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This manual gives basic information on some parasites of different infected fish submitted to the Southeast Asian Fisheries Development Center/ Aquaculture Department (SEAFDEC/AQD) Diagnostic Service Laboratory in Tigbauan, Iloilo, Philippines. It is intended as a reference for those working in aquaculture in the Philippines. It is also designed to be a valuable reference for students, researchers, and enthusiasts eager to delve into the captivating field of parasitology and parasitic disease management.

Since the first occurrence of White Spot Disease (WSD) in 1992, it is still listed as one of the crustacean diseases by the World Organisation for Animal Health in 2022. Horizontal transmission in co-culture systems is one of the usual modes in the spread of the disease. WSD outbreak was recorded during the experimental run of the co-culture of black tiger prawn (Penaeus monodon) and sandfish (Holothuria scabra) in the grow-out phase. In this study, artificial infection through two cohabitation experiments were conducted to determine if H. scabra is a potential non-crustacean vector or reservoir of WSSV. Samples were checked using one-step and nested PCR for increased readout sensitivity of virus infection to investigate the horizontal transmission between prawn and sandfish. During the first cohabitation (5 days) where WSSV (+) prawn were cohabited with WSSV (-) sandfish, 100% of the prawn were one-step PCR positive for WSSV while 100% of the sandfish were nested PCR positive. Subsequently, WSSV (+) sandfish from the first cohabitation were transferred to another tank to be cohabited with WSSV (-) prawn. Sampling of both prawn and sandfish was done every 6 days post-infection (dpi). At 6 to 18 dpi, prawn and sandfish were nested PCR positive. At 25 dpi, there were no prawns left due to mortality and 1 of the 3 remaining sandfish was nested PCR positive. Based on the results, it elucidates the ability of sandfish to bioaccumulate the viral particles when cohabited with WSSV (+) prawn. Results suggest that WSSV is viable in the sandfish confirming its potential as a vector or reservoir due to the consistent nested PCR positive results of the prawn during the second cohabitation. Hence, it can be inferred that sandfish can be a potential non-crustacean vector or reservoir of WSSV for a limited period of time.

Growth performance (specific growth rate [SGR] and biomass yield), carrageenan properties (yield, viscosity, and gel strength), and disease and pest incidence were compared between Kappaphycus alvarezii var. tambalang (brown) tissue-cultured (TC) plantlets and farm-sourced (FS) seedlings. Seedlings used in the trials were propagules (4‒19 g pieces) and these were deployed on lines in sea cages over three separate runs of 60 days. The highest growth rate was recorded in TC plantlets and the SGRs were significantly higher (1.5‒6.5x) in TC than FS after Day of Culture (DOC) 15 (i.e., measurements taken every 15 days in each cycle). The carrageenan properties did not differ between treatments, with the exception of viscosity at DOC 45, where TC was higher than FS. “Ice-ice” disease (IID) was observed in both FS and TC in May‒June and August‒October at DOC 45 to 60, coinciding with changes in environmental conditions (i.e., dry‒wet and wet seasons). IID in FS had a moderate to very high incidence (31.2‒86.2%), while TC IID had a lower range of incidence (12.8‒60.6%). On the other hand, epiphytic filamentous algae (EFA) were observed only in August‒October with a low incidence (≤25%). However, EFA appeared earlier in FS than in TC. Taken together, these results highlight that tissue-cultured seaweeds had higher growth performance without compromising carrageenan quality and may be more resistant to health problems than farm-sourced seedlings. Studies should now assess the long-term viability of using tissue-cultured seaweeds over farm-sourced seedlings scaling to allow testing in standard farm settings, including the size of the initial seed.

We announce the complete genome sequence of Vibrio parahaemolyticus strain PH1273. This strain was collected from a Penaeus vannamei pond in the Philippines in 2015. Genome analysis revealed that it lacks the gene pirAB responsible for causing acute hepatopancreatic necrosis disease but encode multiple secretion systems and the associated effectors.

The growth efficiency, productivity and economic viability of Indian white shrimp (Penaeus indicus) fed with diet containing 40% (CPhigh) and 34% (CPlow) crude protein (CP) levels were determined in a 90-day grow-out culture trial. Growth parameters including average body weight (ABW), specific growth rate (SGR), survival and feed conversion ratio (FCR) were not statistically different. CPlow had lower overall shrimp production (797.53 kg/ha, DOC 90), but was not significantly different from CPhigh (843.78 kg/ha, DOC 90). The mean values for the water parameters of both treatments, such as DO, pH, salinity, temperature, TAN, NO2-N, NO3-N and PO4-P, also showed no significant differences. However, the TAN values of CPhigh were consistently higher from day of culture (DOC) 63 and increased significantly (p = .048) at DOC 90. Furthermore, the total bacterial and Vibrio counts in both treatments were not significant throughout the culture period. The economic analysis revealed that CPlow obtained higher net profits (6093 USD/ha/year), return on investment (72%) and discounted benefit–cost ratio (1.73). The current study found that feeding a low protein diet to P. indicus is economically viable and profitable and results in improved growth efficiency and productivity while also improving water quality and lowering nitrogen discharge.

Long before the COVID-19 pandemic, the Broodstock Facility and Shrimp Hatchery Complex of SEAFDEC Aquaculture Department (AQD) in Tigbauan, Iloilo, Philippines, has already been practicing the best quarantine protocols. The gold standards to ensure the production of diseasefree and high-quality shrimp are being developed by AQD under the program “OPLAN Balik Sugpo” or Operation Plan for Black Tiger Prawn Revival. Initiated in 2017, this Program generally aims to bring back the Penaeus monodon industry of the Philippines and help farmers revive their hopes and venture again into shrimp culture. Under the Program, the disease prevention scheme is currently undergoing verification for responsible management of shrimp broodstock obtained from the natural environment at AQD’s Broodstock Facility, and for the care of the postlarval stage at AQD’s Shrimp Hatchery Complex.

