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			<p class="Type-of-Article">&nbsp;</p>
			<p class="Type-of-Article"><span class="CharOverride-1">Review Article</span></p>
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		<div class="Basic-Text-Frame">
			<p class="title- ParaOverride-1">&nbsp;</p>
			<p class="title- ParaOverride-1">Early Mortality Syndrome (EMS) as new Emerging Threat in Shrimp Industry</p>
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			<p class="Affiliations ParaOverride-1">&nbsp;</p>
			<p class="Affiliations ParaOverride-1"><span class="CharOverride-2">1</span>Iranian Coldwater Fishes Scientific Society (CFS); <span class="CharOverride-2">2</span>Aquatic animal Health &amp; Diseases Dept.; <span class="CharOverride-2">3</span>Shrimp Division in Aquatic animal Health &amp; Diseases Dept., Iranian Fisheries Research Organization (IFRO), Tehran-Karaj Highway, Paykan shahr, Sarve Azad Ave., Botanical National Blvd. Tehran, I.R. Iran.</p>
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			<p class="Authors ParaOverride-1">Mohammad Jalil Zorriehzahra<span class="CharOverride-2">1, 2</span>*, Reza Banaederakhshan<span class="CharOverride-2">3</span></p>
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			<p class="Abstract ParaOverride-1">&nbsp;</p>
			<p class="Abstract ParaOverride-1"><span class="CharOverride-3">Abstract</span> | <span lang="en-US">Early Mortality Syndrome (EMS) also named Acute Hepatopancreatic Necrosis Disease or AHPND should be considered as a new emerging shrimp disease that has been attacked to shrimp farms in Southeast Asia. It was detected in shrimp farms in southern China as first record in 2009 and then in Hainan Island in 2010 and afterward in Viet Nam and Malaysia in 2011 and consequently in the eastern part of Thailand since 2012 and widely spread to other culture areas of Thailand. Its annual losses are estimated more than USD1 billion. Outbreaks of EMS naturally occur in the first 30 days after stocking a freshly arranged shrimp pond, and rate of mortality can pass beyond 70%. The scientists recently found that (EMS/AHPND) could be initiated by a bacterial agent that termed </span><span class="CharOverride-4" lang="en-US">V. parahaemolyticus</span><span lang="en-US"> is transferred through oral and then localizes the shrimp gastrointestinal tract and create a poison that causes tissue devastation and invalidism of the shrimp digestive system known as the hepatopancreas. So far, no official report was recorded in human cells. Therefore, it is unlikely that the specific strain of </span><span class="CharOverride-4" lang="en-US">V. parahaemolyticus</span><span lang="en-US"> will pose any risk when consumed by human beings. It causes some clinical signs which include pale discoloration and atrophy (size reduction) of hepatopancreas, which appears granular when pressed between the fingers, with occasional black streaks. Other clinical signs include pale and empty stomach and gut, reduced growth, movable shell and black discoloration. Also, lethargy, swimming sluggishly along the dikes, spiral swimming and reduced flourish and feeding are observed in EMS/AHPND. More investigation are needed and  should be continue on the development of rapid diagnostic tests for prompt detection of the EMS/AHPND pathogen that will facilitate developed management of hatcheries and ponds, and could be lead to a long-term elucidation for the disease aspects. </span> </p>
		  <p class="Abstract ParaOverride-1">&nbsp;</p>
			<p class="Abstract ParaOverride-1"><span class="CharOverride-3">Keywords </span>| Shrimp, Early mortality syndrome, Acute Hepatopancreatic Necrosis Disease, EMS/ AHPNS, Iran</p>
		  <p class="Abstract ParaOverride-1">&nbsp;</p>
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			<p class="Editor----Citation"><span class="CharOverride-6">Editor</span> | Ruchi Tiwari, College of Veterinary Sciences, Department of Veterinary Microbiology and Immunology Uttar Pradesh Pandit Deen Dayal Upadhayay Pashu Chikitsa, Vigyan Vishvidhyalaya Evum Go-Anusandhan Sansthan (DUVASU), Mathura (U.P.) – 281001, India.</p>
			<p class="Editor----Citation"><span class="CharOverride-6">Special Issue</span> | 2 (2015) “Reviews on Trends and Advances in Safeguarding Terrestrial /Aquatic Animal Health and Production”</p>
			<p class="Editor----Citation"><span class="CharOverride-3">Received</span> | February 27, 2015; <span class="CharOverride-3">Revised</span> | March 06, 2015; <span class="CharOverride-3">Accepted</span> | March 07, 2015; <span class="CharOverride-3">Published</span> | March 13, 2015&#9;&#9;</p>
			<p class="Editor----Citation"><span class="CharOverride-3">*Correspondence</span> | Mohammad Jalil Zorriehzahra, Iranian Fisheries Research Organization (IFRO), Tehran, Iran; <span class="CharOverride-3">Email:</span> zorrieh@yahoo.com</p>
			<p class="Editor----Citation ParaOverride-2"><span class="CharOverride-3">Citation</span> | Zorriehzahra MJ, Banaederakhshan R (2015). Early mortality syndrome (EMS) as new emerging threat in shrimp industry. Adv. Anim. Vet. Sci. 3(2s): 64-72.  </p>
			<p class="Editor----Citation"><span class="CharOverride-3">DOI</span> | <a href="http://dx.doi.org/10.14737/journal.aavs/2015/3.2s.64.72"><span class="Hyperlink">http://dx.doi.org/10.14737/journal.aavs/2015/3.2s.64.72</span></a></p>
			<p class="Editor----Citation"><span class="Editor---Citation CharOverride-3" lang="en-US">ISSN </span><span class="Editor---Citation CharOverride-3" lang="en-US">(</span><span class="Editor---Citation CharOverride-3" lang="en-US">Online</span><span class="Editor---Citation CharOverride-3" lang="en-US">)</span> | 2307-8316; <span class="Editor---Citation CharOverride-3" lang="en-US">ISSN </span><span class="Editor---Citation CharOverride-3" lang="en-US">(Print) </span> | 2309-3331</p>
