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SEAFDEC

International Workshop on Fish Health Management: Accelerating Awareness and Capacity-Building in Southeast Asia 1-2 March 2012 Sarabia Manor Hotel and Convention Center Iloilo City, Philippines

PROGRAM & ABSTRACTS


Table of Contents Overview

2

Organizing Committee

3

Message

4

Scientific Program

5

Abstracts of Plenary Lectures

10

Abstracts of Country Presentations

14

Abstracts of Oral Presentations

26

List of Poster Presentations

41

Abstracts of Poster Presentations

42

Exhibitors

57

Advertisers

62

Index of Presenters

74


Overview Aquaculture is a major food producing sector in Southeast Asia, contributing 43% of the world’s supply. It is a major economic activity, which produces the main source of dietary protein for the expanding populations in the region. However, outbreaks of known and emerging fish diseases continue to threaten aquaculture, bringing considerable economic losses worldwide. Despite technical advances in the diagnosis, prevention & control of fish diseases worldwide, the status of fish health management in many countries of the Association of Southeast Asian Nations (ASEAN) remains generally unsatisfactory. This suggests a lack of effective dissemination of available information to stakeholders. How best to bridge the gap between knowledge and practice is the focus of the current “International Workshop on Fish Health Management” with the theme “Accelerating awareness and capacity building.” Specifically, this international workshop is organized with the following objectives: 1.

To identify the issues and gaps towards accelerating awareness and capacity building on fish health management in Southeast Asia, particularly in small-holder farms in rural communities;

2.

To provide updates on novel fish health management and progress of innovative researches to prevent devastating diseases; and

3.

To optimize the ability of both the fish health practitioners and aquaculturists to control fish disease outbreaks and improve the productivity through healthy and wholesome aquaculture.

2


Organizing Committee CHAIRPERSON Dr. Teruo Azuma

VICE-CHAIRPERSONS Dr. Edgar Amar Dr. Rolando Pakingking, Jr.

MEMBERS Ms. Gregoria Pagador Dr. Ma. Michelle Pe単aranda Dr. Leobert de la Pe単a Ms. Eleanor Tendencia Ms. Kaylin Gonzales-Corre Ms. Milagros Casta単os Ms. Nanette Bantillo Mr. Demy Catedral

3


Message

W

arm greetings and welcome to the SEAFDEC International Workshop on Fish Health Management: Accelerating Awareness through Capacity Building in Southeast Asia. I would like to thank all the participants for providing their ample contribution to this workshop, especially the plenary speakers, country report speakers, and contributed oral paper and poster paper presenters. The Aquaculture Department of the Southeast Asian Fisheries Development Center (SEAFDEC/AQD) initiated the Japanese Trust Fund- supported Fish Disease Projects in response to numerous requests from various sectors for intensified research on fish health-related problems arising in the Southeast Asian region. Phase I (2000-2004) of the said projects focused on technologies to control diseases through timely and accurate recognition, sound diagnostic capabilities, and control measures for various diseases. Phase II (2005-2009) focused on disease surveillance activities based on the results of the earlier program. The importance of accelerating the delivery of information and awareness among aquafarmers and the establishment of disease prevention methods emerged after reviewing the outcomes of the previous two project phases. To attain the above targets, Phase III (2010-2014) is presently focusing on the greater dissemination of knowledge relevant to fish heath management, especially to the SEAFDEC Member Countries whose capacities still need to be developed and improved. At the same time, research and technology development are also implemented as essential activities to sustain SEAFDEC’s role as “A Leading Fish Disease Technology Center in the Region”. An integrated fish-health-care system expected to be established through this project will ensure a holistic approach to a stable supply of safe aquaculture products. As one of the proposed activities crucial to attaining the objectives of the Phase III Project, we have organized this International Workshop on Fish Health Management with a theme entitled “Accelerating awareness and capacity building in Southeast Asia”. This workshop aims to: 1) identify the issues and gaps towards accelerating awareness and capacity building on fish health management in Southeast Asia, particularly in small-holder farms in rural communities; 2) provide updates on novel fish health management and progress of innovative researches to prevent devastating diseases; and, 3) optimize the ability of both fish health practitioners and aquaculturists to control fish disease outbreaks and improve productivity through healthy and wholesome aquaculture. On behalf of the workshop organizing committee, I sincerely wish that this workshop will provide the momentum for the promotion of fish health management awareness and application of appropriate technology, not only among aquafarmers but also among the academe, the government and other stakeholders in order to mitigate the impact of emerging fish diseases and contribute to the establishment of sustainable aquaculture practices in the Southeast Asian region. Finally, I would like to express my sincere gratitude to Dr. Chumnarn Pongsri, SEAFDEC Secretary-General and Dr. Joebert D. Toledo, Chief, SEAFDEC/AQD, for their warm encouragement and support for the hosting of this International Workshop. Many thanks are also due to the members of the workshop Organizing Committee who efficiently did the required preparations leading to the workshop. Lastly, I wish to acknowledge the Fisheries Agency of the Government of Japan for providing financial support for the conduct of this workshop.

Teruo Azuma, Ph. D. Chairperson, Organizing Committee, Deputy Chief & Co-Manager of GOJ-TF Program, SEAFDEC/AQD.

4


Scientific Program Thursday, 1 March AM 7:30-8:30 8:30-9:30

Registration Opening Ceremony (Emcee: Dr. Ma. Rowena Romana-Eguia)

Welcome Messages – Dr. Joebert Toledo Chief, SEAFDEC/ AQD

Dr. Asis Perez National Director, Bureau of Fisheries and Aquatic Resources (BFAR) and SEAFDEC Council Director for the Philippines

Opening Remarks – Dr. Teruo Azuma Deputy Chief, SEAFDEC/AQD AND Co-Manager, GOJ-TF 5 Projects

Keynote Address – Dr. Chumnarn Pongsri Secretary General, SEAFDEC

9:30-10:00

Ribbon Cutting for the Official Opening of the Poster Session/ BREAK

ACCELERATING AWARENESS IN FISH HEALTH MANAGEMENT THROUGH CAPACITY BUILDING 1March AM

SESSION 1 (Moderator: Dr. Erlinda Cruz-Lacierda)

10:00-10:40

Plenary Lecture 1: Accelerating Awareness and Capacity Building in Asia Dr. Eduardo Leaño (Network of Aquaculture Centers in Asia (NACA), Bangkok, Thailand)

10:40-11:10

The Aquatic Animal Health Status in Brunei Darussalam Dr. Wanidawati Tamat (Aquatic Animal Health Services Centre, Department of Fisheries, Ministry of Industry and Primary Resources, Brunei Darussalam)

11:10-11:40

Fish Health Management in Cambodia Dr. So Nam (Inland Fisheries and Research and Development Institute (IFReDI), Fisheries Administration, Pnom Penh, Cambodia)

11:40-12:10

Fish Health Management Program in Indonesia: Fish Disease Status, Monitoring & Surveillance, Fish Health Laboratory, and Capacity Building Taukhid (Fish Health Research Laboratory, Research Institute for Freshwater Aquaculture, Bogor, Indonesia)

12:10-1:30

LUNCH

5


1 March PM

SESSION 2 (Moderator: Dr. Gilda Lio-Po)

1:30-2:00

Status of Fish Health Activities in LAO PDR Vannaphar Tammajedy (Namxouang Aquaculture Development Center, Department of Livestock and Fisheries, Lao PDR)

2:00-2:30

Aquatic Animal Health Management in Malaysian Aquaculture Industry: Current Status & Its Significance to Aquaculture Sustainability Dr. Siti-Zahrah Abdullah (National Fish Health Research Centre, Penang, Malaysia)

2:30-3:00

Accelerating Awareness in Fish Management Through Capacity Building in Myanmar Me Me Thin (Department of Fisheries, Ministry of Livestock and Fisheries, Myanmar)

3:00-3:30

Status of Aquatic Animal Health in the Philippines Dr. Joselito Somga (Fish Health management and Quality assurance Section, Bureau of Fisheries and Aquatic Resources, Quezon City, Philippines)

3:30-3:50

BREAK/ POSTER VIEWING SESSION 3 (Moderator: Dr. Ma. Rowena Romana-Eguia)

3:50-4:20

Status of Aquatic Animal Health in Singapore Low Yilin Agri-Food Veterinary Authority (AVA), Singapore

4:20-4:50

Aquatic Animal Health Activities in Thailand Dr. Temduong Somsiri (Inland Aquatic Animal Health Research Institute, Department of Fisheries, Thailand)

4:50-5:20

Aquatic Animal Health Management in Vietnam Dr. Le Van Khoa (Department of Animal Health, Ministry of Agriculture and Rural Development, Vietnam)

5:20-6:45

WORKSHOP ON ACCELERATING AWARENESS IN FISH HEALTH MANAGEMENT THROUGH CAPACITY BUILDING Moderators: Dr. Edgar Amar and Dr. Celia Lavilla-Pitogo Rapporteurs: Gregoria Pagador and Demy Catedral

7:00-9:00

WELCOME DINNER

6


INNOVATIVE RESEARCH AND NOVEL DIAGNOSTIC TECHNIQUES FOR FISH PATHOGENS

2 March AM

SESSION IV (Moderator: Dr. Jane Geduspan)

8:00-8:40

Plenary Lecture 2: Investigation on Post-Transportation Mortalities is Necessary to Prevent Introduction of Exotic Pathogens Prof. Teruo Miyazaki (Fish Pathology Laboratory, Department of Aquaculture Science, Faculty of Life Science, Graduate school of Bioresources, Mie University, Japan)

8:40-9:00

Fish Health Management on Food Safety Control in the Philippines Dr. Simeona E. Regidor (Fish Health and Quality Assurance Section, Bureau of Fisheries and Aquatic Resources, Quezon city, Philippines)

9:00-9:20

Surveillance of Parasite Fauna of Economically Important Freshwater Fish in Some Southeast Asian Countries Gregoria Pagador (Fish Health Section, SEAFDEC/AQD, Philippines)

9:20-9:40

Impact of Fish Vaccination for Fish Health Management: Examples from Pangasius in Vietnam Vo Thanh Tung (PHARMAQ Vietnam Ltd., Vietnam)

9:40-10:00

BREAK/ POSTER VIEWING SESSION V (Moderator: Dr. Simeona Regidor)

10:00-10:40

Plenary Lecture 3: Managing Health and Sustainability of Specific-PathogenFree Penaeus stylirostris Through Hatchery and Farm Surveillance Dr. Celia R. Lavilla-Pitogo (Integrated Aquaculture International, Bandar Seri Begawan, Brunei Darussalam)

10:40-11:00 11:00-11:20

Silencing White Spot Syndrome Virus and Elucidation of Shrimp-Virus Interaction by RNA Interference Technology Dr. Mary Beth B. Maningas (Research Center for the Natural and Applied Sciences, University of Santo Tomas, Manila, Philippines) Development of Loop-Mediated Isothermal Amplification (LAMP) Assay for the Detection of Philippine Isolate of the Penaeus monodon-Type White Spot Syndrome Virus (WSSV) May Flor J. Sibonga (National Institute of Molecular Biology and Biotechnology (NIMBB), University of the Philippines-Visayas, Philippines)

7


11:20-11:40 11:40-12:00

Loop-Mediated Isothermal Amplification (LAMP) Detection of White Spot Syndrome Virus (WSSV) in Litopenaeus vannamei in Selected Sites of the Philippines Amalea Dulcene D. Nicolasora (Research Center for the Natural and Applied Sciences, University of Santo Tomas, Manila, Philippines)

12:00-1:00

LUNCH

Sandfish (Holothuria scabra): a Potential Carrier of White Spot Syndrome Virus Dr. Leobert de la Pe単a (Fish Health Section, SEAFDEC/ AQD, Philippines)

2 March PM

SESSION VI (Moderator: Dr. So Nam)

1:00-1:20

Development of Practical Shrimp Vaccination Techniques Against White Spot Syndrome Virus in Penaeus monodon: Progress of Tank and Pond Studies Dr. Edgar C. Amar (Fish Health Section, SEAFDEC/ AQD, Philippines)

1:20-1:40

Establishment of an Immunization Regimen for the Prevention of Viral Nervous Necrosis (VNN) in Asian Sea Bass (Lates calcarifer) Broodfish Dr. Rolando V. Pakingking Jr. (Fish Health Section, SEAFDEC/ AQD, Philippines)

1:40-2:00

Cloning, Characterization and Expression Analysis of a cDNA Encoding Granulin Gene in Nile tilapia (Oreochromis niloticus, Linn.) Myat Khine Mar (Laboratory of Aquatic Animal Health Management, Department of Aquaculture, Faculty of Fisheries, Kasetsart University, Bangkok, Thailand)

2:00-2:20

Lectins as Non-Self Recognition Molecules and Their Relevance in Innate Immunity and Fish Health Dr. Lito Argayosa (Institute of Biology, College of Science, University of the Philippines- Diliman, Quezon City, Philippines)

2:20-2:40

Aptamer Technology for Aquaculture Dr. Mudjekeewis D. Santos (Genetic Fingerprinting Laboratory, National Fisheries Research and Development Institute (NFRDI), Quezon City, Philippines)

2:40-3:00

Isolation of Dehalogenase Producing Bacteria from the Gut of Pond-Reared Rohu (Labeo rohita) Juveniles in Myanmar Dr. Fahrul Huyop (Faculty of Biosciences and Bioengineering, Universiti Teknologi Malaysia, Johor Bahru, Malaysia)

3:00-3:20

Early Warning Activities Prevent Disease Outbreak and High Mortality Impact of Overturn on Fish Culture in Jatiluhur Dam, Indonesia Heny Budi Utari (Department of Biology, Faculty of Science, Mahidol University, Bangkok, Thailand)

8


3:20-3:40

BREAK/ POSTER VIEWING

3:40-5:00

WORKSHOP ON INNOVATIVE RESEARCH ON FISH HEALTH MANAGEMENT Moderators: Dr. Rolando Pakingking Jr. and Dr. Erlinda Cruz-Lacierda Rapporteurs: Dr. Ma. Michelle Pe単aranda and Dr. Romualdo Balagapo