The ability of bacteria to metabolize a wide variety of carbon sources has been known to aid in their ability for efficient colonization. Vibrio parahaemolyticus, a known aquatic pathogen has been reported to have the ability to metabolize a number of carbohydrates including D-glucose, D-galactose, L-arabinose, D-mannose, and D-ribose to name a few. Classical isolation of V. parahaemolyticus from other members of the family Vibrionaceae relies on its carbon utilization pattern. Conventionally, V. parahaemolyticus lacks the ability to utilize sucrose and this has been the basis for its isolation using the Thiosulfate-citrate-bile salts-sucrose (TCBS) agar. Reports of V. parahaemolyticus having the ability to utilize sucrose have been presented yet there is paucity of information and detailed study on this phenotype. In this study, we report the V. parahaemolyticus strain PH05 that has the ability to metabolize sucrose. Phenotypic and genotypic characterization of this V. parahaemolyticus strain isolated from Negros Island, Philippines, revealed that V. parahaemolyticus strain PH05 is atypical appearing yellow on TCBS agar plates. It is capable of utilizing sucrose, unlike the majority of V. parahaemolyticus isolates. Genome analyses of this strain revealed the presence of a chromosomally encoded sucrose operon-like gene cluster encoded in chromosome 2 with the following sucrose-utilization associated genes: scrY, ccpA, treP, scrK, and scrB genes coding for sucrose porin, catabolite control protein A, PTS System sucrose-specific EIIBC component, fructokinase, and sucrose-6-phosphate hydrolase. The mode of transmission of these genes to V. parahaemolyticus strain PH05 is still unknown. However, the presence of insertion sequences (IS) and phage elements in the same chromosome suggests horizontal gene transfer events. Taken together, our results point to the possibility that acquired sucrose utilization genes may contribute to the fitness of V. parahaemolyticus strain P

As aquaculture in Southeast Asia rapidly grows, it is being confronted with incidences of aquatic animal diseases threatening its sustainability. In early 2013, the SEAFDEC Member Countries raised concerns regarding the outbreaks of acute hepatopancreatic necrosis disease (AHPND), one of the causative agents of early mortality syndrome (EMS), that affected the shrimp industry of the region, specifically in Viet Nam, Thailand, Malaysia, and the Philippines that led to low production and significant economic losses. In addressing such concerns, the SEAFDEC Council of Directors during its Meeting in 2014, suggested that health management, particularly the control and prevention of transboundary aquatic animal diseases, should be included in the future programs of the Aquaculture Department (AQD) of the Southeast Asian Fisheries Development Center (SEAFDEC). The pressing situation on AHPND in cultured shrimp in the region prompted SEAFDEC/AQD to seek financial support to fund a consultative meeting that would assess the occurrence of EMS/AHPND and other emerging diseases in farmed shrimps in the ASEAN Member States (AMSs). In response, the Bureau of Fisheries and Aquatic Resources (BFAR) of the Philippine Department of Agriculture (DA) collaborated with SEAFDEC/AQD and agreed to co-host the said consultative meeting, while the Government of Japan committed to fund the meeting through the Japan-ASEAN Integration Fund (JAIF). Thus, the Regional Technical Consultation (RTC) on EMS/AHPND and Other Transboundary Diseases for Improved Aquatic Animal Health in Southeast Asia was organized on 22-24 February 2016 in Makati City, Philippines. While assessing the status of EMS/AHPND and other emerging diseases in farmed shrimps, the RTC identified the gaps and priority areas for R&D and potential collaboration, and formulated the regional policy recommendations that centered on emergency preparedness and response systems (EPRS) through early warning, detection and response, for ef

Myanmar is one of the OIE members and the Department of Fisheries (DoF) is highly concerned with transboundary aquatic animal pathogens. Therefore, the Aquatic Animal Health & Disease Control Section has already been formed not only for field diagnostic surveys but also for border control especially at international airport and border trade areas by checking and counter checking export and import of aquatic animals and products. At the moment, the DoF is stressing an issue of some transboundary diseases for finfish such as Gyrodactylus sp., Dactylogyrus sp., Argulus sp., Trichodena sp., Streptococcus sp., Aeromonas sp., and for crustacean are MrNV/XSV and WSSV. In addition, the DoF is facing challenges with parasitic disease and bacterial disease problems due to poor water quality management at culturing fish ponds. For the prevention and control of fish diseases within the country, the DoF is issuing Health Certificates by physical and microbiological examination of fishes and fisheries products. At the same time, Quarterly report on fish disease has being regularly submitted to NACA, OIE since 1998 until now. Although the DoF has no specific law and legislation on the control of quarantine pest and disease of aquatic animal, a good aquaculture practice has been implemented and code of conduct responsible for aquaculture is being followed in the country. The aquatic health management is a challenging issue in aquaculture development. Myanmar is still needing technical assistance to improve quarantine system especially for importation and exportation of live aquatic animals. Moreover, monitoring and surveillance programs with harmonized aquatic emergency preparedness and response system are required to boost up not only for Myanmar but also for effective management of transboundary disease outbreaks in Southeast Asia.