			<p class="Editor----Citation"><span class="CharOverride-6">Copyright </span>© 2015 Zorriehzahra 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>
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			<p class="Heading-1--Introduction----">&nbsp;</p>
			<p class="Heading-1--Introduction----">&nbsp;</p>
			<p class="Heading-1--Introduction----">Introduction</p>
			<p class="Heading-1--Introduction----">&nbsp;</p>
			<p class="Heading-1--Introduction----"><span class="_idGenDropcap-1">T</span>he annual world aquatic animal production has been on growth during last recent decade. These amounts prepare 19Kg of per capita consumption and 17% of animal protein requirements and have a strong roll on food safety. The total aquatic animal production on 2010 was about 148 million Metric tons that 128 million tons of them had been directly consume by man and their remains used for fish meal and other purposes (<a href="#FAO-Fisheries-and-Aquaculture-Report--2013-"><span class="Hyperlink">FAO, 2013</span></a>). Although the portion of fish catching is bigger than aquaculture, but aquaculture contribution increased from 13% on 1990 to 40% on 2010. The roll of aquaculture portion in human consumption was about 47% on 2010. This product prepares about 220 million jobs opportunity in different branches in the world (<a href="#FAO-Fisheries-and-Aquaculture-Report--2013-"><span class="Hyperlink">FAO, 2013</span></a>). One of the most important problems in aquatic animal production especially in aquaculture activity is health and diseases management, so the fish and shrimp farmer lost several millions USD due to diseases each year. According to worldwide extension of aquaculture activity, new emerged diseases and occurrence of other diseases increased year by year. So far, about twenty viral, four bacterial, three fungal and some protozoan diseases had been recognized in crustacean and shrimps (<a href="#Muhammad-F--Zhang-ZF--Shao-MY--Shi-XL--Shafi-M--2012-"><span class="Hyperlink">Muhammad, et al., 201</span></a><span class="Hyperlink">2</span>). Some important viral diseases consist of WSSD and TSV. Recently, a new emerged lethal disease that termed EMS (Early Mortality Syndrome) or AHPND (Acute Hepatopancreatic Necrosis Disease) had been added to list of shrimp diseases during last recent years.</p>
		  <p class="Heading-1--Introduction----">&nbsp;</p>
			<p class="Heading-1--Introduction----">Epidemiology</p>
			<p class="Heading-1--Introduction----">&nbsp;</p>
			<p class="Body-Text ParaOverride-1">This disease had caused mass mortality in China (2009) as first time and then in Viet Nam (2010), afterward in Malaysia (2011) and finally in Thailand (2012) (<a href="#x1-Figure-1-"><span class="Hyperlink">Figure</span><span class="Hyperlink"> 1</span></a>). It named as EMS due to mass mortality during few days after shrimp post larvae stoking. This is an acute disease and affected region in China had lost 80% of their products due to this catastrophic disease. The economic lost was estimated 570000 till 7200000 USD on 2011 and 2012 in Vietnam. This disease decreased the mass production from 70000 tons in 2010 to 40000 and 30000 tons in 2011 and 2012, respectively. About 7% of total shrimp production lost in Thailand especially in coastal area during 2012 (<a href="#Lightner-DV--Redman-CR--Pantoja-BL--Noble-LM--Nunan--Loc-Tran--2013-"><span class="Hyperlink">Lightner et al., 2013</span></a>). This disease had been reported from Western Hemisphere and Mexico during 2013 and made about 118 million USD economic lost in a part of this country (<a href="#Schryver-P--Defoirdt-T--Sorgeloos-P--2014"><span class="Hyperlink">Schryver et al., 2014</span></a>). The spread to the Western Hemisphere is a major concern to shrimp producers in the region. To date, the only peer-reviewed published method for determining whether mortalities are due to AHPND is through histological examination. A novel PCR detection method was employed to assess samples from Mexico in order to confirm the presence of the pathogen in this country. This manuscript details the detection methods used to confirm the presence of AHPND in Mexico. Both immersion and per os challenge studies were used to expose the <span class="CharOverride-4">Penaeus vannamei</span> to the bacteria in order to induce the disease. Histological analysis confirmed AHPND status following the challenge studies. Also provided are the details of the molecular test by PCR that was used for screening candidate <span class="CharOverride-4">V. parahaemolyticus</span> isolates. A rapid PCR assay for detection of AHPND may help with early detection and help prevent the spread of AHPND to other countries (<a href="#Nunan-L--Lightner-D--Pantoja-C--Gomez-Jimenez-S--2014-."><span class="Hyperlink">Nunanr et al., 2014</span></a>). Despite of trying to disease control during last recent years, it is not under control and made severe mortality in some countries such as Vietnam and Thailand in 2014. Although the disease has already been identified and it is transmitted by ingestion is expressed, it seems that the following conditions are included in the incidence and prevalence of disease:</p>
			<ol>
				<li class="Body-Text ParaOverride-3">High stocking density</li>
				<li class="Body-Text ParaOverride-3">Sensitivity of species</li>
				<li class="Body-Text ParaOverride-3">The high salinity of the water in areas near the sea</li>
				<li class="Body-Text ParaOverride-3">The lack of reservoir pond and water treatment in shrimp farms</li>
				<li class="Body-Text ParaOverride-4">Lack of attention to process of Pond preparation</li>
				<li class="Body-Text ParaOverride-3">Stress during transportation or storage of shrimp post larvae</li>
				<li class="Body-Text ParaOverride-3">Low oxygen conditions</li>
				<li class="Body-Text ParaOverride-3">Feed mismanagement (<a href="#Ghavampour--2012-.-A"><span class="Hyperlink">Ghavampour, 2012</span></a>)</li>
			</ol><br>
			
