AQUA Practical Magazine Vol3 Issue 3

www.asianaquaculturenetwork.com

VOL. 3 ISSUE 3 AquaSG’18 S p e c i a l E di t i o n

3-6 October 2018

School of Applied Science Temasek Polytechnic Singapore

INVESTMENT & CURRENT PRACTICES IN AQUACULTURE TECHNOLOGY

AsIAN PACIfIC AquACulTure 2019 AsIAN 2019 AsIAN PACIfIC PACIfIC AquACulTure AquACulTure 2019 This event includes: This This event event includes: includes: Asian Pacific Aquaculture 2019 Asian AsianPacific Pacific Aquaculture Aquaculture 2019 2019 ISTA 2019 ISTA 2019 ISTA 2019 Chennai Trade Center

Chennai ChennaiTrade Trade Center Center

Chennai Tamil Nadu India Chennai Chennai ---Tamil Tamil Nadu Nadu -- India June 19 -21, 2019 June June 19 19 -21, -21, 2019

Aquaculture for Health, Wealth and Happiness Happiness Aquaculture Aquaculturefor forHealth, Health,Wealth Wealth and

JUNE 19 - 21 JUNE JUNE 19 19--21 21 Hosted by: Tamil Nadu Fisheries University Hosted Hostedby: by:Tamil TamilNadu NaduFisheries FisheriesUniversity University Organized by: World Aquaculture Society - Asian Pacific Chapter Organized Organizedby: by:World WorldAquaculture AquacultureSociety Society-- Asian AsianPacific Pacific Chapter for more information contact conference manager: formore moreinformation information contact conference manager: manager: for contact conference P.O. Box 2302 P.O.Box Box 2302 P.O. 2302 Valley Center, CA 92082 usA Valley Center, CA 92082usA usA Valley Center, CA 92082 Tel: +1.760.751.5005 Tel:+1.760.751.5005 +1.760.751.5005 Tel: fax: +1.760.751.5003 fax: +1.760.751.5003 fax: +1.760.751.5003 worldaqua@was.org - www.was.org worldaqua@was.org -www.was.org www.was.org worldaqua@was.org Trade show & sponsorship: Trade show&&sponsorship: sponsorship: Trade show mario@marevent.com mario@marevent.com

COORDINATION OFFICE:

599/27 Ratchadapisek Road, Jatujak. 10900 Thailand Tel: +66 (0) 21921787 to 88 | Fax: +66 (0) 21921733 | info@asianaquaculturenetwork.com

SINGAPORE ADDRESS:

31 Harrison Road #04-02, Food Empire Building, Singapore 369649

{ All right reserved. Reproduction of the magazine, in whole or in part, is prohibited without the prior written consent not unreasonably withheld, of the publisher }

EDITORIAL

Use of the information in this magazine is at your own risk. AAN and its employees do not warrant or make any representation regarding the use, or results of the use, of the information contained herein in terms of its suitability, correctness, accuracy, reliability, currency or otherwise.

The entire risk of the implementation of the information which has been provided to you is assumed by you. All liability or responsibility to any person using the information is expressly disclaimed by AAN and its employees

• Editor in Chief: Dr. Farshad Shishehchian

• Managing Editor: Nathalie Lim

• Marketing Communications: Simona Al-Fadhli

• Technical Supports: Zahra Javidi

• Designer: Pipitpan Lerdpipitpon Itticheat Issalaseree

Disclaimer PUBLISHED BY: ASIAN AQUACULTURE NETWORK (AAN)

Editor's letter In this special issue, we will base our discussion on the theme of this year’s AquaSG Conference, on the importance of investment and innovation in current practices in aquaculture technology. Local and international renowned speakers who specialize in research, nutrition, farming technology and investment in the aquaculture industry are brought together this year. On day 1 of the conference, Dr. Farshad will conduct a workshop session and discuss smart farming technology application of science and AI in aquaculture. The utilization of AI and smart technologies should be implemented into our farming practices in order to improve productivity, quality and efficiency of our operations. Dr Claude Boyd will talk about the role of nitrogen in aquaculture on day 1 as well. There is a growing trend towards increased production intensity in almost all kinds of aquaculture and maintaining acceptable water quality within production systems requires much greater importance as production intensity increases. On day 2 of AquaSG, Dr Jose Domingos will have a conference session to examine the industrial production of high quality eggs and larvae of tropical marine food fish. The availability of high quality, disease free and genetically improved juvenile seed stock is vital for aquaculture business to increase productivity and minimize economic risks. Dr. Giana Gomes will also discuss environmental DNA (eDNA) as a forensic technique to detect pathogens in aquaculture systems. The development of aquaculture and its capacity to ensure food security to our world’s expanding population will depend on our ability to develop innovative technologies to minimize disease risk within this industry. On day 3 of the conference, Georg Baunach will discuss building an ecosystem for external innovation in aquaculture. Being a globally-dispersed industry, the buildup of a strong innovation ecosystem, where entrepreneurs, investors, industry researches and governments can come together has not taken place yet.

AquaSG’18 is in its 3rd year and we strive to be an exceptional platform for industry players

and academics to examine and discuss the current issues and present innovative applications in aquaculture. We hope to continue this trend and together grow our knowledge and practical solutions to the aquaculture industry.

1

CONTENTS Page

CONTENTS

1 EDITORIAL 3

INDUSTRY UPDATES

From the other side of the pond

by Antonio Garza de Yta, Ph.D.