Emergency preparedness is the ability to respond effectively and in a timely fashion to aquatic animal disease emergencies (e.g. disease outbreaks, mass mortalities). It is a key element of a National Strategy on Aquatic Animal Health and an important consideration of the Progressive Management Pathway for improving Aquaculture Biosecurity. The important principles, requirements and elements and components of emergency prepareness and contingency plans are briefly described. The emergency preparedness response system audit is also presented as contingency planning arrangements that can provide useful insights and guidance in improving response action to disease emergencies. The paper concludes that many important lessons and insights learned from dealing with disease epizootics in the early 2000 remains valid after more than two decades when the aquaculture sector continues to be plagued with emerging diseases. Past lessons and more recent experiences demonstrated the value of rapid response, reporting/notification by competent authorities, continuous development of knowledge base and capacities in diagnostics, epidemiology, risk analysis, advanced financial planning and the important roles of governments and producer sectors in co-managing disease outbreak events as they both remain the critical entities responsible for launching rapid response. Skills and knowledge need to be passed on to locals as they are in the frontline of any disease emergency. Share key lessons from experiences by state and non-state actors (producer and academic sectors and other important players in the value chain), the international players that launch emergency responses, disease investigations and field situation assessments as well as financial entities that support these actions need to be continued. However, we also need to do - a stock taking exercise to evaluate what worked, what did not work, what resources are needed and to understand what are the new drivers for aquatic animal

Aquaculture is one of the important sectors in the economy of most Asia-Pacific countries. However, majority of aquaculture farms are small-scale and most often lack the necessary facilities to comply with or are not well informed of the product standards imposed by concerned authorities, especially for international trade. Most countries in the region have a high reliance on aquatic animals as the major source of protein for their populations. In the past 20 years, farming of shrimp and fish for export has become a major employer and revenue earner for many countries in the region. Aquaculture is a major employer, contributes significantly to national economies, assists in poverty reduction, and is an important element in food security and other national development priorities. Aquaculture has developed rapidly in the region and is now a significant component in the national economies of many countries. However, recent disease events in fish and shrimp farming have indicated that preparedness and response measures are lacking, contributing to spread of disease across large areas of the countries involved. The growth of aquaculture in recent decades has been dependent on the international movement of aquatic animals and, in particular, the introduction of non-native species. The movement of live aquatic animals and their products has the potential to spread pathogens from one country or region to another, which may result to disease outbreaks. In shrimps as example, most major disease outbreaks were associated with the movement of live animals (broodstock, nauplii and postlarvae) when the patterns of disease spread were analyzed. Many aquatic animal diseases, once established, are often difficult to treat or to eliminate. Over the past 30 years, the Asia-Pacific region has been swept by a number of devastating diseases of aquatic animals which have caused massive economic and social losses. These include spread and outbreaks of infection with Aphanomyces invadans (E

In Thailand, Department of Fisheries (DoF) is the competent authority for various aspects of aquatic animals including aquatic animal health. There are two principal legislation giving DoF power to apply for aquatic animal disease prevention and control measures in the country as well as import-export control; namely, Royal Ordinance on Fisheries and Animal Epidemic Act. DoF has two national reference laboratories for aquatic animal health, one is Aquatic Animal Health Research and Development Division (AAHRDD) for freshwater aquatic animal disease diagnosis and another is Songkhla Aquatic Animal Health Research Center (SAAHRC) for brackish water aquatic animal disease diagnosis. Both are ISO/IEC 17025 accredited laboratories. Besides, there are 19 regional laboratories of DoF located in different areas of the country. All of 21 laboratories are responsible for performing disease diagnosis service for fish farmers as well as for disease surveillance. There are a number of surveillance and control programs for aquatic animal diseases in Thailand. Passive surveillance: information can be collected from disease reporting and other sources such as scientific research, news, publications, social network, or rumor. Active surveillance: to provide assurance of disease status for trade purposes, DoF has setup nationally active targeted surveillance program for demonstrating a number of diseases free status of country or farm establishment in accordance with OIE Aquatic Animal Health Code. There are several farm standards in Thailand such as Good Aquaculture Practices for Hatchery of Disease Free Pacific White Shrimp, Good Aquaculture Practices for Marine Shrimp Farm, and Aquaculture Establishment for Export of Aquatic Animals. Each standard includes necessary biosecurity practices in order to prevent the introduction of pathogenic agent into or spread within or release from the farm. To control domestic movement of live aquatic animals, in normal situation, Aquatic Animal M

The regulatory authority responsible for the control of aquatic animal diseases in Japan is the Animal Products Safety Division, Food Safety and Consumer Affairs Bureau, Ministry of Agriculture, Forestry, and Fisheries (MAFF). The ministry (Animal Products Safety Division) specifies certain diseases and their host species that are subjects for import quarantine on the basis of the law called Fisheries Resources Protection Act. The MAFF also implements risk control measures for the same diseases within Japan on the basis of another law, Fish Farming Production Maintaining Act. Currently, 24 such diseases are listed. For disease control within Japan, the MAFF issues Guideline for the Control of Aquatic Animal Diseases, which states the roles of different stakeholders, appropriate actions that are to be taken on the occurrences of specified or other diseases, fish health guidelines for fish farmers, or diagnostic methods for specified diseases, etc. Local prefectural governments in Japan are required to place personnel who work on fish health issues at the prefectural fisheries research laboratories. These people usually inspect fish farms, observe cultured aquatic animals, supervise the use of antibiotics or vaccines, and guide fish farmers for disease control. Disease diagnosis for aquatic animals is usually conducted by these local fisheries research labs for free. The Japan Fisheries Resource Conservation Association provides a comprehensive training course on fish diseases including laws or hands-on trainings for the staff of prefectural fisheries research laboratories. The JFRCA also give local fish health personnel the qualification as the fish health expert, if the person passes the examination conducted after the training course. Primary diagnosis for specified diseases is conducted by local fisheries laboratories. On the occurrence of the diseases that are suspected to be one of the specified diseases or OIE listed diseases that have not been reported in Japa