            <div class="pt" > <a href="http://nexusacademicpublishers.com/uploads/figures/20150316002047.jpg" target="new"><img class="img_display" src="http://nexusacademicpublishers.com/uploads/figures/20150316002047.jpg" width="80" height="80"></a>
            
            <p class="Figure--and-Table-Heading ParaOverride-1"><span class="CharOverride-3"><a id="x1-Figure-1-"></a>Figure 1:</span><a href="http://nexusacademicpublishers.com/uploads/figures/20150316002047.jpg"> Geographical distribution of (EMS/AHPND) in the world</a></p>
       </div>

			<p class="Heading-1--Introduction----">&nbsp;</p>
			<p class="Heading-1--Introduction----">Etiology</p>
		  <p class="Heading-1--Introduction----">&nbsp;</p>
			<p class="Body-Text">After occurrence and spreading of EMS/AHPND in Southeast Asia, initial study had been started and focused on recognition of pathogen. This disease had considered as an unknown pathogen (Idiopathic) article and had been believed several parameter like cypermethrin and other insecticide pollution, environmental pollution, some parasites, some unknown parameters in feed, harmful algae, probiotics, family cross breeding, etc. and have a critical role in it (<a href="#FAO-Fisheries-and-Aquaculture-Report--2013-"><span class="Hyperlink">FAO, 2013</span></a>). However, in early 2013, the Aquaculture Pathology Laboratory at the University of Arizona was able to isolate the causative agent of AHPND in pure culture. Immersion challenge tests were employed for infectivity studies, which induced 100% mortality with typical AHPND pathology to experimental shrimp exposed to the pathogenic agent. Subsequent histological analyses showed that AHPND lesions were experimentally induced in the laboratory and were identical to those found in AHPND-infected shrimp samples collected from the endemic areas. Bacterial isolation from the experimentally infected shrimp enabled recovery of the same bacterial colony type found in field samples. In 3 separate immersion tests, using the recovered isolate from the AHPND-positive shrimp, the same AHPND pathology was reproduced in experimental shrimp with consistent results. Hence, AHPND has a bacterial etiology and Koch’s Postulates have been satisfied in laboratory challenge studies with the isolate, which has been identified as a member of the <span class="CharOverride-4">Vibrio harveyi</span> clade, most closely related to <span class="CharOverride-4">V. parahemolyticus</span> (<a href="#Tran-LH--Fitzsimmons-KM--Lightner-DV--2014-."><span class="Hyperlink">Tran et al., 2014</span></a>).</p>
		  <p class="Body-Text">&nbsp;</p>
			<p class="Body-Text ParaOverride-1">Regarding to last investigation bacteriophage virus has a potential role in pathogenicity in this disease (Flegel, unpublished data). They could not find any relationship between virus and bacterial pathogenicity by DNA sequencing but they confirmed that the virus presence over the <span class="CharOverride-4">Vibrio parahaemolyticus</span> by TEM (<a href="#Eduardo-M-Lea-o--Mohan-CV--2012-."><span class="Hyperlink">Eduardo et al., 2012</span></a>). Finally, an especial strain of <span class="CharOverride-4">Vibrio parahaemolyticus </span>that affected by phage virus was responsible of EMS\AHPND (Lightner, unpublished data). They identified it as a unique strain of <span class="CharOverride-4">V. parahaemolyticus </span>after affected by bacteriophage virus can produce a kind of toxin that makes this disease (<a href="#GAA--2013-"><span class="Hyperlink">GAA, 2013</span></a>). An analogous advent occurs in the cholera as human killer disease, where a phage makes the <span class="CharOverride-4">V. cholerae</span> bacterium talented of making a poison that causes cholera’s life-threatening gastroenteritis that may quickly cause dangerous fluid loss in human.</p>
		  <p class="Body-Text ParaOverride-1">&nbsp;</p>
			<p class="Body-Text ParaOverride-1">Researchers found main pathogen in shrimp farms that transmitted to sensitive host by digestive system and located in the gut and then produced the toxin that causes necrosis and degeneration especially in haepathopancreas. This disease cannot transmit to human and it is not considered as a man health problem (<a href="#FAO-Fisheries-and-Aquaculture-Report--2013--1"><span class="Hyperlink">FAO News, 2013</span></a>). Although the main pathogen in this disease is <span class="CharOverride-4">V. parahaemolyticus</span> but there is some reports about <span class="CharOverride-4">V. owensii</span> that report it as a causative agent (<a href="#Shrimp-news-international--December--24---2013-"><span class="Hyperlink">Shrimp news, 2013</span></a>). </p>
		  <p class="Body-Text ParaOverride-1">&nbsp;</p>
			<p class="Body-Text ParaOverride-1">Also, variation of <span class="CharOverride-4">V. parahaemolyticus</span> isolates from a single Thai shrimp farm that experiencing an outbreak of acute hepatopancreatic necrosis disease (AHPND) were investigated and their results revealed the possibility of diversity in isolates of <span class="Emphasis">V. parahaemolyticus</span> that may cause early mortality in shrimp cultivation ponds in Thailand (<a href="#Joshi-J--Srisala-J--Truong-VH--Chen-2014"><span class="Hyperlink">Joshi et al., 201</span></a><span class="Hyperlink">4</span>).</p>
		  <p class="Body-Text ParaOverride-1">&nbsp;</p>
			<p class="Body-Text ParaOverride-1">Polecats worm that used for feeding spowner shrimp in hatcheries can transmit the pathogen to hatchery and affect shrimp post larvae (Flegel, unpublished data). </p>
		  <p class="Body-Text ParaOverride-1">&nbsp;</p>
			<p class="Heading-1--Introduction----">Resistance to physical and chemical action</p>
		  <p class="Heading-1--Introduction----">&nbsp;</p>
			<p class="Body-Text ParaOverride-1">Temperature and Freezing: Attempts to transmit AHPND using infected frozen shrimp have been unsuccessful. Strains of <span class="CharOverride-4">V. parahaemolyticus </span>in seafood are known to be sensitive to freezing (–18°C to –24°C) and reductions in cultural cells to non-detectable levels have been reported after freezing for several weeks. Time of total inactivation depends on the initial number of bacteria and temperature.</p>
		  <p class="Body-Text ParaOverride-1">&nbsp;</p>
			<p class="Body-Text ParaOverride-1"><span class="CharOverride-3">Refrigeration: </span>Strains of <span class="CharOverride-4">V. parahaemolyticus </span>in seafood are known to be sensitive to refrigeration (4°C) and reductions in cultural cells have been reported, but not to non-detectable levels during the storage life of seafood products at this temperature.</p>
		  <p class="Body-Text ParaOverride-1">&nbsp;</p>
			<p class="Body-Text ParaOverride-1"><span class="CharOverride-3">Heating:</span> Strains of <span class="CharOverride-4">V. parahaemolyticus </span>in seafood are known to be sensitive to heating and reductions in cultural cells to non-detectable levels have been reported at 55°C for 5 minutes and 80°C for 1 minute (<a href="#Acute-Hepatopancreatic-Necrosis-Disease-Etiology-Epidemiology-Diagnosis-Prevention-and-Control--OIE"><span class="Hyperlink">OIE, 2013</span></a>) so pelleted feed cannot transmit them due to heath usage in production process (<a href="#Lightner-DV--Redman-CR--Pantoja-BL--Noble-LM--Nunan--Loc-Tran--2013-"><span class="Hyperlink">Lightner  et al., 2013</span></a>).</p>
		  <p class="Body-Text ParaOverride-1">&nbsp;</p>
			<p class="Body-Text ParaOverride-1"><span class="CharOverride-3">Chemicals/Disinfectants:</span> Susceptible to common disinfectants. No cultural cells after exposure to pH 5 for15 minutes.</p>
			<p class="Body-Text ParaOverride-1">&nbsp;</p>
			<p class="Body-Text ParaOverride-1"><span class="CharOverride-3">Survival:</span> Survival to 9 and 18 days in filtered estuarine water and filtered seawater respectively.</p>
			<p class="Body-Text ParaOverride-1">Densities of <span class="CharOverride-4">V. parahaemolyticus </span>in seawater are known to be temperature dependent (<a href="#Acute-Hepatopancreatic-Necrosis-Disease-Etiology-Epidemiology-Diagnosis-Prevention-and-Control--OIE"><span class="Hyperlink">OIE, 2013</span></a>).</p>
		  <p class="Body-Text ParaOverride-1">&nbsp;</p>
			<p class="Heading-2--History-in-MM-">Susceptible species</p>
		  <p class="Heading-2--History-in-MM-">&nbsp;</p>
			<p class="Body-Text ParaOverride-1">This disease had been reported from both two important cultivated shrimps such as black tiger shrimp (<span class="CharOverride-4">P. monodon</span>) and American white leg shrimp (<span class="CharOverride-4">L. vannamei</span>)<span lang="ar-SA"> </span>in Southeast Asia. This disease also reported from (<span class="CharOverride-4">P. chinensis</span>) in that region. It seems that other species are resistant or less susceptible to EMS\AHPND (<a href="#FAO-statistical-yearbook--2013"><span class="Hyperlink">FAO, 2011</span></a> and <a href="#Acute-Hepatopancreatic-Necrosis-Disease-Etiology-Epidemiology-Diagnosis-Prevention-and-Control--OIE"><span class="Hyperlink">OIE, 2013</span></a>).</p>
		  <p class="Body-Text ParaOverride-1">&nbsp;</p>
			<div class="pt" > <a href="http://nexusacademicpublishers.com/uploads/figures/20150316002249.jpg" target="new"><img class="img_display" src="http://nexusacademicpublishers.com/uploads/figures/20150316002249.jpg" width="80" height="80"></a>
            