5 FEATURES 5

Nitrogen Dynamic in Aquaculture

by Claude E. Boyd

7

What role can insects play towards the sustainability of aquaculture?

by Leo Wein, Yingyu Law

9

Urban farming with integrated aquaponics systems

by Khin Mar Cho

10

Indoor culture of freshwater tilapia using biofloc

by Christopher Marlowe A. Caipang

12

From chemotherapeutics to phytobiotics in aquaculture

by Christopher Marlowe A. Caipang, Clara M. Lay-yag and Sakinah Mulyana

14

Indoor Mudcrab Hatchery and Larviculture

by Chan Diana, Khoo Hock Lai William, Wong Yee Man Cynthia, Nadiah Sata, Glendon Teo and Lee Chee Wee

15

Rapid on-site detection of Iridovirus by optical immunoassay (OIA)

by Masato Miyata, Syed Khader Syed Musthaq, Nushahidah Ali, Alden Toh, Sakinah Mulyana, Nadiah Sata, Chan Pek Sian Diana, Padmanabhan Saravanan and Lee Chee Wee

16

Building research capabilities in aquaculture locally

17

Industrial production of high quality eggs and larvae of tropical marine food fish

by Jose A. Domingos, Jennifer M. Cobcroft, Dean R. Jerry and Giana Bastos Gomes

21 ISSUE HIGH LIGHT:

Evaluation of Functional Feed Additives Against Shrimp Pathogens with an emphasis on Vibrio parahaemolyticus and White Spot Syndrome Virus (WSSV) in Litopenaeus vannamei

by Farshad Shishehchian, Krit Khemayan, Zahra Javidi26

26

INTERVIEW

7 Questions with Mr. Ravi Kumar Yellanki

29

NEWS & PRESS:

29 30 32 33

News Around the World : EU to fund Ecuadorean shrimp sustainability initiative Investments in Russian aquaculture on the rise Vietnam poised to become top player in ocean aquaculture ‘Omni-channel’ shrimp tech platform looks to modernize Indian sector

36

EVENTS CALENDAR

INDUSTRY UPDATE

FROM

THE OTHER SIDE OF THE POND Antonio Garza de Yta, Ph.D. Rector

Universidad TecnolĂłgica del Mar de Tamaulipas Bicentenario

The Future of Aquaculture Education

T

oday the world can be divided in two kinds of people: the ones that have adapted to globalization and the ones who had not. Being aquaculturist, its hard not to be fond of internationalization and global trade and markets; as seafood is one of the most traded goods between borders worldwide.

I have observed, with some sadness, how some precursors of globalization have turned away and backpedaled, as the gap between countries becomes smaller and the field gets more leveled; some people are just afraid of losing power. The pro globalization group, where I am included, imagine a world where most people have a global culture while not losing their local identity; the best of all worlds. We dream of a world where breaches of all kinds are minimized, and basic services, education and healthcare are available for everybody. One day, regardless of the reluctancy of the people in power, borders will disappear.

For the world to become one, as John Lennon imagined, there is still a long way to go. Regardless, education, today, is contributing to the world to become a smaller place. We all know that English, whether or not, has become the international language; and accept,that it is a very easy language to learn. I can imagine that if you are reading this article in Asia, English is your second or third language and that it has helped you to communicate between colleagues and has been one of the most useful tools for your professionalization; it is my third language and has been a key factor for my professional development. Taking this into consideration, I think there are three basic steps to establish the foundations for the future of aquaculture education; same steps that we are putting in practice at the UTMarT. The first step is to make sure that your students are completely

proficient in English. How to achieve it? Well, this is not the only model, but it is one that has been used in many places and I consider is working adequately: Start a bilingual program. In the UTMarT we have started a certified Bilingual, International and Sustainable (BiS) program in which students can arrive without any knowledge of English and through a year long transition program we insert them in an all-English environment.

The second step is to create a solid practical international experience. The process of internationalization is key for the students to grasp the global concept. In our program we have two four-month practical internships where students ideally work on the topics that they are interested in. The internationalization process goal is that the percentage of students that go overseas for their internships increases year by year. For this internationalization process we also must be ready to constantly receive visitors from all over the world.

The third and final step is to insert the graduates on a worldwide professional organization that promotes the exchange of ideas and knowledge. is where the World Aquaculture Society takes place. We encourage our students to become part of it.

No research or education center can be an expert on every field of aquaculture. Knowledge and human resources exchange between regions and countries is imperative to achieve the production required to feed the growing world population. We need to start this process ASAP; I invite everyone interested in establishing a partnership with a serious institution in the other side of the world to contact me directly (garzadeyta@utmart. edu.mx). Our job as educators is to open the minds of our students, after we achieve it‌ the world is theirs.

3

FEATURES

NITROGEN DYNAMIC

IN AQUACULTURE Claude E. Boyd Professor Emeritus

School of Fisheries, Aquaculture and Aquatic Sciences Auburn University, Auburn, Alabama 36849, USA

P

lants require 14-18 chemical elements for growth, but only nitrogen and phosphorus commonly limit plant productivity in aquaculture ponds. These two elements also are critical in animal nutrition. Nitrogen is a component of amino acids of which protein consists, and phosphorus is involved in many biochemical reactions including those related to energy use in cells. Calcium phosphate is also the main component of bone.

Nitrogen and phosphorus are intertwined through their importance as plant and animal nutrients, and because they coexist in organic matter added, produced in aquaculture systems.

The most important forms of nitrogen in aquaculture usually are organic and inorganic nitrogen in fertilizers, protein nitrogen in feed, and ammonia nitrogen in water.