The Bureau of Fisheries and Aquatic Resources (BFAR) of the Department of Agriculture as the Competent Authority, develops and implements rules and regulations on aquatic animal health for the Philippines. It establishes the monitoring system for OIE/NACA listed aquatic animal diseases. The disease surveillance and reporting activities are being carried out by the BFAR Fish Health Laboratory of the National Fisheries Laboratory Division and its counterparts at the regional offices. BFAR Fish Health Laboratories have different levels of diagnostic and detection capabilities for aquatic animal diseases. Diagnostic services and technical assistance are rendered to farmers on aquatic animal health. Results of diagnostic services and surveillance by BFAR central and regional offices, and other laboratories (SEAFDEC/AQD-Fish Health, DA-Biotech, Negros Prawn Cooperative) are part of the country s aquatic animal disease reports to the OIE/NACA. BFAR has a Fish Health Network that responds to aquatic animal disease emergencies. It also coordinates and collaborates through networking with research agencies, academe, private sectors and other stakeholders on aquatic animal health. The Fisheries Inspection and Quarantine Division implements the policies on biosecurity, quarantine and health certification for trade and transboundary movement of aquatic animals. It is also responsible for risk analysis on the importation of fish and fishery/aquatic products. Other regulatory requirements for in-country movement include local transport permit for fish and fishery/aquatic products for traceability. Importers and exporters are also registered by BFAR to ensure compliance to sanitary and food safety measures and requirements. BFAR is continuously strengthening its technical capacity, human resources, policies and regulations for a more efficient implementation of aquatic animal health services that includes response to transboundary disease emergencies of aquatic animals.

The overall objective of this technical consultation is to bring together the representatives of ASEAN Member States and technical experts to examine the status of aquatic emergency preparedness and response systems currently being practiced in the region in order to identify gaps and other initiatives for regional cooperation. In the general sense, the RTC is successful in achieving the general objective. As for the specific objectives, (a) to assess existing laws, legislations and standard operating procedures (SOPs), among others had been partially achieved. This is because the consultation didn t assess but was only informed (through the reports of country representatives) of the current situation in ASEAN member countries. The way forward of this is to complete the EPRS audit questionnaire as basis of the more systematic assessment. The second objective is (b) to assess the need for a regional aquatic EPRS in the ASEAN, the participant voted in the affirmative. The way forward of this is to create the ASEAN guidelines including the mechanics. The third objective is to (c) enhance cooperation among Member States, regional/international organizations and other relevant stakeholders on initiatives that support aquatic EPRS for effective management of aquatic animal disease outbreaks. This objective has been achieved. The way forward for this is to get the same people for a planned and proposed consultation II for continuity and for emphasis on more private sector and academe representation.

Singapore s population-dense, urban environment presents a unique context for her increasingly important aquaculture industry. This paper provides an overview of Singapore s existing aquatic emergency preparedness and response systems, which have been constructed and refined by the Agri-Food and Veterinary Authority (AVA) in view of past experience with detections of pathogens of warmwater fish. These systems have been developed to fulfil Singapore s obligations as an OIE member country and AVA s duty to safeguard food security, animal and public health. As a trade and export hub, it is critical for Singapore to have timely detection and reporting of diseases which can have an impact on trade. Singapore also needs to balance the needs and perceptions of the multiple stakeholders using the limited space and resources in our island state. Finally, this paper outlines the current issues and gaps of Singapore s existing aquatic emergency preparedness and response systems.

The purpose of this survey is to obtain information on national capacity and the agencies mandated to implement emergency preparedness and response systems with respect to aquatic animal diseases. The results of this survey will help guide regional and national strategic planning with respect to improving aquatic EPR systems, thereby improving aquatic animal health more broadly and assuring adequate and rational support services to achieve sustainable aquaculture development. This FAO questionnaire on aquatic EPR system capacity and performance is a country level self-assessment survey with four sections: (1) general administration, (2) operational components, (3) support systems and (4) additional information.

Outbreaks of acute hepatopancreatic necrosis disease (AHPND) have caused great economic losses to many shrimp producing countries in Asia since its first appearance in 2009. The causative agent was first reported in 2013 as specific isolates of Vibrio parahaemolyticus (VPAHPND) that were later found to harbor a plasmid (pVA) encoding the Pir-like binary toxin genes PirvpA and PirvpB. More recent information indicates that pVA plasmid and variants occur in many Vibrio parahaemolyticus serotypes and also in other Vibrio species such as V. campbellii, V. harveyi and V. owensii. Information on such genomic and proteomic studies of different VPAHPND isolates from different countries are reviewed. A cohort study carried out in Thailand in 2014 indicated that AHPND outbreaks account for only a portion of the disease outbreaks reported by shrimp farmers as outbreaks of early mortality syndrome (EMS). It is urgent that the etiology of the other EMS-associated mortalities be investigated and not be overlooked. It is recommended that a regional research network and surveillance program for newly-emerging or re-emerging pathogens be established to speed up the process of diagnosis and the implementation of coordinated control measures and to avoid a repeat of the EMS/AHPND scenario.

The World Organisation for Animal Health (OIE) is the intergovernmental organisation responsible for improving animal health worldwide. It is recognised as a reference organisation by the World Trade Organization (WTO) to develop international standards for animal health and zoonoses; as of May 2018, it counts a total of 182 Members. As the global leader for animal health and welfare standards, the OIE plays an influential role in the prevention, control and information sharing of animal diseases including aquatic animal diseases. The objectives of OIE are to: (1) Ensure transparency in the global animal disease situation; (2) Collect, analyse and disseminate veterinary scientific information; (3) Encourage international solidarity in the control of animal diseases; (4) Safeguard World trade by publishing health standards for international, trade in animals and animal products; (5) Improve the legal framework and resources, national veterinary services and aquatic animal health services; and (6) to provide a better guarantee of food of animal origin and to promote animal welfare. As an international standard setting organisation, the OIE Aquatic Animal Health Code (the Aquatic Code) provides standards for the improvement of aquatic animal health worldwide. It also includes standards for the welfare of farmed fish and use of antimicrobial agents in aquatic animals. The sanitary measures in the Aquatic Code provide international standards on importing and exporting countries for early detection, reporting and control of pathogenic agents in aquatic animals (amphibians, crustaceans, fish and molluscs) and to prevent their spread via international trade in aquatic animals and their products, while avoiding unjustified sanitary barriers to trade. In addition, to provide a standardised approach to the diagnosis of the diseases listed in the Aquatic Code and to facilitate health certification for trade in aquatic animals and aquatic animal products, the OIE also developed