            <p class="Figure--and-Table-Heading ParaOverride-1"><span class="CharOverride-3"><a id="x2-Figure-2--1"></a>Figure 2:</span><a href="http://nexusacademicpublishers.com/uploads/figures/20150316002249.jpg"> Gross signs of <span class="CharOverride-4">Penaeus vannamei</span> affected by AHPNS (Left) compared to normal shrimp (right)</a></p>
       </div><p class="Figure--and-Table-Heading ParaOverride-1"><span class="CharOverride-5">Source: </span><span class="CharOverride-9">D. Lightner (1)</span></p><br>
			
			<div class="pt" > <a href="http://nexusacademicpublishers.com/uploads/figures/20150316005550.png" target="new"><img class="img_display" src="http://nexusacademicpublishers.com/uploads/figures/20150316005550.png" width="80" height="80"></a>
            
            <p class="Figure--and-Table-Heading ParaOverride-1"><span class="CharOverride-3"><a id="x1-Figure-3--1"></a>Figure</span> <span class="CharOverride-3">3:</span><a href="http://nexusacademicpublishers.com/uploads/figures/20150316005550.png">  Pale atrophied hepatopancreas of juvenile <span class="CharOverride-4">Penaeus monodon</span> from Vietnam with AHPNS</a></p>
       </div>
			
			
		  <p class="Figure--and-Table-Heading ParaOverride-1"><span class="CharOverride-5">Source: </span><span class="CharOverride-9">D. Lightner (1)</span></p><br>
			<p class="Heading-2--History-in-MM-">Clinical signs  </p>
			<p class="Body-Text ParaOverride-1">This disease specified with mass mortality (up to 100%) 20-30 days after stocking. Most important clinical symptoms consist of lethargy, low speed in growth rate, spiral swimming, as well as empty or interrupted gut. Also infected shrimps constantly reveal an abnormal hepatopancreas (Emaciated, small, distended, pale or black coloration) (<a href="#Lightner-DV--Redman-CR--Pantoja-BL--Noble-LM--Nunan--Loc-Tran--2013-"><span class="Hyperlink">Lightner  et al., 2013</span></a>). Therefore, some of the behavioural changes such as lethargy, swimming sluggishly along the dikes, spiral swimming and reduced preening and feeding are also observed in EMS.</p>
		  <p class="Body-Text ParaOverride-1">&nbsp;</p>
			<p class="Body-Text ParaOverride-1">Affected shrimp sank to the pond bottom and die there. Hepatopacreatic discoloration caused by loses of connective and epithelial tissues pigmentation and black coloration is due to melanization (<a href="#Afsharnasab-M--Banaederakhshan-R--2014-"><span class="Hyperlink">Afsharnasab et al., 2014</span></a>). Affected hepatopancreas doesn’t squash easily by finger that probably caused by increasing of connective tissues or aggregation of haemocyte (<a href="#Lightner-DV--2012-"><span class="Hyperlink">Lightner, 2012</span></a>). Attendance of some bacteria containing <span class="CharOverride-4">V.</span> <span class="CharOverride-4">microsporidians </span>and worm parasite such as nematode has been detected in some affected samples (<a href="#Eduardo-M-Lea-o--Mohan-CV--2012-."><span class="Hyperlink">Eduardo et al., 2012</span></a>). The affected shrimp are smaller than healthy one in size (<a href="#x2-Figure-2--1"><span class="Hyperlink">Figure 2</span></a>, <a href="#x1-Figure-3--1"><span class="Hyperlink">3</span></a> and <a href="#x1-Figure-4--1"><span class="Hyperlink">4</span></a>).</p>
		  <p class="Body-Text ParaOverride-1">&nbsp;</p>
			
            <div class="pt" > <a href="http://nexusacademicpublishers.com/uploads/figures/20150316002659.jpg" target="new"><img class="img_display" src="http://nexusacademicpublishers.com/uploads/figures/20150316002659.jpg" width="80" height="80"></a>
            
            <p class="Figure--and-Table-Heading ParaOverride-1"><span class="CharOverride-3"><a id="x1-Figure-4--1"></a>Figure 4:</span><a href="http://nexusacademicpublishers.com/uploads/figures/20150316002659.jpg">  Atrophied hepatopancrease indicative of AHPNS</a></p>
       </div>
			
		  <p class="Figure--and-Table-Heading ParaOverride-1"><span class="CharOverride-5">Source: </span><span class="CharOverride-9">D. Lightner (4)</span></p>
		  <p class="Figure--and-Table-Heading ParaOverride-1">&nbsp;</p>
			
			
            <div class="pt" > <a href="http://nexusacademicpublishers.com/uploads/figures/20150316001555.png" target="new"><img class="img_display" src="http://nexusacademicpublishers.com/uploads/figures/20150316001555.png" width="80" height="80"></a>
            
            <p class="Figure--and-Table-Heading ParaOverride-1"><span class="CharOverride-3"><a id="x1-Figure-5"></a>Figure</span> <span class="CharOverride-3">5:</span><a href="http://nexusacademicpublishers.com/uploads/figures/20150316001555.png">  Key diagnostic feature of AHPNS-affected hepatopancreas (HP) with medial sloughing of HP cells (right photo), compared with normal HP (left photo) with intact tubules and distinct F, B and R cells</a></p>
       </div>
			
		  <p class="Figure--and-Table-Heading ParaOverride-1"><span class="CharOverride-5">Source:</span><span class="CharOverride-9"> T. Flegel (4)</span></p>
			<p class="Heading-1--Introduction----">&nbsp;</p>
			<p class="Heading-1--Introduction----">Histopathology</p>
			<p class="Heading-1--Introduction----">&nbsp;</p>
			<p class="Body-Text">Hepatopancreas is the main target tissue in this disease. The major lesions should be considered in the hepatopancreas during acute progressive degeneration of the hepatopancreas with initial decrease of R, B and F-cells followed by an obvious reduction of mitotic activity in E-cells. The development of lesion is noticed from proximal to distal with dysfunction of R, B, F, and finally E-cells, with affected hepato pancreas tubule mucosal cells presenting prominent karyomegaly (enlarged nuclei), and rounding and sloughing into the hepatopancreas tubules (<a href="#x1-Figure-5"><span class="Hyperlink">Figure 5</span></a>). The sloughed hepatopancreas cells prepare a substrate for bacterial growth, resulting in massive secondary bacterial infection (putative Vibrio spp.) and complete devastation of hepatopancreas at the fatal phase of the disease. Inter tubular haemocytic aggregation and haemocytes encapsulation of necrotic hepatopancreas tubules and melanisation of the more proximal portions of hepatopancreas tubules is seen in some infected shrimp.  Also, expanding inefficiency of hepatopacreas due to degeneration and incapacitation of tubular epithelium from beginning to end part of them (<a href="#Acute-Hepatopancreatic-Necrosis-Disease-Etiology-Epidemiology-Diagnosis-Prevention-and-Control--OIE"><span class="Hyperlink">OIE, 2013</span></a>) and (<a href="#Praveena-PE--Bhuvaneswari-T--Otta-SK--Raj-NJM--Jithendran-KP--2014-"><span class="Hyperlink">Praveena et al., 2014</span></a>). </p>
		  <p class="Body-Text">&nbsp;</p>
			<p class="Heading-1--Introduction----">Disease diagnosis</p>
		  <p class="Heading-1--Introduction----">&nbsp;</p>
			<p class="Body-Text ParaOverride-1">Although research on rapid diagnosis of the disease continues, clinical signs of the disease can help to initial diagnosis to professionals. Specific clinical signs such as soft and dark shell, wasting, anorexia and discoloration of the hepatopancreas are the major clinical signs (<a href="#Afsharnasab-M--Banaederakhshan-R--2014-"><span class="Hyperlink">Afsharnasab et al., 2014</span></a>). Histopathological studies show some changes such as atrophy, discoloration and growth of bacteria in the hepatopancreas tissue. Rapid diagnosis of this disease and its control has led researchers to combat attempts to develop methods for early diagnosis of this disease, eventually led to the construction of PCR kit for detection of the disease by (<a href="#Lightner-DV--Redman-CR--Pantoja-BL--Noble-LM--Nunan--Loc-Tran--2013-"><span class="Hyperlink">Lightner et al., 2013</span></a>). The main advantages of this new method of diagnosis can be made as follows:</p>
			
