Food and Agriculture Organization of the United Nations (FAO) Early Mortality Syndrome (EMS) in Shrimp Melba B. Reantaso Food and Agriculture Organization of the United Nations (FAO) Melba.Reantaso@fao.org and Bruno Gomez-Gil CIAD, A.C. Unidad Mazatlán bruno@ciad.mx I am going to make this presentation on behalf of Dr Melba B. Reantaso of FAO who sends her regrets for not being able to make it to this conference due to urgent tasks related to Haiyan Typhoon (Yolanda) that has severely devastated many parts of the Philippines. She sends her apologies to the organizers and best wishes for a productive conference.

Outline General background on early mortality syndrome/acute hepatopancreatic necrosis syndrome (EMS/AHPNS) Regional and international initiatives Impacts of EMS FAO technical assistance Status of EMS in Mexico Latest findings The Way Forward Read text as is Courtesy of Prof. Lightner

General background on EMS/AHPN Unknown disease called Early Mortality Syndrome (EMS) affecting Penaeus vannamei, P. monodon, and P. chinensis Earliest publication in GAA Jan/Feb 2012: Prof. Donald Lightner (UAZ) reported the first occurrence of this disease in China and Hainan Island in 2010. Role of bacteria was suggested to be secondary; bacterial colonization was prominent at the latter stage of the disease. The role of bacteria Vibrio parahaemolyticus came up on several occasions. Vibrio parahaemolyticus was consistently isolated from EMS-infected shrimp. Based on the work done in China (Zhang et al., 2012), Chinese researchers reported on a virulent strain of V. parahaemolyticus isolated from P. vannamei suffering from this early mortality disease in 2010 in Guangxi Province. Read text as is

General background EMS/AHPN The potential involvement of a phage (a virus infecting bacteria) was highlighted, during the FAO TCP/VIE/3304 First Joint Technical Workshop conducted in Hanoi, Viet Nam from 5–8 November 2012, by Prof. Timothy W. Flegel of Mahidol University (MU). Studies at MU also reported on the isolation of phage from two cases: (i) most probably from V. parahaemolyticus samples from Rayong, Thailand, and (ii) from V. fluvialis (anonymous source). The breakthrough finding of Prof. Lightner’s team pointing to a strain of V. parahaemolyticus as the causative agent of EMS/AHPNS Global Aquaculture Alliance and FAO, May 2012 Diseases of Aquatic Organisms, June 2013 (Tran et al., 2013) Read text as is

EMS/APHND Early Mortality Syndrome (EMS) not an appropriate name Acute Hepatopancreatic Necrosis Disease (APHND) Significant atrophy of the hepatopancreas (HP) Destruction of the HP epithelial cells that round off and detach Massive secondary bacterial infections

EMS en Asia 2011 2009 Año de aparición 2010 2009 2012 2013

Impacts of EMS/AHPNS GAA estimate: Asian losses 1 billion USD Shortage of supply = impact on shrimp prices Public health concerns Private sector Closure of farms (non-stocking) Thai Frozen Foods: annual output to half in 2013 to 300 000 t CP Foods: 70% drop in operating profit (USD 109 M) Thai Union Frozen Products: decline by 30% this year Impacts on trade several countries have suspended or banned the importation of live shrimps and/or all forms of shrimp products Read text as is

Findings of the FAO TCP/VIE/3304 Bacterial aetiology: found by UAZ Mixed infections with WSSV, MBV, HPV, gregarine-like entities, fungi, and other bacterial species BLASTn analysis of these mass sequences gave 99 percent identity to V. proteolyticus and V. parahaemolyticus, respectively. The results suggested that further research should be focused on these two bacterial species. Transmission of EMS/AHPNS occurred via water (16/60 tested shrimp) and cohabitation (27/60 tested shrimp) of healthy shrimp in EMS/AHPNS-infected ponds. Read text as is CMC-AH Mission: July 2011 Emergency TCP Project Formulation Mission: September 2011 TCP/VIE/3304: April 2012- August 2013

Findings of TCP/VIE/3304 Human pathogens: tdh and trh genes (marker genes for human pathogens) not detected Environmental studies: assessment of toxins in feed, sediment, and water Findings did not show an association between environmental conditions (water quality, sediment condition, pollution, or feed ingredients) and EMS/AHPNS-status of shrimp. Read text as is

Findings of TCP/VIE/3304 At the farm level: Risk factors identified during the cross-sectional studies At the farm level: characteristics of AHPND-affected farms larger farm water supply from brackish water semi-closed system factors reducing the risk of AHPND ratio of water reservoir and culture area more than 30 percent holding of water longer than 35 days before use in ponds feed control through use of feeding tray Read text as is

Findings of TCP/VIE/3304 At the pond level: factors increasing the risk of AHPND: larger pond size extreme weather conditions source of post-larvae. factors reducing the risk of AHPND culture of shrimp in ponds deeper than 1.8 meters use of sufficient aeration decreased feeding rates during first month after stocking lower density stocking of PL application of probiotics in the pond during culture period Read text as is

Hanoi EMS Technical Consultation12-point EMS Actions and Risk Management measures “Complacency in the shrimp aquaculture sector resulting to laxity, during a period of trouble-free shrimp production, led to vulnerability of the sector to any newly emerging pathogen that might arise unexpectedly, as is the case of EMS/AHPNS. Poor management practices, weak compliance with standard good biosecurity, and good aquaculture practices still evident. Shrimp aquaculture sector needs to improve and continue to implement responsible and science-based farming practices.” The June 2013 technical consultation came up with a 12-point EMS actions and risk management measures pertaining to various aspects and various actors/players in the shrimp aquaculture sector. The general feeling of the experts was that... Read text as is (Complacency.....)