Ammonia nitrogen consists of nitrogen in ammonia (NH3) and ammonium (NH4+). The two forms of ammonia nitrogen are in a pH and temperature dependent equilibrium:

NH3 + H+ ↔ NH4+ Increased temperature and pH increase the proportion of NH3 relative to NH4+. Ammonia (NH3) is the toxic form of ammonia nitrogen, but ammonium is essentially non-toxic.

Fertilizers include livestock manures and other agricultural waste that decompose releasing ammonia nitrogen or chemical fertilizers such as urea, ammonium sulfate, and ammonium phosphate that dissolved to release plant-available nitrogen.

Nutrients from fertilizers stimulate phytoplankton productivity, the base of the food web culminating in aquaculture biomass.

In semi-intensive and intensive aquaculture, manufactured feeds allow much greater shrimp and fish production than possible with fertilizers alone. Aquaculture feeds contain around 4.0-7.2% nitrogen (25-45% crude protein), but only 20-40% of nitrogen in feed is converted to protein nitrogen in aquaculture biomass. The balance of feed nitrogen is converted to ammonia nitrogen and released into the water through microbial decomposition of uneaten feed and feces and excretion of ammonia as a metabolic waste by culture animals.

The amounts of nitrogen added to aquaculture production units and the ammonia waste load are related to production objectives.

The ammonia nitrogen load to aquaculture systems increases in response to greater production intensity. Moreover, at a given stocking density, the ammonia nitrogen load progressively increases as culture animals grow and biomass increases.

Phosphorus and nitrogen stimulate phytoplankton productivity, and organic matter from dead plankton decomposes and ammonia is released. Of course, some organic matter (and the nitrogen contained in it) accumulate in pond bottom soils.

Almost all readers will have heard of the global nitrogen cycle (Fig. 1). Most of the pathways of nitrogen transformations in this cycle occur in aquaculture production systems. These transformations influence aquaculture production and should be understood by aquaculturists.

Organic matter decomposition rate increases with greater water temperature, but the composition of the organic matter itself influences decomposition rate. Simple organic compounds such as sugar and starch decompose quickly; but, the more complex a compound, the slower it will decompose. Cellulose and protein decompose slower than sugar and starches but faster than fats and waxes, and especially faster than lignin and tannin. As a given organic residue decomposes, the easily decomposed portion (labile fraction) decreases faster than the more resistant fraction (the recalcitrant fraction), and the ratio of the labile fraction to the recalcitrant fraction decreases as decomposition progresses.

Organic fertilizers decompose slower than fish and shrimp feces, dead phytoplankton, and especially uneaten feed. As a general rule, the lower the carbon/nitrogen (C/N) ratio in an organic residue, the faster will be its rate of decomposition. The C/N ratio of organic fertilizers usually range from 25:1-100:1, and these residues decompose slowly. For residues at the higher end of the C/N ratio range, much of the nitrogen in the residue will be retained in biomass of the microorganisms of decay. As a result, the C/N ratio decreases as a particular organic residue decomposes and eventually stabilizes at around 8:1-12:1. The C/N ratios of feeds and dead plankton usually are less than 10.

5

FEATURES

Their decomposition is rapid, and much ammonia is released into the water.

The major form of nitrogen in most culture systems is dinitrogen gas (N2). This gas comprises about 78% of atmospheric gases, and it is soluble in water. The equilibrium concentration (or ŕš? solubility) of dinitrogen gas in water at 28 C and standard atmospheric pressure at sea level is 12.99 mg/L in freshwater and 10.51 mg/L in ocean water. However, dinitrogen is largely inert and has little or no effect on culture animals under normal conditions.

Some cyanobacteria (frequently called blue-green algae) and some other species of bacteria conduct nitrogen fixation. In this process, cellular energy is used to reduce dinitrogen to ammonia, and ammonia is used to synthesize protein. When nitrogen-fixing microorganisms die, they are decomposed with release of ammonia. Many aquaculturists are of the opinion that nitrogen fixation is a major source of nitrogen in aquatic environments. However, nitrogen fixation by microbial cells is an anaerobic process, and the presence of dissolved oxygen in pond water impedes nitrogen fixation. In addition, nitrogen fixation is depressed when plenty of ammonia nitrogen or nitrate is available. Nitrogen fixation probably is of little importance in most aquaculture systems.

Bacteria are about 10% nitrogen while fungi are around 5% nitrogen; thus, considerable nitrogen is bound in microbial biomass during decomposition. Decomposition of organic residues with a high C/N ratio may be limited by the quantity of nitrogen available from the residue. In such instances, microbes must die so that ammonia nitrogen released through their decomposition will support continuing decomposition of the organic residues. Alternatively, if there is ammonium nitrogen or nitrate in the water, microbes may use this nitrogen in decomposition of residues with high C/N ratios.

In feed-based aquaculture, feed and other nitrogen inputs have a low C/N ratio. Organic residues found in these systems tend to decompose rapidly with release of appreciable ammonia

nitrogen. Culture animals also excrete considerable ammonia.

Ammonia nitrogen concentration may become so great that there is a sufficient concentration of ammonia (NH3) to cause toxic effects.

Fortunately, several processes remove ammonia. It is lost to the air by diffusion, a process greatly favored by high pH, because the ratio of NH3 to total ammonia nitrogen increases as pH rises. Windy conditions and mechanical aeration also enhance ammonia loss by diffusion. Nevertheless, diffusion is a minor nitrogen loss in most instances.

Phytoplankton can remove large amounts of ammonia N from pond water for use in protein synthesis, and nitrifying bacteria can oxidize large amounts of ammonia N to non-toxic nitrate.