The Department of Fisheries (DoF) Malaysia is the custodian of the Fisheries Act 1985, which serves as the main legislative source for subsidiary regulations, including aquaculture and fish health management. It has established Emergency Disease Task Force Committee for any emergency related to disease outbreak as well as standard operating procedures for massive fish kill. This committee consists of taskforce teams from federal and/or state fisheries and oversee the operations of the task force. Fisheries Biosecurity Division under DoF Malaysia holds the primary responsibility for managing the country s emergency preparedness and response system for aquatic animal diseases. As for early detection system, Fisheries Biosecurity Division has established official control and official analysis for targeted diseases listed under OIE and National Listed Diseases. Fish health monitoring programmes are conducted every six months and samples are analyzed by accredited laboratories. Quarterly and half year reports are submitted to representative offices for the health status of targeted disease. Apart from the targeted fish health monitoring program, epidemiology on common and emerging diseases are conducted by National Fish Health Research Division (NaFisH) which is the only research and development arm under DoF. Laboratories under Fisheries Biosecurity Division are responsible for organizing and coordinating surveillance programs for diseases in the OIE list while NaFisH is responsible for conducting research and development on aquatic diseases that cause high losses in industry since 2002. Currently, the DoF has four servicing laboratories under Fisheries Biosecurity Division and one NaFisH laboratory under Fisheries Research Institute for fish health diagnosis in Malaysia.

Tilapia lake virus (TiLV) is a novel RNA virus resembling Orthomyxovirus. It has been recently re-classified to Tilapia tilapinevirus species, under Tilapinevirus genus, Amnoonviridae family (ICTV, 2018). Since the first discovery in Israel in 2014, so far TiLV has been reported from 14 countries in three continents (Asia, Africa, and South America). Thailand is one of the affected countries that reported emergence of this virus in 2017. Initially, we employed nested RT-PCR primer sequences previously published for TiLV diagnosis. However, the resulting amplification of nonspecific fish genes led us to modify the nested RT-PCR protocols into a semi-nested RT-PCR by omitting a non-specific primer to avoid false positive results. Subsequently, our molecular work together with histopathology and sequence analysis confirmed the presence of TiLV infection in Thailand. Prior to the publication of our manuscript, we informed the Thai Department of Fisheries of our discovery of TiLV in Thailand. Our publication was preceded by a brief article at the website of the Network of Aquaculture Centers in Asia-Pacific in which we warned of the spread of TiLV and offered free use of a newly improved, semi-nested RT-PCR method and positive control plasmid for detection of TiLV. To date, we have provided positive controls in response to 44 requests from 24 countries who have expressed their appreciation for our attempt to help in emergent controlling the spread of this fish pathogen. Our current study focuses on genetic diversity of TiLV and development of detection method that covers all genetic variants.

Viet Nam is one of the top worldwide producers of aquaculture products which accounts for about 22 percent of total agricultural GDP of Viet Nam. Recently, diseases have become the biggest challenge for global aquaculture development therefore the Vietnamese government has paid close attention to develop an effective aquatic emergency preparedness and response system to timely deal with disease introduction and outbreaks. The Department of Animal Health (DAH), under the Ministry of Agriculture and Rural Development (MARD), which is the competent authority of aquatic animal health management. To monitor transboundary diseases (especially the OIE-listed diseases), the current Vietnamese regulations only allow import of aquatic animals and its products which are certified as disease-free by competent authority of exporting country, and export aquatic animals and its products complying with importing conditions of importing country. Regional Animal Health Offices (belong to DAH) shall carry out sampling for testing pathogens and isolation for imported aquatic animals and its products as regulated in Circular 26/2016/TT-BNNPTNT dated 30 June 2016 before granting permit to import or export. For domestic transportation of aquatic animals, provincial sub DAH is responsible for monitoring infectious pathogens to certify disease-free status of aquatic animals before issuing health certificate for movement. In addition, a reporting and response system to aquatic animal diseases was established in the country from farm level to central level (DAH). Early detection and warning of diseases is critical for disease prevention and control, thus since 2014, the DAH has implemented national surveillance programs focusing on dangerous diseases in the key farming species (brackish-water shrimps, pangasius catfish) according to Circular 04/2016/TT-BNNPTNT dated 10 May 2016 of MARD and support exportation of aquatic animals and its products complying with international regulations and imp

The Competent Authority (CA) on aquatic animal health in Indonesia is the Directorate Aquaculture Area Development and Fish Health of the Directorate General of Aquaculture (DGA) under the Ministry of Marine Affairs. CA is supported by national reference laboratories and professional human resources that are capable of fish disease diagnosis; as well as an established network with the farmers, trading association, and relevant stakeholders which are actively involved in national meetings, conferences, socialization of emerging diseases and policy and regulation. To control transboundary fish disease at national level, the government of Indonesia has a National Strategy on Aquatic Animal Health and Environment, which was developed by FAO under project of TCP/INS/3402: 2013-2015) collaboration with DGA, Ministry of Marine Affairs and Fisheries (MMAF). To strenghten the implementation of aquaculture in the country, the Indonesian Government issued President regulation Number 28 in 2017 which includes the implementation of fish health management as well as emergency response (Section VI, Article 60). To support the implementation of EPRS, Special Task Force Teams are formed by the Director General of Aquaculture. Stakeholders' participation is very important in the implementation of EPRS, such as a prompt report by fish farmers and extension officer to the upper level fisheries officers at district, provincial, and national level of any observed unusual mortality that indicates disease outbreaks. EPRS activities consist of emergency response on early warning (disease information, disease preventing guideline and regulation); early detection (surveillance, appointed diagnostic laboratory); and early response (collecting information, task force formation, public awareness). Standard Operational Procedures, and detection and control were done based on published scientific information available and guidelines from World Organisation for Animal Health (OIE), Network of Aquac