5:00-5:30

Open Forum/ Discussion

5:30-6:00

Closing Ceremony

Impressions on the Conference/ Workshop Dr. Yukio Maeno Director of Pathology Division, National Research Institute of Aquaculture, Japan Dr. Clarissa L. Marte Former Senior Scientist and Research Division Head, SEAFDEC/AQD, Philippines Closing Remarks Dr. Relicardo M. Coloso Research Division Head, SEAFDEC/AQD, Philippines

9


Abstracts Of Plenary Lectures

10


Accelerating Awareness and Capacity Building in Asia-Pacific Eduardo M. Leaño*, C.V. Mohan Network of Aquaculture Centres in Asia-Pacific,Bangkok, Thailand (*email: eduardo@enaca.org) Aquatic animal health is one of the major hurdles facing the aquaculture sector. The epidemic spread and devastating impacts of several aquatic animal diseases in the region have clearly demonstrated the vulnerability of the different aquaculture systems. The increasing globalization and trade volume of the aquaculture sector has created new mechanisms by which pathogens and diseases are introduced or spread to new areas. The Network of Aquaculture Centers in Asia Pacific (NACA), an intergovernmental organization of 18 governments in the Asia-Pacific, works on the principle of cooperation and collaboration. Addressing aquatic animal health is one of the key program areas of NACA, with the purpose of assisting member governments to “reduce the risks of aquatic animal diseases impacting the livelihoods of aquaculture farmers, national economies, trade, environment, and human health”. Development and adoption of the FAO/NACA’s Asia regional technical guidelines (TG) for responsible movement of live aquatic animals by 21 Asia-Pacific governments is a major outcome facilitated by NACA. The framework provided by the TG is rather comprehensive and includes all major requirements for managing risks associated with the movement of live aquatic animals and trans-boundary pathogens. A network of 21 National Coordinators has been guiding the process of development and implementation of national aquatic animal health strategies under the NACA’s Regional Aquatic Animal Health Programme. Key activities under this programme include: • Quarterly aquatic animal disease (QAAD) reporting system which was initiated since the second quarter of 1998 as a joint activity among NACA, FAO and OIE Regional Representation for Asia and the Pacific (Tokyo); • Asia Regional Advisory Group on Aquatic Animal Health (AG) which was constituted by NACA’s Governing Council in 2001, in cooperation with OIE and FAO. Other activities include: (a) capacity building on diagnostics, epidemiology, surveillance, risk analysis; (b) development of resource materials; (c) provision of technical assistance to individual countries; (d) development of standard operating procedures; and more recently, (g) promoting adoption of better aquatic animal health management practices in some of the member countries. Through more than 10 years of the Aquatic Animal Health Programme of NACA, it can be confidently said that the region as a whole is now in a much better state of preparedness in dealing with aquatic animal disease outbreaks and emergencies. However, the region can’t be complacent, and should ensure strong national commitment and continuous awareness and capacity building at producer, disease support and decision making levels for effective implementation of aquatic animal health management strategy and improve biosecurity. Keywords: NACA, capacity building, aquatic animal health, diagnostics, epidemiology, surveillance, risk analysis, technical guidelines

11


Investigation on Post-Transportation Mortalities is Necessary to Prevent Introduction of Exotic Pathogen Teruo Miyazaki Professor of Fish Pathology Laboratory, Department of Aquaculture Science, Faculty of Life Science, Graduate school of Bioresources, Mie University, 1577 Kurimamachiya-cho, Tsu, Mie 514-8507, Japan (email: miyazaki@bio.mie-u.ac.jp)

We often transport fishes domestically or import fishes internationally. After transportation, mass mortalities often occur. We usually understand that mortality is caused by transportation stress. However, single transportation stress is not a definitive factor for mortality. The transportation stress just promotes mortality of fishes that have been infected with some pathogens in their country of origin. If you do not perform any investigation on the weakened or dead fish after transportation, you would miss to find the pathogen and allow the entrance of exotic pathogens. Here, I introduce some cases of mass mortality caused by exotic pathogens. 1. Megalocytivirus (=Madai iridovirus, Red sea bream iridovirus, RSIV). This virus caused mass mortalities of a large quantity of red sea bream juveniles (2-10 million) and marketable size (2milion) in all over Japan in 1991. More than 30 species of maricultured fishes were involved in this virus infection in 1995. Miyazaki revealed that this virus originated from tropical fishes living in the South China Sea. Japanese fish farmers imported 10 million juveniles of amber jack and sea bass every year from mid 1980s from Hainan Island, China. Few effort of quarantine allowed the entrance and rooting of Megalocytivirus in Japan. Megalocytivirus has spread in Australia, Canada, USA, UK, Thailand, Malaysia, Indonesia, Israel etc. with transportation and trade of fishes. 2. Akoyavirus. In Japan, mass mortalities occurred in the Japanese pearl shell (400 million shells) causing large economical losses of 50 billion Japanese Yen in 1997. Chinese pearl shells that were imported from southern China carried the Akoyavirus. 3. Ranavirus (Epizootic Hematopoietic Necrosis Virus: EHNV). Redfin perch which were introduced from England carried EHNV to Australia.

Keywords: post-transportaion mortalities, exotic pathogens, megalocytivirus, madai iridovirus, red sea bream iridovirus, RSIV, akoyavirus, ranavirus, epizootic hematopoeietic

12


Managing Health and Sustainability of Specific-Pathogen-Free Penaeus stylirostris Through Hatchery and Farm Surveillance Wanidawati Tamat1, Dayangku Siti Norhaziyah Pengiran Haji Halim1, Hajijah Mohammad Said1, Rahimah Haji Mohammad Tahir1, Celia R. Lavilla-Pitogo2* Aquatic Animal Health Services Centre, Department of Fisheries, Ministry of Industry and Primary Resources, Jalan Menteri Besar, BB3910,Brunei Darussalam 2 Integrated Aquaculture International, Block A Bangunan Lim Seng Kok, Simpang 628, JalanTutong, Bandar Seri Begawan BF 1120, Brunei Darussalam (email: celia.pitogo@fulbrightmail.org) 1

Shrimp aquaculture in Brunei Darussalam is currently based on the use of specific-pathogenfree (SPF) Penaeus stylirostris stocks, whose breeding program is sustained through a cooperative effort between the Department of Fisheries and the private sector. Health surveillance is conducted at all levels of production to improve and protect the health of the SPF stocks with the ultimate goal of helping producers maximize production and profitability. The Aquatic Animal Health Services Centre (AAHSC) plays a key role in managing the health of SPF stocks by setting health policies and surveillance schedule in hatcheries and farms. These are complimented with biosecurity measures to avoid the entrance of infection into a facility, control its dissemination within the unit, and avoid its spread to other units or farms. Regular surveillance involves analysis of 9 viruses that affect shrimp for each sample, namely: White spot syndrome virus, Infectious hypodermal and haematopoietic necrosis virus, Hepatopancreatic parvovirus, Yellowhead virus, Gill associated virus, Taura syndrome virus, Infectious myonecrosis virus, Mourilyn virus, and Monodon baculovirus. Histology is conducted to obtain supplementary information on other diseases affecting the stocks as well as to provide an indicator of future problems by focusing on key structures like the lymphoid organ, gills, hepatopancreas and antennal organ. More than four years of monitoring has shown that the SPF P. stylirostris broodstock and hatchery-reared postlarvae have remained free of the 9 viruses. The outbreak of WSSV in the farms in February to March 2011 led DOF to embark on a containment and eradication program followed by stoppage of shrimp farming for 5 months. Surveillance results by PCR and histopathology in hatcheries and farms will be presented as well as eradication and biosecurity measures that were implemented in order to contain WSSV infection.

Keywords: Penaeus stylirostris, specific-pathogen-free shrimp, surveillance, shrimp viruses

13


Abstracts Of Country Presentations

14


The Aquatic Animal Health Status in Brunei Darussalam Wanidawati Tamat Aquatic Animal Health Services Centre, Department of Fisheries, Ministry of Industry and Primary Resources, Jalan Menteri Besar, BB3910 Brunei Darussalam Fisheries have been identified as one of the potential sector that could contribute to the diversification of the Brunei Darussalam’s economy, away from the reliance on oil and gas sector. The aquaculture industry in Brunei Darussalam consists of cage culture of marine fishes and pond culture of freshwater and marine shrimps. The Department of Fisheries has the vision for the development of the aquaculture industry into a sustainable source of income. The shrimp industry has contributed significantly to the aquaculture production since the industry started in 1989 with a contribution of 80% of the total national aquaculture production. In order to assure the sustainability of the industry and safeguard the investments from both the government and private sector, it is imperative that effective means of control and management of incursions of exotic diseases into the country be in place. The Department of Fisheries has developed its Aquatic Animal Health Strategy which comprises of the mechanism, action plan and management system in order to strengthen the aquatic animal health and the biosecurity system in Brunei. To start off a SPF shrimp breeding program, a state-ofthe-art aquatic animal health laboratory with molecular, histological, and microbiological methods capable of detecting all known shrimp diseases was established. The laboratory was crucial in disease screening to develop SPF populations that serve as foundation of a nucleus breeding program where the shrimp are propagated in biosecure facilities. Patterned after monitoring and surveillance activities for shrimp, DOF also actively monitors fish cage culture farms. Information from the surveillance program and data collected are used for the OIE reporting. Several local and international training and workshops have been held to increase the knowledge and update information on health management of aquaculture. However, staff development needs to continue in order to meet the demands of aquaculture for quality and timely disease diagnosis.

Keywords: aquatic animal health, status, Brunei Darussalam, Department of Fisheries, shrimp, SPF breeding program, biosecurity

15


Fish Health Management in Cambodia So Nam Inland Fisheries Research and Development Institute (IFReDI), Fisheries Administration, Phnom Penh, Cambodia (email: so_nam@hotmail.com) About 86% of Cambodia’s land area is within the Mekong catchment, and about 20% of the Mekong River’s catchment is within Cambodia containing high minerals and nutrients. This has made Cambodia ranked number four in inland fisheries productivities (approx. 400,000 tons/year) after China, India and Bangladesh. The coastal area also has a high potential for fisheries productivities (Approx. 75,000 tons/year). Cambodian fisheries resources play a very important role in contributing to national food and nutrition security, national economy, and people’s livelihoods and culture. The total marine and freshwater capture fisheries production is estimated at about 400,000 to 500,000 tons per year, and the catch of other aquatic animals (OAAs) such as shrimps, crabs, snails, frogs, freshwater edible insects, snakes, and turtles are at least 60,000-100,000 tons per year. As a result, aquaculture development is not significant in Cambodia, although show a rapid growth over the past two decades and increased from 1,610 tons in 1984 to 13,857 tons in 2004 and 60,000 tons in 2010, representing a 37.3-time increase or a growth of around 18% per year for the past ten years, ahead of annual growth rate (10%) of world aquaculture production, and 7th rank in the world in terms of growth rate. It represented around 8% of total inland fisheries production in 2004 and around 11% in 2010. Recently, the Inland Fisheries Research and Development Institute (IFReDI) in cooperation with WorldFish Center under the Project FIS/2010/031 Fish Supply and Demand in the Mower Mekong Basin funded by Australia Center for International Agriculture Research (ACIAR) have identified seven main aquaculture systems in Cambodia (WFC/IFReDI/ACIAR, 2011): (1) Smallholderlow input pond culture (Extensive pond polyculture of carps, tilapia, silver barb and pangasiids); (2) Smallholder- high input pond culture (Intensive pond polyculture and monoculture of carps, tilapia, silver barb and river pangasiid catfish); (3) SME semi-intensive pond culture (Intensive pond monoculture of river pangasiid catch, hybrid clariid catch or snakeheads); (4) Freshwater cage culture (Intensive polyculture of pangasiids and carps, and intensive monoculture of snakeheads); (5) Marine cage culture (Intensive monoculture of sea bass or grouper); (6) Rice-fish systems (Extensive, low input system of carps, with concurrent rice and fish culture); and (7) Brackish water extensive large ponds (Extensive, low input system of fish and shrimps). Based on the past few years of IFReDI capacity building, monitoring and networking program over the country, there have been no detections of clinical signs of disease outbreaks caused by pathogens, including bacterial and viral agents, in fish cultured in the above seven culture systems and in wild fish species due to some extents of a lack of fish health R & D and diagnostic capability. However, in the capacity of IFReDI, there are seven genera of parasites detected in fish, including Lernaea, Trichodina, Argulus, Itchthyophthiosis, Alitropus, Gyrodactylus and Nematoda. Therefore capacity building and strengthening in fish health (including human capacity: i.e. research personnel, and facility and equipment capacity: i.e. field and laboratory diagnostic capacity) as highlighted in the draft National Strategic Framework for Aquatic Animal Health in Cambodia are needed to manage aquatic animal health in Cambodia in order to reduce risks of aquatic animal diseases impacting on livelihoods of fish farmers and fishers, national economy, trade and human health. Keywords: fish health management, Cambodia, Inland Fisheries Research and Development Institute, aquaculture systems, fish diseases

16


Fish Health Management Program in Indonesia: Fish Disease Status, Monitoring and Surveillance, Fish Health Laboratory, and Capacity Building Taukhid*, Hambali Supriyadi Fish Health Research Laboratory, Research Institute for Freshwater Aquaculture, JI. Sempur No. 1, Bogor 16154, Indonesia (email: taukhid_as@yahoo.co.id) Aquaculture in Indonesia has been growing rapidly; it plays an important role in rural development, source for export earning, and has been a leading sector in economic growth. Due to the flat tendency from capture fisheries production, Indonesia will depend on aquaculture in the future. Intensification of aquaculture has led to remarkable improvements in productivity; on the other hand, it is also associated with disease epidemics. Fish diseases are a major risk and a primary constraint to the growth of the aquaculture in Indonesia. The increasing fish transportation and trade volume has created new mechanisms by which pathogens may be introduced or spread to new areas. Many of infectious diseases (motile aeromonad septicemia, streptococcosis, koi herpes virus disease, viral nervous necrosis, white spot syndrome virus disease, Taura syndrome virus disease, and infectious myonecrosis virus disease) have been realized to be a serious constraint on the production of economically important fish commodities, such as catfish, common carp, tilapia, groupers, and penaeid shrimps. The National Fish Health Commission (NFHC) in 2008 has classified “The Most Significant and Potential Fish Diseases in Indonesia.” Previously, control of fish disease in Indonesian aquaculture mainly relied on the use of chemicals/antibiotics. The government has issued strict regulations concerning the supply, distribution, use of drug, and control of these materials in fish production. Today, fish disease control focuses on prevention employing biological approaches: use of specificpathogen-free (SPF) broodstock/population, improvement of husbandry techniques and good sanitation (biosecurity), vaccination, and application of immunostimulants and probiotics. In connection with fish health management, the government develops a program namely “National Strategy on Fish Health Management”, which focuses on ten priorities: (1) Surveillance, monitoring and reporting of fish disease; (2) Fish quarantine; (3) Institutional framework; (4) Fish disease control and contingency planning; (5) Building public awareness; (6) Research and development; (7) Rules and regulation; (8) Regional and international fish health collaboration network; (9) Fisheries resources and environment management for aquaculture; and (10) Funding the national strategy. In addition, progress and implementation of these activities, especially monitoring & surveillance, current status of fish health laboratory and its diagnostic capability (diagnostic methods), and capacity building will be presented.