Nitrification is a two-step process in which ammonia nitrogen is first oxidized to nitrite by one group of bacteria and nitrite is then oxidized to nitrate by a second group of bacteria. Under certain conditions, the first step progresses faster than the second step, and nitrite accumulates in the water. Nitrite is potentially toxic to culture animals.

In anaerobic areas of pond bottoms, denitrifying bacteria convert nitrate to dinitrogen gas which diffuses into the water column and then into the air. Under certain conditions, denitrification may stop at nitrite without further reduction to dinitrogen. This can result in potentially toxic nitrite entering the water column.

Some organic matter entering ponds does not decompose quickly and accumulates in bottom soil. Decomposition of this organic matter is a continuing source of ammonia nitrogen to the water column. Nevertheless, decomposition of organic matter that has accumulated in pond bottoms from previous crops usually does not contribute nearly as much nitrogen to the water column as does decomposition of fresh organic matter resulting from the crop in progress. Many shrimp farmers remove sediment from bottoms of culture systems after each crop. This practice is likely not as important as thought by many, but it does result in removal of organic matter (and nitrogen).

In heterotrophic biofloc aquaculture production systems, ammonia nitrogen concentration may become very high in spite of nitrification. A pure carbohydrate source such as sugar or molasses may be added to the systems. Bacteria will remove ammonia nitrogen from the water to decompose the nitrogenfree carbohydrate. This removes ammonia nitrogen, and it also produces biofloc that can serve as food for the culture animals.

In highly-intensive raceway, tank, and cage culture, ammonia nitrogen is flushed from the grow-out units by water flowing through them. However, in water re-circulating aquaculture systems, ammonia nitrogen must be removed through nitrification within a biofilter.

Fig. 1. The nitrogen cycle.

6

This brief discussion of nitrogen dynamics reveals the importance of nitrogen in aquaculture systems. Those interested in more specific information on nitrogen dynamics can find a wealth of information on the topic in the scientific literature and from postings on the internet.

THE SUSTAINABILITY OF AQUACULTURE? Leo Wein, Yingyu Law Founder & CEO, Research Director Protenga Pte Ltd

A

s the world’s population and wealth become more concentrated in greedily expanding cities, so too are nutrients from all over the world funnelled more and more into these centres of consumption in the form of protein.

Significant amounts (30% and more) of protein are discarded and wasted along the food value chain and then buried in landfills or incinerated, where they degrade and leach out into the environment to pollute waterways and the atmosphere.

The world can’t afford these protein losses, given the rapacious global appetite for protein that grows with increasing population and wealth, and that traditional agricultural practises are at their production limits or in decline.

Aquaculture is a promising growth sector that is projected to help meet this shortfall in protein supply. However, aquaculture also heavily relies on protein for aquafeed. These proteins currently derive predominately from fishmeal and soy, which both have significant environmental footprints, limited expansion potential and rising prices, are produced by only a handful of major producer countries scattered across the world and for which there is competition in the form of human consumption.

The increased demand for fresh, high-quality, traceable, uncontaminated seafood from mostly urban, affluent markets has developed strong value propositions for aquaculture to move closer and closer to (or even into) cities. These factors have inadvertently created an opportunity to close the protein loop for aquaculture feed using insects.

Insects, such as the black soldier fly larvae (BSF, Hermetia Illucens), have the natural ability to extract and concentrate valuable protein, fats and minerals from waste nutrients along

7

FEATURES

WHAT ROLE CAN INSECTS PLAY TOWARDS

FEATURES the food value chain and demonstrate better feed conversion rates than most farmed animals. This ability can be leveraged in combination with engineered systems and technology, such as the Insect Bioconversion System we have developed at Protenga, to upcycle waste nutrients into high-quality, nutrient-packed feed ingredients for aquaculture. Insects, as a sustainable feed ingredient and alternative to fish meal, has been heralded for at least the last 5 plus years. However, to seize on the opportunity to build local, resilient nutrient cycling mechanisms from food waste streams to aquafeed that fulfils sustainable, highquality aquaculture production for urban markets, the insect bioconversion system needs to satisfy several criteria. It needs to be scalable, modular, resource-efficient, produce adequate volumes for the target consumers and most importantly be able to produce cost-competitive, consistent, safe and high-quality insect products.

While insects, such as BSF and mealworms can be used as live feeders for fish and shrimp on a small scale, there are many reasons – such as required volume, shelf-life, handling and transport, digestibility, feed safety and hygiene, inclusion in existing feed formulation and feeding processes, etc. – why insects should be processed into actual feed ingredients in order to impact aquaculture at scale. The two key feed ingredient products from the Black Soldier Fly Larvae (and most farmed feed insects) are insect protein meal and insect oil. While the main product characteristics are comparable, it is worth noting that the actual product quality and attributes will vary depending on the insect species, insect bioconversion system, process parameters, feedstock and downstream processing line used.

8

Generally, biosafety of BSF bioconversion even on problematic or artificially contaminated feedstock has been demonstrated in many research studies; however specifically, producers should design their bioconversion systems around biosafety, traceability and hygiene, particularly in the downstream processing. For Black Soldier Fly larvae, a typical processing line after the larvae have been harvested from their feedstock substrate may include washing, sterilizing, drying, oil separation, milling and packaging. Defatted BSF meal’s crude protein content ranges from 55% to 65%, while fat content is reduced to below 10%.

Our tests and numerous other academic and commercial trials have repeatably demonstrated that the proteins are a very good source of essential amino acids with high digestibility in fish and shrimp. Physical characteristics are finely granular and free-flowable for use in existing feed mixing and extrusion lines similar to fishmeal, although it has a more pleasant, nutty smell. When packaged and stored well, it possesses long shelflife (~12 months). The BSF oil provides an easily metabolizable energy-source particularly for early growth stages and is high in antimicrobial and immune system-boosting lauric acids. The oil can also be used as an excellent attractant and increases feed palatability.