Effective monitoring of hazardous incidents for timely dissemination of notifications and warnings involves a thoughtful mixture and application of information, technology and intuitional processes. It starts with the identification of the right data - data to be used in decision making processes - from the right sources - authoritative sources that can be trusted and relied upon. Processes must then be developed to routinely and swiftly acquire, process, and ingest these data into an early warning system (EWS). Decision criteria - sometime referred to as business rules - must be established to transform these data into actionable information, including for the dissemination of warning messages. Finally, the warning messages must be quickly and securely transmitted to the intended recipients, often via redundant mechanisms to insure receipt. Of course, warning messages themselves, even if timely, accurate, and actionable, are not sufficient without an overall context in which to assess them as well as pre-established processes for taking actions, sometimes referred to as Standard Operating Procedures (SOPs). However, even the best SOPs will be ineffective if their users are not adequately skilled and knowledgeable. This generally means that a training and exercise program must be a key component of any successful monitoring and warning system. These elements of effective monitoring - and strategies for their implementation - are described and illustrated via the Pacific Disaster Center s DisasterAWARE all-hazards monitoring, early warning and decision support system.

Importation of live fish to Brunei Darussalam have incurred a major biosecurity risk to the aquaculture industry. Preventing disease incursions through quarantine, legislation and education is currently the most cost-effective management approach in Brunei. Once an incursion has occurred, national emergency response system arrangements are implemented to facilitate immediate response actions for containment and eradication. Brunei Darussalam has a list of legislation and policies to aid in the immediate response of disease outbreak. However, fisheries staff lack basic emergency response training and there are few skilled staff and resources available. Simulation exercise to review the effectiveness of the AEPR system needs to be addressed.

A few decades ago, Cambodia was rich in both freshwater and marine fisheries resources because of its favorable geographical area. However, the fisheries resources have depleted and were unable to totally fulfill the increasing fish demands of its people. This was caused by various factors including unfavorable climate changes, increase in population, improper agricultural production practices, and other human affecting activities. In this sense, aquaculture development in Cambodia becomes increasingly important in order to reduce the fishing pressure on its natural resources which are mainly for food security and economy of Cambodian people. Aside from this, aquatic animals in the country are vulnerable to infectious aquatic transboundary diseases as a result of insufficient and low transboundary diseases monitoring capacity. Neither the official list of aquatic transboundary diseases was created nor are the emergency preparedness and response systems for effective management of transboundary disease outbreaks in Cambodia has been well-established. Nonetheless, the government fisheries officers of both central and provincial levels have conducted fish health monitoring and undertake sample collection from fish farmers since 2016 in 10 targeted provinces as funded by the European Union s Programme. Regarding the capacity of the diagnostic laboratory, officers can perform level I and II but not for all species and diseases. Level III diagnoses cannot be effectively performed yet due to the lack of facilities, skills, and knowledge. The Marine Aquaculture Research and Development Center (MARDeC) is the only main laboratory for aquatic animal health diagnosis in the country. To minimize the spread of aquatic transboundary diseases in freshwater and seawater, the Ministry of Agriculture, Forestry and Fisheries has been moving to initiate and establish policies regarding: (1) the registrations, licensing, and law enforcement; (2) the inspection of sites; (3) and the issu

The information presented in this paper were taken from several key FAO documents. The objective is to continuously raise awareness about the concept of risk analysis and its application to the aquaculture sector. The paper provides information in response to several key risk questions, e.g.: (1) what is risk versus hazard, (2) what is risk analysis, (3) who uses risk analysis, (4) why do countries need to be able to use risk analysis? An overview of the risks in aquaculture is also provided in terms of the process and approaches; and the different risk sectors in aquaculture. The paper concludes with some key points and challenges. Risk analysis is a decisionmaking tool that contributes to protecting national health and welfare. It can also contribute to sustainable aquaculture and the success of individual aquaculture businesses and operations. Risk analysis does not stand alone - it supports and is supported by other components of a National Strategy on Aquatic Animal Health. A basic strength of the risk analysis process is its flexibility - it is adaptable to almost any sector/system where risk and uncertainty occur. Countries will often be confronted with a lack of scientific information, both quality and quantity, to support the risk analysis process. Nevertheless, governments must often act under these uncertainties as well as make decisions in the face of a great deal of complexity, significant variability, and multiple management goals.

The susceptibility of different weight ranges of orange-spotted grouper (Epinephelus coioides) to piscine nodavirus was studied by experimental infection. Size ranges of 1-2 g, 6-8 g, 10-13 g, 30-60 g, and 90-120 g were intramuscularly injected with 50 µL 106 TCID50/mL of the cell culture supernatant. Clinical signs were observed and mortality occurred from 4 days post infection (dpi) for weight range 1-2 g to 7 dpi for weight range 30-60 g. Fish with weight ranges 1-2 g and 6-8 g showed highest susceptibility with mortality rate of 80% while fish with weight range 90-120 g were no longer susceptible. Mortality rates for weight ranges 10-13 g and 30-60 g were 50% and 20%, respectively. No mortality was observed in the 90-120 g weight range and the control group. These data show that the susceptibility of orange-spotted grouper to VNN is weight-dependent.

Mortalities of up to 60% were observed in pond-cultured Litopenaeus vannamei in Bohol, Philippines. Histopathological examination revealed typical acute hepatopancreatic necrosis disease (AHPND) pathology. PCR test generated 1,269 bp and 230 bp amplicons confirmative for the toxin-producing AHPND strain of Vibrio parahaemolyticus among shrimp sampled from eight ponds. The same samples were subjected to PCR analyses for the presence of other viruses, namely WSSV, IHHNV, IMNV, and TSV. The samples were negative for the viruses except WSSV, which was detected after one-step PCR in six out of eight ponds. These results suggested that shrimp were infected dually with AHPND V. parahaemolyticus and WSSV.

