-M--Tsukamoto-K--Hihara-H--Yuasa-N--1999"><span class="Hyperlink">Imai et al., 1991</span></a>; <a href="#McNeilly-F--Smyth-JA--Adair-BM--McNulty-MS--1995-."><span class="Hyperlink">McNeilly et al., 1995</span></a>), or adenovirus (<a href="#Toro-H--Gonzalez-2000"><span class="Hyperlink">Toro et al., 2000</span></a> and <a href="#Toro-H--Gonzalez-2001"><span class="Hyperlink">2001</span></a>) have resulted in increased morbidity and mortality. CAV infection in poultry has also been associated with poor efficacy of vaccines against Marek’s disease, Newcastle disease and infectious laryngotracheitis (<a href="#Box-PG--Holmes-HC--Bushell-AC--Finney-PM--1988-."><span class="Hyperlink">Box et al., 1988</span></a>; <a href="#Otaki-Y--Nunoya-T--Tajima-M--Kato-A--Nomura-Y--1998-."><span class="Hyperlink">Otaki et al., 1988</span></a>; <a href="#Cloud-SS--Rosenberger-JK--Lillehoj-HS--1992-"><span class="Hyperlink">Cloud et al., 1992</span></a>; <a href="#Liu-ZG--Zheng-SM--Yang-LP--Xu-YB--Zhou-ZY--2001"><span class="Hyperlink">Liu et al., 2001</span></a>) Precipitation and/ or increase in mortality due to these diseases by concomitant CAV infection owes to immunosuppression caused by CAV (<a href="#McNulty-MS--1991-"><span class="Hyperlink">McNulty, 1991</span></a>; <a href="#Dhama-K--Mahe-2008"><span class="Hyperlink">Dhama et al., 2008</span></a>; <a href="#Pope-CR--1991"><span class="Hyperlink">Pope, 1991</span></a>).</p>
		  <p class="Body-Text">&nbsp;</p>
			<p class="Body-Text ParaOverride-1">CAV has been reported from poultry flocks of India based on clinical signs, virus isolation, serological and molecular detection studies (<a href="#Natesan-S--Kataria-JM--Dhama-K--Rahul-S--Baradhwaj-N--2006"><span class="Hyperlink">Natesan et al., 2003</span></a>; <a href="#Dhama-K--2004"><span class="Hyperlink">Dhama et al., 2004</span></a>; <a href="#Bhatt-P--Shukla-SK--Mahendran-M--Dhama-K--Chawak-MM--Kataria-JM--2011"><span class="Hyperlink">Bhatt et al., 2011</span></a>; <a href="#Wani-MY--Dhama-2013"><span class="Hyperlink">Wani et al., 2013</span></a>), however, detailed molecular epidemiological studies, immunosuppression, clinical presentation and association of the virus with other infectious pathogens/diseases of poultry is lacking. Applications of molecular tools like PCR along with RE analysis and sequencing have emerged as effective confirmatory tools for diagnosis and studying the molecular epidemiology of poultry pathogens including CAV, which help in designing and adapting appropriate disease prevention and control programme. However, applications of clinical diagnostic methods based on haemato-biochemical parameters help in devising the effective treatment strategies. Present study describes the molecular epidemiology of CAV and its role in precipitating important infectious diseases of poultry in commercial flocks of northern Indian states (Uttarakhand and Uttar Pradesh) and Nepal.</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-">Sample Collection </p>
			<p class="Body-Text">Epidemiological data of different infectious diseases from the poultry farms of Uttarakhand state (Udham Singh Nagar, Nainital districts), Uttar Pradesh (Bareilly, Pilibhit and Rampur districts), India and Nepal (Mahendra Nagar district) were recorded during the period of September-2012 to April-2013 (<a href="#Table-1-"><span class="Hyperlink">Table 1</span></a>). Blood samples were collected from affected birds of all the flocks under investigation at the beginning of different disease outbreaks. Five apparently unhealthy birds were selected from each flock, and 2 ml of blood was collected in heparinised vials (10-20 IU/ml) for haemato-biochemical studies and in sterilized test tubes for separation of serum. Post-mortem examination of at least three dead birds from each farm was carried out. Detailed gross post-mortem lesions were recorded and tentative diagnosis was made. Tissue samples from liver, thymus, spleen, bursa of fabricius and bone marrow were collected in sterilized vial using aseptic conditions and kept in air tight containers at -20°C for PCR detection of CAV genome.</p>
		  <p class="Body-Text">&nbsp;</p>
			<p class="Heading-2--History-in-MM-">PCR Detection of CAV </p>
			<p class="Body-Text">For the detection of CAV by PCR in clinical samples, whole tissue DNA was extracted from thymus, bursa, spleen, liver and bone marrow using GeneiPure<span class="CharOverride-9">TM </span>Mammalian Genomic DNA Purification Kit (Bangalore Genei) as per manufacturer’s recommendations. The absorbance of the extracted DNA was measured at 260 nm and 280 nm in UV-VIS spectrophotometer. The ratio of OD<span class="CharOverride-10">260</span> and OD<span class="CharOverride-10">280</span> was calculated to check the purity of extracted DNA. The DNA samples having ratio of ≥ 1.8 were used for PCR. The amplification of CAV VP2 gene was carried out using sense VP2F: 5‘ATG CAC GGG AAC GGC GGA C3’ and antisense VP2R: 5‘TCA CAC TAT ACG TAC CGG GG3’ primers as described previously (<a href="#Basaraddi-MS--Dhama-K--Wani-M--Sawant-PM--Tiwari-R--Tiwari-R--Kumar-D--Singh-SD--Singh-R--2013"><span class="Hyperlink">Basaraddi et al., 2013</span></a>). The PCR products were confirmed by running in l % (w/v) agarose gel in 0.5X TBE buffer with DNA bands stained with ethidium bromide (0.5µg/ml) as per the method of <a href="#Sambrook-J--Fristch-EF--Miniatus-TC--1989"><span class="Hyperlink">Sambrook et al. (1989)</span></a>, and visualized and documented using Gel documentation system (Alpha Imager).</p>
		  <p class="Body-Text">&nbsp;</p>
			<p class="Heading-2--History-in-MM-">Restriction Endonuclease (RE) Analysis </p>
			<p class="Body-Text ParaOverride-1">Full genome of the virus was amplified by using primers viz., FGF: 5’-GAA TTC CGA GTG GTT ACT ATT CCA TCA-3 and FGR: ‘5’-GAT AGT GCG ATA AAT CTA TTT TCT GCG T-3’. The amplified products were subjected to restriction endonuclease (RE) enzyme digestion using <span class="CharOverride-6">Hin</span>d III, <span class="CharOverride-6">Pst </span>I, <span class="CharOverride-6">Nhe </span>I and <span class="CharOverride-6">Mbo </span>II restriction enzymes<span class="CharOverride-6"> </span>as per the manufacturer’s protocol, using specific assay conditions and the buffers supplied (Merck Genei). The molecular weights of various fragments of the digested viral DNA were determined using Fortran Software and expressed in base pairs (bp).</p>
		  <p class="Body-Text ParaOverride-1">&nbsp;</p>
			<p class="Heading-2--History-in-MM-">Haemato-biochemical Analysis</p>
			<p class="Body-Text ParaOverride-1">The various haematological parameters viz., haemoglobin (Hb) content, packed cell volume (PCV), total erythrocyte count (TEC), total leukocyte count (TLC) and differential leukocyte count (DLC) were estimated by standard procedures. For biochemical parameters, aspartate aminotransferase (AST), alanine aminotransferase (ALT), alkaline phosphtase (ALP) and uric acid (UA) were estimated using commercially available reagent kits.</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="Heading-2--History-in-MM-">Prevalence of CAV and Co-infections</p>
			<p class="Body-Text ParaOverride-1">During the disease investigation in the affected poultry flocks, birds were found suffering from chicken infectious anaemia– gangrenous dermatitis syndrome (CIA-GDS), Marek’s disease, infectious bursal disease and inclusion body hepatitis (<a href="#Figure-1-"><span class="Hyperlink">Figure 1</span></a> and <a href="#Table-2-"><span class="Hyperlink">Table 2</span></a>). In the PCR based screening of a total of 185 tissue samples collected from twelve different flocks, 145 samples (78.38%) were found positive for CAV (95/95), Marke’s disease (15/15), infectious bursal disease (20/20) and inclusion body hepatitis (15/15), while 40 samples tested negative for fowl pox, CRD and gout. The CAV positive flocks showed virus positive PCR amplicons of 651 bp (VP2 gene) and ~2.3 kb (complete genome) (<a href="#Figure-2-"><span class="Hyperlink">Figure 2</span></a>). The detailed results of the molecular surveillance of CAV in the affected poultry flocks are presented in <a href="#Table-3-"><span class="Hyperlink">Table 3</span></a>.</p>
			<p class="Body-Text ParaOverride-1">&nbsp;</p>
		  <p class="Body-Text ParaOverride-1">&nbsp;</p>
			<p class="Figure--and-Table-Heading ParaOverride-1"><span class="CharOverride-5"><a id="Table-1-"></a>Table 1: </span>Details of poultry flocks</p>
			<table width="659" height="324" class="Table-Style-1" id="table-1">
				<colgroup>
					<col />
					<col />
					<col />
					<col />
					<col />
					<col />
				</colgroup>
				<tbody>
					<tr class="_idGenTableRowColumn-1">
						<td width="51">
							<p class="Basic-Paragraph"><span class="CharOverride-11">Sr. No.</span></p>
						</td>
						<td width="188">
							<p class="Basic-Paragraph"><span class="CharOverride-11">Farm Code/Area</span></p>
						</td>
						<td width="47">
							<p class="Basic-Paragraph"><span class="CharOverride-11">Code No.</span></p>
						</td>
						<td width="108">
							<p class="Basic-Paragraph"><span class="CharOverride-11">System of management</span></p>
						</td>
						<td width="106">
							<p class="Basic-Paragraph"><span class="CharOverride-11">Age of Birds</span></p>
						</td>
						<td width="131">
							<p class="Basic-Paragraph"><span class="CharOverride-11">Total Flock Strength </span></p>
						</td>
					</tr>
					<tr class="_idGenTableRowColumn-2">
						<td>
							<p class="Basic-Paragraph"><span class="CharOverride-12">1.</span></p>
						</td>
						<td>
							<p class="Basic-Paragraph"><span class="CharOverride-12">N (Baazpur, UK</span><span class="CharOverride-13">*</span><span class="CharOverride-12">)</span></p>
						</td>
						<td>
							<p class="Basic-Paragraph"><span class="CharOverride-12">N</span></p>
						</td>
						<td>
							<p class="Basic-Paragraph"><span class="CharOverride-12">Cage</span></p>
						</td>
						<td>
							<p class="Basic-Paragraph"><span class="CharOverride-12">14 weeks</span></p>
						</td>
						<td>
							<p class="Basic-Paragraph"><span class="CharOverride-12">6000</span></p>
						</td>
					</tr>
					<tr class="_idGenTableRowColumn-1">
						<td>
							<p class="Basic-Paragraph"><span class="CharOverride-12">2.</span></p>
						</td>
						<td>
							<p class="Basic-Paragraph"><span class="CharOverride-12">SP (Baazpur, UK)</span></p>
						</td>
						<td>
							<p class="Basic-Paragraph"><span class="CharOverride-12">SP</span></p>
						</td>
						<td>
							<p class="Basic-Paragraph"><span class="CharOverride-12">Cage</span></p>
						</td>
						<td>
							<p class="Basic-Paragraph"><span class="CharOverride-12">12 weeks</span></p>
						</td>
						<td>
							<p class="Basic-Paragraph"><span class="CharOverride-12">12000</span></p>
						</td>
					</tr>
					<tr class="_idGenTableRowColumn-2">
						<td>
							<p class="Basic-Paragraph"><span class="CharOverride-12">3.</span></p>
						</td>
						<td>
							<p class="Basic-Paragraph"><span class="CharOverride-12">KP (Kashipur, UK)</span></p>
						</td>
						<td>
							<p class="Basic-Paragraph"><span class="CharOverride-12">KP</span></p>
						</td>
						<td>
							<p class="Basic-Paragraph"><span class="CharOverride-12">Cage</span></p>
						</td>
						<td>
							<p class="Basic-Paragraph"><span class="CharOverride-12">23 weeks</span></p>
						</td>
						<td>
							<p class="Basic-Paragraph"><span class="CharOverride-12">9000</span></p>
						</td>
					</tr>
					<tr class="_idGenTableRowColumn-1">
						<td>
							<p class="Basic-Paragraph"><span class="CharOverride-12">4.</span></p>
						</td>
						<td>
							<p class="Basic-Paragraph"><span class="CharOverride-12">NP (Jaffarpur, UK)</span></p>
						</td>
						<td>
							<p class="Basic-Paragraph"><span class="CharOverride-12">NP</span></p>
						</td>
						<td>
							<p class="Basic-Paragraph"><span class="CharOverride-12">Cage</span></p>
						</td>
						<td>
							<p class="Basic-Paragraph"><span class="CharOverride-12">17 weeks</span></p>