				<li class="Body-Text ParaOverride-5">Rapid diagnosis AHPND / EMS</li>
				<li class="Body-Text ParaOverride-5">Economical advantage of this method</li>
				<li class="Body-Text ParaOverride-5">No need for time-consuming and costly tests of histopathology</li>
		  <li class="Body-Text ParaOverride-5">Preparation of the test on a large number of shrimp samples (<a href="#Shrimp-news-international--December-13---2013-"><span class="Hyperlink">Shrimp news, 2013</span></a>)</li>
			
			<p class="Body-Text ParaOverride-1">According to this method, had made a commercial diagnosis test kite that called IQ2000 (<a href="#Shrimp-news-international--December--24---2013-"><span class="Hyperlink">Shrimp news, 2013</span></a>). This method makes a new, rapid and inexpensive method called IQ Plus TM AHPND/EMS Kit (POCKIT) additional to PCR kit to disease diagnosis. The new method is able to evaluate the hepatopancreas, stool, post larvae, water and food samples (<a href="#Simon-C--2014"><span class="Hyperlink">Simon, 2014</span></a>).</p>
		  <p class="Body-Text ParaOverride-1">&nbsp;</p>
			<p class="Body-Text ParaOverride-1">Also, a new PCR test kit had been made for EMS\AHPND diagnosis (<span class="Hyperlink">Flegel, 2013</span>). According to this research, acute strain of <span class="CharOverride-4">V. parahaemolyticus</span> genome contains a special gene that it is invisible in other strain. They design their PCR test kit based on these genes. This method contains 2 primer that called AP1 and AP2. In case of EMS\AHPND and acute strain of <span class="CharOverride-4">V. parahaemolyticus</span> these two primers activate and help to case recognition (<a href="#Shrimp-news-international--January-7---2014-"><span class="Hyperlink">Shrimp news, 2014</span></a>).</p>
		  <p class="Body-Text ParaOverride-1">&nbsp;</p>
			<p class="Body-Text ParaOverride-1">In June 2014, a new and improved PCR method (called the AP3 primer method) for detection of AHPND bacteria has been developed from the laboratory test results in screening AHPND bacterial isolates using this AP3 primer method, the team recommended that the previously announced AP1 and AP2 primer methods be replaced with this new method (<span class="Hyperlink">NACA, 2014</span>).</p>
		  <p class="Body-Text ParaOverride-1">&nbsp;</p>
			<p class="Heading-1--Introduction----">Disease prevention and treatment</p>
		  <p class="Heading-1--Introduction----">&nbsp;</p>
			<p class="Body-Text ParaOverride-1">Due to identify the cause of disease and its bacterial agent possibility of authorized types of antibiotics are effective against Gram-negative bacteria are available but some limitation such as antibiotic resistant, public health problems disease  prevention and health management are the best methods for disease control. On the other hand antibiotic usage killed the useful bacteria in the pond that increased disease potency (<a href="#Schryver-P--Defoirdt-T--Sorgeloos-P--2014"><span class="Hyperlink">Schryver, et al., 2014</span></a>). </p>
		  <p class="Body-Text ParaOverride-1">&nbsp;</p>
			<p class="Heading-2--History-in-MM-">Notes in shrimp hatcheries</p>
			
				<li class="Body-Text ParaOverride-5">Pay attention to health, water and feed management for high quality and health shrimp post larvae production as the first way to disease control (<a href="#Afsharnasab-M--Banaederakhshan-R--2014-"><span class="Hyperlink">Afsharnasab et al., 2014</span></a>).</li>
				<li class="Body-Text ParaOverride-5">Continual control of different parts of hatchery and shrimp larvae for detection of vibrio bacteria especially <span class="CharOverride-4">V. parahaemolyticus</span> for decreasing of bacterial load and disease prevention (<a href="#Ministry-of-Agriculture-and-Cooperatives--MAC--in-Thailand--Department-of-Fisheries--2013-."><span class="Hyperlink">MAC, 2013</span></a>).</li>
				<li class="Body-Text ParaOverride-5">Post larvae health control and certified by responsible organs for presence of vibrio and other organisms and their quality control (<a href="#Ministry-of-Agriculture-and-Cooperatives--MAC--in-Thailand--Department-of-Fisheries--2013-."><span class="Hyperlink">MAC, 2013</span></a> and <a href="#Shrimp-news-international--December--24---2013-"><span class="Hyperlink">Shrimp news, 2013</span></a>).</li>
		  <li class="Body-Text ParaOverride-5">Attention to faster growth rate ability and inbreeding, caused disease sensitive production (<a href="#Shrimp-news-international--December--24---2013-"><span class="Hyperlink">Shrimp news, 2013</span></a>).</li>
				<li class="Body-Text ParaOverride-5">Also eye stalk oblation in hatcheries increase the product quantity but decrease the quality and disease tolerance (<a href="#Shrimp-news-international--December--24---2013-"><span class="Hyperlink">Shrimp news, 2013</span></a>).  </li>
			<p class="Heading-2--History-in-MM-">&nbsp;</p>
			<p class="Heading-2--History-in-MM-">Notes in shrimp farms</p>
			