Keywords: fish health management, status, capacity building, Indonesia, shrimp viruses, fish pathogens, National Fish Health Commission, National Strategy on Fish Health Management

17


Status of Fish Health Activities in LAO PDR Vannaphar Tammajedy Namxouang Aquaculture Development Center, Department of Livestock and Fisheries, Ministry of Agriculture and Forestry, Vientiane, Lao PDR Inland capture fisheries and aquaculture in Lao PDR are based mainly on water resources ecosystems consisting of rivers and their basins, hydropower and irrigation reservoirs, temporary or permanent derivation weirs, gates and dykes, small water bodies, flood plains and wet season rice fields. The total water resources for capture fisheries are believed to be more than 1.2 million ha. The estimated yield of inland fish in Lao PDR is approximately 30,900 tons per year while other aquatic animals are estimated at 82,100 tons per year. These estimated yields are conservatively valued at almost US$ 150 million per year. The people of Lao PDR, especially in the rural communities which account for more than 75%, still rely on fishes and other aquatic animals as their reliable sources of healthy animal protein intake. More than 481fish species have been identified in Lao PDR including 22 fish species identified as exotic species. To date, the Fisheries Division is under the umbrella of the Department of Livestock and Fisheries (DLF) of the Ministry of Agriculture and Forestry (MAF). The Fisheries Division employs 39 persons country-wide which is insufficient to provide effective technical support throughout the provinces and districts. The MAF has recently designated the Namxouang Aquaculture Development Center as the country’s prime laboratory for fish disease diagnosis. However, the level of fish disease diagnosis in the aforestated laboratory is currently categorized as level I in spite of the available equipment suited for a level III fish disease diagnostic laboratory. This discrepancy is chiefly attributed to lack of qualified fish pathologists. Because freshwater fish aquaculture in Lao PDR is still relatively extensive and fish farmers are fewer compared with neighboring countries like Thailand, serious outbreaks of diseases in various cultured freshwater fish species are apparently scarce. However, intensification of small scale/ large scale freshwater fish aquaculture shall be anticipated in the coming years to meet the increasing demand of freshwater fish for local consumption and export. Alongside with this vision, introduction of novel freshwater fish aquaculture practices coupled with the occurrences of infectious diseases will become inevitable. Thus to aptly address future fish disease problems and improve the current marginal level of fish disease diagnosis in LAO PDR, continued upgrading of laboratory facilities and recruitment of qualified fish pathologists or perhaps identification of existing fisheries officers to undertake intensive hands-on training on fish pathology is prudently needed.

Keywords: fish health activities, status, LAO PDR, Department of Livestock and Fisheries, Ministry of Agriculture and Forestry, Namxouang Aquaculture Development Center, aquaculture

18


Aquatic Animal Health Management in Malaysian Aquaculture Industry: Current Status and Its Significance to Aquaculture Sustainability Siti-Zahrah, A.1 , Kua, B.C1., Iftikar, A. A. R.1, Nik Haiha, N.Y.2, Zulkafli, A.R.3, Hussin, M.A.2, Raihan, S.A.4 National Fish Health Research Centre (NaFisH) FRI Batu Maung, 11960,Penang 2 FRI Tg Demong, Terengganu, 3 FRI Glami Lemi, Jelebu, N.Sembilan 4 Department Of Fisheries (DOF), Wisma Tani, Putrajaya, Kuala Lumpur

1

Fish health, precisely aquatic animal health has been a factor of concern lately especially so when quality aquaculture production is expected. Two decades ago, in early 1990s in Malaysia, emphasis was given mainly on aspects of increasing production without considering related aspects of health management to maximise output. However, when disease outbreaks occurred in mid and late 1990s, most aquaculturists suffered high economic losses. The industry operators, stakeholders and directly DOF thus realized the importance of aquatic animal health management inputs for the benefits of the industry. Accordingly, National Fish Health Research Centre (NaFisH) was formed in 1994 dealing with R&D in aquatic animal health to support the industry. This was practically not sufficient however, when ornamental fish trade is involved. Biosecurity division in DOF was immediately formed in 2008, after the exported koi from Malaysia to EU was reported to be infected with KHV in 2006, banning further exports. Such government initiative is meant to ensure that standard guidelines on Aquatic Animal Health (AAH) procedures outlined by OIE in farmed and for exports of aquaculture products can be followed without fail. Since then, NaFisH with Biosecurity division have complimented each other in combined effort to tackle related issues of aquatic animal health to ensure improved and better rearing practices to be followed and continuously managed by farmers to produce quality fish products for both domestic consumption and exports. Aquatic animal health programmes for both aquaculture practitioners and stakeholders were given serious attention and support to help improved better awareness of AAH importance to the industry. Aggressive approach by the competent authority (CA) to educate and improved their human capability in those aspects of aquatic animal health in the last 5 years was made to ensure smooth implementation of better management practices in aquaculture industry and be knowledgeable on technical matters concerning aquatic animal health aspects so as standard guidelines of both OIE could be followed and understand fully. This paper will try to present our limited experience, the limitations and constraints in implementing steps and measures for aquatic animal health programmes within the industry as well as its other strategies and way forward for better sustainable aquaculture.

Keywords: aquatic animal health, status, Malaysia, National Fish Health Research Centre, biosecurity

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Accelerating Awareness in Fish Health Management through Capacity Building in Myanmar Me Me Thin Department of Fisheries, Ministry of Livestock and Fisheries, Myanmar The fishery sector in Myanmar is the most important sector next to agriculture. It aims to fulfill the protein requirement of the people, provide food security, and as well offers opportunity for employment to a large number of fishery communities and rural dwellers. Myanmar is endowed with rich natural resources both in freshwater and marine fisheries. At present, more than 20 species of indigenous and exotic freshwater fish species including rohu (Labeo rohita), catla (Catla catla), mrigal (Cirrhinus mrigala), and butter fish (Silondia spp.), and high value marine fish species such as grouper, snapper, and sea bass are the commonly cultured fish species. Aside from the giant freshwater prawn (Macrobrachium rosenbergii) which is the most common and prioritized species, culture of Penaeus vannamei has been recently introduced in Myanmar. However, the Department of Fisheries (DoF), the sole competent authority of fisheries sector, fully understands that P. vannamei may carry exotic viruses such as the Taura Syndrome Virus (TSV). At present 4 private farms are trying experimental culture of P. vannamei. Mud crab aquaculture has also become a booming industry as domestic consumption and export demands are growing rapidly. In particular, soft shell mud crab farming has become popular as it commands high price. However, supply of crab juveniles from nature is decreasing due to over exploitation, habitat deterioration, and climate change. In Myanmar, aquatic animal health management is being undertaken by the Aquatic Animal Health and Disease Control Section (AAHDCS) of the DoF. The AAHDCS has assigned mobile teams to conduct monitoring and surveillance of fish ponds in strategic areas. Fish disease information is gathered by active and passive reporting systems. Quarterly reports on fish disease have been regularly sent to NACA, OIE since 1998 to date. Because the DoF is highly aware of transboundary aquatic animal pathogens, it expanded its disease control unit into a new set up of organization to conduct field survey diagnostics and assigned officials in airports and sea ports and border trade areas to closely check every export and import commodity. Generally, few diseases have been detected in aquatic animals both in capture and culture fisheries. In 1985, Epizootic Ulcerative Syndrome (EUS) occurred in Myanmar. It occurred only for a short period and was brought under control before it became widespread. Fish and shrimp parasitic diseases are not common. Unfortunately, occurrence of White Spot Syndrome Virus disease and Yellow Head Disease in tiger shrimp farms in Rakhine State, Yangon and Ayeyarwaddy Regions had caused serious economic losses. Thus to accelerate awareness on fish health management, the DoF recommends the following: (a) Introduction of alien Aquatic species should only be made prior to consideration for safeguarding natural resources and ecosystems; (b) If some species, that may not cause negative impact to conservation of fishery resources and ecosystem, are decided to be introduced, awareness on prior and reliable reporting should be conducted; (c) Culture of alien aquatic species should be facilitated through good aquaculture practice (GAP) and/ on environment friendly aquaculture practices; (d) Introduced alien species should be genetically upgraded through high health management and screening method so as to sustain specific pathogen free (SPF) parent stock; (e) Moving culture system and feeding system from traditional to best management practice (BMP) can control feed quantity and reduce the costs, and importantly control the occurrence of diseases. Keywords: accelerating awareness, capacity building, fish health management, Myanmar

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Status of Aquatic Animal Health in the Philippines Joselito R. Somga*, Simeona E. Regidor, Maria Abegail A. Albaladejo, Sonia S. Somga Fish Health Management and Quality Assurance Section, Bureau of Fisheries and Aquatic Resources, 860 Quezon Avenue, Quezon City, Philippines (email: josomga@yahoo.com) The aquaculture sector contributes significantly to the country’s total fisheries production, employment and foreign exchange generation. Majority of the aquaculture production comes from milkfish, tilapia and shrimp. The aquatic animal species are being cultured either in ponds or cages in freshwater, brackishwater and marine environment using different production system. The intensification of culture system and diversification of farmed species has led to the rapid growth of the aquaculture industry. Maintaining the optimum condition of the aquatic environment as well as good husbandry and health management practices for productivity and sustainability becomes a big challenge for the industry. As such disease outbreaks and occurrences in fish and shrimp farming were considered as one of the major constraints to successful aquaculture production. The Bureau of Fisheries and Aquatic Resources (BFAR) under the Department of Agriculture is the agency mandated by law to be responsible in leading the implementation of legislations regarding the protection, conservation and management of the country’s fishery and aquatic resources. Recognizing the importance of aquatic animal health, BFAR through the Fish Health Management and Quality Assurance Section (FHMQAS) has established fish health laboratories in different regions of the country in order to provide accessible diagnostic services and technical assistance needed by the industry. These laboratories are manned by designated regional fish health officers with the technical supervision and guidance from the FHMQAS of the Central Office. They have different level of capabilities depending on the needs in the areas of their jurisdiction. Through the Fish Health Network, coordination among and between the central and regional offices in the implementation of various programs and activities has been strengthened and harmonized. Major programs and activities on fish health include disease detection and surveillance, disease reporting, management of disease outbreaks, quarantine and health certification as well as national residue monitoring and control program for aquaculture products. The Network has been an effective means in disseminating information on disease occurrences and fish health issues and concerns to the stakeholders either through trainings, seminars and meetings. BFAR has been involved and participated in various programs on fish health initiated and implemented by international and regional organizations such as FAO, OIE, NACA, SEAFDEC, JICA, IDRC, AusAID, and others. The technical assistance and funding support from these organizations and other external sources has been instrumental in capacity building of BFAR on fish health. At present, BFAR is actively participating in the aquatic animal disease reporting system by the OIE and NACA. BFAR will continuously strengthen aquatic animal health programs and services in line with international standards to further improve aquatic animal health, food safety and aquaculture sustainability. Keywords: aquatic animal health, surveillance, disease outbreaks, quarantine, health certification, BFAR, Philippines

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Status of Aquatic Animal Health in Singapore Low Yilin Agri-Food Veterinary Authority (AVA), Singapore Aquaculture in Singapore consists of both the foodfish and ornamental aquaculture industry. Currently, as of 2010, there are 113 coastal fish farms, 9 land-based food fish farms and 74 ornamental aquaiurm fish farms licensed by the Agri-food & Veterinary Authority of Singapore (AVA). The foodfish aquaculture industry currently produces about 5% of Singapore’s annual local fish consumption. Since 2011, there has been a target to increase this percentage to 15%. For ornamental aquaculture industry, Singapore is known as one of the top exporters of ornamental fish globally. In 2010, Singapore exported ornamental fish with an approximate value of $79.4 million to over 80 countries. In order to maintain our position as one of the top exporters of ornamental fish globally and in order to meet Singapore’s target of increasing local food finfish production, AVA has recognized the importance of building capabilities of all stakeholders involved, including diagnostic staff, field/extension officers, farmers and other industry players. This is done through multiple platforms via training courses, meetings and dialogues, of which some will be mentioned in the country report.