						</td>
						<td>
							<p class="Basic-Paragraph"><span class="CharOverride-12">8000</span></p>
						</td>
					</tr>
					<tr class="_idGenTableRowColumn-2">
						<td>
							<p class="Basic-Paragraph"><span class="CharOverride-12">5.</span></p>
						</td>
						<td>
							<p class="Basic-Paragraph"><span class="CharOverride-12">IP (Punjabnagar, UP</span><span class="CharOverride-13">*</span><span class="CharOverride-12">)</span></p>
						</td>
						<td>
							<p class="Basic-Paragraph"><span class="CharOverride-12">IP</span></p>
						</td>
						<td>
							<p class="Basic-Paragraph"><span class="CharOverride-12">Deep litter</span></p>
						</td>
						<td>
							<p class="Basic-Paragraph"><span class="CharOverride-12">2 weeks</span></p>
						</td>
						<td>
							<p class="Basic-Paragraph"><span class="CharOverride-12">1000</span></p>
						</td>
					</tr>
					<tr class="_idGenTableRowColumn-1">
						<td>
							<p class="Basic-Paragraph"><span class="CharOverride-12">6.</span></p>
						</td>
						<td>
							<p class="Basic-Paragraph"><span class="CharOverride-12">DP (Ramnagar, UK)</span></p>
						</td>
						<td>
							<p class="Basic-Paragraph"><span class="CharOverride-12">DP</span></p>
						</td>
						<td>
							<p class="Basic-Paragraph"><span class="CharOverride-12">Deep litter</span></p>
						</td>
						<td>
							<p class="Basic-Paragraph"><span class="CharOverride-12">3 weeks</span></p>
						</td>
						<td>
							<p class="Basic-Paragraph"><span class="CharOverride-12">1200</span></p>
						</td>
					</tr>
					<tr class="_idGenTableRowColumn-2">
						<td>
							<p class="Basic-Paragraph"><span class="CharOverride-12">7.</span></p>
						</td>
						<td>
							<p class="Basic-Paragraph"><span class="CharOverride-12">AP (Bilaspur, UP)</span></p>
						</td>
						<td>
							<p class="Basic-Paragraph"><span class="CharOverride-12">AP</span></p>
						</td>
						<td>
							<p class="Basic-Paragraph"><span class="CharOverride-12">Deep litter</span></p>
						</td>
						<td>
							<p class="Basic-Paragraph"><span class="CharOverride-12">3 weeks</span></p>
						</td>
						<td>
							<p class="Basic-Paragraph"><span class="CharOverride-12">2400</span></p>
						</td>
					</tr>
					<tr class="_idGenTableRowColumn-1">
						<td>
							<p class="Basic-Paragraph"><span class="CharOverride-12">8.</span></p>
						</td>
						<td>
							<p class="Basic-Paragraph"><span class="CharOverride-12">ZP (Pilibhit, UP)</span></p>
						</td>
						<td>
							<p class="Basic-Paragraph"><span class="CharOverride-12">ZP</span></p>
						</td>
						<td>
							<p class="Basic-Paragraph"><span class="CharOverride-12">Deep litter</span></p>
						</td>
						<td>
							<p class="Basic-Paragraph"><span class="CharOverride-12">5 weeks</span></p>
						</td>
						<td>
							<p class="Basic-Paragraph"><span class="CharOverride-12">2000</span></p>
						</td>
					</tr>
					<tr class="_idGenTableRowColumn-2">
						<td>
							<p class="Basic-Paragraph"><span class="CharOverride-12">9.</span></p>
						</td>
						<td>
							<p class="Basic-Paragraph"><span class="CharOverride-12">PN (Golapar, UK)</span></p>
						</td>
						<td>
							<p class="Basic-Paragraph"><span class="CharOverride-12">PN</span></p>
						</td>
						<td>
							<p class="Basic-Paragraph"><span class="CharOverride-12">Deep litter</span></p>
						</td>
						<td>
							<p class="Basic-Paragraph"><span class="CharOverride-12">4 weeks</span></p>
						</td>
						<td>
							<p class="Basic-Paragraph"><span class="CharOverride-12">1200</span></p>
						</td>
					</tr>
					<tr class="_idGenTableRowColumn-1">
						<td>
							<p class="Basic-Paragraph"><span class="CharOverride-12">10.</span></p>
						</td>
						<td>
							<p class="Basic-Paragraph"><span class="CharOverride-12">AH (Baazpur, UK)</span></p>
						</td>
						<td>
							<p class="Basic-Paragraph"><span class="CharOverride-12">AH</span></p>
						</td>
						<td>
							<p class="Basic-Paragraph"><span class="CharOverride-12">Cage</span></p>
						</td>
						<td>
							<p class="Basic-Paragraph"><span class="CharOverride-12">16 weeks</span></p>
						</td>
						<td>
							<p class="Basic-Paragraph"><span class="CharOverride-12">9000</span></p>
						</td>
					</tr>
					<tr class="_idGenTableRowColumn-2">
						<td>
							<p class="Basic-Paragraph"><span class="CharOverride-12">11.</span></p>
						</td>
						<td>
							<p class="Basic-Paragraph"><span class="CharOverride-12">PI (Pilibhit, UP)</span></p>
						</td>
						<td>
							<p class="Basic-Paragraph"><span class="CharOverride-12">PI</span></p>
						</td>
						<td>
							<p class="Basic-Paragraph"><span class="CharOverride-12">Deep litter</span></p>
						</td>
						<td>
							<p class="Basic-Paragraph"><span class="CharOverride-12">6 weeks</span></p>
						</td>
						<td>
							<p class="Basic-Paragraph"><span class="CharOverride-12">1300</span></p>
						</td>
					</tr>
					<tr class="_idGenTableRowColumn-1">
						<td>
							<p class="Basic-Paragraph"><span class="CharOverride-12">12.</span></p>
						</td>
						<td>
							<p class="Basic-Paragraph"><span class="CharOverride-12">PF (Mahendra Nagar, Nepal)</span></p>
						</td>
						<td>
							<p class="Basic-Paragraph"><span class="CharOverride-12">PF</span></p>
						</td>
						<td>
							<p class="Basic-Paragraph"><span class="CharOverride-12">Deep litter</span></p>
						</td>
						<td>
							<p class="Basic-Paragraph"><span class="CharOverride-12">2 weeks</span></p>
						</td>
						<td>
							<p class="Basic-Paragraph"><span class="CharOverride-12">1800</span></p>
						</td>
					</tr>
					<tr class="_idGenTableRowColumn-2">
						<td>
							<p class="Basic-Paragraph"><span class="CharOverride-12">13.</span></p>
						</td>
						<td>
							<p class="Basic-Paragraph"><span class="CharOverride-12">MP (Bareilly, UP)</span></p>
						</td>
						<td>
							<p class="Basic-Paragraph"><span class="CharOverride-12">MP</span></p>
						</td>
						<td>
							<p class="Basic-Paragraph"><span class="CharOverride-12">Deep litter</span></p>
						</td>
						<td>
							<p class="Basic-Paragraph"><span class="CharOverride-12">7 weeks</span></p>
						</td>
						<td>
							<p class="Basic-Paragraph"><span class="CharOverride-12">900</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-7">* </span><span class="CharOverride-15">UK</span><span class="CharOverride-7"> = Uttarakhand (India), </span><span class="CharOverride-15">UP</span><span class="CharOverride-7"> = Uttar Pradesh (India)</span></p>
			<p class="Figure--and-Table-Heading ParaOverride-1">&nbsp;</p>
		  <p class="Figure--and-Table-Heading ParaOverride-1">&nbsp;</p>
			<br>
			
          <div class="pt" > <a href="http://nexusacademicpublishers.com/uploads/figures/20150619153426.png" target="new"><img class="img_display" src="http://nexusacademicpublishers.com/uploads/figures/20150619153426.png" width="80" height="80"></a>
            
     	<p class="Figure--and-Table-Heading ParaOverride-1"><span class="CharOverride-5"><a id="Figure-1-"></a>Figure 1: </span><a href="http://nexusacademicpublishers.com/uploads/figures/20150619153426.png">Necropsy findings of the suspected CAV infected poultry birds collected from different farms</a></p>
       </div>

		  <p class="Figure--and-Table-Heading ParaOverride-1">(<span class="CharOverride-5">a</span>) Moist haemorrhages on the skin of CIA-GDS affected bird; (<span class="CharOverride-5">b</span>) Nodular appearance of liver in Marek’s disease affected bird; (<span class="CharOverride-5">c</span>) Swollen liver with necrotic foci in CIA-GDS affected bird; (<span class="CharOverride-5">d</span>) Urate deposits in Gout affected bird; (<span class="CharOverride-5">e</span>) Thickened proventriculus in Marek’s disease affected bird; (f) Caseous plug on opening of trachea in Fowl pox affected bird; (<span class="CharOverride-5">g</span>) Haemorrhages on mucosal surface of bursa in IBD affected bird; (<span class="CharOverride-5">h</span>) Pericarditis and perihepatitis in CCRD affected bird and (<span class="CharOverride-5">i</span>) Punctate haemorrhages on the liver of IBH affected bird</p>
		  <p class="Figure--and-Table-Heading ParaOverride-1">&nbsp;</p>
			
			
             <div class="pt" > <a href="http://nexusacademicpublishers.com/uploads/figures/20150619155427.png" target="new"><img class="img_display" src="http://nexusacademicpublishers.com/uploads/figures/20150619155427.png" width="80" height="80"></a>
            
     	<p class="Figure--and-Table-Heading ParaOverride-1"><span class="CharOverride-5"><a id="Figure-2-"></a>Figure 2: </span><a href="http://nexusacademicpublishers.com/uploads/figures/20150619155427.png">Amplification of CAV-DNA using PCR assay using VP2 specific primer (<span class="CharOverride-5">A</span>) and by using both VP2 and full length genome primers (<span class="CharOverride-5">B</span>)</a></p>
       </div>
       <br>
			
		  <div class="pt" > <a href="http://nexusacademicpublishers.com/uploads/figures/20150619155529.png" target="new"><img class="img_display" src="http://nexusacademicpublishers.com/uploads/figures/20150619155529.png" width="80" height="80"></a>
            
     		<p class="Figure--and-Table-Heading ParaOverride-1"><span class="CharOverride-5"><a id="Figure-3-"></a>Figure 3: </span><a href="http://nexusacademicpublishers.com/uploads/figures/20150619155529.png">Restriction fragment length polymorphism (RFLP) pattern after RE digestion of amplified full length CAV genome with four different enzymes and their combinations</a></p>
       </div>
       
			
			
		  <p class="Heading-2--History-in-MM- ParaOverride-1">&nbsp;</p>
		  <p class="Heading-2--History-in-MM- ParaOverride-1">Restriction Enzyme (RE) Analysis of Full Genome CAV Isolates </p>
			<p class="Body-Text ParaOverride-1">RE analysis of full genome amplicon (~2.3 kb) of all CAV isolates with <span class="CharOverride-6">HindIII</span>, <span class="CharOverride-6">PstI</span>, <span class="CharOverride-6">NheI</span> and <span class="CharOverride-6">MboII</span> restriction enzymes revealed identical RFLP patterns (<a href="#Figure-3-"><span class="Hyperlink">Figure 3</span></a>, <a href="#Table-4-"><span class="Hyperlink">Table 4</span></a>). All amplicons contained three <span class="CharOverride-6">HindIII</span> sites, one <span class="CharOverride-6">PstI</span> site, one <span class="CharOverride-6">NheI</span> site and one <span class="CharOverride-6">MboII</span> site. </p>
		  <p class="Body-Text ParaOverride-1">&nbsp;</p>
			<p class="Heading-2--History-in-MM-">Analysis of Haematological Parameters   </p>
			<p class="Body-Text ParaOverride-1">Haematological parameters of CAV positive and negative groups revealed a significant (P&lt;0.05) decline in the Hb, PCV, TEC, mean Hb and mean PCV in the CAV positive groups as compared to the control group (<a href="#Table-5-"><span class="Hyperlink">Table 5</span></a>). In comparison to healthy control, a significant (P&lt;0.05) reduction in the mean TEC values of CAV positive flocks was observed except the one farm (KP). In the myeloid lineages also, a significant (P&lt;0.05) decline in the TLC, percent lymphocyte count (PLC) and mean PLC was observed in all the CAV positive birds as compared to the negative group. The mean TLC was found significantly (P&lt;0.05) lower in the all the CAV positive flocks except in one positive flock (MP) which was suspected for IBH based on gross lesions. All the CAV positive flocks showed a significant (P&lt;0.05) increase in the mean percent heterophil count as compared to the negative control group.</p>
		  <p class="Body-Text ParaOverride-1">&nbsp; </p>
			<p class="Heading-2--History-in-MM-">Analysis of Biochemical parameters </p>
			<p class="Body-Text ParaOverride-1">All serum samples showed significantly (P&lt;0.05) higher activities of AST, ALT and ALP enzymes in all the CAV positive flocks (<a href="#Table-6-"><span class="Hyperlink">Table 6</span></a>). The mean uric acid value was found significantly (P&lt;0.05) higher in the CAV positive flocks except in one CAV positive farm (KP) when compared with the negative group. </p>