While these two insect-based core products are an interesting alternative and addition to the existing ingredient portfolio on a macronutrient-level, we are also looking forward to connect with interested parties in new product development and application testing and new use-case developments, for which you can contact me directly (leo@protenga.com). Together we can close the missing link towards feed sustainability in aquaculture using local insect protein and resilient nutrient cycles powered by insects.

WITH INTEGRATED AQUAPONICS SYSTEMS Khin Mar Cho

Centre for Aquaculture & Veterinary Science

A

bout 1% (738 hectares) of the local land area is used for agriculture, of which more than 90% of leafy vegetables are grown from soil culture (41 farms for soil cultivation, 3 organic farms, and 6 hydroponics farms). Currently, the local vegetable farms are able to supply 10-12% of the local consumption needs. In order to enhance vegetable production for food security, intensive and integrated farming systems would need to be developed to suit urban farming.

Aquaponics is an integrated fish farming with plants growing in a combined cultured system. Aquaponics set-up aims at growing vegetables using recycled spent fish water containing nutrients from faecal waste and uneaten feed via the recirculating aquaculture system (RAS). Tilapias are often used in aquaponics.

Water coming from the fish tank is filtered through physical and biological filtering processes, whereby Nitrosomonas bacteria will convert ammonia to nitrite after which Nitrobacter

School of Applied Science, Temasek Polytechnic, Singapore

bacteria will convert nitrite (NO2-) to nitrate (NO3-) by bacterial nitrification. The filtered, nutrient–rich water is then circulated to the hydroponic growing units. Plant roots will serve as a natural biofilter in the aquaponics system whereby nitrates in the water are taken up.

The research team at the Agrotechnology Domain of the School of Applied Science is currently looking into optimizing fish stocking density, fish feed nutrition, and water quality as well as the optimum hydroponic culture conditions for growing Chinese leafy vegetables under intensive conditions. Once the conditions are established for urban farming, it would be able to provide an environment-friendly and sustainable food source of leafy vegetables.

NO2

NO3

NH3

NO2

๐

Filtration System

๐

๐

Integrated Aquaculture and Hydroponics Growing Systems

9

FEATURES

URBAN FARMING

FEATURES

INDOOR CULTURE

OF FRESHWATER TILAPIA USING BIOFLOC Christopher Marlowe A. Caipang Centre for Aquaculture & Veterinary Science

School of Applied Science, Temasek Polytechnic, Singapore

S

tudies have indicated that the animals utilized in aquaculture usually assimilate approximately 20–25 % of protein in the feed and the remaining amounts are discharged into the water in the form of waste (ammonia) and uneaten feeds. This condition if left unchecked will result in reduced water quality that poses serious effects on the cultured animals. In the aquaculture context, Biofloc Technology (BFT) relies on the highly efficient use of nutrient inputs in the aquaculture facility with limited or zero water exchange. This technology operates on the principle of recycling nutrients through maintenance of a high carbon-to-nitrogen (C:N) ratio in the water. A number of studies have shown that a C:N ratio of 10-20 are able to promote and sustain biofloc production in the water. The presence of a high C:N ratio in the rearing water allows the growth of heterotrophic bacteria, which convert ammonia into microbial biomass. This microbial biomass forms aggregates with other microorganisms and particles that are suspended in the water, resulting in the formation of “flocs”.

These “flocs” are eventually consumed by the cultured animals or they can be harvested and processed as feed ingredients.

Because these “flocs” are living components, they are known as “bioflocs”.

Feed + C source Biofloc

C source Inorganic N How bioflocs are produced and maintained in an aquaculture system

10

The foodfish aquaculture industry in Singapore currently produces roughly about 6% of the estimated 100,000 MT of the annual foodfish requirement. The main bulk of foodfish production comes from marine coastal farms and to a certain extent from some land-based foodfish farms. With limited land for agricultural purposes and sea space available for fish farming, Singapore depends heavily on importation of fresh seafood. In spite of these limitations in land and water resources, aquaculture can still be carried out in the urban areas via the closed-containment aquaculture systems or through vertical farming using aquaponics. BFT suits well in closed- containment systems where water, either fresh- or seawater, is recycled throughout the rearing period or this can be done under limited water exchange.

There are two basic types of biofloc systems that are suitable for urban aquaculture: those that are exposed to natural light and those that are not. Examples of biofloc systems that are exposed to natural light include outdoor ponds or tanks that are used for the culture of shrimp or tilapia. In this system, a complex mixture of algal and bacterial processes helps to control the water quality. This is commonly known as "greenwater" biofloc as a result of the green coloration of the water due to the presence of microalgae. On the other hand, some biofloc systems are not exposed to natural light and are installed indoors. This type of system is known as the "brownwater" biofloc system, where bacterial processes mostly control the water quality in the system. At the Aquaculture Research Facility of the Centre for Aquaculture and Veterinary Science (CAVS) in Temasek Polytechnic, we utilized a “brown-water” biofloc system, because the rearing tanks of tilapias were located indoors.

The nutritional quality of biofloc was sufficient to augment the growth requirements of freshwater tilapias as shown by the comparable growth rates of the fish reared using bioflocs and those fed commercial feed pellets. The increased populations of heterotrophic bacteria in the rearing water of fish with biofloc, could potentially improve water quality by the constant utilization and recycling of nutrients in the water of the culture tanks. A clear understanding on the various microbiological aspects including characterization of biofloc and manipulation of microbial community is crucial towards the successful design and implementation of such technology in the indoor farming of freshwater tilapias.