Keywords: status, aquatic animal health, Singapore

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Aquatic Animal Health Activities in Thailand Temdoung Somsiri Inland Aquatic Animal Health Research Institute, Department of Fisheries, Thailand

Effective programs of aquatic animal health control are necessary to prevent spread of disease agents and sustain aquaculture in Thailand. Department of Fisheries (DoF) has set up the program for finfish and crustacean health based on risk of aquatic animal diseases and international trade requirements. The program was developed and implemented by DoF as well as private sectors. Surveillance and monitoring of aquatic animal diseases, import requirement for risk species, health certificate for export of aquatic animals and emergency disease plan have been developed under national legislations and regulations. There are 36 aquatic animal diseases that have been listed in the Animal Epidemic Act. The national listed diseases will be updated following announcement of OIE. Thailand has 2 regional reference laboratories for aquatic animal diseases which have been accredited to ISO/IEC 17025. Diagnostic techniques using molecular applications and cell culture were employed for almost all diseases. Quarterly report of disease status in Thailand is submitted to OIE and Network of Aquaculture Centers in Asia Pacific (NACA). Most of the production farms have been registered by the DoF. Disease monitoring program has been conducted particularly on susceptible species. Some marine shrimp and Koi carp farms have employed biosecurity management to prevent diseases and reduce economic losses. In addition, DoF is going to develop proficiency testing program for shrimp viruses and koi herpes virus. Because the number of current staff working on aquatic animal health is not sufficient compared to their workload due to routine diagnostic service, farm monitoring, quarantine inspection and issuing health certificate, R&D capability has been consequently reduced; hence, fewer research outputs have been published. Moreover, the number of government field personnel is also insufficient to give proper service to the farmers. Therefore, it is the responsibility of DoF to increase its number of aquatic animal health staff to serve the need of aquatic animal health control activities and services. The Aquatic Animal Health Research Institute (AAHRI) has not received any internal or international support to strengthen the capacity and capability of the staff on aquatic animal health for more than 10 years. Training on import risk analysis and aquatic animal epidemiology are needed to improve national disease surveillance program to meet the international standards and approval of authorized importing countries. Research collaboration between ASEAN member countries should be the way forward to improve research activities of aquatic animal heath institutions in Thailand.

Keywords: aquatic animal health, Thailand, Department of Fisheries, OIE, biosecurity, shrimp viruses, koi herpesvirus

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Aquatic Animal Health Management in Vietnam Le Van Khoa Department of Animal Health, Ministry of Agriculture and Rural Development, Vietnam This paper describes issues pertaining to the status of aquaculture, structure of aquatic animal health management, and emerging diseases of aquatic animals in Vietnam. Vietnam is endowed with an abundant supply of water resources that are ideal for fisheries and aquaculture. It has 3,260 km long coastline, 112 estuaries, 1 million km2 of Exclusive Economic Zone (EEZ) and more than 4,000 islands which form many bays, straits and lagoons. The total aquaculture area which accounted 1,066,000 m2 produced 2,707,000 tons of aquaculture species, mainly Pangasius and shrimp (Penaeus monodon and Litopenaeus vannamei) approximated at 1.2 million tons and 0.45 million tons, respectively. Productivity of Pangasius has reached 350 tons per ha. Other important cultured species include hard clam, cobia, grouper, sweet snail, tilapia, snakehead and seaweeds. These aquaculture species are reared in different aquaculture systems including small scale family fish pond, rice-cum fish, traditional floating cages and intensive shrimp pond, and modern marine cages. The total export earnings from the fisheries and aquaculture sector in 2011 were over US$ 6 billion. The Aquatic Animal Health Management was formerly run by the National Fisheries Quality Assurance and Veterinary Drugs (NAFIQAVED). Since January 2008, with the merging of the Ministry of Fisheries (MoFi) and Ministry of Agriculture and Rural Development (MARD), functions of these agencies have been shifted to the Department of Animal Health (DAH). The National Agro Forestry and Fisheries Quality Assurance Department (NAFIQAD) and the Aquaculture Directorate (D-Fish) are responsible for some of the tasks. Details of both the advantages and disadvantages of the structural rearrangement will be presented in this paper. Vietnamese Government strongly claimed that aquaculture is a tool to attain food security and alleviate poverty as well as foreign currency return. Species diversification and intensification have however resulted in disease outbreaks and water pollution that caused great losses along with the increase in production. White Spot Disease has been a major disease that caused serious losses in P. monodon and L. vannamei cultured in most of the principal production areas (southern provinces) of the country. Mortality accounts approximately 20% (varies from 10 – 30%) annually. The disease occurs mainly in extensive and improved extensive system due to lack of pond management practices and low quality seed. In April 2011, serious shrimp mortalities reaching up to 65% in all aquaculture scales, i.e. intensive, semi-intensive and improved-extensive shrimp farms, were encountered in 6 major aquaculture provinces in Mekong delta. Clinical signs of the early stages of mortality were almost not clear. Black tiger shrimp P. monodon and white leg shrimp L. vannamei respectively died at the age of 20-30 days post stocking (35-45 day old) and 30-35 days (45-50 day old). Shrimp showed slow growth rate, swollen and soft liver, loose shells, and discoloration. In some cases, diseased shrimps showed atrophied hepatopancreas and occasionally white spot on shells combined with signs of hepatopancreatic necrosis.   Necrotizing hepatopancreatitis (NHP), Spiroplasma penaei and Enterocytozoon hepatopenaei (microsporidian) were not detected by PCR. No lesions diagnostic of infection by the NHP-bacteria or by microsporidians were detected by histological method. Initial epidemiological investigation showed that the problem occurred 3-4 years ago, caused death in bigger size shrimps weighing 10-15g, and shrimp mortalities occurred in several localities but did not spread widely. So far, the causative agent of the disease is still unknown. Other diseases that pose some problems to Vietnam aquaculture include Monodon Baculovirus (MBV), Milky Lobster Disease, Enteric Septicemia in Pangasius, Streptococosis in Tilapia, and Viral nervous necrosis (VNN) in marine species.

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The government of Vietnam has been supportive in implementing a comprehensive health management strategy in order to sustain the rapidly growing aquaculture sector. Developing a simple and practical national aquatic animal health management surveillance system that effectively utilizes the existing resources of national expertise is being prioritized. Thus, key elements considered are (a) Country list of aquatic animal diseases; (b) National level surveillance system; (c) Regional and international reporting; (d) Emergency responses; (e) Health certification for domestic and international movement of aquatic animals; (f) Drug management; (g) Hygienic practices; (h) Capacity building; (i) Legal framework; and (j) Contributions to the international standards (e.g. OIE code and diagnostic manuals). In the long run, DAH with support from a team of OIE experts has carried out a “Performing of Veterinary Services (PVS)� study on aquatic animal health in Vietnam. The results have been wealthily assisted to develop a national strategy for five year plan (2011 to 2015) focusing on the following objectives/issues: (1) to build up human capacities for professional and technical staff on aquatic animal health; (2) to improve technical authority and capability on laboratory diagnosis, risk analysis, quarantine and inspection, epidemiology, control and warning; (3) to strengthen cooperation between DAH and other national institutions/organizations such as NAFIQAD and D-Fish on aquatic animal health management; and (4) to strengthen international cooperation and private sector involvements on reaction to new/emerging diseases and trade barriers.

Keywords: aquatic animal health management, Vietnam, Department of Animal Health, surveillance, shrimp and fish diseases

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Abstracts Of Oral Presentations

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Fish Health Management on Food Safety Control in the Philippines Simeona E. Regidor*, Joselito R. Somga, Sonia S. Somga, Martha C. L. Funtillar, Romualdo R. Balagapo II Fish Health and Quality Assurance Section, Bureau of Fisheries and Aquatic Resources, 860 Quezon Avenue, Quezon city, Philippines (*simeona03@yahoo.com) Fish health management activities are now taking a bigger role in the determination of safety of the cultured fish and fishery products. There is now a shift of direction in the culture from maximum production to sustainable development considering the effect of aquaculture on the environment, welfare and health of the fish. Most often inputs like pro-biotics, vitamins, and immune enhancers are added to maintain the health of the fish until harvest. Chemicals and veterinary medicines are also used to prevent or cure diseases, like antibiotics, antiparasitic, antifungal, disinfectants and vaccines. These inputs sometimes leave harmful residues in meat even after harvest. Residues in meat which cannot be removed by processing most often are food safety issues. Hence, nowadays trading partners requires certification that farm to fork food safety programs are being judiciously implemented by trading partner. Documentation and procedures for inspection certification and laboratory analysis must follow international standards. In the Philippines, the National residues Control Program are handled by the Fish Health Management and Quality Assurance Section. The Section is also responsible for the disease monitoring and surveillance for disease occurrences and reporting system. This document will provide the summary of the food safety program at the primary production, results of analysis for the residues in fish and shrimp meat and report on parasitic diseases recently encountered.

Keywords: fish health management, food safety, National Residues Control Program, Quality Assurance

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Surveillance of Parasite Fauna of Economically Important Freshwater Fish in Some Southeast Asian Countries Gregoria Erazo-Pagador*, Rolando V. Pakingking Jr Aquaculture Department, Southeast Asian Fisheries Development Center, Tigbauan 5021, Iloilo, Philippines (email: gpagador@seafdec.org.ph). Fish-borne zoonotic parasites are a significant public health problem in Southeast Asia. In the present study, surveillance of fish-borne zoonotic parasites from various freshwater fish species was conducted in Myanmar, Lao PDR, and Philippines. Specifically, the presence of fish-borne trematode (FZT) in fish samples was examined using the muscle compression and artificial pepsin digestion methods. A total of 60 freshwater fish species including rohu (Labeo rohita), catla (Catla catla), carp (Cyprinus carpio), and catfish (Pangasius sutchi) were collected from earthen ponds in Myanmar in 2010. Only catla were positive for the presence of FZT metacercaria, specifically Clonorchis sp., with a prevalence rate of 6.5%. The surveillance on fish borne zoonotic parasites in freshwater fish species was also conducted in Lao PDR and the Philippines in 2011. A total of 315 tilapia (Oreochromis spp.) samples collected from earthen ponds in Region II (Isabela), Region III (Nueva Ecija, Pamapanga, Bulacan), Region IV (Batangas, Laguna), and Region VI (Negros Occidental) were negative for FZT. In addition, freshwater fish species including silver bard (Puntius gonionotus) and tilapia (Tilapia niloticus) obtained from the local market (n=40) and catfish (Clarias spp.), common carp (Cyprinus carpio), silver carp (Hypoththalmicthys molitrix), and grass carp (Ctenopharyngodon idellus) obtained from earthen ponds (n=80) in Lao PDR were also examined for FZT. An unidentified FZT metacercariae was present at low prevalence (2.5%) in silver barb from the local market. On contrary, no FZT metacercariae were recovered in all fish samples collected from different fish ponds.

Keywords: fish-borne zoonotic parasite, surveillance, freshwater fish, Southeast Asia

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Impact of Fish Vaccination for Fish Health Management: Examples from Pangasius in Vietnam Vo Thanh Tung1*, Pham Cong Thanh1, Tu Thanh Dung2, Kjersti Gravningen1 PHARMAQ Vietnam Ltd., 2 Fisheries Department of Can Tho University, Vietnam (*Vo-thanh.tung@pharmaq.no; pham-cong.thanh@pharmaq.no; ttdung@ctu.edu.vn; kjersti. gravningen@pharmaq.no) 1

During the last decade, pangasius farming in Vietnam has grown from a small industry to one of the biggest suppliers of white fish in the global seafood market. The annual production of pangasius in Vietnam was 1.1 million metric tons in 2010. Fish health management is an important tool for sustainable fish farming. Fish vaccines are an important tool in prophylactic health management. PHARMAQ AS is the world’s largest fish vaccine supplier. Based on the experience with salmon, PHARMAQ initiated development of vaccines for pangasius in 2006. The formalin inactivated oil-based injection vaccine against Edwardsiella ictaluri for pangasius, ALPHA JECT Panga1, has been developed. Our laboratory study with pangasius fingerlings (14 ¹ 1 g) showed good and long lasting protection after they were challenged with E. ictaluri at 2, 4, 10 and 20 weeks post-vaccination. In addition, significantly high levels of antibodies were detected in the sera of these vaccinated fish by direct agglutination. The result of our laboratory experiment was further confirmed in a field trial conducted at 3 different farms. A total of 360,000 fingerlings (28-58g) were injected with the vaccine. The same number of fish not injected with the vaccine was kept in the same pond conditions to serve as control. Data on vaccine safety was determined in at least 30 fish collected every 10 days within the first 60 days, and thereafter every 30 days within the 170-day period of the field trial. Results of the field trial revealed that the vaccine did not induce any negative effect on the growth of vaccinated fish. In addition to the above results, the impact and advantages of fish vaccination to the environment and food safety will also be presented.

Keywords: vaccination, pangasius, Edwardsiella ictaluri, PHARMAQ, ALPHA JECT Panga1, field trial

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Silencing White Spot Syndrome Virus and Elucidation of Shrimp-Virus Interaction by RNA Interference Technology Rod Russel R. Alenton,1 Jassy Mary S. Lazarte,1,4 Hidehiro Kondo,5 Ikuo Hirono,5 Mary Beth B. Maningas1,2,3 The Graduate School, University of Santo Tomas, Manila, Philippines College of Science Biological Sciences, University of Santo Tomas, Manila, Philippines 3 Research Center for the Natural and Applied Sciences, University of Santo Tomas, Manila, Philippines 4 University of the Philippines, Manila, Philippines 5 Tokyo University of Marine Sciences and Technology, Japan (email: marybethmaningas@yahoo.com/mbmaningas@mnl.ust.edu.ph) 1

2

White Spot Syndrome Virus (WSSV) remains the most widespread and devastating infectious agent that hit the shrimp aquaculture industry worldwide. To date, there are no available cost-effective remedies yet for WSSV infection. Hence, functional studies on genes critical for viral infection, is essential in elucidating shrimp-virus interaction. Here we report two genes: one from WSSV Viral Protein 9 (VP9), a newly identified nonstructural protein suspected to be involved in viral transcription, and contig-23, a contiguous sequence first identified in Marsupenaeus japonicum that was found to have high homology with gene sequences of WSSV. This study therefore utilized a gene knock-down technology through RNA interference (RNAi), to elucidate the functions of VP9 and contig-23 in shrimp-virus interaction, using both viral (VP9) and host gene (Contig 23). Twenty-two (22) Macrobrachium rosenbergii daqueti shrimps were injected for each treatment of VP9-, Contig 23-, GFP-dsRNA, and PBS (Phospahte buffered saline). After 24 hrs, shrimps were challenge with WSSV and survival rate was recorded. Three (3) shrimps were sampled on 0, 1, 3, and 7 days post-infection (d.p.i) for gene expression analysis by RT-PCR. The VP9-, c23-, and GFP-dsRNA injected shrimps showed a significant survival rate of 80% ,100%, and 100%, respectively compared to that of the PBS injected shrimp which decreased to 20% after day 14 post infection (pi). Silencing of VP9 and contig-23 genes was observed on day 0 and 3 pi, respectively, followed by the suppression of viral load starting 1 d.p.i. The significant increase of survival rate of dsRNA treated shrimps underscores the critical function of VP9 and contig 23 in WSSV infection. It further indicates that VP9, a full-length nonstructural protein, play a key-role in viral replication. Moreover, contig 23 which is a host gene homologous to WSSV, is indeed involved in shrimp-virus interaction. It would be interesting to study the potency of this gene silencing technology and assess its application in the management of WSSV infection in the shrimp aquaculture industry.