		  <p class="Body-Text ParaOverride-1">&nbsp;</p>
			<p class="Figure--and-Table-Heading ParaOverride-1"><span class="CharOverride-5"><a id="Table-2-"></a>Table 2: </span>Necropsy findings and molecular surveillance of suspected twelve poultry flocks for CAV with their respective tentative diagnosis</p>
			<table width="657" height="1009" class="Table-Style-1" id="table-2">
				<colgroup>
					<col />
					<col />
					<col />
					<col />
					<col />
					<col />
					<col />
					<col />
					<col />
				</colgroup>
				<tbody>
					<tr class="_idGenTableRowColumn-1">
						<td width="24" rowspan="2">
							<p class="Basic-Paragraph"><span class="CharOverride-11">Sr. No.</span></p>
						</td>
						<td width="36" rowspan="2">
							<p class="Basic-Paragraph"><span class="CharOverride-11">Farm Code </span></p>
						</td>
						<td width="264" rowspan="2">
							<p class="Basic-Paragraph"><span class="CharOverride-17">Necropsy findings</span></p>
						</td>
						<td width="62" rowspan="2">
							<p class="Basic-Paragraph"><span class="CharOverride-11">Tentative diagnosis</span></p>
						</td>
						<td width="36" rowspan="2">
							<p class="Basic-Paragraph"><span class="CharOverride-11">Mort ality </span><span class="CharOverride-11">(%)</span></p>
						</td>
						<td width="55" rowspan="2">
							<p class="Basic-Paragraph"><span class="CharOverride-11">Type of tissue   sample</span></p>
						</td>
						<td colspan="3">
							<p class="Basic-Paragraph"><span class="CharOverride-11">No. of samples screened for CAV</span></p>
						</td>
					</tr>
					<tr class="_idGenTableRowColumn-2">
						<td width="35">
							<p class="Basic-Paragraph"><span class="CharOverride-12">Total </span></p>
						</td>
						<td width="51">
							<p class="Basic-Paragraph"><span class="CharOverride-12">Positive </span></p>
						</td>
						<td width="54">
							<p class="Basic-Paragraph"><span class="CharOverride-12">Negative</span></p>
						</td>
					</tr>
					<tr class="_idGenTableRowColumn-1">
						<td>
							<p class="Basic-Paragraph"><span class="CharOverride-12">1</span></p>
						</td>
						<td>
							<p class="Basic-Paragraph"><span class="CharOverride-12">SP</span></p>
						</td>
						<td>
							<p class="Basic-Paragraph"><span class="CharOverride-18">- Haemorrhagic and necrotic lesions on the wings and breast region</span></p>
							<p class="Basic-Paragraph"><span class="CharOverride-18">- Severe atrophy of thymus and bursa</span></p>
							<p class="Basic-Paragraph"><span class="CharOverride-18">- Bone marrow aplasia with yellowish discolouration</span></p>
							<p class="Basic-Paragraph"><span class="CharOverride-18">- Pale discolouration of liver</span></p>
						</td>
						<td>
							<p class="Basic-Paragraph"><span class="CharOverride-12">Chicken Infectious Anaemia  Gangren Dermatitis Syndrome</span></p>
						</td>
						<td>
							<p class="Basic-Paragraph"><span class="CharOverride-12">5.72</span></p>
						</td>
						<td>
							<p class="Basic-Paragraph"><span class="CharOverride-12">Thymus, Bursa,</span></p>
							<p class="Basic-Paragraph"><span class="CharOverride-12">Bone marrow, Spleen, Liver</span></p>
						</td>
						<td>
							<p class="Basic-Paragraph"><span class="CharOverride-12">20</span></p>
						</td>
						<td>
							<p class="Basic-Paragraph"><span class="CharOverride-12">20</span></p>
						</td>
						<td>
							<p class="Basic-Paragraph"><span class="CharOverride-12">-</span></p>
						</td>
					</tr>
					<tr class="_idGenTableRowColumn-2">
						<td>
							<p class="Basic-Paragraph"><span class="CharOverride-12">2</span></p>
						</td>
						<td>
							<p class="Basic-Paragraph"><span class="CharOverride-12">KP</span></p>
						</td>
						<td>
							<p class="Basic-Paragraph"><span class="CharOverride-18">- Discolouration of liver with presence of nodules</span></p>
							<p class="Basic-Paragraph"><span class="CharOverride-18">- Marked thickening of proventriculus</span></p>
						</td>
						<td>
							<p class="Basic-Paragraph"><span class="CharOverride-12">Marek’s Disease</span></p>
						</td>
						<td>
							<p class="Basic-Paragraph"><span class="CharOverride-12">2.67</span></p>
						</td>
						<td>
							<p class="Basic-Paragraph"><span class="CharOverride-12">-do-</span></p>
						</td>
						<td>
							<p class="Basic-Paragraph"><span class="CharOverride-12">15</span></p>
						</td>
						<td>
							<p class="Basic-Paragraph"><span class="CharOverride-12">15</span></p>
						</td>
						<td>
							<p class="Basic-Paragraph"><span class="CharOverride-12">-</span></p>
						</td>
					</tr>
					<tr class="_idGenTableRowColumn-1">
						<td>
							<p class="Basic-Paragraph"><span class="CharOverride-12">3</span></p>
						</td>
						<td>
							<p class="Basic-Paragraph"><span class="CharOverride-12">NP</span></p>
						</td>
						<td>
							<p class="Basic-Paragraph"><span class="CharOverride-18">- Cheesy plug on opening of trachea</span></p>
							<p class="Basic-Paragraph"><span class="CharOverride-18">- Diptheritic membrane with white nodules along mucosal lining of trachea, upper esophagus and nasal cavity</span></p>
						</td>
						<td>
							<p class="Basic-Paragraph"><span class="CharOverride-12">Fowl Pox</span></p>
						</td>
						<td>
							<p class="Basic-Paragraph"><span class="CharOverride-12">1.20</span></p>
						</td>
						<td>
							<p class="Basic-Paragraph"><span class="CharOverride-12">-do-</span></p>
						</td>
						<td>
							<p class="Basic-Paragraph"><span class="CharOverride-12">15</span></p>
						</td>
						<td>
							<p class="Basic-Paragraph"><span class="CharOverride-12">-</span></p>
						</td>
						<td>
							<p class="Basic-Paragraph"><span class="CharOverride-12">15</span></p>
						</td>
					</tr>
					<tr class="_idGenTableRowColumn-2">
						<td>
							<p class="Basic-Paragraph"><span class="CharOverride-12">4</span></p>
						</td>
						<td>
							<p class="Basic-Paragraph"><span class="CharOverride-12">IP</span></p>
						</td>
						<td>
							<p class="Basic-Paragraph"><span class="CharOverride-18">- Chalky white deposition of urate crystals on heart, liver, kidneys and under the skin</span></p>
							<p class="Basic-Paragraph"><span class="CharOverride-18">- Swollen kidneys with urate deposits in ureters giving white chord appearance</span></p>
						</td>
						<td>
							<p class="Basic-Paragraph"><span class="CharOverride-12">Gout</span></p>
						</td>
						<td>
							<p class="Basic-Paragraph"><span class="CharOverride-12">3.50</span></p>
						</td>
						<td>
							<p class="Basic-Paragraph"><span class="CharOverride-12">-do-</span></p>
						</td>
						<td>
							<p class="Basic-Paragraph"><span class="CharOverride-12">15</span></p>
						</td>
						<td>
							<p class="Basic-Paragraph"><span class="CharOverride-12">-</span></p>
						</td>
						<td>
							<p class="Basic-Paragraph"><span class="CharOverride-12">15</span></p>
						</td>
					</tr>
					<tr class="_idGenTableRowColumn-1">
						<td>
							<p class="Basic-Paragraph"><span class="CharOverride-12">5</span></p>
						</td>
						<td>
							<p class="Basic-Paragraph"><span class="CharOverride-12">DP</span></p>
						</td>
						<td>
							<p class="Basic-Paragraph"><span class="CharOverride-18">- Haemorrhagic and necrotic lesions on the wings, breast and thighs</span></p>
							<p class="Basic-Paragraph"><span class="CharOverride-18">- Thymus and bursal atrophy</span></p>
							<p class="Basic-Paragraph"><span class="CharOverride-18">- Bone marrow hypoplasia with presence of pink colour</span></p>
						</td>
						<td>
							<p class="Basic-Paragraph"><span class="CharOverride-12">Chicken Infectious Anaemia Gangren Dermatitis Syndrome</span></p>
						</td>
						<td>
							<p class="Basic-Paragraph"><span class="CharOverride-12">11.25</span></p>
						</td>
						<td>
							<p class="Basic-Paragraph"><span class="CharOverride-12">-do-</span></p>
						</td>
						<td>
							<p class="Basic-Paragraph"><span class="CharOverride-12">15</span></p>
						</td>
						<td>
							<p class="Basic-Paragraph"><span class="CharOverride-12">15</span></p>
						</td>
						<td>
							<p class="Basic-Paragraph"><span class="CharOverride-12">-</span></p>
						</td>
					</tr>
					<tr class="_idGenTableRowColumn-2">
						<td>
							<p class="Basic-Paragraph"><span class="CharOverride-12">6</span></p>
						</td>
						<td>
							<p class="Basic-Paragraph"><span class="CharOverride-12">AP</span></p>
						</td>
						<td>
							<p class="Basic-Paragraph"><span class="CharOverride-18">- Haemorrhages on thigh muscles</span></p>
							<p class="Basic-Paragraph"><span class="CharOverride-18">- Enlarged bursa with presence of haemorrhages on mucosal surface</span></p>
							<p class="Basic-Paragraph"><span class="CharOverride-18">- Swollen kidneys </span></p>
						</td>
						<td>
							<p class="Basic-Paragraph"><span class="CharOverride-12">Infectious Bursal Disease</span></p>
						</td>
						<td>
							<p class="Basic-Paragraph"><span class="CharOverride-12">4.00</span></p>
						</td>
						<td>
							<p class="Basic-Paragraph"><span class="CharOverride-12">-do-</span></p>
						</td>
						<td>
							<p class="Basic-Paragraph"><span class="CharOverride-12">20</span></p>
						</td>
						<td>
							<p class="Basic-Paragraph"><span class="CharOverride-12">20</span></p>
						</td>
						<td>
							<p class="Basic-Paragraph"><span class="CharOverride-12">-</span></p>
						</td>
					</tr>
					<tr class="_idGenTableRowColumn-1">
						<td>
							<p class="Basic-Paragraph"><span class="CharOverride-12">7</span></p>
						</td>
						<td>
							<p class="Basic-Paragraph"><span class="CharOverride-12">ZP</span></p>
						</td>
						<td>
							<p class="Basic-Paragraph"><span class="CharOverride-18">- Haemorrhagic and necrotic lesions on the wings, breast and thighs</span></p>
							<p class="Basic-Paragraph"><span class="CharOverride-18">- Thymus and bursal atrophy</span></p>
							<p class="Basic-Paragraph"><span class="CharOverride-18">- Pink coloured marrow cavity of femur</span></p>
							<p class="Basic-Paragraph"><span class="CharOverride-18">- Slight enlargement of liver with pale discolouration</span></p>
						</td>
						<td>
							<p class="Basic-Paragraph"><span class="CharOverride-12">Chicken Infectious Anaemia  Gangren Dermatitis Syndrome</span></p>
						</td>
						<td>
							<p class="Basic-Paragraph"><span class="CharOverride-12">4.15</span></p>
						</td>
						<td>
							<p class="Basic-Paragraph"><span class="CharOverride-12">-do-</span></p>
						</td>
						<td>
							<p class="Basic-Paragraph"><span class="CharOverride-12">15</span></p>
						</td>
						<td>
							<p class="Basic-Paragraph"><span class="CharOverride-12">15</span></p>
						</td>
						<td>
							<p class="Basic-Paragraph"><span class="CharOverride-12">-</span></p>
						</td>
					</tr>
					<tr class="_idGenTableRowColumn-2">
						<td>
							<p class="Basic-Paragraph"><span class="CharOverride-12">8</span></p>
						</td>
						<td>
							<p class="Basic-Paragraph"><span class="CharOverride-12">PN</span></p>
						</td>
						<td>
							<p class="Basic-Paragraph"><span class="CharOverride-18">- Haemorrhagic lesions on wings</span></p>
							<p class="Basic-Paragraph"><span class="CharOverride-18">- Marked atrophy of thymus with mild bursal atrophy</span></p>
							<p class="Basic-Paragraph"><span class="CharOverride-18">- Pinkish discolouration of bone marrow</span></p>
							<p class="Basic-Paragraph"><span class="CharOverride-18">- Discolouration of liver</span></p>
						</td>
						<td>
							<p class="Basic-Paragraph"><span class="CharOverride-12">Chicken Infectious Anaemia Gangren Dermatitis Syndrome</span></p>
						</td>
						<td>
							<p class="Basic-Paragraph"><span class="CharOverride-12">3.75</span></p>