			  <li class="Body-Text ParaOverride-5">Full compliance with the principles of pond preparation (washing, spraying lime, plowing, etc.) (<a href="#Afsharnasab-M--Banaederakhshan-R--2014-"><span class="Hyperlink">Afsharnasab et al., 2014</span></a> and <a href="#Ministry-of-Agriculture-and-Cooperatives--MAC--in-Thailand--Department-of-Fisheries--2013-."><span class="Hyperlink">MAC, 2013</span></a>).</li>
				<li class="Body-Text ParaOverride-5">Due to the increasing incidence of disease in high pH water, decreasing or pH control can be considered as a strategy for disease prevention. However, in this case, hydrogen sulfide toxicity possibility increases (<a href="#Afsharnasab-M--Banaederakhshan-R--2014-"><span class="Hyperlink">Afsharnasab et al., 2014</span></a> and <a href="#Shrimp-news-international--December--24---2013-"><span class="Hyperlink">Shrimp news, 2013</span></a>).</li>
				<li class="Body-Text ParaOverride-5">The use of probiotic containing Bacillus in the soil during the pond preparation and during the culture period for the acidic environment can useful for disease control (<a href="#Ministry-of-Agriculture-and-Cooperatives--MAC--in-Thailand--Department-of-Fisheries--2013-."><span class="Hyperlink">MAC, 2013</span></a>).</li>
				<li class="Body-Text ParaOverride-5">Stocking of healthy and high quality post larvae and preparing them from good and authentic hatcheries (<a href="#Afsharnasab-M--Banaederakhshan-R--2014-"><span class="Hyperlink">Afsharnasab et al., 2014</span></a> and <a href="#Shrimp-news-international--December--24---2013-"><span class="Hyperlink">Shrimp news, 2013</span></a>). </li>
				<li class="Body-Text ParaOverride-5">Avoiding from high stocking density for stress decreasing and diseases prevention (<a href="#Afsharnasab-M--Banaederakhshan-R--2014-"><span class="Hyperlink">Afsharnasab et al., 2014</span></a>).</li>
				<li class="Body-Text ParaOverride-5">Continuous soil and water sampling and testing for detection of infectious Vibrio (<a href="#Ministry-of-Agriculture-and-Cooperatives--MAC--in-Thailand--Department-of-Fisheries--2013-."><span class="Hyperlink">MAC, 2013</span></a>).</li>
				<li class="Body-Text ParaOverride-5">Use from high quality and safe food and immune stimulant material for improving shrimp immunity (<a href="#Afsharnasab-M--Banaederakhshan-R--2014-"><span class="Hyperlink">Afsharnasab et al., 2014</span></a>).</li>
				<li class="Body-Text ParaOverride-5">Reduction of environmental stress with optimum water and feeding management (<a href="#Ministry-of-Agriculture-and-Cooperatives--MAC--in-Thailand--Department-of-Fisheries--2013-."><span class="Hyperlink">MAC, 2013</span></a> and <a href="#Shrimp-news-international--December--24---2013-"><span class="Hyperlink">Shrimp news, 2013</span></a>).</li>
				<li class="Body-Text ParaOverride-5">Use of water storage ponds (reservoir) (<a href="#Muhammad-F--Zhang-ZF--Shao-MY--Shi-XL--Shafi-M--2012-"><span class="Hyperlink">Muhammad, et.al, 2012</span></a>).</li>
				<li class="Body-Text ParaOverride-5">Reducing and keeping water salinity (based on observations of low incidence in low salinity) (<a href="#Afsharnasab-M--Banaederakhshan-R--2014-"><span class="Hyperlink">Afsharnasab et al., 2014</span></a>).</li>
				<li class="Body-Text ParaOverride-5">Use from aeration in ponds and prevention from oxygen drop and stress (<a href="#Ministry-of-Agriculture-and-Cooperatives--MAC--in-Thailand--Department-of-Fisheries--2013-."><span class="Hyperlink">MAC, 2013</span></a>).</li>
				<li class="Body-Text ParaOverride-5">Reporting to responsible institutions in case of mortality during first 45 days after stocking for dead shrimp sampling and testing (<a href="#Ministry-of-Agriculture-and-Cooperatives--MAC--in-Thailand--Department-of-Fisheries--2013-."><span class="Hyperlink">MAC, 2013</span></a>).</li>
				<li class="Body-Text ParaOverride-5">In case of disease water must disinfect with high dose chlorine and dead shrimp must keep in 60<span class="CharOverride-2">o</span>C<span class="CharOverride-2"> </span>for minimum 15 minutes after collection (<a href="#Ministry-of-Agriculture-and-Cooperatives--MAC--in-Thailand--Department-of-Fisheries--2013-."><span class="Hyperlink">MAC, 2013</span></a>).</li>
				<li class="Body-Text ParaOverride-5">Utilizing the Biofloc techniques significantly increased the survival rate of infected farms (<a href="#FAO-Fisheries-and-Aquaculture-Report--2013-"><span class="Hyperlink">FAO, 2013</span></a> and <a href="#Seafood-Source-electronic-Daily-News--2013-."><span class="Hyperlink">Seafood Source News, 2013</span></a>)   </li>
				<li class="Body-Text ParaOverride-5">Using the poly culture system(shrimp and filter feeding marine fish) prevent from disease and\or increase the survival rate in affected system (<a href="#Tran-LH--Fitzsimmons-KM--Lightner-DV--2014-."><span class="Hyperlink">Tran et al., 2014</span></a>) for some reasons as follow:</li>
			