Keywords: white spot syndrome virus, shrimp-virus interaction, RNA interference technology, Macrobrachium rosenbergii, viral protein 9, contig-23

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Development of Loop-Mediated Isothermal Amplification (Lamp) Assay for the Detection of Philippine Isolate of the Penaeus monodon-Type White Spot Syndrome Virus (WSSV) May Flor J. Sibonga1, Christopher Marlowe A. Caipang2, Jane S. Geduspan3, Carlo C. Lazado2, Jumelee L. Moleta1 National Institute of Molecular Biology and Biotechnology, University of the Philippines Visayas, Miag-ao 5023, Iloilo, Philippines 2 Faculty of Biosciences and Aquaculture, University of Nordland, Bodø 8049, Norway 3 College of Arts and Sciences, University of the Philippines Visayas, Miag-ao 5023, Iloilo, Philippines (email: mayz_cinth@yahoo.com) 1

White spot syndrome virus (WSSV) is one of the major viral pathogens in shrimp, particularly Penaeus monodon. The speed and accuracy of detection of the pathogen especially at an early stage of infection will ensure effective management procedures are undertaken. In this study, loop-mediated isothermal amplification (LAMP), a novel method of nucleic acid amplification with high degree of sensitivity, specificity and rapidity was developed for detection of Philippine isolate of WSSV. The assay is performed under isothermal condition with a set of four primers which was designed based on WSSV endonuclease gene. Amplification of the target DNA was visualized on an agarose gel to detect the presence of laddering bands. Another way to determine the presence of LAMP amplicon is by addition of ethidium bromide to the reaction tube and visualization under UV light after staining. The optimized time and temperature conditions for the LAMP assay were 60 minutes at 650C respectively. The WSSV LAMP primers were highly specific in detecting WSSV in DNA samples and did not gave positive results for other pathogens including monodon baculovirus (MBV), infectious hypodermal and hematopoietic necrosis virus (IHHNV) and hepatopancreatic parvovirus (HPV). The detection sensitivity of LAMP was 5pg of DNA ml-1 or 5 fg of viral DNA per LAMP reaction. This sensitivity threshold was 10 times more sensitive than conventional PCR in detecting WSSV from infected shrimp. Due to its high degree of sensitivity, accuracy and speed of assay, LAMP has a tremendous potential for field use particularly in the routine diagnosis of WSSV infections in shrimp aquaculture in the Philippines.

Keywords: loop-mediated isothermal amplification, LAMP, Penaeus monodon, white spot syndrome virus, WSSV

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Loop-Mediated Isothermal Amplification (LAMP) Detection of White Spot Syndrome Virus (WSSV) in Litopenaeus vannamei in Selected Sites of the Philippines Amalea Dulcene D. Nicolasora1, Christopher Marlowe Caipang2, Benedict A. Maralit3, Paul F. Cordero1, Mudjekeewis D. Santos3, Mary Beth B. Maningas1* Research Center for the Natural and Applied Sciences, Thomas Aquinas Research Complex/ Biological Sciences Department, College of Science, University of Santo Tomas, Philippines 1008 2 Faculty of Biosciences and Aquaculture, University of Nordland, Bodø 8049, Norway 3 National Fisheries Research and Development Institute, Quezon Avenue, Quezon City Philippines 1103 (*email: marybethmaningas@yahoo.com) 1

White spot syndrome virus (WSSV) is the major culprit in the decline of shrimp production in the Philippines. The uncontrollable occurrence of this disease is the main problem of the aquaculture industry today. Rapid diagnosis at the aquaculture site is crucial to enable timely treatment that will mitigate possible heavy mortality and spread of WSSV. PCR technique is commonly used in WSSV detection however the method is found to be expensive and inaccessible to small shrimp farmers. Thus, there is a need for a development of a practical alternative that will benefit the aquaculture industry. Loop-mediated isothermal amplification is a novel technique that proves to be cheaper and more sensitive than the traditional PCR. It is an assay that amplifies nucleic acid with high specificity under isothermal condition. A single LAMP assay can produce approximately 109 copies of target nucleic acid at 60° - 65° C within an hour. This study, thus aims to investigate the occurrence of WSSV in Litopenaeus vannamei from Samar, Batangas and Zambales using LAMP. Likewise, this study also aims to compare LAMP protocol with PCR method for further verification of the efficiency of LAMP in shrimp pathogen detection. Using the set of primers designed from the conserved region of WSSV, the LAMP protocol demonstrated that L. vannamei from Samar, Batangas and Zambales are WSSVinfected. Conventional PCR resulted in the amplification of a 200-bp amplicon, thus confirming the detection of WSSV. Results showed that LAMP method can serve as a practical alternative to the use of expensive PCR method in detecting shrimp pathogens, including WSSV.

Keywords: loop-mediated isothermal amplification, LAMP, white spot syndrome virus, WSSV, Litopenaeus vannamei

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Sandfish (Holothuria scabra): a Potential Carrier of White Spot Syndrome Virus Leobert D. de la Pe単a, Dieyna B. Caber and Satoshi Watanabe Aquaculture Department, Southeast Asian Fisheries Development Center, TIgbauan 5021, Iloilo, Philippines White spot syndrome virus (WSSV) outbreak in black tiger shrimp Penaeus monodon polycultured with sandfish Holuthoria scabra was encountered during the experimental run in the grow-out phase. To know the potential of sandfish as WSSV carrier, cohabitation trials were conducted. The presence of WSSV was determined using polymerase chain reaction (PCR) methodology on DNA extracted from the gills of black tiger shrimp and coelom, intestine, tentacle and muscle of sandfish. Before the cohabitation trials, both shrimp and sandfish were confirmed to be negative of WSSV by PCR. To be able to infect the shrimp to be used in the first cohabitation, all were fed with WSSV infected tissues twice a day for three days with a feed rate of 3% body weight. For the first cohabitation, infected shrimp cohabited with sandfish on the fourth day until all the infected shrimp died on the seventh day. Then, the contaminated sandfish from the first cohabitation were transferred to another tank with another group of shrimp for the second cohabitation. In the first cohabitation, the infected shrimp were confirmed to be one-step positive for WSSV. During the second cohabitation, both the shrimp and the sandfish were sampled for the detection of WSSV using PCR every 6 d from 0 d to 25 d. During the sampling at 0 h, all of the shrimp were found one-step positive while most of the sandfish were found nested positive for WSSV. On the 6th d of sampling, all shrimp and only 1 sandfish were nested positive for WSSV. On the 12th day of sampling, majority of the shrimp were nested positive and none of the sandfish. On the 18th day, the shrimp were all negative and one sandfish became nested positive for WSSV. On the 25th day, there were no shrimp left due to mortality and only a few of the sandfish were nested positive for WSSV. Based from these results, it was elucidated that the virus is viable in the sandfish body due to the consistent nested positive results in the shrimp during the second cohabitation. Therefore, sandfish can be a potential carrier of WSSV only for a limited period of time.

Key words: white spot syndrome virus (WSSV), polyculture, PCR, cohabitation, sandfish

33


Development of Practical Shrimp Vaccination Techniques Against White Spot Syndrome Virus in Penaeus monodon: Progress of Tank and Pond Studies Edgar C. Amar*, Joseph P. Faisan, Jr., Rolando S. Gapasin, Jocelyn M. Ladja Aquaculture Department, Southeast Asian Fisheries Development Center, TIgbauan 5021, Iloilo, Philippines (email:  eamar@seafdec.org.ph) A safe, effective, and inexpensive vaccine capable of inducing protective immunity is required to limit the impact of WSSV and other shrimp viruses. This study was conducted to develop practical shrimp vaccination technologies applicable to pond grow-out conditions. Vaccination with inactivated white spot syndrome virus (WSSV) was tested in a field efficacy trial. Growth and survival in brackish water ponds were markedly improved with the use of the inactivated virus when administered orally through the feed every 15 days. A vaccine was subsequently produced using recombinant techniques. VP28 gene from a Philippine strain of WSSV was PCR-amplified, cloned in TOPO vector and sequenced. TOP10 cells were transformed with a heat shock technique and were used to store the recombinant plasmid until ready for expression, whereas BL21 cells were used to produce the VP28 inclusion bodies and protein. Prior to the experiment with carriers, inactivated recombinant BL21 cells, inactivated untransformed BL21 cells and PBS were injected into Penaeus monodon juveniles to determine the protective effect of the recombinant protein present in inactivated bacteria. Following the procedure in previous trials, plasmids containing the VP28 inserts as well as VP28 inclusion bodies were successfully produced from IPTG-induced BL21 cells. Experimental challenge showed that cumulative mortality recorded daily for 10 days was markedly lower for the inactivated recombinant BL21 cells consequently eliciting a significantly higher RPS when compared to the other treatments. A practical delivery system for recombinant VP28 vaccine by encapsulation in microparticle carriers such as liposome, alginate, and chitosan is being developed. Encapsulation with carriers may improve immunogenicity of the vaccine but a non-encapsulated form resistant to degradation might also be used.

Keywords: vaccine, Penaeus monodon, white spot syndrome virus, WSSV, field trial

34


Establishment of an Immunization Regimen for the Prevention of Viral Nervous Necrosis (VNN) in Asian Sea Bass (Lates calcarifer) Broodfish Rolando Pakingking Jr.*, Ofelia Reyes and Evelyn Grace de Jesus-Ayson Aquaculture Department, Southeast Asian Fisheries Development Center, Tigbauan 5021, Iloilo, Philippines Outbreaks of viral nervous necrosis (VNN) in Asian sea bass (Lates calcarifer) larvae remain as a major deterrent in the successful production of this fish species in hatcheries. We previously demonstrated that subsequent exposure of sea bass juveniles earlier vaccinated with the formalininactivated Philippine strain of nervous necrosis virus (NNV) to the homologous virus effectively propelled the upregulation of anamnestic responses as evidenced by the reversion of low titer levels of NNV-neutralizing antibodies (NNV-Nab) to high levels and abrogation of NNV replication in the sera and brains of these fish, respectively, indicating the feasibility of using the inactivated-NNV vaccine in establishing an immunization regimen against VNN in sea bass broodfish. In this study, sea bass juveniles with a mean body weight (MBW) of 5 g were primarily injected (intraperitoneal) with the above vaccine and the NNV-Nab titers in the sera of these fish were monitored at scheduled intervals. Fish injected with L-15 medium served as control. NNV-Nab titers in vaccinated fish peaked at 2 months (titer: 1:4480) but thereafter declined and significantly dropped (titer: 1:380) at 12 months post-vaccination. Booster injection and vaccination respectively of unvaccinated and vaccinated sea bass with L-15 and inactivated-vaccine followed by monitoring of NNV-Nab titers in the sera of these fish were similarly conducted 12 months after the 1st booster vaccination. Similarly, NNV-Nab titers in the sera of booster vaccinated (BVac) sea bass suddenly increased at 1 month (1:12800), persisted until 3 months (1:11520) but thereafter gradually declined and significantly dropped at 12 months (1:480) post-BVac. The annual booster vaccination and L-15 injection were continued successively for 4 years until some of these fish became sexually matured. To test the protective effects of NNV-Nab in BVac fish, matured fish from both groups were induced to spawn using luteinizing hormone releasing hormone at a ratio of 1 female to 2 males. Spawned eggs from both BVac and UVac fish were subjected to NNV detection by RT-PCR amplification and cell culture isolation. Although both samples were negative for NNV by cell culture isolation, eggs from UVac fish were RT-PCR positive (nested). Furthermore, eggs from BVac and UVac broodfish subjected to seroneutralization assay showed neutralizing antibody titers of 1:192 and <1: 40, respectively, suggesting that our current immunization regimen is a practical approach to prevent the horizontal transmission of NNV to broodfish and vertically to their offspring.