						</td>
						<td>
							<p class="Basic-Paragraph"><span class="CharOverride-12">-do-</span></p>
						</td>
						<td>
							<p class="Basic-Paragraph"><span class="CharOverride-12">15</span></p>
						</td>
						<td>
							<p class="Basic-Paragraph"><span class="CharOverride-12">15</span></p>
						</td>
						<td>
							<p class="Basic-Paragraph"><span class="CharOverride-12">-</span></p>
						</td>
					</tr>
					<tr class="_idGenTableRowColumn-1">
						<td>
							<p class="Basic-Paragraph"><span class="CharOverride-12">9</span></p>
						</td>
						<td>
							<p class="Basic-Paragraph"><span class="CharOverride-12">AH</span></p>
						</td>
						<td>
							<p class="Basic-Paragraph"><span class="CharOverride-18">- Thymus and bursal atrophy</span></p>
							<p class="Basic-Paragraph"><span class="CharOverride-18">- Bone marrow hypoplasia with presence of pink</span></p>
							<p class="Basic-Paragraph"><span class="CharOverride-18">- Enlargement and mottled appearance of liver</span></p>
							<p class="Basic-Paragraph"><span class="CharOverride-18">- Haemorrhagic and necrotic lesions on wings and breast</span></p>
						</td>
						<td>
							<p class="Basic-Paragraph"><span class="CharOverride-12">Chicken Infectious Anaemia Gangren Dermatitis Syndrome</span></p>
						</td>
						<td>
							<p class="Basic-Paragraph"><span class="CharOverride-12">6.39</span></p>
						</td>
						<td>
							<p class="Basic-Paragraph"><span class="CharOverride-12">-do-</span></p>
						</td>
						<td>
							<p class="Basic-Paragraph"><span class="CharOverride-12">15</span></p>
						</td>
						<td>
							<p class="Basic-Paragraph"><span class="CharOverride-12">15</span></p>
						</td>
						<td>
							<p class="Basic-Paragraph"><span class="CharOverride-12">-</span></p>
						</td>
					</tr>
					<tr class="_idGenTableRowColumn-2">
						<td>
							<p class="Basic-Paragraph"><span class="CharOverride-12">10</span></p>
						</td>
						<td>
							<p class="Basic-Paragraph"><span class="CharOverride-12">PI</span></p>
						</td>
						<td>
							<p class="Basic-Paragraph"><span class="CharOverride-18">- Haemorrhagic and necrotic lesions on wings and breast region</span></p>
							<p class="Basic-Paragraph"><span class="CharOverride-18">- Severe atrophy of thymus and moderate bursal atrophy</span></p>
							<p class="Basic-Paragraph"><span class="CharOverride-18">- Bone marrow aplasia with yellowish discolouration</span></p>
							<p class="Basic-Paragraph"><span class="CharOverride-18">- Pale discolouration of liver</span></p>
						</td>
						<td>
							<p class="Basic-Paragraph"><span class="CharOverride-12">Chicken Infectious Anaemia  Gangren Dermatitis Syndrome</span></p>
						</td>
						<td>
							<p class="Basic-Paragraph"><span class="CharOverride-12">45.69</span></p>
						</td>
						<td>
							<p class="Basic-Paragraph"><span class="CharOverride-12">-do-</span></p>
						</td>
						<td>
							<p class="Basic-Paragraph"><span class="CharOverride-12">15</span></p>
						</td>
						<td>
							<p class="Basic-Paragraph"><span class="CharOverride-12">15</span></p>
						</td>
						<td>
							<p class="Basic-Paragraph"><span class="CharOverride-12">-</span></p>
						</td>
					</tr>
					<tr class="_idGenTableRowColumn-1">
						<td>
							<p class="Basic-Paragraph"><span class="CharOverride-12">11</span></p>
						</td>
						<td>
							<p class="Basic-Paragraph"><span class="CharOverride-12">PF</span></p>
						</td>
						<td>
							<p class="Basic-Paragraph"><span class="CharOverride-18">- Fibrinous pericarditis and perihepatitis</span></p>
							<p class="Basic-Paragraph"><span class="CharOverride-18">- Presence of caseous exudates in the air sac</span></p>
							<p class="Basic-Paragraph"><span class="CharOverride-18">- Catarrhal inflammation of nasal passage, trachea and bronchi</span></p>
							<p class="Basic-Paragraph"><span class="CharOverride-18">- Cheesy exudate around the nostrils and eyes</span></p>
						</td>
						<td>
							<p class="Basic-Paragraph"><span class="CharOverride-12">Chronic Complex Respirat Disease</span></p>
						</td>
						<td>
							<p class="Basic-Paragraph"><span class="CharOverride-12">3.33</span></p>
						</td>
						<td>
							<p class="Basic-Paragraph"><span class="CharOverride-12">-do-</span></p>
						</td>
						<td>
							<p class="Basic-Paragraph"><span class="CharOverride-12">10</span></p>
						</td>
						<td>
							<p class="Basic-Paragraph"><span class="CharOverride-12">-</span></p>
						</td>
						<td>
							<p class="Basic-Paragraph"><span class="CharOverride-12">10</span></p>
						</td>
					</tr>
					<tr class="_idGenTableRowColumn-2">
						<td>
							<p class="Basic-Paragraph"><span class="CharOverride-12">12</span></p>
						</td>
						<td>
							<p class="Basic-Paragraph"><span class="CharOverride-12">MP</span></p>
						</td>
						<td>
							<p class="Basic-Paragraph"><span class="CharOverride-18">- Swelling of the liver with presence of punctiform haemorrhagic foci throughout the surface</span></p>
							<p class="Basic-Paragraph"><span class="CharOverride-18">- Pale coloured marrow cavity of the femur</span></p>
							<p class="Basic-Paragraph"><span class="CharOverride-18">- Enlarged and pale kidneys</span></p>
							<p class="Basic-Paragraph"><span class="CharOverride-18">- Mild bursal atrophy</span></p>
						</td>
						<td>
							<p class="Basic-Paragraph"><span class="CharOverride-12">Inclusion Body Hepatitis</span></p>
						</td>
						<td>
							<p class="Basic-Paragraph"><span class="CharOverride-12">6.22</span></p>
						</td>
						<td>
							<p class="Basic-Paragraph"><span class="CharOverride-12">-do-</span></p>
						</td>
						<td>
							<p class="Basic-Paragraph"><span class="CharOverride-12">15</span></p>
						</td>
						<td>
							<p class="Basic-Paragraph"><span class="CharOverride-12">15</span></p>
						</td>
						<td>
							<p class="Basic-Paragraph"><span class="CharOverride-12">-</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-5"><a id="Table-3-"></a>Table 3: </span>Disease-wise analysis of samples tested by PCR for detection of CAV DNA</p>
			<table width="657" height="249" class="Table-Style-1" id="table-3">
				<colgroup>
					<col />
					<col />
					<col />
					<col />
					<col />
				</colgroup>
				<tbody>
					<tr class="_idGenTableRowColumn-1">
						<td>
							<p class="Basic-Paragraph"><span class="CharOverride-11">Sr. No.</span></p>
						</td>
						<td>
							<p class="Basic-Paragraph"><span class="CharOverride-11">Suspected Disease (Based on gross lesions)</span></p>
						</td>
						<td>
							<p class="Basic-Paragraph"><span class="CharOverride-11">No. of samples tested</span></p>
						</td>
						<td>
							<p class="Basic-Paragraph"><span class="CharOverride-11">No. of positive samples</span></p>
						</td>
						<td>
							<p class="Basic-Paragraph"><span class="CharOverride-11">No. of negative samples</span></p>
						</td>
					</tr>
					<tr class="_idGenTableRowColumn-2">
						<td>
							<p class="Basic-Paragraph"><span class="CharOverride-12">1.</span></p>
						</td>
						<td>
							<p class="Basic-Paragraph"><span class="CharOverride-12">Chicken Infectious Anaemia – Gangrenous Dermatitis Syndrome</span></p>
						</td>
						<td>
							<p class="Basic-Paragraph"><span class="CharOverride-12">95</span></p>
						</td>
						<td>
							<p class="Basic-Paragraph"><span class="CharOverride-12">95</span></p>
						</td>
						<td>
							<p class="Basic-Paragraph"><span class="CharOverride-12">-</span></p>
						</td>
					</tr>
					<tr class="_idGenTableRowColumn-1">
						<td>
							<p class="Basic-Paragraph"><span class="CharOverride-12">2.</span></p>
						</td>
						<td>
							<p class="Basic-Paragraph"><span class="CharOverride-12">Marek’s Disease</span></p>
						</td>
						<td>
							<p class="Basic-Paragraph"><span class="CharOverride-12">15</span></p>
						</td>
						<td>
							<p class="Basic-Paragraph"><span class="CharOverride-12">15</span></p>
						</td>
						<td>
							<p class="Basic-Paragraph"><span class="CharOverride-12">-</span></p>
						</td>
					</tr>
					<tr class="_idGenTableRowColumn-2">
						<td>
							<p class="Basic-Paragraph"><span class="CharOverride-12">3.</span></p>
						</td>
						<td>
							<p class="Basic-Paragraph"><span class="CharOverride-12">Fowl Pox</span></p>
						</td>
						<td>
							<p class="Basic-Paragraph"><span class="CharOverride-12">15</span></p>
						</td>
						<td>
							<p class="Basic-Paragraph"><span class="CharOverride-12">-</span></p>
						</td>
						<td>
							<p class="Basic-Paragraph"><span class="CharOverride-12">15</span></p>
						</td>
					</tr>
					<tr class="_idGenTableRowColumn-1">
						<td>
							<p class="Basic-Paragraph"><span class="CharOverride-12">4.</span></p>
						</td>
						<td>
							<p class="Basic-Paragraph"><span class="CharOverride-12">Gout</span></p>
						</td>
						<td>
							<p class="Basic-Paragraph"><span class="CharOverride-12">15</span></p>
						</td>
						<td>
							<p class="Basic-Paragraph"><span class="CharOverride-12">-</span></p>
						</td>
						<td>
							<p class="Basic-Paragraph"><span class="CharOverride-12">15</span></p>
						</td>
					</tr>
					<tr class="_idGenTableRowColumn-2">
						<td>
							<p class="Basic-Paragraph"><span class="CharOverride-12">5.</span></p>
						</td>
						<td>
							<p class="Basic-Paragraph"><span class="CharOverride-12">Infectious Bursal Disease</span></p>
						</td>
						<td>
							<p class="Basic-Paragraph"><span class="CharOverride-12">20</span></p>
						</td>
						<td>
							<p class="Basic-Paragraph"><span class="CharOverride-12">20</span></p>
						</td>
						<td>
							<p class="Basic-Paragraph"><span class="CharOverride-12">-</span></p>
						</td>
					</tr>
					<tr class="_idGenTableRowColumn-1">
						<td>
							<p class="Basic-Paragraph"><span class="CharOverride-12">6.</span></p>
						</td>
						<td>
							<p class="Basic-Paragraph"><span class="CharOverride-12">Chronic Complex Respiratory Disease</span></p>
						</td>
						<td>
							<p class="Basic-Paragraph"><span class="CharOverride-12">10</span></p>
						</td>
						<td>
							<p class="Basic-Paragraph"><span class="CharOverride-12">-</span></p>
						</td>
						<td>
							<p class="Basic-Paragraph"><span class="CharOverride-12">10</span></p>
						</td>
					</tr>
					<tr class="_idGenTableRowColumn-2">
						<td>
							<p class="Basic-Paragraph"><span class="CharOverride-12">7.</span></p>
						</td>
						<td>
							<p class="Basic-Paragraph"><span class="CharOverride-12">Inclusion Body Hepatitis</span></p>
						</td>
						<td>
							<p class="Basic-Paragraph"><span class="CharOverride-12">15</span></p>
						</td>
						<td>
							<p class="Basic-Paragraph"><span class="CharOverride-12">15</span></p>
						</td>
						<td>
							<p class="Basic-Paragraph"><span class="CharOverride-12">-</span></p>
						</td>
					</tr>
					<tr class="_idGenTableRowColumn-1">
						<td height="21" colspan="2">
							<p class="Basic-Paragraph"><span class="CharOverride-12">Total</span></p>
						</td>
						<td>
							<p class="Basic-Paragraph"><span class="CharOverride-12">185</span></p>
						</td>
						<td>
							<p class="Basic-Paragraph"><span class="CharOverride-12">145</span></p>
						</td>
						<td>
							<p class="Basic-Paragraph"><span class="CharOverride-12">40</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-5"><a id="Table-4-"></a>Table 4: </span>Digestion products of RE analysis with different enzymes</p>
			<table width="657" height="165" class="Table-Style-1" id="table-4">
				<colgroup>
					<col />
					<col />
					<col />
				</colgroup>
				<tbody>
					<tr class="_idGenTableRowColumn-1">
						<td>
							<p class="Basic-Paragraph"><span class="CharOverride-11">Sr. No.</span></p>
						</td>
						<td>
							<p class="Basic-Paragraph"><span class="CharOverride-11">Restriction Enzymes</span></p>