The abdominal segment deformity disease (ASDD) is a new shrimp disease reported only in cultured Penaeus vannamei in Thailand. Shrimp with ASDD have deformed abdominal segment, jagged gut line and bumpy surfaces. Similar signs were observed in cultured P. indicus in the Philippines. However, aside from the signs described for ASDD, some P. indicus showing abdominal segment deformity syndrome (ASDS) had more severe deformities up to the extent that the number of body segments was reduced due to fusion. Shrimp with fused body segment deformity (FBSD) had four instead of five pairs of legs. To account the prevalence of the deformities in P. indicus, shrimp were classified into grossly normal shrimp (NS), shrimp with abdominal segment deformity syndrome (ASDS) and shrimp with fused segments (FBSD). Out of the shrimp sampled, 83.4 ± 5.4% was NS, 10.9 ± 6.2% was ASDS and 5.7 ± 3.0% was FBSD. Morphometric characteristics of the shrimp were measured. There was no significant difference in body weight (BW) among male and female NS, ASDS and FBSD. In both sexes, total length (TL) of FBSD was significantly shorter compared to NS and ASDS. Shrimp samples were also screened to be negative for known infectious viral diseases including white spot syndrome virus (WSSV), infectious hypodermal and haematopoietic necrosis virus (IHHNV), infectious myonecrosis virus (IMNV), P. vannamei nodavirus (PvNV), Macrobrachium rosenbergii nodavirus (MrNV) and Taura syndrome virus (TSV). Occurrence of ASDS and FBSD in post-larvae (PL) produced from captive and wild spawners were also determined. Based on a tank experiment, no significant difference was detected between the percentages of ASDS in PL produced from wild or captive spawners but FBSD was only noted in PL produced from the latter. Deformities generally did not affect the size of P. indicus except for the reduced length of shrimp with FBSD which when coupled with missing pleopods could lead to major economic loss for shrimp farmers if n

An extension manual that describes the clinical signs, behavioral changes, geographical distribution, and species affected by the virus; detection, prevention and control methods especially in the hatchery phase are also included.

Acute hepatopancreatic necrosis disease (AHPND) has recently emerged as a serious disease of cultured shrimp. It has also been described as early mortality syndrome (EMS) due to mass mortalities occurring within 20 to 30 d after stocking of ponds with postlarvae. Here, Penaeus vannamei and Penaeus monodon from shrimp farms in the Philippines were examined for the toxin-producing strain of Vibrio parahaemolyticus due to AHPND-like symptoms occurring in marketable size shrimp. In the P. vannamei, histology revealed typical AHPND pathology, such as sloughing of undifferentiated cells in the hepatopancreatic tubule epithelium. Analysis using the IQ2000 AHPND/EMS Toxin 1 PCR test generated 218 bp and 432 bp amplicons confirmative of the toxin-producing strain of V. parahaemolyticus among shrimp sampled from 8 of 9 ponds. In the P. monodon, histology revealed massive sloughing of undifferentiated cells of the hepatopancreatic tubule epithelium in the absence of basophilic bacterial cells. PCR testing generated the 2 amplicons confirmatory for AHPND among shrimp sampled from 5 of 7 ponds. This study confirms the presence of AHPND in P. vannamei and P. monodon farmed in the Philippines and suggests that the disease can also impact late-stage juvenile shrimp.

Viral nervous necrosis (VNN) caused by piscine nodavirus is a devastating disease affecting mainly marine finfish. In the Philippines, VNN was first reported in hatchery-reared grouper (Epinephelus coioides) broodstock in 2001. These broodstock are usually fed with trash fish. It is therefore suspected that contaminated trash fish may be the source of VNN transmission to the broodstock. To confirm the source of contamination, periodic monitoring of the VNN prevalence using RT-PCR was done on different species of trash fish available in the Iloilo Fishing Port Complex and on the wild-caught fish in Panay Gulf. Results showed that most of the trash fish and wild-caught fish were sub-clinically infected or carriers of VNN, and that the virus might have already been established in the environment where they were living. These findings provide strong evidence that trash fish could be the main source of viral contamination in broodstock since they are identified as the only major input in the culture systems. To prevent the transmission of VNN to broodstock through contaminated trash fish, a shift to a broodstock pelleted feed is highly recommended.

Illegally imported koi carp were confiscated at the Ninoy Aquino International Airport (NAIA), Manila, Philippines by the Fisheries Quarantine and Inspection Service Officers of the Bureau of Fisheries and Aquatic Resources (BFAR). The confiscated fish were turned over to the BFAR Fish Health Laboratory where they were held for observation at a water temperature of 28 degree C. After 5 days, some fish were showing abnormal swimming behavior and some had died. The most prominent disease signs in the freshly dead and moribund fish were body ulcerations and pale gills showing white necrotic patches, consistent with the clinical signs of KHV infection. Gills were dissected and fixed in 95% ethanol. All of the samples tested positive for KHV in a 1-step PCR assay. This paper reports the first case of KHV associated mortalities in illegally important koi carp confiscated by the Fisheries Quarantine and Inspection Service Officers of BFAR. This highlights the importance of the quarantine and inspection service s role in preventing the illegal entry of fish into the country and the introduction of exotic aquatic diseases.

Preventing the transboundary movement of fish viral pathogens in a global environment requires active surveillance. This study examined the presence of three emerging viral pathogens among koi, common, grass, and silver carp in Cambodia, Lao PDR, Myanmar, Philippines, and Vietnam. The studied viruses included koi herpesvirus (KHV), spring viremia of carp virus (SVCV), and grass carp reovirus (GCRV). Detection methods consisted of virus isolation by cell culture, infection assay in naive fish, polymerase chain reaction (PCR), and reverse-transcriptase PCR (RT-PCR). Tissues were collected and pooled from 193 fish samples in Dec. 2004 to Feb. 2005, 406 in Sep. 2005 to Feb. 2006, and 1302 in Oct. 2006 to Feb. 2007. For cell culture, tissue filtrates were prepared from pooled spleens, kidneys, livers, and gills and inoculated onto koi fin (KF-1), grass carp kidney (GCK), and fat head minnow (FHM) cells. For infection assay, tissue filtrates were injected intraperitoneally to healthy, naive common carp. No virus was detected after three cell culture passages and the infection bioassays. One-step and nested-step PCR was used to detect KHV in gills of fish samples. One-step and semi-nested RT-PCR was used to detect SVCV and GCRV in the spleens, kidneys, and livers of fish samples. Samples from all three years from all five countries yielded negative results for all three viruses, indicating that KHV, SVCV, and GCRV were not present in these five countries during the period of the study although KHV outbreaks had been detected in Indonesia, Taiwan, Japan, Thailand, China, and Malaysia.