Keywords: immunization regimen, viral nervous necrosis, VNN, sea bass, broodstock, neutralizing antibodies, titer

35


Cloning, Characterization and Expression Analysis of a cDNA Encoding Granulin Gene in Nile tilapia (Oreochromis niloticus, Linn.) Myat Khine Mar Laboratory of Aquatic Animal Health Management, Department of Aquaculture, Faculty of Fisheries, Kasetsart University, Bangkok 10900, Thailand Granulins are a group of highly conserved growth factors that have been described from variety of organisms spanning the metazoan. It has been shown to have roles in multiple processes involved in cell growth, development of inflammation, wound healing and tissue remodeling. In the present study, the full-length cDNA encoding Nile tilapia granulin gene was cloned by in silico cloning and 5’Rapid Amplification Complementary DNA Ends (RACE) techniques. The complete cDNA sequence of this gene consisted of 1,270 bp containing an open reading frame (ORF) of 618 bp (206 amino acid residues), including 44 and 608 bp of 5’ and 3’ untranslated regions, respectively. Amino acid sequence analysis showed that the Nile tilapia granulin had two conserved-cystein granulin motifs CX5CX5CCX8CCX6CCX5CCX5CX6C similar to those of vertebrates. Amino acid and nucleotide sequences alignment indicated that Nile tilapia granulin was closely similar to Mozambique granulin I and II. Phylogenetic analysis strongly supported that Nile tilapia granulin was clustered in the tiny sister group of vertebrate granulin genes closed to Mozambique granulins. Expression analysis using RT-PCR revealed that the highest expression levels of granulin mRNAs were present in tissues obtained from head kidney, and moderate expression levels in trunk kidney, spleen and liver. Quantitative real-time PCR revealed that fish injected with Streptococcus agalactiae could clearly enhance the up-regulation levels of granulin mRNAs in head kidney, especially at hour 24 after injection.

Keywords: cloning, granulin gene, tilapia, Oreochromis mosambicus, head kidney

36


Lectins as Non-Self Recognition Molecules and Their Relevance in Innate Immunity and Fish Health Anacleto M. Argayosa Institute of Biology, College of Science, University of the Philippines, Diliman, Quezon City 1101 Non-self recognition molecules such as fucose and mannose-binding lectins were isolated from the serum of cultured teleosts species such as Nile tilapia (Oreochromis niloticus), African catfish (Clarias gariepinus), milkfish (Chanos chanos), and orange-spotted-grouper (Epinephelous coioides). Physico-chemical and biological activities of the lectins were determined. Protein subunit molecular weights range from 20-40 kDa while native multimeric size seems to occur above 200 kDa. Isoelectric points obtained show slightly acidic charges from 5-6 pI. Ruthenium red staining of protein bands indicates calcium-binding properties. The ability of lectins from Nile tilapia, African catfish, milkfish, and orange-spotted grouper to agglutinate fish bacterial pathogen Aeromonas hydrophila (BIOTECH 10089) and yeast (Candida albicans) were shown in this study. Agglutination of O. niloticus on human erythrocytes showed specificity towards blood type O. The N-terminal sequences of the fucose binding lectins were obtained and searched for alignments in the protein databases using basic local search alignment tool (BLAST). Alignments of the 23 kDa fucose-binding protein (TFBP23) with the zebra fish interferon-inducible antiviral protein and immunoglobulin light chain were obtained. Alignments with complement protein member (C1q) cDNA of the surfperch (Neoditrema ransonnetii) with O. niloticus fucose-binding properties were also found. Lectin probes comprised of horseradish peroxidase-labeled neoglycoproteins were developed to detect lectins from various tissues of Nile tilapia. Results showed that liver, kidney, spleen, and gut express high levels of the fucose-binding proteins. Identification of gut fucose-binding proteins relevant to mucosal immunity is being undertaken. These studies highlight the isolation and characterization of lectins from the economically important teleosts which could serve as a biomarker in innate immunity for fish health management.

Keywords: lectins, fucose-binding protein, tilapia, Oreochromis niloticus

37


Aptamer Technology for Aquaculture Mudjekeewis D. Santos1*, Ikuo Hirono2, Takashi Aoki2 Genetic Fingerprinting Laboratory, National Fisheries Research and Development Institute, 940 Kayumanggi Bldg., Quezon Ave., Quezon City 1103, Philippines 2 Laboratory of Genome Science, Tokyo University of Marine Science and Technology, Konan 4-5-7, Minato-ku, Tokyo 108-8477 Japan

1

Aptamer technology is a new and promising method for diagnostics and therapeutics. It revolves around the use of an aptamer molecule, an artificial ligand (nucleic acid or protein), which has the capacity to recognize target molecules with high affinity and specificity. Here, we review the existing applications for aptamer technology in the biomedical and life sciences field and show some possible applications to aquaculture, particularly in fish immunology and health management. We likewise present our study on the selection of RNA aptamers using the Systematic Evolution of Ligands by Exponential Enrichment (SELEX), which can specifically bind to and inhibit virulence of a fish Viral hemorrhagic septicemia virus (VHSV) in vitro.

Keywords: aptamer, RNA, ligand, fish immunology, SELEX, VHSV

38


Isolation of Dehalogenase Producing Bacteria from the Gut of Pond-Reared Rohu (Labeo rohita) Juveniles in Myanmar Fahrul Huyop1*, Rolando Pakingking Jr.2, Eleanor Abel1, Gregoria Pagador2, May Thanda Wint3 Faculty of Biosciences and Bioengineering, Universiti Teknologi Malaysia, 81310 Johor Bahru, Malaysia 2 Aquaculture Department, Southeast Asian Fisheries Development Center, Tigbauan 5021, Iloilo, Philippines 3 Fish Health Laboratory, Department of Fisheries, Yangoon, Myanmar (Email: fzhutm@gmail.com; fahrul@fbb.utm.my) 1

Unwarranted accumulation of halogenated compounds in the rivers and streams has arisen in recent years due to the widespread use of agricultural pesticides. The presence of these halogenated compounds in the water does not only suppress the immune system of fish but adversely induce serious morbidity and mortality among cultured stocks. Importantly, gradual accumulation of these compounds in the system of cultured and wild freshwater fish species reared in ponds and floating netcages in dams and rivers, respectively, poses threats to humans, the end users. So far, microorganisms capable of producing dehalogenases were mainly isolated from the soil and scarcely from aquatic animals and their environments. In this study, we isolated eight bacterial strains from the gut of pond-reared rohu (Labeo rohita) juveniles in Myanmar, identified these bacterial strains by biochemical test and 16s rDNA sequencing, screened the isolated bacteria for the presence of dehalogenase gene using oligonucleotide primers designed from group I dehalogenase, and subsequently tested the ability of these isolated bacteria to degrade a halogenated compound, 2,2-dichloropropionic acid (2,2DCP), in vitro. The eight bacterial strains studied were identified as Enterobacter mori (strain: MK121001), Enterobacter cloacae (MK121003, MK121004, MK121010), Ralstonia solanacearum (MK121002), Acinetobacter baumannii (MK121007), Chromobacterium violaceum (MK121009) and Pantoea vagans (MK121011). Although dehalogenase gene was amplified in all of the bacterial strains tested, only Ralstonia solanacearum (MK121002), Acinetobacter baumannii (MK121007) and Chromobacterium violaceum (MK121009) were able to degrade 2,2-dichloropropionic acid (2,2DCP) in vitro, indicating that the putative dehalogenase gene in E. mori (MK121001), E. cloacae (MK121003, MK121004, MK121010) and Pantoea vagans (MK121011) were silenced. To our knowledge, this is the first report on the isolation of dehalogenase-producing bacteria from the gut of pond-reared freshwater fish, L. rohita, in Myanmar.

Keywords: dehalogenase, bacterial isolation, rohu, Labeo rohita

39


Early Warning Activities Prevent Disease Outbreak and High Mortality Impact of Overturn on Fish Culture in Jatiluhur Dam, Indonesia Heny Budi Utari1,3*, Hendi2, Supriyadi2, Maleeya Kruatrachue1, Timothy W. Flegel3,4 Department of Biology, Faculty of Science, Mahidol University, Rama VI Road, Bangkok, 10400, Thailand 2 Department of Aquatic Animal Health Laboratory, Aquabiotech and R&D Division, Central Proteinaprima, Jakarta, Indonesia 3 Center of Excellence for Shrimp Molecular Biology and Biotechnology (Centex Shrimp), Faculty of Science, Mahidol University, Rama VI Rd., Bangkok, 10400, Thailand 4 Department of Biotechnology, Faculty of Science, Mahidol University, Rama VII Rd., Bangkok 10400, Thailand (Email: heny.budi@gmail.com) 1

Early warning activities (EWA) was conducted in Jatiluhur dam, West Java province, Indonesia, after serious mortality happened in fish species cultured in floating net cages. Fish diseases and overturn were the main causes of the problem. The EWA was undertaken by routinely monitoring water quality, fish health, and feeding management. The generated data were rapidly distributed and presented to the fish farmers for field applications. Twenty percent of the 8,300 ha total area of Jatiluhur dam consists of 21,000 fish cages. The cages were set in three layers; the upper layer (1.5 m deep) were stocked with common carp and koi (Cyprinus carpio), the middle layer of 2.0 to 2.5 m deep were stocked with tilapia (Tilapia nilotica), while the deepest of 2.5 m (lower layer) were stocked with Catfish (Pangasius sp). Since 2007 estimated losses attributed to fish diseases and overturn was more than 12 million US dollars. Field applications of EWA resulted in a significant reduction of 5 and 10% respectively in the prevalence rate of koi herpes virus (KHV-type2) and bacterial infections due to Aeromonas hydrophila and Streptococcus sp. and these positive results corresponded with the reduction of stocking density, disease treatment, installations of improvised aerators during disease outbreak, and appropriate waste disposal management. In addition, current data on EWA indicate that fish mortality due to diseases and overturn in Jatiluhur dam could be avoided through reduction in the number of floating fish cages.

Keywords: early warning activities, prevention, water quality, fish health, feeding management, koi herpes virus, Aeromonas hydrophila, Streptococcus sp.

40


List of Poster Presentations P1

Diseases of Marine Fishes Cultured in Floating Cages in Brunei Darussalam Wanidawati Tamat, Dayangku Siti Norhaziyah Pengiran Haji Halim, and Celia R. Lavilla-Pitogo

P2

Detection of Vibrio harveyi in Asian seabass, Lates calcarifer by Conventional PCR CMA Caipang, RV Pakingking Jr., CC Lazado, MJA Amar

P3

Aqueous Extracts from Skin of Tilapia, Oreochromis sp. Inhibit Luminous Vibrio CC Lazado, CMA Caipang, CJ Jaspe

P4

Molecular Analysis and Detection of the Philippine Isolate of Monodon Baculovirus (Mbv) Mary Jane S. Apines-Amar, Christopher Marlowe A. Caipang, Carlo C. Lazado, May Flor Sibonga, Jane Geduspan

P5

Inactivated Escherichia Coli Expressing Recombinant Viral Envelope Protein VP28 from a Philippine Isolate of White Spot Syndrome Virus (WSSV): a Potential ‘Vaccine’ Against WSSV Infection in Black Tiger Shrimp Peneaus monodon Edgar C. Amar, Joseph P. Faisan, Jr.

P6

Antibacterial Activity of Extracts from Panay Seaweed (Ulva pertusa Kjellman) Rolando Pakingking Jr, Anne Jinky Villacastin, Gregoria Pagador

P7

Effect of Greenwater Technology on Water Quality and Occurrence of White Spot Syndrome Virus (WSSV) in Pond Cultured Penaeus monodon Eleonor A. Tendencia, Roel H. Bosma, Marc C. Verdegem, Johan A.J. Verreth

P8

Detection of Luminous Vibrio and White Spot Syndrome Virus in Shrimp by Duplex PCR Christopher M.A. Caipang, Mary Paz N. Aguana, Carlo C. Lazado

P9

Parasitic and Shell Diseases of Abalone (Haliotis asinina) in the Philippines Gregoria Erazo-Pagador, Vincent Encena ll, Rolando V. Pakingking Jr

P10

Can We Use the Autochthonous Bacteria of the Gastrointestinal Tract of Atlantic cod Gadus morhua as Probiotics? Carlo C. Lazado and Christopher Marlowe A. Caipang

P11

Bacterial Diversity and Algal Community Structure in Biofilms of Settlement Plates for Abalone Haliotis asinina Larvae Leobert D. de la Peña, Celia Lavilla-Pitogo, Demy D. Catedral, Milagros R. de la Peña, Nestor C. Bayona, Ellen Grace T. Tisuela

P12

Status and Needs of Primary Aquatic Animal Health Care Small-Scale Aquaculture Edgar C. Amar, Rolando PakingKing Jr., Eleanor Tendencia, Leobert dela Peña, Gregoria Pagador

P13

Pathological Effects of Swine Effluents with Varying Levels of Some Physico- Chemical Properties on Bighead Carps (Aristhycthtys nobilis, Richardson 1845) Rosalina R. Atos, Benjamin Reuel G. Marte, Francis Andrew Eugene M. Bernardo, Joseph S. Masangkay

P14

Potential of Sargassum as Antibacterial Agent and as Immunostimulant for Crustacean and Finfish Aquaculture Francis N. Baleta, Liberato V. Laureta, Mary Jane Pines-Amar, Philip Ian P. Padilla, Gerald F. Quinitio

41


Abstracts Of Poster Presentations

42


Diseases of Marine Fishes Cultured in Floating Cages in Brunei Darussalam Wanidawati Tamat1, Dayangku Siti Norhaziyah Pengiran Haji Halim1*, Celia R. Lavilla-Pitogo2 Aquatic Animal Health Services Centre, Department of Fisheries, Ministry of Industry and Primary Resources, Jalan Menteri Besar, BB3910Brunei Darussalam 2 Integrated Aquaculture International, Block A Bangunan Lim Seng Kok, Simpang 628, Jalan Tutong, Bandar Seri Begawan BF 1120, Brunei Darussalam (*email: haziyah.h@gmail.com ) 1

Culture of marine fish in floating cages in Brunei is largely based on high value commodities like sea bass (Lates calcarifer), several species of groupers (Epinephelus spp.), snapper (Lutjanus sp.), cobia (Rachycentron sp.) and varieties of saline tilapia. Juveniles and fingerlings for cage culture are mostly imported from Malaysia, Taiwan, Thailand and the Philippines thus highlighting the need for health certification prior to their importation and upon arrival. In addition, guidelines are provided for setting up of offshore cages based on physico-chemical and other factors. During the holding period and nursery of juveniles and fingerlings, significant disease problems that have been encountered are mostly parasitic in nature. Infestation with trichodinid protozoans and monogeneans have caused serious problems during culture and considered the most commonly encountered disease problems. In grow-out culture, infestation with monogeneans, leeches and parasitic copepods are significant problems. Various management and husbandry problems have also resulted to mass mortalities. These problems threaten the success and profitability of cage culture operations and the need to implement a disease surveillance system similar to the one established for shrimp is being planned for implementation to make finfish aquaculture more successful and sustainable.