						</td>
						<td>
							<p class="Basic-Paragraph"><span class="CharOverride-11">Digestion products (bp)</span></p>
						</td>
					</tr>
					<tr class="_idGenTableRowColumn-2">
						<td>
							<p class="Basic-Paragraph"><span class="CharOverride-12">1.</span></p>
						</td>
						<td>
							<p class="Basic-Paragraph"><span class="CharOverride-19">Hin</span><span class="CharOverride-12">d III</span></p>
						</td>
						<td>
							<p class="Basic-Paragraph"><span class="CharOverride-12">966, 791, 562</span></p>
						</td>
					</tr>
					<tr class="_idGenTableRowColumn-1">
						<td>
							<p class="Basic-Paragraph"><span class="CharOverride-12">2.</span></p>
						</td>
						<td>
							<p class="Basic-Paragraph"><span class="CharOverride-19">Pst </span><span class="CharOverride-12">I</span></p>
						</td>
						<td>
							<p class="Basic-Paragraph"><span class="CharOverride-12">1743, 576</span></p>
						</td>
					</tr>
					<tr class="_idGenTableRowColumn-2">
						<td>
							<p class="Basic-Paragraph"><span class="CharOverride-12">3.</span></p>
						</td>
						<td>
							<p class="Basic-Paragraph"><span class="CharOverride-19">Nhe</span><span class="CharOverride-12">I</span></p>
						</td>
						<td>
							<p class="Basic-Paragraph"><span class="CharOverride-12">1191, 1128</span></p>
						</td>
					</tr>
					<tr class="_idGenTableRowColumn-1">
						<td>
							<p class="Basic-Paragraph"><span class="CharOverride-12">4.</span></p>
						</td>
						<td>
							<p class="Basic-Paragraph"><span class="CharOverride-19">Mbo </span><span class="CharOverride-12">II</span></p>
						</td>
						<td>
							<p class="Basic-Paragraph"><span class="CharOverride-12">1522, 797</span></p>
						</td>
					</tr>
					<tr class="_idGenTableRowColumn-2">
						<td>
							<p class="Basic-Paragraph"><span class="CharOverride-12">5.</span></p>
						</td>
						<td>
							<p class="Basic-Paragraph"><span class="CharOverride-19">Nhe </span><span class="CharOverride-12">I + </span><span class="CharOverride-19">Pst </span><span class="CharOverride-12">I</span></p>
						</td>
						<td>
							<p class="Basic-Paragraph"><span class="CharOverride-12">1743, 1191, 1128, 615,  576</span></p>
						</td>
					</tr>
					<tr class="_idGenTableRowColumn-1">
						<td height="21">
							<p class="Basic-Paragraph"><span class="CharOverride-12">6.</span></p>
						</td>
						<td>
							<p class="Basic-Paragraph"><span class="CharOverride-19">Nhe </span><span class="CharOverride-12">I + </span><span class="CharOverride-19">Mbo </span><span class="CharOverride-12">II</span></p>
						</td>
						<td>
							<p class="Basic-Paragraph"><span class="CharOverride-12">1522, 1191, 1128, 797, 331</span></p>
						</td>
					</tr>
				</tbody>
			</table>
			<p class="Heading-1--Introduction----">&nbsp;</p>
			<p class="Heading-1--Introduction----">DISCUSSION</p>
		  <p class="Heading-1--Introduction----">&nbsp;</p>
			<p class="Body-Text ParaOverride-1">CAV infection has been incriminated as the cause in several cases where infectious anaemia occurred in chicken flocks of up to 2-4 weeks of age (<a href="#McNulty-MS--1991-"><span class="Hyperlink">McNulty, 1991</span></a>; <a href="#Dhama-K--Mahe-2008"><span class="Hyperlink">Dhama et al., 2008</span></a>; <a href="#Schat-KA--2009"><span class="Hyperlink">Schat, 2009</span></a>; <a href="#Pope-CR--1991"><span class="Hyperlink">Pope, 1991</span></a>; <a href="#Rosenberger-JK--Cloud-SS--1989"><span class="Hyperlink">Rosenberger and Cloud, 1989</span></a>). Age related resistance occurs in chicks as clinical disease does not occur in chicks above 4 weeks of age (<a href="#Wani-MY--Dhama-K--Barathidasan-R--Gowthaman-V"><span class="Hyperlink">Wani et al., 2014</span></a>). Co-infections of CAV with other pathogens can overcome the effects of age and maternal antibody related resistance, thus increasing the susceptible age period for CAV and its persistency (Yuasa et al., 1980; <a href="#Rosenberger-JK--Cloud-SS--1989"><span class="Hyperlink">Rosenberger and Cloud, 1989</span></a>; <a href="#Toro-H--Gonzalez-2000"><span class="Hyperlink">Toro et al., 2000</span></a>). In the field, CAV infection seems to cause few signs of disease, however, dual infections generally occur which are more serious in terms of economic losses to poultry producers (<a href="#Dhama-K--Mahe-2008"><span class="Hyperlink">Dhama et al., 2008</span></a>; <a href="#Pope-CR--1991"><span class="Hyperlink">Pope, 1991</span></a>; <a href="#Imai-K--Mase-M--Tsukamoto-K--Hihara-H--Yuasa-N--1999"><span class="Hyperlink">Imai et al., 1999</span></a>). Therefore, it becomes necessary to determine the existence of CAV in association with other pathogens in poultry population of any country. Although, postmortem findings act as first hand tentative diagnostic method for many infections in poultry, the association of CAV in mixed infections and its diagnosis is difficult especially during subclinical infection. Applications of molecular tools have paved a way for the rapid and convenient diagnosis in such mixed infections.</p>
		  <p class="Body-Text ParaOverride-1">&nbsp;</p>
			<p class="Body-Text ParaOverride-1">In India, chicken infectious anemia (CIA) has been suspected since 1990’s after the emergence of virulent strains of IBDV and on the basis of clinical symptoms, lesions and its detection by immunoperoxidase test (<a href="#Venugopalan-AT--Elankumaran-S--Raj-GD--Manohar-BM--Thangavelu-A--1994"><span class="Hyperlink">Venugopalan et al., 1994</span></a>). Presently, the disease has been confirmed from many states using virus isolation, PCR detection and serological studies (<a href="#Wani-MY--Dhama-2013"><span class="Hyperlink">Wani et al., 2013</span></a>; <a href="#Natesan-S--Kataria-JM--Dhama-K--Rahul-S--Baradhwaj-N--2006"><span class="Hyperlink">Natesan et al., 2003</span></a>; <a href="#Dhama-K--2004"><span class="Hyperlink">Dhama et al., 2004</span></a>; <a href="#Bhatt-P--Shukla-SK--Mahendran-M--Dhama-K--Chawak-MM--Kataria-JM--2011"><span class="Hyperlink">Bhatt et al., 2011</span></a>). The molecular techniques including PCR have been well employed by several workers (<a href="#Wani-MY--Dhama-2013"><span class="Hyperlink">Wani et al., 2013</span></a>; <a href="#Todd-D--Mawhinney-KA--McNulty-MS--1992"><span class="Hyperlink">Todd et al., 1966</span></a>; <a href="#Dhama-K--2004"><span class="Hyperlink">Dhama et al., 2004</span></a>; <a href="#Bhatt-P--Shukla-SK--Mahendran-M--Dhama-K--Chawak-MM--Kataria-JM--2011"><span class="Hyperlink">Bhatt et al., 2011</span></a>; <a href="#Rozypal-TL--Skeeles-JK--Dash-JK--Anderson-EJ--Beasley-JN--1997"><span class="Hyperlink">Rozypal et al., 1997</span></a>; <a href="#Chowdhury-SM--Omar-AR--Aini-I--Hair-BM--Jamaluddin-AA--Kono-Y--Darus-A--Yatim-HM--2002"><span class="Hyperlink">Chowdhury et al., 2002</span></a>; <a href="#Miller-MM--Ealey-KA--Oswald-WB--Schat-KA--2003-"><span class="Hyperlink">Miller et al., 2003</span></a>) for detection of CAV-DNA in clinical samples. However, detailed molecular epidemiological status, immunosuppressive nature and clinical/field interactions of the virus with other infectious pathogens/diseases of poultry have not been performed earlier in the country.</p>
		  <p class="Body-Text ParaOverride-1">&nbsp;</p>
			<p class="Body-Text ParaOverride-1">In the present study, molecular surveillance of the CAV, IBD, IBH, fowl pox, gout, CRD and MD in suspected poultry flocks from north Indian states  and Mahendra Nagar area of Nepal with the history of different disease outbreak, viz. bacterial, viral, protozoal, etc. was undertaken along with clinical, pathological and haemato-biochemical parameters studies. High prevalence of CAV recorded in present study is in accordance with the earlier reports from India (<a href="#Wani-MY--Dhama-2013"><span class="Hyperlink">Wani et al., 2013</span></a>; <a href="#Bhatt-P--Shukla-SK--Mahendran-M--Dhama-K--Chawak-MM--Kataria-JM--2011"><span class="Hyperlink">Bhatt et al., 2011</span></a>). Amongst the nine CIA positive flocks, three flocks were suffering with MD, IBD and IBH based on necropsy examination results. <a href="#Jin-WJ--Cui-ZZ--Liu-YL--Qin-AJ--2001"><span class="Hyperlink">Jin et al. (2001)</span></a> found co-infection of MDV, CAV and REV in some tissue samples of bursa from the chicken flocks suspected to be infected by IBDV according to clinical symptoms and gross lesions by dot hybridization technique in some parts of China. Later, <a href="#Jiang-SJ--Meng-SS--Cui-ZZ--Tian-FL--Wang-ZF--2005"><span class="Hyperlink">Jiang </span><span class="Hyperlink">et al. (2005)</span></a> also found co-infection of MDV, CAV and REV in tissue samples of 828 chickens from 42 different chicken flocks suspected to be infected by immunosuppressive diseases in 11 provinces of China.</p>
		  <p class="Body-Text ParaOverride-1">&nbsp;</p>
			
		  <p class="Figure--and-Table-Heading ParaOverride-1"><span class="CharOverride-5"><a id="Table-5-"></a>Table 5: </span>Mean values of haemoglobin, PCV, TEC, TLC and DLC in different flocks</p>
			<table width="659" height="670" class="Table-Style-1" id="table-5">
				<colgroup>
					<col />
					<col />
					<col />
					<col />
					<col />
					<col />
					<col />
					<col />
					<col />
					<col />
				</colgroup>
				<tbody>
					<tr class="_idGenTableRowColumn-1">
						<td width="24" rowspan="2">
							<p class="Basic-Paragraph"><span class="CharOverride-11">Sr. No.</span></p>
						</td>
						<td width="37" rowspan="2">
							<p class="Basic-Paragraph"><span class="CharOverride-11">Farm Code </span></p>
						</td>
						<td width="50" rowspan="2">
							<p class="Basic-Paragraph"><span class="CharOverride-11">Haemo globin (Hb) </span><span class="CharOverride-11">(g/L)</span></p>
						</td>
						<td width="73" rowspan="2">
							<p class="Basic-Paragraph"><span class="CharOverride-11">Packed Cell Volume (PCV) </span><span class="CharOverride-11">(L/L)</span></p>
						</td>
						<td width="77" rowspan="2">
							<p class="Basic-Paragraph"><span class="CharOverride-11">Total Erythrocyte Count</span></p>
							<p class="Basic-Paragraph"><span class="CharOverride-11">(TEC) (x10</span><span class="CharOverride-21">9</span><span class="CharOverride-11">/L)</span></p>
						</td>
						<td width="82" rowspan="2">
							<p class="Basic-Paragraph"><span class="CharOverride-11">Total Leukocyte Count</span></p>
							<p class="Basic-Paragraph"><span class="CharOverride-11">(TLC) (x10</span><span class="CharOverride-21">6</span><span class="CharOverride-11">/L)</span></p>
						</td>
						<td colspan="4">
							<p class="Basic-Paragraph"><span class="CharOverride-11">Differential Leukocyte Count (DLC)</span></p>
						</td>
					</tr>
					<tr class="_idGenTableRowColumn-2">
						<td width="63">
							<p class="Basic-Paragraph"><span class="CharOverride-12">Heterophil</span></p>
							<p class="Basic-Paragraph"><span class="CharOverride-12">(%)</span></p>
						</td>
						<td width="75">
							<p class="Basic-Paragraph"><span class="CharOverride-12">Lymphocyte</span></p>
							<p class="Basic-Paragraph"><span class="CharOverride-12">(%)</span></p>
						</td>
						<td width="62">
							<p class="Basic-Paragraph"><span class="CharOverride-12">Monocyte</span></p>
							<p class="Basic-Paragraph"><span class="CharOverride-12">(%)</span></p>
						</td>
						<td width="70">
							<p class="Basic-Paragraph"><span class="CharOverride-12">Eosinophil</span></p>
							<p class="Basic-Paragraph"><span class="CharOverride-12">+ Basophil</span></p>
							<p class="Basic-Paragraph"><span class="CharOverride-12">(%)</span></p>
						</td>
					</tr>
					<tr class="_idGenTableRowColumn-1">
						<td>
							<p class="Basic-Paragraph"><span class="CharOverride-12">1.</span></p>
						</td>
						<td>
							<p class="Basic-Paragraph"><span class="CharOverride-12">N</span></p>
						</td>
						<td>
							<p class="Basic-Paragraph"><span class="CharOverride-12">118.33 ±0.24</span></p>
						</td>
						<td>