		  <ol>
				<li class="Body-Text ParaOverride-3">Zooplanktons use from the bacteria as feed and decrease the bacterial load in water and fish by zooplanktons feeding decrease the bacterial total count indirectly</li>
				<li class="Body-Text ParaOverride-3">Blue-green algae population decrease due to fish feed and after that decrease the bacterial population</li>
				<li class="Body-Text ParaOverride-3">The antibacterial effect of fish mucosa decrease the density of bacteria in the ponds (<a href="#FAO-Fisheries-and-Aquaculture-Report--2013-"><span class="Hyperlink">FAO, 2013</span></a>).</li>
			</ol>
			<p class="Heading-1--Introduction----">&nbsp;</p>
		  <p class="Heading-1--Introduction----">Last status of EMS\AHPND in IR. Iran</p>
			<p class="Body-Text ParaOverride-1">&nbsp;</p>
			<p class="Body-Text ParaOverride-1"><span class="CharOverride-4">V. parahaemolyticus</span> is one of the natural bacterial floras in Iran’s costal area. Fortunately, there isn’t any official report about occurrence or distribution of mentioned disease in Iran. There is some similarity between Iran and affected area such as: same species, shrimp culture complex are near the sea, high water salinity, Inaccessibility to fresh water, high pH and temperature, incomplete pond preparation and increases the disease incidence. For these reasons government association must increase their activity especially on live animals import and transporting (quarantine policy, health certificate and biosecurity operations) (<a href="#Afsharnasab-M--Banaederakhshan-R--2014-"><span class="Hyperlink">Afsharnasab et al., 2014</span></a>). In comparison Indonesia could be considered as free of this disease due to live animal import limitation (Lightner, unpublished data).</p>
		  <p class="Body-Text ParaOverride-1">&nbsp;</p>
			<p class="Heading-1--Introduction----">Conclusion</p>
		  <p class="Heading-1--Introduction----">&nbsp;</p>
			<p class="Body-Text ParaOverride-1">(Early mortality syndrome) or (Acute Hepatopancreatic Necrosis Disease), (EMS/AHPND) is a new emerging shrimp disease that affected several countries in eastern and western hemisphere such as China, Vietnam, Malaysia, Thailand and Mexico from 2009. Economic losses of this disease were estimated more than 1bilion US$ in this countries and they lost about 80% of their products in some regions.  As regards, this is a new emerging disease so initial investigation had been started and focused on recognition of causative pathogen. Finally, Dr. Lightner identified it as a unique strain of <span class="CharOverride-4">V. parahaemolyticus</span> after affected by bacteriophage virus that can be produced a kind of toxin that makes this disease. They found the causative pathogen that can be transmitted to sensitive hosts by digestive system and located in the gut and then produced the toxin that causes necrosis and degeneration especially in haepathopancreas and death was happen due to this function . This disease had been reported from <span class="CharOverride-4">P.monodon</span>, <span class="CharOverride-4">L.vannamei</span> and <span class="CharOverride-4">P. chinensis</span> in Southeast Asia and Mexico.</p>
		  <p class="Body-Text ParaOverride-1">&nbsp;</p>
			<p class="Body-Text ParaOverride-1">According to current study Hepatopancreas should be considered as the target tissue in this disease. This disease specified with mass mortality (up to 100%) 20-30 days after stocking. Clinical signs consist of slow growth, corkscrew swimming, loose shells, pale coloration, empty or interrupted gut. Affected shrimps also consistently show an abnormal hepatopancreas (shrunken, small, swollen, discoloration or black coloration) (<a href="#Lightner-DV--Redman-CR--Pantoja-BL--Noble-LM--Nunan--Loc-Tran--2013-"><span class="Hyperlink">Lightner et al., 2013</span></a>). Affected shrimp sank to the pond bottom and die there. Affected hepatopancreas doesn’t squash easily by finger (<a href="#Lightner-DV--2012-"><span class="Hyperlink">Lightner, 2012</span></a>).<span class="CharOverride-12"> </span>Presence of some microbes including Vibrio spp, microsporidians and nematode has been observed in some samples (<a href="#Eduardo-M-Lea-o--Mohan-CV--2012-."><span class="Hyperlink">Eduardo et al., 2012</span></a>). The affected shrimps are smaller than healthy one in size.</p>
		  <p class="Body-Text ParaOverride-1">&nbsp;</p>
			<p class="Body-Text ParaOverride-1">Histological examination showed the following lesions: Lack of mitotic activity in generative E cells of the HP, dysfunction of central hepatopancreatic B, F and R cells, karyomegaly and massive sloughing of central HP tubule epithelial cells, massive inter tubular hemocytic aggregation followed by secondary bacterial infections (<a href="#Eduardo-M-Lea-o--Mohan-CV--2012-."><span class="Hyperlink">Eduardo et al., 2012</span></a>), Expanding inefficiency of hepatopacreas due to degeneration and incapacitation. Although clinical signs of the disease can help to initial diagnosis to professionals but histopathological studies are the best and routine diagnosis tests. For rapid diagnosis of this disease several PCR kites were designed and produced till now. These PCR kites contain AP1, AP2 and/or AP3 primer for detection of specific acute gene in the bacterial genome. These techniques are faster, more special and cheaper than others. </p>
		  <p class="Body-Text ParaOverride-1">&nbsp;</p>
			<p class="Body-Text ParaOverride-1">(EMS/AHPND) should be assumed as bacterial disease and it caused by <span class="CharOverride-4">V. parahaemolyticus</span>, and several parameters have positive roll on its happen and spreading such as: high stocking density, species sensibility, high water salinity, lack of reservoir pond and water treatment in grow out pond, lack of attention to the process of ponds preparation, post larvae poor quality, stress problem specially during transportation and stocking, low oxygen condition, high water pH and weak feeding management.</p>
		  <p class="Body-Text ParaOverride-1">&nbsp;</p>
			<p class="Body-Text ParaOverride-1">According to the pathogen property, using from antibiotics is suitable way but some public health problems (drug residual) make it useless. So, prevention from pathogen and reducing the risk factors could be nominated as the best actions (goal). </p>
		  <p class="Body-Text ParaOverride-1">&nbsp;</p>
			<p class="Body-Text ParaOverride-1">Restricting in the transportation of alive shrimp and good quarantine management will reduce the disease transports risk. High qualified pond preparation (drying, washing, spraying lime, plowing, etc.) are basic activities to reduce the disease transfer risk such as EMS. Using of reservoir and treatment pond can reduce the disease incidence by chemical materials and\or time spending. Since the high pH can increase the EMS prevalence, the water pH control and reduce the shrimps gut pH can decrease the disease occurrence. Bacillus content probiotic usage increases the water pH and control the disease as above, meanwhile bacillus concentrate can control the water Vibrio population.</p>
		  <p class="Body-Text ParaOverride-1">&nbsp;</p>
			<p class="Body-Text ParaOverride-1">Aeration system can control the DO concentration in the pond and reduce the shrimp stress and disease incidences. Also, good feed management reduces the bottom sediment and increases their conditions to control of stress and diseases. High quality feed specially immunostimulant enriched feed, can help to disease prevention. Utilizing the Biofloc techniques significantly increased the survival rate of infected farms (<a href="#FAO-Fisheries-and-Aquaculture-Report--2013-"><span class="Hyperlink">FAO, 2013</span></a> and <a href="#Seafood-Source-electronic-Daily-News--2013-."><span class="Hyperlink">Seafood Source News, 2013</span></a>). Alongside above materials for controlling the disease using the poly culture system (shrimp and filter feeding marine fish) could be useful in diseases prevention and\or increase the survival rate in affected system. It is necessary to stocking the pond with high quality and high health shrimp post larvae with authentic health certificate. </p>
		  <p class="Body-Text ParaOverride-1">&nbsp;</p>
			<p class="Body-Text ParaOverride-1">A good relationship between private sectors (farms and hatcheries), governmental council and professional laboratory as vital triangle can help to disease control. In case of occurrence of suspicious mortality it must be reported to any responsible organs as soon as possible and must increase biosecurity actions in the affected farms and quarantine affaires would be done immediately.   </p>
		  <p class="Body-Text ParaOverride-1">&nbsp;</p>
			<p class="Body-Text ParaOverride-1">In case of disease observation in region first of all farms water must be disinfected with high dose chlorine and then dead shrimp must kept in 60oC for minimum 15 minutes after collection (<a href="#Ministry-of-Agriculture-and-Cooperatives--MAC--in-Thailand--Department-of-Fisheries--2013-."><span class="Hyperlink">MAC, 2013</span></a>).</p>
		  <p class="Body-Text ParaOverride-1">&nbsp;</p>
			<p class="Body-Text ParaOverride-1">Fortunately, there isn’t any official report about occurrence or distribution of mentioned disease in I.R. Iran but concerning to identification of <span class="CharOverride-4">V. parahaemolyticus</span> as a natural flora and presence of <span class="CharOverride-4">L.vannamei</span> as main cultivated species in the country, so it is possible to occur of EMS in the future. It seems restriction in alive shrimp importation and control actions have critical roles in disease control. Also pH control in grow out pond (the pH of pond normally are high in Iran), biosecurity activities in farms and hatcheries, stocking with high quality and health post larvae, reduce the stress, more attention to native species, use from high quality feed and immunostimulant enriched feed, good water and feed management, using of reservoir ponds and reinforcement of health supervision and developing of health laboratory can help to disease prevention in I.R. Iran.</p>
		  <p class="Body-Text ParaOverride-1">&nbsp;</p>
			<p class="Heading-1--Introduction----">References</p>
		  <p class="Heading-1--Introduction----">&nbsp;</p>
			