Wheat Flour Biofloc

FEATURES

and bacilli populations in the rearing water of tilapias. However, the levels of the nitrogenous wastes (ammonia, nitrite and nitrate) in the rearing water with bioflocs were within “safe� levels for the fish.

Control

Bioflocs in freshwater tanks using wheat flour as carbon source

We used freshwater tilapias to assess the feasibility of using BFT for indoor aquaculture with limited water exchange. Tilapias are good candidates to be grown in a biofloc system because they eat a wide variety of food organisms, which are also present in the rearing water that contains bioflocs. Moreover, tilapias are able to efficiently utilize microbial proteins that are produced through bioconversion of ammonia by the heterotrophic bacterial community. A number of earlier studies demonstrated the beneficial effects of bioflocs in fish: improvement in production, low feed conversion, better nutrition and health.

In addition, BFT systems also result in maintaining optimum levels of the water quality in the rearing water of the fish. Our preliminary studies showed that bioflocs that are produced using wheat flour as carbon source at a C:N ratio of 16:1 resulted in increased concentrations of suspended solids and biochemical oxygen demand (BOD), as well as total heterotrophic bacteria

Parameters

Control

Biofloc

Final Weight(g)

35.2+6.3

34.4+7.4

ADG(g/day)

0.55+0.12

0.55+0.13

SGR(% d-1)

3.7+0.32

3.6+0.40

Biomass(g)

930.5+145.4

987.0+219.8

Survival rate (%)

85.75+4.03

92.85+2.05

FCR

0.92+0.14

0.54+0.12

Production of tilapias in tanks reared with and without bioflocs

11

FEATURES

FROM CHEMOTHERAPEUTICS TO PHYTOBIOTICS

IN AQUACULTURE Clara M. Lay-yag, Sakinah Mulyana and Christopher Marlowe Caipang Centre for Aquaculture & Veterinary Science

School of Applied Science, Temasek Polytechnic, Singapore

T

he aquaculture sector is expanding at a rapid rate to meet the food demands of the growing population. Recent estimates show that the aquaculture industry is increasing at an annual rate of more than 9%, in contrast with the annual increase of almost 1.5% for capture fisheries and almost 3% for terrestrial farmed meat production systems.

these plants; thus, these have potential use in the culture of fish and shellfish when incorporated as feed additives without having to worry about environmental and biological problems.

These natural and plant-derived substances that are used as feed additives to improve animal productivity are known as phytogenics or phytobiotics.

Diseases are a crucial factor that inhibits the expansion of aquaculture. With rapid intensification in aquaculture, the incidence of infectious diseases has also increased resulting in significant economic losses. To curb these problems, various chemotherapeutants have been used for treating or preventing diseases in the aquaculture facility. Even though they yield positive effects, they cannot be recommended due to their residual and other side effects. For example, the use of these chemicals and drugs has resulted in more resistant bacterial strains. These resistant bacterial strains pose more serious negative impacts when treating future fish disease problems as well as to the environment and even public health.

Phytobiotics are composed of a wide range of substances and have been classified according to plant origin, processing and composition. These plant-based feed additives include herbs, which are non-woody flowering plants and possess medicinal properties; spices, which are herbs having intense smell or taste and are commonly added to human food; essential oils, which are aromatic oils derived from plant materials through hydrodistillation; and oleoresins, which are extracts derived from plants through the use of non-aqueous solvents.

Medicinal plants have been known as immunostimulants for thousands of year, and these are widely used in veterinary and human medicine. Because these plants are used to develop natural products, these are not only safe for consumers but are also widely available. Hence, medicinal plants show great potential as alternatives to antibiotics and chemical immunoprophylactics. Several studies have demonstrated the positive effects of using plants for livestock and aquaculture operations. A wide range of these plants possesses growthpromoting ability and improves the immune system. They can stimulate the appetite of the animal, induce maturation, and to a certain extent exert antimicrobial activity. There were experiments carried out that showed anti-stress properties of

12

Phytobiotics Botanicals Herbs

Spices

Plant extracts Essential

oils

Oleoresins

Classification of phytobiotics for aquaculture

0.0125 g ml-1

0.00625 g ml-1

Control

Lysozyme (Innate immunity)

Day 3

Day 7

Control 5

FEATURES

0.25 g ml-1

Lysozyme Units

0.05 g ml-1

Scavenging Activity

0.1 g ml-1

3.5 3 2.5 2 1.5 1 0.5 0

Day 15

Treatment

Anti-oxidant activity

4 3 2 1 0 Day 3

Control

Day 7

Day 15

Treatment

In-vitro inhibition of a pathogenic Vibrio at different concentrations of cinnamon extract

Modulation of the fish immune response during feeding with cinnamon extract

The whole plant or its parts, for example, roots, leaves, seeds, flowers or barks can be used. The extraction process is simple: ethanol, methanol or water are commonly used as extracting solvents. The chemicals that are used to extract compounds from plants may have different effects on the fish or shellfish when used as feed additives. Application methods can be either single or in combination, or even in a mixture with other feed additives. The dosages and duration of time of oral administration vary and the optimal levels for the host are species-dependent.

their anti-oxidant activities, which could have contributed to their better survival when exposed to the bacterial pathogen.

Although the effects look promising, more evidence is needed to confirm the apparent beneficial effects on the performance of fish and shellfish when these phytobiotics are added to the feeds on a regular basis. In addition, although these phytobiotics are considered natural products, their use and administration in the feeds need to be carefully assessed for potential interactions with other ingredients that could result in potentially negative effects to the host.