							<p class="Basic-Paragraph"><span class="CharOverride-12">0.32±0.00</span></p>
						</td>
						<td>
							<p class="Basic-Paragraph"><span class="CharOverride-12">3.20±0.05</span></p>
						</td>
						<td>
							<p class="Basic-Paragraph"><span class="CharOverride-12">21.47±0.81</span></p>
						</td>
						<td>
							<p class="Basic-Paragraph"><span class="CharOverride-12">33.67 ±0.58</span></p>
						</td>
						<td>
							<p class="Basic-Paragraph"><span class="CharOverride-12">60.00 ±0.33</span></p>
						</td>
						<td>
							<p class="Basic-Paragraph"><span class="CharOverride-12">3.67 ±0.33</span></p>
						</td>
						<td>
							<p class="Basic-Paragraph"><span class="CharOverride-12">2.67±0.33</span></p>
						</td>
					</tr>
					<tr class="_idGenTableRowColumn-2">
						<td>
							<p class="Basic-Paragraph"><span class="CharOverride-12">2.</span></p>
						</td>
						<td>
							<p class="Basic-Paragraph"><span class="CharOverride-12">SP</span></p>
						</td>
						<td>
							<p class="Basic-Paragraph"><span class="CharOverride-12">66.00 ±0.12*</span></p>
						</td>
						<td>
							<p class="Basic-Paragraph"><span class="CharOverride-12">0.21±0.01*</span></p>
						</td>
						<td>
							<p class="Basic-Paragraph"><span class="CharOverride-12">1.92±0.04*</span></p>
						</td>
						<td>
							<p class="Basic-Paragraph"><span class="CharOverride-12">11.56±1.74*</span></p>
						</td>
						<td>
							<p class="Basic-Paragraph"><span class="CharOverride-12">53.00 ±1.53*</span></p>
						</td>
						<td>
							<p class="Basic-Paragraph"><span class="CharOverride-12">41.00 ±1.53*</span></p>
						</td>
						<td>
							<p class="Basic-Paragraph"><span class="CharOverride-12">3.33 ±0.33</span></p>
						</td>
						<td>
							<p class="Basic-Paragraph"><span class="CharOverride-12">2.67±0.33</span></p>
						</td>
					</tr>
					<tr class="_idGenTableRowColumn-1">
						<td>
							<p class="Basic-Paragraph"><span class="CharOverride-12">3.</span></p>
						</td>
						<td>
							<p class="Basic-Paragraph"><span class="CharOverride-12">KP</span></p>
						</td>
						<td>
							<p class="Basic-Paragraph"><span class="CharOverride-12">116.00 ±0.12</span></p>
						</td>
						<td>
							<p class="Basic-Paragraph"><span class="CharOverride-12">0.30±0.01*</span></p>
						</td>
						<td>
							<p class="Basic-Paragraph"><span class="CharOverride-12">3.15±0.14</span></p>
						</td>
						<td>
							<p class="Basic-Paragraph"><span class="CharOverride-12">17.73±0.18*</span></p>
						</td>
						<td>
							<p class="Basic-Paragraph"><span class="CharOverride-12">42.00 ±1.15*</span></p>
						</td>
						<td>
							<p class="Basic-Paragraph"><span class="CharOverride-12">53.00 ±1.53*</span></p>
						</td>
						<td>
							<p class="Basic-Paragraph"><span class="CharOverride-12">2.66 ±0.67</span></p>
						</td>
						<td>
							<p class="Basic-Paragraph"><span class="CharOverride-12">2.33±0.33</span></p>
						</td>
					</tr>
					<tr class="_idGenTableRowColumn-2">
						<td>
							<p class="Basic-Paragraph"><span class="CharOverride-12">4.</span></p>
						</td>
						<td>
							<p class="Basic-Paragraph"><span class="CharOverride-12">NP</span></p>
						</td>
						<td>
							<p class="Basic-Paragraph"><span class="CharOverride-12">109.00 ±0.25</span></p>
						</td>
						<td>
							<p class="Basic-Paragraph"><span class="CharOverride-12">0.32±0.01</span></p>
						</td>
						<td>
							<p class="Basic-Paragraph"><span class="CharOverride-12">2.74±0.09*</span></p>
						</td>
						<td>
							<p class="Basic-Paragraph"><span class="CharOverride-12">19.93±0.18</span></p>
						</td>
						<td>
							<p class="Basic-Paragraph"><span class="CharOverride-12">55.67 ±0.88*</span></p>
						</td>
						<td>
							<p class="Basic-Paragraph"><span class="CharOverride-12">39.33 ±0.88*</span></p>
						</td>
						<td>
							<p class="Basic-Paragraph"><span class="CharOverride-12">3.00 ±0.58</span></p>
						</td>
						<td>
							<p class="Basic-Paragraph"><span class="CharOverride-12">2.00±0.00</span></p>
						</td>
					</tr>
					<tr class="_idGenTableRowColumn-1">
						<td>
							<p class="Basic-Paragraph"><span class="CharOverride-12">5.</span></p>
						</td>
						<td>
							<p class="Basic-Paragraph"><span class="CharOverride-12">IP</span></p>
						</td>
						<td>
							<p class="Basic-Paragraph"><span class="CharOverride-12">114.67 ±0.18</span></p>
						</td>
						<td>
							<p class="Basic-Paragraph"><span class="CharOverride-12">0.33±0.00</span></p>
						</td>
						<td>
							<p class="Basic-Paragraph"><span class="CharOverride-12">2.99±0.12</span></p>
						</td>
						<td>
							<p class="Basic-Paragraph"><span class="CharOverride-12">20.47±1.05</span></p>
						</td>
						<td>
							<p class="Basic-Paragraph"><span class="CharOverride-12">56.67 ±0.88*</span></p>
						</td>
						<td>
							<p class="Basic-Paragraph"><span class="CharOverride-12">37.67 ±1.45*</span></p>
						</td>
						<td>
							<p class="Basic-Paragraph"><span class="CharOverride-12">3.33 ±0.33</span></p>
						</td>
						<td>
							<p class="Basic-Paragraph"><span class="CharOverride-12">2.33±0.33</span></p>
						</td>
					</tr>
					<tr class="_idGenTableRowColumn-2">
						<td>
							<p class="Basic-Paragraph"><span class="CharOverride-12">6.</span></p>
						</td>
						<td>
							<p class="Basic-Paragraph"><span class="CharOverride-12">DP</span></p>
						</td>
						<td>
							<p class="Basic-Paragraph"><span class="CharOverride-12">68.00 ±0.12*</span></p>
						</td>
						<td>
							<p class="Basic-Paragraph"><span class="CharOverride-12">0.24±0.01*</span></p>
						</td>
						<td>
							<p class="Basic-Paragraph"><span class="CharOverride-12">2.10±0.10*</span></p>
						</td>
						<td>
							<p class="Basic-Paragraph"><span class="CharOverride-12">9.53±0.42*</span></p>
						</td>
						<td>
							<p class="Basic-Paragraph"><span class="CharOverride-12">50.00 ±0.58*</span></p>
						</td>
						<td>
							<p class="Basic-Paragraph"><span class="CharOverride-12">44.67 ±0.88*</span></p>
						</td>
						<td>
							<p class="Basic-Paragraph"><span class="CharOverride-12">3.33 ±0.67</span></p>
						</td>
						<td>
							<p class="Basic-Paragraph"><span class="CharOverride-12">2.33±0.33</span></p>
						</td>
					</tr>
					<tr class="_idGenTableRowColumn-1">
						<td>
							<p class="Basic-Paragraph"><span class="CharOverride-12">7.</span></p>
						</td>
						<td>
							<p class="Basic-Paragraph"><span class="CharOverride-12">AP</span></p>
						</td>
						<td>
							<p class="Basic-Paragraph"><span class="CharOverride-12">72.67 ±0.07*</span></p>
						</td>
						<td>
							<p class="Basic-Paragraph"><span class="CharOverride-12">0.27±0.02*</span></p>
						</td>
						<td>
							<p class="Basic-Paragraph"><span class="CharOverride-12">2.72±0.05*</span></p>
						</td>
						<td>
							<p class="Basic-Paragraph"><span class="CharOverride-12">9.60±0.38*</span></p>
						</td>
						<td>
							<p class="Basic-Paragraph"><span class="CharOverride-12">50.33 ±1.20*</span></p>
						</td>
						<td>
							<p class="Basic-Paragraph"><span class="CharOverride-12">44.00 ±1.15*</span></p>
						</td>
						<td>
							<p class="Basic-Paragraph"><span class="CharOverride-12">3.33 ±0.67</span></p>
						</td>
						<td>
							<p class="Basic-Paragraph"><span class="CharOverride-12">2.67±0.33</span></p>
						</td>
					</tr>
					<tr class="_idGenTableRowColumn-2">
						<td>
							<p class="Basic-Paragraph"><span class="CharOverride-12">8.</span></p>
						</td>
						<td>
							<p class="Basic-Paragraph"><span class="CharOverride-12">ZP</span></p>
						</td>
						<td>
							<p class="Basic-Paragraph"><span class="CharOverride-12">68.00 ±0.12*</span></p>
						</td>
						<td>
							<p class="Basic-Paragraph"><span class="CharOverride-12">0.22±0.00*</span></p>
						</td>
						<td>
							<p class="Basic-Paragraph"><span class="CharOverride-12">1.95±0.08*</span></p>
						</td>
						<td>
							<p class="Basic-Paragraph"><span class="CharOverride-12">10.80±0.81*</span></p>
						</td>
						<td>
							<p class="Basic-Paragraph"><span class="CharOverride-12">49.00 ±1.00*</span></p>
						</td>
						<td>
							<p class="Basic-Paragraph"><span class="CharOverride-12">44.00 ±1.15*</span></p>
						</td>
						<td>
							<p class="Basic-Paragraph"><span class="CharOverride-12">3.67 ±0.33</span></p>
						</td>
						<td>
							<p class="Basic-Paragraph"><span class="CharOverride-12">2.67±0.33</span></p>
						</td>
					</tr>
					<tr class="_idGenTableRowColumn-1">
						<td>
							<p class="Basic-Paragraph"><span class="CharOverride-12">9.</span></p>
						</td>
						<td>
							<p class="Basic-Paragraph"><span class="CharOverride-12">PN</span></p>
						</td>
						<td>
							<p class="Basic-Paragraph"><span class="CharOverride-12">68.00 ±0.12*</span></p>
						</td>
						<td>
							<p class="Basic-Paragraph"><span class="CharOverride-12">0.24±0.01*</span></p>
						</td>
						<td>
							<p class="Basic-Paragraph"><span class="CharOverride-12">2.09±0.09*</span></p>
						</td>
						<td>
							<p class="Basic-Paragraph"><span class="CharOverride-12">9.73±0.48*</span></p>
						</td>
						<td>
							<p class="Basic-Paragraph"><span class="CharOverride-12">50.67 ±0.88*</span></p>
						</td>
						<td>
							<p class="Basic-Paragraph"><span class="CharOverride-12">44.67 ±1.45*</span></p>
						</td>
						<td>
							<p class="Basic-Paragraph"><span class="CharOverride-12">3.00 ±0.58</span></p>
						</td>
						<td>
							<p class="Basic-Paragraph"><span class="CharOverride-12">1.67±0.33</span></p>
						</td>
					</tr>
					<tr class="_idGenTableRowColumn-2">
						<td>
							<p class="Basic-Paragraph"><span class="CharOverride-12">10.</span></p>
						</td>
						<td>
							<p class="Basic-Paragraph"><span class="CharOverride-12">AH</span></p>
						</td>
						<td>
							<p class="Basic-Paragraph"><span class="CharOverride-12">68.67 ±0.18*</span></p>
						</td>
						<td>
							<p class="Basic-Paragraph"><span class="CharOverride-12">0.23±0.00*</span></p>
						</td>
						<td>
							<p class="Basic-Paragraph"><span class="CharOverride-12">2.05±0.04*</span></p>
						</td>
						<td>
							<p class="Basic-Paragraph"><span class="CharOverride-12">9.70±0.46*</span></p>
						</td>
						<td>
							<p class="Basic-Paragraph"><span class="CharOverride-12">49.67 ±0.67*</span></p>
						</td>
						<td>
							<p class="Basic-Paragraph"><span class="CharOverride-12">43.67 ±0.88*</span></p>
						</td>
						<td>
							<p class="Basic-Paragraph"><span class="CharOverride-12">4.00 ±0.58</span></p>
						</td>
						<td>
							<p class="Basic-Paragraph"><span class="CharOverride-12">3.00±0.58</span></p>
						</td>
					</tr>
					<tr class="_idGenTableRowColumn-1">
						<td>
							<p class="Basic-Paragraph"><span class="CharOverride-12">11.</span></p>
						</td>
						<td>
							<p class="Basic-Paragraph"><span class="CharOverride-12">PI</span></p>
						</td>
						<td>
							<p class="Basic-Paragraph"><span class="CharOverride-12">65.33 ±0.37*</span></p>
						</td>
						<td>
							<p class="Basic-Paragraph"><span class="CharOverride-12">0.19±0.01*</span></p>
						</td>
						<td>
							<p class="Basic-Paragraph"><span class="CharOverride-12">1.69±0.22*</span></p>
						</td>
						<td>
							<p class="Basic-Paragraph"><span class="CharOverride-12">9.87±0.35*</span></p>
						</td>
						<td>
							<p class="Basic-Paragraph"><span class="CharOverride-12">49.00 ±1.73*</span></p>
						</td>
						<td>
							<p class="Basic-Paragraph"><span class="CharOverride-12">43.67 ±2.40*</span></p>
						</td>
						<td>
							<p class="Basic-Paragraph"><span class="CharOverride-12">4.00 ±0.58</span></p>
						</td>
						<td>
							<p class="Basic-Paragraph"><span class="CharOverride-12">2.67±0.33</span></p>
						</td>
					</tr>
					<tr class="_idGenTableRowColumn-2">
						<td>
							<p class="Basic-Paragraph"><span class="CharOverride-12">12.</span></p>