			  <li class="References ParaOverride-6" lang="en-US"><span class="Hyperlink CharOverride-7"><a id="Acute-Hepatopancreatic-Necrosis-Disease-Card--NACA-publication--updated-June-2014-"></a>Acute Hepatopancreatic Necrosis Disease Card, NACA publication (updated June 2014). Disease library, available at: </span><a href="http://www.enaca.org/modules/library/publication.php?tag_id=244&amp;label_type=1&amp;title=disease-library"><span class="Hyperlink">http://www.enaca.org/modules/</span><span class="Hyperlink">library/publication.php?tag_id=244&amp;label_</span><span class="Hyperlink">type=1&amp;title=disease-library</span></a></li>
				<li class="References ParaOverride-6" lang="en-US"><a id="Acute-Hepatopancreatic-Necrosis-Disease-Etiology-Epidemiology-Diagnosis-Prevention-and-Control--OIE"></a>Acute Hepatopancreatic Necrosis Disease Etiology Epidemiology Diagnosis Prevention and Control, OIE Technical Disease Cards (2013). Available <span class="Hyperlink CharOverride-7">at</span>: <a href="http://www.oie.int/fileadmin/Home/eng/Internationa_Standard_Setting/docs/pdf/Aquatic_Commission/AHPND_DEC_2013.pdf "><span class="Hyperlink">http://www.oie.int/fileadmin/Home/eng/Internationa_Standard_Setting/docs/pdf/Aquatic_Commission/AHPND_DEC_2013.pdf</span></a></li>
				<li class="References ParaOverride-6" lang="en-US"><span class="Hyperlink CharOverride-7"><a id="Afsharnasab-M--Banaederakhshan-R--2014-"></a>Afsharnasab M, Banaederakhshan R (2014). Early Mortality Syndrome, a new threat for Iran’s shrimp industry. No.56, Iranian Veterinary Councils Magazine, 2014 (Persian language).</span></li>
				<li class="References ParaOverride-6" lang="en-US"><span class="Hyperlink CharOverride-7"><a id="Asia-Pacific-Emergency-Regional-Consultation-o-2012"></a>Asia Pacific Emergency Regional Consultation on the Emerging Shrimp Disease: Early Mortality Syndrome (EMS) / Acute Hepatopancreatic Necrosis Syndrome (AHPNS), Bangkok, Thailand, 9-10 August 2012.   </span></li>
				<li class="References ParaOverride-6" lang="en-US"><a id="GAA--2013-"></a><span class="Hyperlink CharOverride-7">Culprit behind massive shrimp die-offs in Asia unmasked. FAO News Article, (2013). available at: </span><a href="http://www.fao.org/news/story/en/item/175416/icode/"><span class="Hyperlink">http://www.fao.org/news/story/en/item/175416/icode/</span></a></li>
				<li class="References ParaOverride-6" lang="en-US"><a id="Eduardo-M-Lea-o--Mohan-CV--2012-."></a>Eduardo M Leaño, Mohan CV (2012). Emerging threat in the Asian Shrimp Industry: Early Mortality Syndrome (EMS)/Acute Hepatopancreatic Necrosis Syndrome (AHPNS). Network of Aquaculture Centres in Asia-Pacific. Asian fisheries society. Fish health section. Electronic newsletter. No.10.</li>
				<li class="References ParaOverride-6" lang="en-US"><a id="FAO-Fisheries-and-Aquaculture-Report--2013-"></a><a id="FAO-Fisheries-and-Aquaculture-Report--2013--1"></a>FAO Fisheries and Aquaculture Report (2013). No. 1053. Hanoi, Vietnam, p.126-130.  </li>
				<li class="References ParaOverride-6" lang="en-US"><a id="FAO-statistical-yearbook--2013"></a><a id="FAO-statistical-yearbook--2013--"></a>FAO statistical yearbook (2013), p.146-150.</li>
				<li class="References ParaOverride-6" lang="en-US"><span class="Hyperlink CharOverride-7"><a id="Flegel-TW--2013-."></a><a id="Flegel-TW--2013-"></a>Flegel TW (2013). Center of Excellence for Shrimp Molecular Biology and Biotechnology, Faculty of Science, Mahidol University, available at: </span><a href="http://www.shrimpnews.com/FreeReportsFolder/NewsReportsFolder/ThailandTaiwanFreeEMStests.html"><span class="Hyperlink">http://www.shrimpnews.com/</span><span class="Hyperlink">FreeReportsFolder/NewsReportsFolder/</span><span class="Hyperlink">ThailandTaiwanFreeEMStests.html</span></a></li>
				<li class="References ParaOverride-6" lang="en-US"><a id="GAA--2013-."></a>GAA (2013). Cause of EMS Shrimp Disease Identified, The global aquaculture alliance, available <span class="Hyperlink CharOverride-7">at</span>: <a href="http://www.gaalliance.org/newsroom/news.php?Cause-Of-EMS-Shrimp-Disease-Identified-107"><span class="Hyperlink">http://www.gaalliance.org/newsroom/news.php?Cause-Of-EMS-Shrimp-Disease-Identified-107</span></a></li>
				<li class="References ParaOverride-6" lang="en-US"><a id="Ghavampour--2012-.-A"></a>Ghavampour (2012). A. Epidemiology and risk factors on Early Mortality Syndrome (EMS), available <span class="Hyperlink CharOverride-7">at</span> : <a href="http://abzistan.persianblog.ir/"><span class="Hyperlink">http://abzistan.persianblog.ir/</span></a> (Persian language)</li>
				<li class="References ParaOverride-6" lang="en-US"><a id="Joshi-J--Srisala-J--Truong-VH--Chen-2014"></a>Joshi J, Srisala J, Truong VH, Chen IT, Nuangsaeng B, Suthienkul O, Lo CF, Flegel TW, Sritunyalucksana K, Thitamadee S (2014). Variation in <span class="CharOverride-4">Vibrio parahaemolyticus</span> isolates from a single Thai shrimp farm experiencing an outbreak of acute hepatopancreatic necrosis disease (AHPND). Aquaculture. (428–429): 297-302. <a href="http://dx.doi.org/10.1016/j.aquaculture.2014.03.030"><span class="Hyperlink">http://dx.doi.org/10.1016/j.aquaculture.2014.03.030</span></a></li>
				<li class="References ParaOverride-6" lang="en-US"><a id="Lightner-DV--2012-"></a>Lightner DV (2012). Biology and pathology of early mortality syndrome of shrimp, Global outlook for aquaculture leadership, Bangkok, p.40.</li>
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