Asian sea bass, Lates calcarifer (Bloch), exhibited strong immune responses against a single injection of the formalin-inactivated red-spotted grouper nervous necrosis virus (RGNNV), a betanodavirus originally isolated in Japan. Fish produced neutralizing antibodies at high titre levels from days 10 (mean titre 1:480) to 116 (1:1280), with the highest titre at day 60 post-vaccination (1:4480). When fish were challenged with the homologous RGNNV at day 54 post-vaccination, there were no mortalities in both the vaccinated and unvaccinated control fish. However, a rapid clearance of the virus was observed in the brains and kidneys of vaccinated fish, followed by a significant increase in neutralizing-antibody titres. Furthermore, the vaccine-induced antibodies potently neutralized Philippine betanodavirus isolates (RGNNV) in a cross-neutralization assay. The present results indicate the potential of the formalin-inactivated RGNNV vaccine against viral nervous necrosis (VNN) of Asian seabass.

Three regions of koi herpesvirus (KHV) genomic DNA were compared for 34 samples from Japan, six from Indonesia, two from Taiwan, one from the Philippines, 13 from the Netherlands, one from the UK, one from the USA and one from Israel. The analyzed genomic regions included known PCR-detection targets (SphI-5, 9/5 and the thymidine kinase gene). The KHVs from Asian countries were very homogeneous, although two variants were noted based on a single nucleotide polymorphism. In contrast, seven variants were found in KHVs from outside of Asia, and although closely related to one another, they were clearly distinct from those from Asian. The results suggest that a clear genetic distinction exists between Asian and European (including each single isolate from the USA and Israel) types of KHV, and that unique types of KHV were independently introduced or emerged in the respective geographic locations.

Viral nervous necrosis caused by betanodaviruses is one of the most devastating diseases in cultured marine finfish. In the Philippines, mass mortalities occurred in sea bass, Lates calcarifer larvae and grouper, Epinephelus coioides broodstock. The virus was isolated using SSN-1 fish cell line and confirmed by PCR. Cytopathic effect started to develop in the cell line 2 days post infection (p.i) with tissue filtrates until the cells completely disintegrated and detached from the flask at 5 days p.i. and the viral protein was detected by immunofluorescence. Sequence analysis revealed that VNN isolated from the brain of grouper broodstock and sea bass larvae were 98.6% similar. Sequence analysis between the Philippine isolates and red-spotted grouper nervous necrosis virus (RGNNV) genotype is 96.9% similar as compared to 72.0% and 64.0% similar with the barfin flounder nervous necrosis virus (BFNNV) and tiger puffer nervous necrosis virus (TPNNV) genotypes, respectively. These results confirm that the Philippine isolates belong to RGNNV genotype.

The black tiger shrimp (Penaeus monodon) is an ecologically and economically important penaeid species and is widely distributed in the Indo-Pacific region. Here we investigated the genetic diversity of P. monodon (n = 355) from eight geographical regions by genotyping at 10 microsatellite loci. The average observed heterozygosity at various loci ranged from 0.638 to 0.743, indicating a high level of genetic variability in this region. Significant departures from Hardy–Weinberg equilibrium caused by heterozygote deficiency were recorded for most loci and populations. Pairwise FST and RST values revealed genetic differentiation among the populations. Evidence from the assignment test showed that the populations in the West Indian Ocean were unique, whereas other populations examined were partially admixed. In addition, the non-metric multidimensional scaling analysis indicated the presence of three geographic groups in the Indo-Pacific region, i.e. the African populations, a population from western Thailand and the remaining populations as a whole. We also sequenced and analysed the mitochondrial control region (mtCR) in these shrimp stocks to determine whether the nuclear and mitochondrial genomes show a similar pattern of genetic differentiation. A total of 262 haplotypes were identified, and nucleotide divergence among haplotypes ranged from 0.2% to 16.3%. Haplotype diversity was high in all populations, with a range from 0.969 to 1. Phylogenetic analysis using the mtCR data revealed that the West Indian Ocean populations were genetically differentiated from the West Pacific populations, consistent with the microsatellite data. These results should have implications for aquaculture management and conservation of aquatic diversity.

SEAFDEC Aquaculture Department (AQD) pioneered fish disease work in the Philippines and developed diagnostic tools through research. Its Diagnostic Service Laboratory was established in the late 1970s to serve the budding aquaculture industry. Through the assistance of SEAFDEC AQD, this service has been replicated by both private and government agencies involved in shrimp aquaculture, thus, data on disease occurrence and prevalence are already available in databases in various forms. Laboratory analysis of hatchery-reared shrimp postlarvae has become an important tool for marketing using both physical and health attributes as gauges for acceptance or rejection of specific batches. Through the years, the diagnostic tools have evolved from mere wet mount microscopy to molecular diagnostic techniques by means of polymerase chain reaction (PCR) to detect viruses. Despite this development, however, disease information is still patchy and difficult to use as decision-support tools because it remains in highly technical and in difficult to visualize information spreadsheets and tables. GIS is a tool that translates complex data in tables and spreadsheets into maps that provide visual displays of information in both spatial and temporal forms. It shows disease trends that are not presently seen and understood by all stakeholders. This paper will highlight the evolution of shrimp health monitoring as a marketing tool in the Philippines and how the application of GIS has helped in understanding disease patterns in the shrimp industry.