						</td>
						<td>
							<p class="Basic-Paragraph"><span class="CharOverride-12">PF</span></p>
						</td>
						<td>
							<p class="Basic-Paragraph"><span class="CharOverride-12">114.00 ±0.23</span></p>
						</td>
						<td>
							<p class="Basic-Paragraph"><span class="CharOverride-12">0.32±0.00</span></p>
						</td>
						<td>
							<p class="Basic-Paragraph"><span class="CharOverride-12">2.89±0.15</span></p>
						</td>
						<td>
							<p class="Basic-Paragraph"><span class="CharOverride-12">25.67±0.70*</span></p>
						</td>
						<td>
							<p class="Basic-Paragraph"><span class="CharOverride-12">58.00 ±0.57*</span></p>
						</td>
						<td>
							<p class="Basic-Paragraph"><span class="CharOverride-12">34.33 ±1.20*</span></p>
						</td>
						<td>
							<p class="Basic-Paragraph"><span class="CharOverride-12">5.33 ±0.33*</span></p>
						</td>
						<td>
							<p class="Basic-Paragraph"><span class="CharOverride-12">2.33±0.33</span></p>
						</td>
					</tr>
					<tr class="_idGenTableRowColumn-1">
						<td>
							<p class="Basic-Paragraph"><span class="CharOverride-12">13.</span></p>
						</td>
						<td>
							<p class="Basic-Paragraph"><span class="CharOverride-12">MP</span></p>
						</td>
						<td>
							<p class="Basic-Paragraph"><span class="CharOverride-12">78.00 ±0.23*</span></p>
						</td>
						<td>
							<p class="Basic-Paragraph"><span class="CharOverride-12">0.26±0.01*</span></p>
						</td>
						<td>
							<p class="Basic-Paragraph"><span class="CharOverride-12">2.81±0.10*</span></p>
						</td>
						<td>
							<p class="Basic-Paragraph"><span class="CharOverride-12">22.80±1.36</span></p>
						</td>
						<td>
							<p class="Basic-Paragraph"><span class="CharOverride-12">38.33 ±1.20*</span></p>
						</td>
						<td>
							<p class="Basic-Paragraph"><span class="CharOverride-12">53.00 ±1.00*</span></p>
						</td>
						<td>
							<p class="Basic-Paragraph"><span class="CharOverride-12">5.67 ±0.33*</span></p>
						</td>
						<td>
							<p class="Basic-Paragraph"><span class="CharOverride-12">2.67±0.33</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-7">* Significantly (P&lt;0.05) different</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-5"><a id="Table-6-"></a>Table 6: </span>Mean value of AST, ALT, ALP, Uric Acid in different flocks</p>
			<table width="658" height="325" class="Table-Style-1" id="table-6">
				<colgroup>
					<col />
					<col />
					<col />
					<col />
					<col />
					<col />
				</colgroup>
				<tbody>
					<tr class="_idGenTableRowColumn-1">
						<td>
							<p class="Basic-Paragraph"><span class="CharOverride-11">Sr. No.</span></p>
						</td>
						<td>
							<p class="Basic-Paragraph"><span class="CharOverride-11">Farm Code </span></p>
						</td>
						<td>
							<p class="Basic-Paragraph"><span class="CharOverride-11">AST (IU/L)</span></p>
						</td>
						<td>
							<p class="Basic-Paragraph"><span class="CharOverride-11">ALT (IU/L)</span></p>
						</td>
						<td>
							<p class="Basic-Paragraph"><span class="CharOverride-11">ALP (IU/L)</span></p>
						</td>
						<td>
							<p class="Basic-Paragraph"><span class="CharOverride-11">Uric acid (UA) (mg/dl)</span></p>
						</td>
					</tr>
					<tr class="_idGenTableRowColumn-2">
						<td>
							<p class="Basic-Paragraph"><span class="CharOverride-12">1.</span></p>
						</td>
						<td>
							<p class="Basic-Paragraph"><span class="CharOverride-12">N</span></p>
						</td>
						<td>
							<p class="Basic-Paragraph"><span class="CharOverride-12">174.83±0.66</span></p>
						</td>
						<td>
							<p class="Basic-Paragraph"><span class="CharOverride-12">34.00±0.31</span></p>
						</td>
						<td>
							<p class="Basic-Paragraph"><span class="CharOverride-12">485.50±1.55</span></p>
						</td>
						<td>
							<p class="Basic-Paragraph"><span class="CharOverride-12">5.53±0.05</span></p>
						</td>
					</tr>
					<tr class="_idGenTableRowColumn-1">
						<td>
							<p class="Basic-Paragraph"><span class="CharOverride-12">2.</span></p>
						</td>
						<td>
							<p class="Basic-Paragraph"><span class="CharOverride-12">SP</span></p>
						</td>
						<td>
							<p class="Basic-Paragraph"><span class="CharOverride-12">322.33±3.60*</span></p>
						</td>
						<td>
							<p class="Basic-Paragraph"><span class="CharOverride-12">70.97±0.84*</span></p>
						</td>
						<td>
							<p class="Basic-Paragraph"><span class="CharOverride-12">586.13±6.23*</span></p>
						</td>
						<td>
							<p class="Basic-Paragraph"><span class="CharOverride-12">7.00±0.11*</span></p>
						</td>
					</tr>
					<tr class="_idGenTableRowColumn-2">
						<td>
							<p class="Basic-Paragraph"><span class="CharOverride-12">3.</span></p>
						</td>
						<td>
							<p class="Basic-Paragraph"><span class="CharOverride-12">KP</span></p>
						</td>
						<td>
							<p class="Basic-Paragraph"><span class="CharOverride-12">256.83±1.55*</span></p>
						</td>
						<td>
							<p class="Basic-Paragraph"><span class="CharOverride-12">59.91±0.78*</span></p>
						</td>
						<td>
							<p class="Basic-Paragraph"><span class="CharOverride-12">558.93±1.08*</span></p>
						</td>
						<td>
							<p class="Basic-Paragraph"><span class="CharOverride-12">5.90±0.17</span></p>
						</td>
					</tr>
					<tr class="_idGenTableRowColumn-1">
						<td>
							<p class="Basic-Paragraph"><span class="CharOverride-12">4.</span></p>
						</td>
						<td>
							<p class="Basic-Paragraph"><span class="CharOverride-12">NP</span></p>
						</td>
						<td>
							<p class="Basic-Paragraph"><span class="CharOverride-12">178.30±2.16</span></p>
						</td>
						<td>
							<p class="Basic-Paragraph"><span class="CharOverride-12">34.54±0.83</span></p>
						</td>
						<td>
							<p class="Basic-Paragraph"><span class="CharOverride-12">488.13±1.30</span></p>
						</td>
						<td>
							<p class="Basic-Paragraph"><span class="CharOverride-12">5.70±0.04</span></p>
						</td>
					</tr>
					<tr class="_idGenTableRowColumn-2">
						<td>
							<p class="Basic-Paragraph"><span class="CharOverride-12">5.</span></p>
						</td>
						<td>
							<p class="Basic-Paragraph"><span class="CharOverride-12">IP</span></p>
						</td>
						<td>
							<p class="Basic-Paragraph"><span class="CharOverride-12">279.67±2.03*</span></p>
						</td>
						<td>
							<p class="Basic-Paragraph"><span class="CharOverride-12">66.90±0.85*</span></p>
						</td>
						<td>
							<p class="Basic-Paragraph"><span class="CharOverride-12">572.80±1.64*</span></p>
						</td>
						<td>
							<p class="Basic-Paragraph"><span class="CharOverride-12">12.27±0.99*</span></p>
						</td>
					</tr>
					<tr class="_idGenTableRowColumn-1">
						<td>
							<p class="Basic-Paragraph"><span class="CharOverride-12">6.</span></p>
						</td>
						<td>
							<p class="Basic-Paragraph"><span class="CharOverride-12">DP</span></p>
						</td>
						<td>
							<p class="Basic-Paragraph"><span class="CharOverride-12">263.43±2.75*</span></p>
						</td>
						<td>
							<p class="Basic-Paragraph"><span class="CharOverride-12">64.23±0.35*</span></p>
						</td>
						<td>
							<p class="Basic-Paragraph"><span class="CharOverride-12">567.87±3.46*</span></p>
						</td>
						<td>
							<p class="Basic-Paragraph"><span class="CharOverride-12">6.76±0.44*</span></p>
						</td>
					</tr>
					<tr class="_idGenTableRowColumn-2">
						<td>
							<p class="Basic-Paragraph"><span class="CharOverride-12">7.</span></p>
						</td>
						<td>
							<p class="Basic-Paragraph"><span class="CharOverride-12">AP</span></p>
						</td>
						<td>
							<p class="Basic-Paragraph"><span class="CharOverride-12">241.03±4.36*</span></p>
						</td>
						<td>
							<p class="Basic-Paragraph"><span class="CharOverride-12">54.80±0.97*</span></p>
						</td>
						<td>
							<p class="Basic-Paragraph"><span class="CharOverride-12">537.10±3.40*</span></p>
						</td>
						<td>
							<p class="Basic-Paragraph"><span class="CharOverride-12">11.47±0.57*</span></p>
						</td>
					</tr>
					<tr class="_idGenTableRowColumn-1">
						<td>
							<p class="Basic-Paragraph"><span class="CharOverride-12">8.</span></p>
						</td>
						<td>
							<p class="Basic-Paragraph"><span class="CharOverride-12">ZP</span></p>
						</td>
						<td>
							<p class="Basic-Paragraph"><span class="CharOverride-12">254.20±1.97*</span></p>
						</td>
						<td>
							<p class="Basic-Paragraph"><span class="CharOverride-12">57.23±1.28*</span></p>
						</td>
						<td>
							<p class="Basic-Paragraph"><span class="CharOverride-12">558.10±2.64*</span></p>
						</td>
						<td>
							<p class="Basic-Paragraph"><span class="CharOverride-12">6.83±0.08*</span></p>
						</td>
					</tr>
					<tr class="_idGenTableRowColumn-2">
						<td>
							<p class="Basic-Paragraph"><span class="CharOverride-12">9.</span></p>
						</td>
						<td>
							<p class="Basic-Paragraph"><span class="CharOverride-12">PN</span></p>
						</td>
						<td>
							<p class="Basic-Paragraph"><span class="CharOverride-12">251.13±1.50*</span></p>
						</td>
						<td>
							<p class="Basic-Paragraph"><span class="CharOverride-12">56.13±1.86*</span></p>
						</td>
						<td>
							<p class="Basic-Paragraph"><span class="CharOverride-12">552.47±4.22*</span></p>
						</td>
						<td>
							<p class="Basic-Paragraph"><span class="CharOverride-12">6.73±0.04*</span></p>
						</td>
					</tr>
					<tr class="_idGenTableRowColumn-1">
						<td>
							<p class="Basic-Paragraph"><span class="CharOverride-12">10.</span></p>
						</td>
						<td>
							<p class="Basic-Paragraph"><span class="CharOverride-12">AH</span></p>
						</td>
						<td>
							<p class="Basic-Paragraph"><span class="CharOverride-12">265.07±1.68*</span></p>
						</td>
						<td>
							<p class="Basic-Paragraph"><span class="CharOverride-12">64.23±0.32*</span></p>
						</td>
						<td>
							<p class="Basic-Paragraph"><span class="CharOverride-12">572.27±2.16*</span></p>
						</td>
						<td>
							<p class="Basic-Paragraph"><span class="CharOverride-12">6.96±0.09*</span></p>
						</td>
					</tr>
					<tr class="_idGenTableRowColumn-2">
						<td>
							<p class="Basic-Paragraph"><span class="CharOverride-12">11.</span></p>
						</td>
						<td>
							<p class="Basic-Paragraph"><span class="CharOverride-12">PI</span></p>
						</td>
						<td>
							<p class="Basic-Paragraph"><span class="CharOverride-12">284.03±6.85*</span></p>
						</td>
						<td>
							<p class="Basic-Paragraph"><span class="CharOverride-12">54.73±1.36*</span></p>
						</td>
						<td>
							<p class="Basic-Paragraph"><span class="CharOverride-12">571.43±1.76*</span></p>
						</td>
						<td>
							<p class="Basic-Paragraph"><span class="CharOverride-12">6.96±0.09*</span></p>
						</td>
					</tr>
					<tr class="_idGenTableRowColumn-1">
						<td>
							<p class="Basic-Paragraph"><span class="CharOverride-12">12.</span></p>
						</td>
						<td>
							<p class="Basic-Paragraph"><span class="CharOverride-12">PF</span></p>
						</td>
						<td>
							<p class="Basic-Paragraph"><span class="CharOverride-12">243.90±2.41*</span></p>
						</td>
						<td>
							<p class="Basic-Paragraph"><span class="CharOverride-12">70.20±0.54*</span></p>
						</td>
						<td>
							<p class="Basic-Paragraph"><span class="CharOverride-12">542.57±1.81*</span></p>
						</td>
						<td>
							<p class="Basic-Paragraph"><span class="CharOverride-12">5.82±0.04</span></p>
						</td>
					</tr>
					<tr class="_idGenTableRowColumn-2">
						<td>
							<p class="Basic-Paragraph"><span class="CharOverride-12">13.</span></p>
						</td>
						<td>
							<p class="Basic-Paragraph"><span class="CharOverride-12">MP</span></p>
						</td>
						<td>
							<p class="Basic-Paragraph"><span class="CharOverride-12">324.33±3.60*</span></p>
						</td>
						<td>
							<p class="Basic-Paragraph"><span class="CharOverride-12">74.53±0.54*</span></p>
						</td>
						<td>
							<p class="Basic-Paragraph"><span class="CharOverride-12">603.37±2.59*</span></p>