12-point EMS Actions and Risk Management measures Nomenclature: from EMS to AHPND Diagnosis: rapid and reliable pond-side Reporting/notification: OIE Feb. 2014 International trade: live shrimp, products, live Countries affected by AHPND: proactive reporting Countries not yet affected by AHPND: notification/reporting Read text as is..

12-point EMS Actions and Risk Management measures Farm facilities: good aquaculture practices (GAP) Hatchery facilities: good hatchery practices (GHP) Pharmaceutical and feed companies and shrimp producers: no know treatment Knowledge and capacity development: training Outbreak investigation/emergency response: authorities and public and private sectors. Targeted AHPND research: epidemiology, diagnostics, pathogenicity and virulence, public health, mixed infections, control measures, environment measures, polyculture technologies Read text as is..

Centro de Investigación en Acuicultura y Desarrollo, A.C. Unidad Mazatlán en Acuicultura y Manejo Ambiental México Mortalidades atípicas inician en abril 2013 en Nayarit, quizá ya desde fin de marzo. 1ª semana de abril se reportan en Sinaloa y Sonora. La enfermedad se presenta de manera muy parecida a la reportada recientemente en Asia EMS/APHNS que afecta a P. vannamei y P. monodon (FAO 2013). Descenso de hasta 90% de la producción de camarón: Pérdidas estimadas de 3 mil millones de pesos. Pérdidas aproximadamente 13 mil empleos directos y 67 mil indirectos.

Estados afectados Sonora Sinaloa Nayarit Centro de Investigación en Acuicultura y Desarrollo, A.C. Unidad Mazatlán en Acuicultura y Manejo Ambiental Estados afectados Sonora Sinaloa Nayarit

Sinaloa Juntas de Sanidad Acuícola Ahome Los Mochis Guasave Nte. Guasave sur Angostura Navolato Nte. Culiacán Navolato sur Eldorado Cospita Elota Mazatlán Mazatlán Rosario Escuinapa

Casos reportados en Sinaloa Junta local marzo abril mayo junio julio agosto septiembre octubre 1 2 3 4 Ahome   9 7 12 6 5 Guasave norte 8 13 11 18 Guasave sur Angostura Navolato norte Navolato sur Eldorado 50 24 32 19 26 Cospita 29 Elota Mazatlán Rosario Escuinapa norte sur datos: CESASIN

Porcentaje de granjas afectadas en Sinaloa datos: CESASIN

Correlación porcentaje de granjas afectadas con las hectáreas afectadas por junta local en Sinaloa datos: CESASIN

Peso de camarones al momento del brote en Sinaloa datos: CESASIN

Producción (ton) -70%

Valor (millones pesos)

Identificación del agente causal Centro de Investigación en Acuicultura y Desarrollo, A.C. Unidad Mazatlán en Acuicultura y Manejo Ambiental Identificación del agente causal México

Postulados Koch El microorganismo debe estar presente en todos los organismos enfermos Debe aislarse del organismos enfermo y cultivarlo en forma pura Inocularlo en un organismo sano y reproducir la enfermedad Re aislarlo del organismo enfermo

Postulado 1. Agente causal presente en organismos enfermos Se obtuvieron camarones enfermos de varias granjas Se aislaron mas de 300 cepas de vibrios Se obtuvieron 37 cepas de Vibrio parahaemolyticus 10 fueron PCR+ a EMS (D. Lightner, Univ. Arizona- OIE) De diferentes camarones y de agua

Postulado 2. Cultivado en forma pura Se obtuvieron al menos seis cepas de Vp patógenas a b c d e f

Postulado 3. Causar la enfermedad en organismos sanos Mortalidad causada por tres cepas de Vp en camarones sanos inoculados por inmersión (108 cel-ml) Horas post infección

Postulado 4. Re-aislarlo del organismo enfermo Rep-PCR de aislados de camarones infectados en bioensayo Cam3 HP Agua Cam2 ES Cam2 HP Ctrl M0603

Latest findings/developments (November 2013) PCR Rapid Detection Kit for EMS/AHPND released to GeneReach Biotechnology Corp. on October 17, 2013 by UAZ – may take 2-3 months before marketing Rapid detection Cost effective without the need for histology Testing of much larger quantities of shrimp EMS in India: news reports of suspicion but yet to be confirmed Additional species: news reports of V. owensii association with EMS but yet to be confirmed Read text as is

The Way Forward Diagnostic kit will be available soon (in 2-3 months) Still many things are unknown about EMS/AHPND Risk management measures and research identified (FAO report); in addition (V. Alday): Ecology of vibrios, similarity with cholera Microbial ecosystem studies in shrimp farming areas Exclusion health strategy Extreme caution in the use of antibiotics Monitor presence of pathogen in frozen products More detailed studies on risk factors identified Trans boundary aquatic diseases FAO Inter-regional TCP on EMS (pipeline): LAC and Asia – will provide additional platform to better understand the disease and practical measures to control. Read text as is

gracias Centro de Investigación en Acuicultura y Desarrollo, A.C. Unidad Mazatlán en Acuicultura y Manejo Ambiental EMS/APHND gracias