Keywords: marine fishes, cage culture, parasites, disease

43


Detection of Vibrio harveyi in Asian seabass, Lates calcarifer by PCR Christopher M.A. Caipang1, Rolando V. Pakingking Jr.2, Carlo C. Lazado1, Mary Jane A. Amar3 Faculty of Biosciences and Aquaculture, University of Nordland, Bodø 8049, Norway 2 Aquaculture Department, Southeast Asian Fisheries Development Center, Tigbauan 5021, Iloilo, Philippines 3 Institute of Aquaculture, College of Fisheries and Ocean Sciences, University of the PhilippinesVisayas, Miag-ao 5023, Iloilo, Philippines 1

Partial sequence of the dnaJ gene of Vibrio harveyi that was isolated from diseased juvenile Asian seabass, Lates calcarifer was identified. The dnaJ gene fragment of V. harveyi was 447 bp and shared at least 80% identity at the nucleotide level with the dnaJ gene of other Vibrios. It was closely related with the dnaJ gene of V. rotiferianus and V. campbellii having at least 90% nucleotide identity. PCR primers targeting this gene were designed to detect the pathogen in Asian seabass. The assay was highly specific to V. harveyi and the limit of detection was 100 pg of genomic DNA ml-1 or 100 fg of bacterial genomic DNA in a PCR reaction. This corresponded to a sensitivity of approximately 20 genome equivalents (GE) of the bacterial pathogen. Taken together our results indicate that the dnaJ gene is a good candidate to develop primers for the PCR assay in detecting V. harveyi in fish.

Keywords: PCR, Vibrio harveyi, dnaJ gene, Lates calcarifer, seabass

44


Aqueous Extracts from Skin of Tilapia, Oreochromis sp. Inhibit Luminous Vibrio Christopher M.A. Caipang1, Carlo C. Lazado1, Cecilia J. Jaspe2 Faculty of Biosciences and Aquaculture, University of Nordland, Bodø 8049, Norway 2 Institute of Aquaculture, College of Fisheries and Ocean Sciences, University of the Philippines Visayas, Miag-ao 5023, Iloilo, Philippines 1

The effects of aqueous skin extracts obtained from both high saline and freshwater tilapia, Oreochromis sp. on luminous Vibrio harveyi were conducted using the well-diffusion and co-incubation assays. Luminous V. harveyi was inhibited until the 10-2 dilution in aqueous extracts from both the freshwater and high saline tilapia strains, although higher frequency of inhibition was observed in the latter. Both skin extracts inhibited the number of luminous V. harveyi in co-incubation assays; however, the concentration used for the extracts from the high saline tilapia was one-hundred times lower than what was used from the freshwater strain. These results suggest that the aqueous extracts from the skin of tilapia from different environments could inhibit the growth the luminous Vibrio and the effects were more apparent in fish obtained from the high-saline rearing environments.

Keywords: luminous vibriosis, skin extracts, tilapia, Oreochromis sp.

45


Molecular Analysis and Detection of the Philippine Isolate of Monodon Baculovirus (Mbv) Mary Jane S. Apines-Amar1, Christopher Marlowe A. Caipang2, Carlo C. Lazado2, May Flor Sibonga1, Jane Geduspan1 University of the Philippines Visayas, Miag-ao 5023, Iloilo, Philippines 2 University of Nordland, 8049 Bodø, Norway

1

Monodon baculovirus (MBV) is a DNA virus that infects postlarvae and early juveniles of shrimp, Penaeus monodon. Nucleotide analysis of its genomic DNA revealed the presence of several variants of this virus. Partial genomic DNA sequences of a Philippine isolate of MBV was identified. The two partial sequences of the MBV genomic DNA showed 100% homology at the nucleotide level, indicating the occurrence of a single strain. It shared at least 87% homology of their nucleotides with the partial sequences of MBV isolates from Taiwan and India. The sequences of the genomic DNA from the Philippine isolates clustered together and were distinct from both the Taiwan and Indian isolates. To facilitate specific detection of the Philippine isolate of MBV, both the one-step PCR and the loop-mediated isothermal amplification (LAMP) assay for this viral pathogen was developed. The onestep PCR primers yielded an amplicon size of 193-bp. Its sensitivity was comparable to the published PCR assays. On the other hand, the LAMP assay was optimized at 60 min and 63oC. The sensitivity threshold of LAMP was 10 times more than conventional PCR in detecting MBV from infected shrimp. Both detection methods were highly specific for MBV and did not amplify other shrimp viruses. These assays could be used for regular monitoring and surveillance of MBV in shrimp as well as to trace the movement of the Philippine MBV in shrimp farms in different geographic regions. As a possible field application, both LAMP and conventional PCR were able to detect the virus from infected shrimp postlarvae. However, in practical terms, the LAMP assay is more advantageous for field use than conventional PCR because the former is easier and more sensitive in detecting the virus in shrimps during the early stages of infection.

Keywords: monodon baculovirus, Philippine isolate, one-step PCR, loop-mediated isothermal amplification, LAMP

46


Inactivated Escherichia Coli Expressing Recombinant Viral Envelope Protein VP28 from a Philippine Isolate of White Spot Syndrome Virus (WSSV): a Potential ‘Vaccine’ Against WSSV Infection in Black Tiger Shrimp Peneaus monodon Edgar C. Amar, Joseph P. Faisan, Jr. Aquaculture Department, Southeast Asian Fisheries Development Center, Tigbauan 5021, Iloilo, Philippines White spot syndrome virus (WSSV) contains at least five major virion proteins, one of which is the viral envelope protein VP28, which plays a key role in systemic WSSV infection. A recombinant VP28 was produced as a potential ‘vaccine’ to alleviate WSSV infection in Penaeus monodon. Genomic DNA was extracted from WSSV infected gills of shrimp collected from a local outbreak, the VP28 gene was amplified with primers designed from the VP28 open reading frame, and visualized by agarose gel electrophoresis. The purified PCR product was cloned in TOPO vector and the recombinant plasmid was used for transformation of Escherichia coli TOP10 competent cells. Ten clones were selected on Ampicillin-containing LB plates (100 µg ml-1), purified and screened by PCR using the above mentioned primers. Positive transformants subjected to sequencing and BLAST search displayed high homology with VP28 sequences from different WSSV strains submitted to the Genebank. Two clones (C6 and C7) showed 99% and 8 clones (C1, C2, C3, C4, C5, C8, C9 and C10) showed 100% homology with several Asian strains. E. coli BL21 (DE3) strain was then transformed with the C5 V28-harboring plasmid DNA and the expression was confirmed by SDS-PAGE analysis. The protein, expressed in positive transformants, showed a band size of 32 kDa. For preliminary trial, naïve black tiger shrimp juveniles were injected intramuscularly with inactivated VP28-expressing BL21 and untransformed E. coli as ‘vaccinated’ treatments and PBS only for ‘unvaccinated’ treatment. Three days post-‘vaccination’, the shrimp were challenged with active WSSV and cumulative mortality was recorded for 15 days. Lower cumulative mortality was observed in VP28-‘vaccinated’ treatment (60%) compared to E. coli (75%) and ‘unvaccinated’ treatment (85%). The results indicate that intramuscular application of the ‘vaccine’ induced a markedly higher resistance of shrimp against the virus. Thus, inactivated VP28expressing E. coli could be a potential effective ‘vaccine’ against WSSV.

Keywords: shrimp ‘vaccination’, inactivated recombinant Escherichia coli, VP28, WSSV, viral resistance, Penaeus monodon.

47


Antibacterial Activity of Extracts from Panay Seaweed (Ulva pertusa Kjellman) Rolando Pakingking Jr.*, Anne Jinky Villacastin, Gregoria Pagador Aquaculture Department, Southeast Asian Fisheries Development Center, Tigbauan 5021, Iloilo, Philippines (email: rpakingking@seafdec.org.ph) In the current study, the antibacterial activity of the crude extract of Panay seaweed (Ulva pertusa Kjellman) collected from floating fish net cages in Igang Marine Station, Guimaras, were tested against Escherichia coli (ATCC 25922), Staphylococcus aureus (ATCC 25923), Pseudomonas aeruginosa (ATCC 27853), Vibrio alginolyticus (isolated from eye of diseased pompano), and Vibrio harveyi (isolated from kidney of diseased sea bass) using the cup-well method. Varying doses of the extract ranging from 100 mg to 3.1 mg were used together with Amoxicillin (10 Âľg) that served as control. Aqueous extract at 100 mg showed maximum inhibition zone of 31 mm against S. aureus, higher than the antibacterial activity (28 mm) of the control (Amoxicillin). Growth of S. aureus was otherwise inhibited when exposed to 50 mg (27 mm), 25 mg (24 mm), 12.5 mg (20 mm), 6.3 mg (12 mm), and 3.1 mg (4 mm). Similarly, V. alginolyticus showed inhibition zones of 32 mm and 29 mm when exposed to 100 mg and 50 mg of aqueous extracts, respectively. In addition, a minimal inhibition zone of 6 mm was noted in V. harveyi exposed to the highest dose of the aqueous extract tested. On the contrary, no antibacterial activity was noted in E. coli and P. aeruginosa exposed to the highest dose of the extract.

Keywords: antibacterial activity, extract, seaweed, Ulva pertusa

48


Effect of Greenwater Technology on Water Quality and Occurrence of White spot Syndrome Virus (WSSV) in Pond Cultured Penaeus monodon Eleonor A. Tendencia1,2, Roel H. Bosma2, Marc C. Verdegem2, Johan A.J. Verreth2 Aquaculture Department, Southeast Asian Fisheries Development Center, Tigbauan, Iloilo 5021, Philippines 2 Aquaculture and Fisheries, Wageningen University, the Netherlands

1

White spot syndrome virus (WSSV) has caused severe production drops in the shrimp industry. Despite the bulk of literatures related to WSSV epidemiology, reports on minimizing disease outbreaks through ecological means are rare. Industry stakeholders resorted to various innovative techniques to revive the industry and recover after suffering heavy economic losses. Some shrimp farmers in the Philippines claimed that â&#x20AC;&#x153;greenwaterâ&#x20AC;? (GW) technology could prevent disease outbreak due to WSSV. The efficiency of the GW technology was evaluated by comparing three farms using the GW culture technique to three farms not using it. Lower total suspended solids (TSS) and nutrient levels despite the higher FCR; higher plankton counts; greater shrimp mean daily weight gain; and higher shrimp survival were observed in GW farms, suggesting that the use of the GW technique to culture Paneus monodon improves water quality. Furthermore, WSSV was detected in the GW farms but no WSSV disease outbreak occurred.

Keywords: greenwater, water quality, white spot syndrome virus, WSSV, Penaeus monodon

49


Detection of Luminous Vibrio and White Spot Syndrome Virus in Shrimp by Duplex PCR Christopher M.A. Caipang1, Mary Paz N. Aguana2, Carlo C. Lazado1 Faculty of Biosciences and Aquaculture, University of Nordland, Bodø 8049, Norway Institute of Aquaculture, College of Fisheries and Ocean Sciences, University of the Philippines Visayas, Miag-ao 5023, Iloilo, Philippines 1

2

Heavy mortalities associated with luminous vibriosis and white spot syndrome virus (WSSV) infections have been the major problems faced by the shrimp culture industry in the Philippines. We optimized published PCR protocols for the diagnosis of vibriosis and white spot syndrome virus (WSSV) disease in shrimp that could be suited in the country. Genomic DNAs of Vibrio spp. that exhibited luminescence as well as those that grew on thiosulfate citrate bile salt sucrose agar (TCBS) were used for the PCR amplification of the ribonuclease P (RNase P) gene. There was differential amplification of the RNase P gene based on the phenotypic characters of the Vibrio spp. Direct colony PCR of bacterial colonies yielded similar results. White spot syndrome virus was also detected in the infected shrimp and there were differences in the detection frequency relative to the tissues used for PCR amplification. Duplex PCR was also optimized for the simultaneous detection of these pathogens in shrimp.

Keywords: duplex PCR, shrimp, luminous vibriosis, Vibrio sp., white spot syndrome virus, WSSV

50


Parasitic and Shell Diseases of Abalone (Haliotis asinina) in the Philippines Gregoria Erazo-Pagador, Vincent Encena ll, Rolando V. Pakingking Jr Aquaculture Department, Southeast Asian Fisheries Development Center, Tigbauan 5021, Iloilo, Philippines The donkeyâ&#x20AC;&#x2122;s ear abalone Haliotis asinina is a very promising aquaculture species. However, wild and sea cage abalone are being plagued by parasitic and shell diseases. In the current study, abalone samples were periodically collected from two sites in Igang Marine Station and Sabang, Guimaras from January to December 2011. Macroscopic and histological analyses were carried out in the abalonesâ&#x20AC;&#x2122; shells and tissues. Majority of the abalone samples collected from the grow-out sea cages were infested with shell-boring polychaetes belonging to Serpulidae, Spionidae (Polydora sp.) and Dorveillidae with prevalence rates of 49%, 55% and 76%, respectively. Similarly, abalone samples collected from the wild were infested with polychaetes belonging to Serpulidae (Spirorbis sp., Pomatoceros sp.), Dorveillidae, and Spionidae with prevalence rates of 58%, 30%, and 20%, respectively. Rickettsia-like organisms (RLO) were observed in low numbers in the oesophagus or digestive tubules of seven cultured (prevalence rate: 5%) and five wild (3%) abalone samples examined. Ciliates were present in the gills of cultured and wild stocks with prevalence rates of 40% and 6%, respectively. In addition, metacestodes or plerocercoids (Tylocephalum sp.) were detected in cultured (15%) and wild (8%) abalone samples. Lastly, gregarine-like organisms (Nematopsis sp.) were likewise observed in cultured and wild abalone samples with prevalence rates of 13% and 5%, respectively. To our knowledge, this is the first report on parasitic and shell diseases in wild and cultured H. asinina in the Philippines.