At the Aquaculture Research Facility of the Centre for Aquaculture and Veterinary Science (CAVS) of Temasek Polytechnic, we have tested several of these phytobiotics to enhance growth, survival and immunity of tilapias and ornamental fish. We have tested a variety of herbs and spices including garlic, turmeric, cinnamon, coriander and annatto as feed additives for these fish. These phytobiotics demonstrated anti-bacterial activity. When used as feed additives, they modulated the immune response, enhanced the anti-oxidant activity and to a certain extent improved survival of the fish during infection with a bacterial pathogen.

From our preliminary trials, the beneficial effects that were observed on the fish fed with these phytobiotics were mostly on the improvement of the immune responses particularly on the innate immunity of the fish as well as enhancement of

13

FEATURES

INDOOR MUDCRAB HATCHERY AND LARVICULTURE Khoo Hock Lai William, Wong Yee Man Cynthia, Nadiah Sata, Glendon Teo, Chan Pek Sian Diana and Lee Chee Wee Centre for Aquaculture & Veterinary Science

School of Applied Science, Temasek Polytechnic, Singapore

M

ud crab is one of the most desired seafood items especially in Asia with the greatest demand shown in China and Japan (soft-shell crabs). The crustacean is an important ingredient in Singapore’s iconic seafood dishes such as chilli crab and black pepper crab. Crab farming and food industry-supply is still heavily dependent on wild catch and this has led to depleting numbers in the wild, thus creating a major constraint in the development of this crustacean industry. The crabs harvested for consumption have also been observed to be smaller in size. Overharvesting of adult mud crabs is also a serious conservation concern as it could lead to a loss of the species population, biodiversity and ecosystem functions.

Zoea Stage 1

Zoea Stage 4

Although there are mangrove crab hatcheries that have been established in some parts of Asia, majority of them rely on flow-through water system or open pond culture. In Temasek Polytechnic, our research team aims to develop and optimize indoor controlled conditions for producing Scylla sp. crablets so as to reduce wild harvest; reduce importation cost and to create a sustainable local supply in Singapore.

In this study, berried mangrove crabs, Scylla serata and Scylla olivacea, were kept under controlled environmental conditions in a closed containment system at the Aquaculture Research Facility, Center for Aquaculture & Veterinary Science in Temasek Polytechnic. Successful spawning has been performed sustainably for the past few batches of mud crabs with successful rearing of zoeas until they reach juvenile crab stage in captivity.

Normal husbandry and health monitoring of the crabs, zoeas and juveniles were carried out regularly. Currently, the project team has been able to work out the growth requirements of the different stages from zoea, megalopa to the crablet stages.

The juvenile crablets are reared in closed recirculating tanks and study is underway in improving the growth and feeding conditions for the different stages of growth and development.

14

Scylla Serata crablet

FEATURES

RAPID ON-SITE DETECTION

OF IRIDOVIRUS BY OPTICAL IMMUNOASSAY (OIA) Masato Miyata, Syed Khader Syed Musthaq, Nushahidah Ali, Alden Toh, Sakinah Mulyana, Nadiah Sata, Chan Pek Sian Diana, Padmanabhan Saravanan and Lee Chee Wee Centre for Aquaculture & Veterinary Science

School of Applied Science, Temasek Polytechnic, Singapore

T

here is a greater demand to intensify aquaculture practices for foodfish production. The demand is driven by the projected population growth to hit 9 billion by 2030.

Singapore imports more than 90% of our food fish consumption (~ 100,000 tonnes a year). One of Agri-Food and Veterinary Authority's (now known as Singapore Food Agency) objective is to meet 15% of Singapore’s food consumption through increased productivity of aquaculture in local farms. Due to the growing emphasis on intensification of aquaculture practices in which fish is grown in close contact at high density within a confined space, there is a potentially high risk of an infectious disease outbreak. Infectious diseases are a primary constraint to the growth of many aquaculture species and is responsible for severely impeding both economic and socio-economic development in many countries of the world.

Iridoviruses (Figure 1) are a significant cause of mortality in farmed seabass, grouper and more than 30 other species of cultured marine and fresh water fishes. Affected fish become lethargic, exhibit severe anaemia, petechiae of the gills, and enlargement of the spleen. The principal mode of transmission of this virus is horizontal via the water. There is a need for rapid onsite detection of iridoviral disease in farms to enable increased farm biosecurity and to prevent and control iridovirus outbreaks in farm settings. This could enable early disease risk management, as well as, monitoring of vaccinated fishes in terms of the protective efficacy of oral and injectable vaccines for the prevention and control of fish diseases.

The project team has successfully developed an onsite sample preparation methodology and point-of-care test (POCT) device to detect iridoviral disease in fishes. Concentration of targets was achieved by immunomagnetic bead-based sample extraction from tissue samples collected on-site using a tissue punch (Figure 2). The optical immunoassay (OIA) device works on the

Figure 1:

Figure 2:

principle of thin film biosensor. The immunobinding events on the chip surface leads to interference of reflected light leading to a change in colour from gold to purple blue (Figure 3). The limit of detection achieved for iridovirus was found to be 5000 particles per millilitre of processed tissue extract. For screening of antibody response to iridovirus fish blood sample (plasma) was obtained via filtration through a hydrophobic surface and diluted 1:40 before applying a drop onto the chip surface. OIA performance was evaluated with the gold standard polymerase chain reaction (PCR) method subjecting 30 samples each of positive and negative in PCR. The sensitivity and specificity of OIA was observed to be >90% and 100% respectively. The operational advantages of OIA includes field deployable, rapid and sensitive detection, user friendly instructions, non-powered, less logistics load and visual readout.