						</td>
						<td>
							<p class="Basic-Paragraph"><span class="CharOverride-12">7.14±0.08*</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-7">* Mean values (±SE) are significantly different (P&lt;0.05)</span></p>
			<p class="Body-Text">&nbsp;</p>
			<p class="Body-Text">&nbsp;</p>
			<p class="Body-Text">In the present investigation, all the PCR amplified full genome DNA amplicons from CAV positive flocks showed identical RFLP pattern with restriction enzymes (<span class="CharOverride-6">Hin</span>d III, <span class="CharOverride-6">Pst </span>I, <span class="CharOverride-6">Nhe </span>I and <span class="CharOverride-6">Mbo </span>II), signifying ciruclation of single serotype of the virus. Previous reports have shown pathogenic variations among the different isolates (<a href="#Kim-HR--Kwon-YK--Bae-YC--Oem-JK--Lee-OS--2010-."><span class="Hyperlink">Kim et al., 2010</span></a>; <a href="#Natesan-S--Kataria-JM--Dhama-K--Rahul-S--Baradhwaj-N--2006"><span class="Hyperlink">Natesan et al., 2003</span></a>; <a href="#Yuasa-N--Imai-K--1986-"><span class="Hyperlink">Yuasa et al., 1986</span></a>; <a href="#McNulty-MS--Connor-TJ--McNeilly-F--McLoughlin-MF--Kirkpatrick-KS--1990"><span class="Hyperlink">McNulty et al., 1990</span></a>; <a href="#Adair-BM--2000-"><span class="Hyperlink">Adair, 2000</span></a>; <a href="#Connor-TJ--McNeilly-F--Firth-GA--McNulty-MS--1991"><span class="Hyperlink">Connor et al., 1991</span></a>; <a href="#Davidson-I--Kedem-M--Borochovitz-H--Kass-N--Ayali-G--Hamzani-E--Perelman-B--Smith-B--Perk-S--2004"><span class="Hyperlink">Davidson et al., 2004</span></a>; <a href="#Oluwayelu-DO--Todd-D--Ball-NW-Scott-AN--Oladele-OA--Emikpe-BO--Fagbohun-OA--Owoade-AA--Olaleye-OD--2"><span class="Hyperlink">Oluwayelu et al., 2005</span></a>). A study from the USA reported an antigenically different CAV isolate (<a href="#Spackman-E--Cloud-SS--Rosenberger-JK--2002"><span class="Hyperlink">Spackman et al., 2002</span></a>), which could be a prototype virus of serotype 2. It has been highlighted that due to recombination mechanisms the new genetic variants are arising (<a href="#Eltahir-YM--Qian-K--Jin-W--Qin-A--2011-"><span class="Hyperlink">Eltahir et al., 2011</span></a>). Recently, genetically similar variants of gyroviruses have been detected in the human populations, alarming their importance in human health (<a href="#Biagini-P--B-darida-S--Touinssi-M--Galicher-V--de-Micco-P--2013"><span class="Hyperlink">Biagini et al., 2013</span></a>; <a href="#Smuts-HE--2014-."><span class="Hyperlink">Smuts, 2014</span></a>). <a href="#Todd-D--Mawhinney-KA--McNulty-MS--1992"><span class="Hyperlink">Todd et al. (1992)</span></a> assigned 14 CAV isolates of seven countries in to seven groups by RE analysis of the PCR amplified DNA with <span class="CharOverride-6">Hae </span>III, <span class="CharOverride-6">Hinf </span>I and <span class="CharOverride-6">Hpa </span>II enzymes. <a href="#Noteborn-MHM--Verschueren-CAJ--Van-Roozelaar-DJ-"><span class="Hyperlink">Noteborn et al. (1992)</span></a> used the enzymes, viz., <span class="CharOverride-6">Eco</span>RI, <span class="CharOverride-6">Acc</span>I, <span class="CharOverride-6">Bgl</span>II, <span class="CharOverride-6">Hin</span>dIII, <span class="CharOverride-6">Sst</span>I, <span class="CharOverride-6">Bam</span>HI, <span class="CharOverride-6">Xba</span>I and noticed some interesting minor differences by <span class="CharOverride-6">Acc</span>I, <span class="CharOverride-6">Hin</span>dIII and <span class="CharOverride-6">Eco</span>R I enzymes in spite of the overall high degree of similarity. Similarly, <a href="#Imai-K--Mase-M--Tsukamoto-K--Hihara-H--Yuasa-N--1999"><span class="Hyperlink">Imai et al. (1998)</span></a> analyzed 14 CAV amplified DNA products to <span class="CharOverride-6">Bgl </span>II, <span class="CharOverride-6">Hin</span>d III, <span class="CharOverride-6">Pst </span>I and <span class="CharOverride-6">Sac </span>I which showed the amplified region as highly conserved among all the isolates . However, in one of our study (<a href="#Dhama-K--2004"><span class="Hyperlink">Dhama et al., 2004</span></a>) we found some variations among the six Indian CAV isolates on the basis of RE analysis of PCR amplified products. Similarly, <a href="#Natesan-S--Kataria-JM--Dhama-K--Rahul-S--Baradhwaj-N--2006"><span class="Hyperlink">Natesan et al. (2003)</span></a> reported differences among Indian isolates of CAV by using three restriction enzymes (<span class="CharOverride-6">Hha </span>I, <span class="CharOverride-6">Dde </span>I and <span class="CharOverride-6">Hae </span>III). The previous reports are based on RE analysis of partial CAV genome segments and the novelty in RE digestion in this present study is that PCR products covering the whole viral genome were employed rather than any CAV gene specific smaller region PCR products, and such studies are lesser revealing RFLP pattern of the whole genome sequence, thus providing a useful molecular data regarding CAV RE analysis and comparison with other isolates of worldwide origin. Further studies including digestion with other specific restriction enzymes, sequence analysis and phylogenetic analysis of CAV genome are recommended for the differentiation and molecular characterization of all the CAV isolates.</p>
		  <p class="Body-Text">&nbsp;</p>
			<p class="Body-Text ParaOverride-1">During acute phases of the CIA especially in young chicks, CAV causes destructive effects on erythroid and myeloid tissues of bone marrow leading to suppression of differentiation and proliferation of haemopoietic precursor cells. This drastically affects erythropoiesis and myelopoiesis, leading to anaemia and (<a href="#McNulty-MS--1991-"><span class="Hyperlink">McNulty, 1991</span></a>; <a href="#Dhama-K--Mahe-2008"><span class="Hyperlink">Dhama et al., 2008</span></a>). In the present study, a significant (P&lt;0.05) decline in the Hb, PCV and TEC was observed in the CAV positive groups. In the myleiod lineages also, a significant (P&lt;0.05) decline in the TLC and percent lymphocyte count was observed in all CAV positive birds as compared to the selected healthy group. Such haematological studies could be useful in determining the haematological effects of CAV in co-infections with other pathogens, for which purpose explorative studies are suggested.</p>
		  <p class="Body-Text ParaOverride-1">&nbsp;</p>
			<p class="Body-Text ParaOverride-1">It is well documented that AST, ALT and ALP are found in the liver and ALP is found in the kidney also (<a href="#Kaneko-JJ--Harvey-JW--Bruss-ML--1997"><span class="Hyperlink">Kaneko et al., 1997</span></a>). CAV has a wide spread distribution in the body resulting in damage and focal necrosis of liver, kidney and spleen. Haemorrhages in the proventricular mucosa and subcutaneous tissues and muscular haemorrhages within wing tips are sometimes associated with severe anaemia (<a href="#Dhama-K--Mahe-2008"><span class="Hyperlink">Dhama et al., 2008</span></a>; <a href="#Yuasa-N--Imai-K--1986-"><span class="Hyperlink">Yuasa et al., 1986</span></a>; <a href="#Engstrom-BE--Fossum-O--Luthman-M--1998-."><span class="Hyperlink">Engstrom et al., 1988</span></a>). The  present study demonstrated  significant (P&lt;0.05) elevation in the activities of ALT, AST and ALP enzymes, and uric acid of CAV positive birds, which can be associated with the damage to the organs like liver, kidneys and muscles of CAV affected chicks. Elaborative studies in this direction could make understand the pathology and pathogenesis of the CIA more clearly.</p>
		  <p class="Body-Text ParaOverride-1">&nbsp;</p>
			<p class="Body-Text ParaOverride-1">CAV has been found to be associated with both clinical and sub-clinical infections and have substantial effects particularly on commercial broiler performance. Economic losses stem from increased mortality, condemnations, the cost of antibiotics used to control secondary bacterial infections due to immunosuppression, vaccine failures and poor production performance (<a href="#McNulty-MS--1991-"><span class="Hyperlink">McNulty, 1991</span></a>; <a href="#Dhama-K--Mahe-2008"><span class="Hyperlink">Dhama et al., 2008</span></a>; <a href="#McIlroy-SG--McNulty-MS--Bruce-DW--Smyth-JA--Goodall-EA--Alcorn-MJ--1992-"><span class="Hyperlink">McIlroy et al., 1992</span></a>). In the present study, all the CAV positive flocks showed significantly lower value of ND HI titres as compared to the normal healthy flocks. This outcome suggested that CAV is responsible for lower immune response against ND vaccination. The destructive effect of CAV due to suppression of the population of both helper (CD4<span class="CharOverride-9">+</span>) and cytotoxic (CD8<span class="CharOverride-9">+</span>) T-Lymphocytes in the thymus as suggested by <a href="#Hu-LB--Lucio-B--Schat-KA--1993"><span class="Hyperlink">Hu et al. (1993)</span></a> and <a href="#Adair-BM--2000-"><span class="Hyperlink">Adair (2000)</span></a> could be the reason for poor but protective immune response in CAV inoculated chicks. The low antibody titer in CAV infected birds and marked depression of humoral immune response has also been reported by other (<a href="#Bhatt-P--Shukla-2013"><span class="Hyperlink">Bhatt et al., 2013</span></a>; <a href="#Cloud-SS--Rosenberger-JK--Lillehoj-HS--1992-"><span class="Hyperlink">Cloud et al., 1992</span></a>; <a href="#Liu-ZG--Zheng-SM--Yang-LP--Xu-YB--Zhou-ZY--2001"><span class="Hyperlink">Liu et al., 2001</span></a>).</p>
		  <p class="Body-Text ParaOverride-1">&nbsp;</p>
			<p class="Body-Text ParaOverride-1">Overall, findings of the present study highlight the high prevalence of CAV in Indian poultry flocks with confirmation of its immunosuppressive role with other pathogens in field conditions. Furthermore, the active presence of CIA viral DNA in the tissue samples of diseased birds along with clinico-pathological and haemato-biochemical findings in the disease outbreaks investigated confirmed the involvement of CAV and its association with precipitation of other poultry diseases through its marked immunosuppressive potential. Though, this appears to be the first report from India indicating association of CAV in precipitating other secondary pathogens/diseases in commercial poultry flocks, additional studies are required to find out the subclinical infection of CAV in commercial poultry flocks by suitable diagnostic assays and its association with other immunosuppressive and opportunistic pathogens in causing significant economic losses to poultry population. The usage of full length viral genome based PCR-RFLP for studying molecular epidemiology of this emerging and economically important virus is the most significant finding of the present study. The recent reports regarding CAV related variants in human population and its potent role in precipitating other infectious diseases in birds further warrants in depth molecular epidemiological studies, antigenic and genomic characterization of viral isolates, so that an appropriate and effective strategy could be designed and adopted for the prevention and control of this important avian pathogen, and alleviate economic losses being suffered in poultry population.</p>
		  <p class="Body-Text ParaOverride-1">&nbsp;</p>
			<p class="Figure--and-Table-Heading ParaOverride-1"><span class="CharOverride-23">CONFLICT OF INTEREST </span></p>
		  <p class="Figure--and-Table-Heading ParaOverride-1">&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----">ACKNOWLEDGEMENTS</p>
		  <p class="Heading-1--Introduction----">&nbsp;</p>
			<p class="Body-Text ParaOverride-1">The authors are thankful to the Director, Experiment Station, Dean, College of Veterinary and Animal Sciences, Dean, Post Graduate Studies, G.B. Pant University of Agriculture and Technology, Pantnagar, Uttarakhand, and Director, Indian Veterinary Research Institute, Izatnagar, Uttar Pradesh for providing necessary facilities to carry out this research work.</p>
		  <p class="Body-Text ParaOverride-1">&nbsp;</p>
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