Keywords: parasites, shell disease, abalone, Haliotis asinina

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Can We Use the Autochthonous Bacteria of the Gastrointestinal Tract of Atlantic Cod Gadus morhua as Probiotics? Carlo C. Lazado, Christopher Marlowe A. Caipang Faculty of Biosciences and Aquaculture, University of Nordland, Bodø 8049, Norway Two probiotic candidates, GP21 (Pseudomonas sp.) and GP12 (Psychrobacter sp.) were identified from the gastrointestinal tract of Atlantic cod, Gadus morhua. Both bacteria have a wide range of antagonism against Vibrio anguillarum and Aeromonas salmonicida subsp. salmonicida. The probiotic candidates could produce siderophores, had high tolerance to acidic environment and the bile salts and had the ability to form biofilms. The bacteria could adhere on the intestinal epithelial cells and they interfered with the adhesion of pathogens in different mechanisms. Incorporation of these bacteria in the diet showed a significant influence on the digestive physiology of cod as some intestinal enzymatic activities were enhanced. Further, significant changes in the transcription of immunerelevant genes were observed distinctly in the blood than in the intestine. Given these characteristics, these two autochthonous bacteria of the gastrointestinal tract are potential host-derived probiotics that could be used as an alternative health management strategy in cod aquaculture.

Keywords: Atlantic cod, probiotics, intestinal bacteria, immune response

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Bacterial Diversity and Algal Community Structure in Biofilms of Settlement Plates for Abalone Haliotis asinina Larvae Leobert D. de la Peña, Celia Lavilla-Pitogo, Demy D. Catedral, Milagros R. de la Peña, Nestor C. Bayona, Ellen Grace T. Tisuela Aquaculture Department, Southeast Asian Fisheries Development Center, Tigbauan 5021, Iloilo, Philippines Larval settlement plates for abalone develop a complex community of microorganisms and epiphytic unicellular primary producers that provide nourishment for developing larvae on the surfaces while they remain suspended in the culture tanks. Past studies have shown that bacterial and algal communities found in the biofilm on these plates can affect the integrity of biofilms and the resulting settlement, growth and survival of mollusk larvae. But roles of these microbes play in early feeding postlarvae are poorly understood. Thus, this on-going study is being done to characterize the serial changes in community structure that occur from plate preparation until 90 days of rearing and to compare the bacterial diversity on settlement plates for larval abalone in relation with the dominant and prevailing algal community that proliferates out of diatom slurry seeding. This will establish information on potential pathogens, microbial species that may boost abalone production, or define their synergistic action. In this study, biofilms from uniform-sized areas (2 x 2 inches) of the same plates are being scraped off regularly from day 1 of suspension up to harvest of young abalone after one hatchery cycle (90 days). Separate samplings are being done for the dry season (March– June) and wet season (November–February) to determine season variation of microbial community. Total bacteria are then enumerated on nutrient agar, while bacterial group populations are identified by using selective media for vibrios and pseudomonads. Remaining suspension of biofilm materials are then fixed in formalin and Lugol’s iodine for identification of their algal components. Bacterial and algal components were identified to genus level by microscopy and biochemical tests. The initial results of this study are the following: no observable trend was observed if bacterial counts were examined based on the stages of abalone culture; for water samples, luminous bacterial count (LBC) were observed to be generally higher in dry season than in wet season; for biofilm samples on larval settlement plates, bacterial counts were generally higher by 1–2 logs in dry season than in wet season. For identification of bacterial isolates, most were identified as Vibrio vulnificus (32.7%) and Vibrio alginolyticus (18.2%). Others were identified as Chryseobacterium indologenes (10.9%), Aeromonas spp., (5.5%) Moraxella lacunata (5.5%), Photobacterium damsela (5.5%), Brevundimonas vesicularis (3.6%), Comamonas testosteroni/Pseudomonas alcaligenes (3.6%), and Pasteurella multocida (3.6%). Algal communities from the settlement plates are still to be identified. So far, these microbial community data are coming from successful larval rearing runs of abalone in the hatchery. Microbial community data associated with unsuccessful larval rearing runs are yet to be determined.

Keywords: bacterial diversity, algal community, biofilm, settlement plates, abalone, Haliotis asinina

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Status and Needs of Primary Aquatic Animal Health Care Small-Scale Aquaculture Edgar C. Amar, Rolando PakingKing Jr., Eleanor Tendencia, Leobert dela Pe単a, Gregoria Pagador Aquaculture Department, Southeast Asian Fisheries Development Center, Tigbauan 5021, Iloilo, Philippines A need for better understanding of issues affecting disease and their control can promote the livelihood of small-holder fish farmers specifically in rural communities, where awareness of fish health management remains generally poor despite significant advances in the diagnosis, prevention and control of fish diseases. Awareness of fish health concepts will translate to increased productivity of food products that are safe for consumers. A survey was conducted in target countries in Southeast Asia to identify the gaps and needs in order to equip small farmers and other industry stakeholders with capability to monitor disease, identify and prioritize important small-scale aquaculture commodities, conduct audits of selected farms to identify key issues affecting fish health and production, boost staff capability as well as translate learning materials, conduct on-site and other training courses and guided research and information dissemination. About 15, 40, and 25 small-scale farmer respondents, were respectively interviewed in Myanmar, Philippines, and Lao PDR. Tilapia was the main species cultured in the Philippines. Hatchery, fingerling production, and grow-out farms were represented. There was a wide variation in awareness of fish health management concepts among farmers in Luzon, Philippines. Carps and tilapia were the predominant species cultured in Vientiane while native catfish was the most common species cultured in Vang Vieng Province. Compared to Myanmar and the Philippines, small-scale farmers in Lao PDR had low level of awareness of fish health management and food safety, and other issues affecting fish production in ponds. There is a need to strengthen the expertise of government fish health staff in Myanmar, the Philippines, and Lao PDR in order to affectively disseminate available information on fish health management.

Keywords: status and needs survey, primary aquatic animal health care, small-scale aquaculture

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Pathological Effects of Swine Effluents with Varying Levels of Some Physico- Chemical Properties on Bighead Carps (Aristhycthtys nobilis, Richardson 1845) Rosalina R. Atos1, Benjamin Reuel G. Marte2, Francis Andrew Eugene M. Bernardo2, Joseph S. Masangkay2 School of Veterinary Medicine, Aklan State University, Banga, Aklan, Philippines College Of Veterinary Medicine, University of the Philippines, Los BaĹ&#x2C6;os College, Los BaĹ&#x2C6;os, Laguna, Philippines (email add - rosalinaatos@yahoo.com.ph.) 1

2

This study was conducted to determine the effects of varying levels of some physico-chemical properties of swine effluents (SE) on the behavior, mortality rate, survival time and gross and histopathological lesions on juvenile bighead carps (Aristhycthtys nobilis). Result showed that swine effluents can be toxic to juvenile Bighead carps and toxicity will depend on its quantity in the holding water. In SE with unacceptable Biological Oxygen Demand (BOD), Total Kjelhdal Nitrogen (TKN), Dissolved oxygen (DO) and Total Solids (TS), an active jumping, highly erratic swimming activities, floating while piping with wide open opercula, then sinking and death where the behavioral changes observed. One hundred percent mortality rate with shorter survival time and death intervals was noted with the absence of significant gross and microscopic lesion in the vital organs of the fishes. In SE wherein all parameters were within acceptable limits except for BOD and suboptimal DO levels, none to minimal changes in swimming behavior was noted initially, however, later, carps swim in an upside down position and breath slowly with wide operculum then died after long hours of inactivity. Mortality rate of 95% with longer survival time and interval between deaths were observed with more severe and well defined gross and histopathological lesions on the gills, liver, kidney, intestine and spleen.

Keywords: swine effluents, physico-chemical properties, Aristhycthtys nobilis

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Sargassum oligocystum as Antibacterial Agent and as Immunostimulant for Pacific White Shrimp (Litopenaeus vannamei) and Summer Flounder (Paralichthys dentatus) Francis N. Baleta1,2*, Liberato V. Laureta Jr1, Mary Jane S. Apines-Amar1, Philip Ian P. Padilla3, and Gerald F. Quinitio4 Institute of Aquaculture, College of Fisheries and Ocean Sciences (CFOS), University of the Philippines Visayas (UPV), Miagao, Iloilo, Philippines 2 Provincial Institute of Fisheries, Isabela State University â&#x20AC;&#x201C; Roxas, Roxas, Isabela 3 Department of Biological Sciences, College of Arts and Sciences, UPV, Miagao, Iloilo, Philippines 4 Institute of Marine Fisheries and Oceanography, CFOS,UPV, Miagao, Iloilo, Philippines 1

Studies were conducted to determine the potentials of a brown seaweed Sargassum oligocystum collected from the coastal area of Sta. Ana, Cagayan, Philippines as antibacterial agent and as immunostimulant for cultured crustaceans and finfish. The first study determined the in vitro antibacterial activity of S. oligocystum organic solvent and aqueous extracts against six aquaculture pathogenic bacteria (Vibrio spp., Flavobacterium aurantiacum BIOTECH 1492, Streptococcus faecalis BIOTECH 1072), and Pseudomonas aeruginosa. The methanol extracts of S. oligocystum showed strong antibacterial activity against V. harveyi, S. faecalis and P. aeruginosa and moderate activity against the rest of the test pathogens. V. harveyi was the most susceptible bacterial strain. Subsequently, the non-specific immune response and disease resistance of Pacific white shrimp Litopenaeus vannamei immersed in S. oligocystum aquaeous extract were investigated. Immune parameters such as total haemocyte count (THC), granular cell count (GC), hyaline cell count (HC), phenoloxidase (PO) activity, respiratory burst (RB) activity, superoxide dismutase (SOD) activity, gluthathione peroxidase (GPx) activity, and lysozyme acitivity were measured after immersion for 0.5, 1, 3, and 4 h in 100, 300, and 500 mg l-1 of the extracts. Significant increases in these parameters were noted with increases in extract concentration and immersion time. A significant increase in the phagocytic activity and clearance efficiency against V. alginolyticus was observed after immersion of the shrimp in all the test concentrations for 3-4 h. Survival rates of shrimp at 300 and 500 mg l-1 were significantly higher than that of the control group 120 h post immersion challenge with V. alginolyticus. Finally, the effects of S. oligocystum hot-water extract on the non-specific immune response and disease resistance of summer flounder Paralichthys dentatus were evaluated. Fish were either injected with 1 or 5 mg ml-1, or immersed in 100 or 500 mg l-1 of the extracts. The non-specific humoral (lysozyme, plasma protein, bactericidal activity) and cellular (respiratory burst, hematocrit) parameters and disease resistance against V. harveyi infection were determined. Results showed significant increases in plasma protein level, lysozyme, and bactericidal activities obtained in most of the periodic sampling days. Upon challenge with V. harveyi, mortality was significantly reduced in all treated groups, indicating the potential of S. oligocystum as an immunoprophylactic agent for finfish aquaculture. Keywords: Sargassum oligocystum, antibacterial activity, immunostimulant, bacterial pathogens

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Index of Presenters Presenter

Page/s

Amar EC Amar MJA Argayosa AM Atos RR Baleta FN Caipang CMA de la Pe単a LD Erazo-Pagador G Faisan JP Halim DSNPH Huyop F Khoa LV Lavilla-Pitogo CR Lea単o EM Maningas MB Mar MK Miyazaki T Nam S Nicolsora AMD Pakingking RV Jr. Regidor SE Santos MD Sibonga MFJ Siti-Zarah Somga JR Somsiri T Tamat W Tammajedy V Taukhid Tendencia EA Thin MM Tung VT Utari HB Yilin L

34, 54 46 37 55 56 44, 45, 50, 52 33, 53 28, 51 47 43 39 24 13 11 30 36 12 16 32 35, 48 27 38 31 19 21 23 15 18 17 49 20 29 40 22

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About SEAFDEC The Southeast Asian Fisheries Development Center (SEAFDEC) is a regional treaty organization established in December 1967 to promote fisheries development in the region. The member-countries are Brunei Darussalam, Cambodia, Indonesia, Japan, Lao PDR, Malaysia, Myanmar, Philippines, Singapore, Thailand, and Vietnam. The policy-making body of SEAFDEC is the Council of Directors, made-up of representatives of the member countries. SEAFDEC has four departments that focus on different aspects of fisheries development: • The Training Department (TD) in Samut Prakan, Thailand (1967) for training in marine capture fisheries • The Marine Fisheries Research Department (MFRD) in Singapore (1967) for post-harvest technologies • The Aquaculture Department (AQD) in Tigbauan, Iloilo, Philippines (1973) for aquaculture research and development • The Marine Fishery Resources Development & Management Department (MFRDMD) in Kuala Terengganu, Malaysia (1992) for the development and management of fishery resources in the exclusive economic zones of SEAFDEC member countries

Tigbauan Main Station

Dumangas Brackishwater Station

AQD is mandated to: • Conduct scientific research to generate aquaculture technologies appropriate for Southeast Asia • Develop managerial, technical and skilled manpower for the aquaculture sector • Produce, disseminate and exchange aquaculture information AQD maintains four stations: the Tigbauan Main Station and Dumangas Brackishwater Station in Iloilo province; the Igang Marine Station in Guimaras province; and the Binangonan Freshwater Station in Rizal province. AQD also has a Manila Office in Quezon City. SEAFDEC Aquaculture Department Tigbauan Main Station Tigbauan 5021 Iloilo, Philippines

Igang Marine Station

Email: aqdchief@seafdec.org.ph Tel. (63 33) 511-9170; 511-9171 Fax (63 33) 511-8709; 511-9070

www.seafdec.org.ph

Binangonan Freshwater Station


seafdec-abstract-proceedings