OIA Device

Reacted positive

Figure 3:

Figure 4:

15

FEATURES

BUILDING RESEARCH CAPABILITIES IN AQUACULTURE LOCALLY James Cook University Singapore

T

ropical regions of the world face unprecedented challenges due to population growth and an associated demand for high-quality seafood. This population expansion will continue to pressure wild fisheries resources and dictate ever higher efficiencies from aquaculture production. In Singapore specifically, the average amount of fish consumed by each Singaporean is 22kg. This is more than the global average of 20kg. Yet, only 8% of all fish are produced by farms in the country.

Capitalising on its location right in the middle of Asia, James Cook University in Singapore (JCUS) has research expertise that specialises in the sustainable production of tropical aquaculture species. With world-class experts in aquaculture genetics, nutrition, hatchery production, husbandry and aquatic animal health, researchers are ready to partner with commercial industry, government institutions, universities, polytechnics and other stakeholders, to conduct high-quality scientific R&D in Singapore to meet the growing demand locally.

The team is led by Dean of Research Professor Dean Jerry, a recognised leader in the application of genetic and genomic solutions for the aquaculture industry. He leads one of the largest research teams globally devoted specifically to the application of genetic technologies to improve the productivity of aquaculture species.

An area of research that the university is currently working on is environmental DNA (eDNA). It is a front-line detection method that is useful in identifying genetic material discharged into the environment by both micro and macro organisms. Even

16

when there is no visible trace of the organism when doing the sampling, this technique is sensitive enough to be effective.

The research is led by Dr Giana Bastos Gomes who originally started her career in aquaculture after graduating in veterinary medicine in Brazil. In the aquaculture context, she is researching the use of eDNA as a biosecurity tool to detect pathogens and aquaculture pests. JCUS is currently establishing a laboratory for eDNA detection specifically for aquaculture. The university laboratory aims to provide an efficient, non-invasive and most importantly cost-effective service for pathogen surveys for the industry both locally and regionally.

While JCUS already has a world-class team of experts working on projects. The long term goal is to build capabilities locally. With the Bachelor of Business and Environmental Science (Majoring in Aquaculture), the university aims to prepare a workforce with the core knowledge and training in the application of business and environmental principles, with particular attention to aquaculture.

Through this multi-disciplinary program, students will learn how to manage the delicate balance between profit, policy, conservation and aquaculture. Bridging the gap between business and science. Once there is a sufficiently trained workforce, it is hoped that we can work with industry, government and other stakeholders to start on capacity building to scale up and make a larger impact on aquaculture as a whole.

For further information on how to collaborate with JCUS, please send an email to researchsupport-singapore@jcu.edu.au.

FEATURES

INDUSTRIAL PRODUCTION OF HIGH QUALITY EGGS AND LARVAE OF TROPICAL MARINE FOOD FISH Jose A. Domingos, Jennifer M. Cobcroft, Dean R. Jerry and Giana Bastos Gomes Affiliation: Tropical Futures Institute, James Cook University Singapore and Centre for Sustainable Tropical Fisheries and Aquaculture, James Cook University Australia.

Correspondence: J. A. Domingos (jose.domingos1@jcu.edu.au)

I

n agriculture, seed quality and vigour are critical determining factors of crop quality and yield. Although the cost of seed represents a small fraction (<3%) of the total production costs, the quality of seed has a significant direct impact on the profitability of the farming enterprise. This concept is not different for livestock or aquaculture businesses. However, many fish farmers, in particular within tropical regions, still rely on juveniles of variable and unpredictable quality produced from eggs spawned in sea cages from broodstock fed on trash fish, along with larvae reared in extensive pond systems. This traditional production method, which attempts to mimic the fish species natural ecosystem, has been in the early establishment of many now mature aquaculture industries which have then evolved to use intensive and biosecure propagation practices.

For a number of tropical marine fish species though, natural maturation and pond-based larval rearing is still the means to produce eggs, rear larvae and produce juveniles, as is their biology and intensive culture requirements are not fully understood.

These activities are normally carried out by small-scale family based backyard hatcheries and represent an important income generation opportunity for lower income farmers throughout Southeast Asia, and are effective in producing low cost fry.

However, traditional production methods have limited industrial applicability due to low and variable productivity, and the use of relatively extensive areas within coastal regions (highly sought after by estate developers) for the production of negligible fish biomass per unit of area.

The high variability in survival and larval quality obtained from open cage broodstock and extensive larviculture pond systems is due to a variety of reasons, starting with an inadequate â&#x20AC;&#x153;trash fishâ&#x20AC;? diet for the broodstock. If specific vitamins, fatty or amino acids are lacking, limited or in inadequate ratios within the diet, this will translate into poor quality of eggs and sperm.

The problem is then passed into the hatcheries which must deal with high percentages of unfertilized eggs, poor hatching rates and continuous mortalities of larvae. This scenario is well known among hatchery managers who rightfully prefer to discard batches with poor egg fertilization (<65%) than to deal with a cascade of negative effects in attempts to save weak larvae, a situation where their best efforts in hatchery practices are not resulting in large numbers of quality larvae for stocking.

In open systems, there is little to no control over water quality parameters, flow rates, lighting, etc. to provide animals with a stable and stress-free environment, tailored to match their optimal physiological conditions. In pond-based larval systems, for instance, fish larvae will rely on the natural phytoplankton and zooplankton productivity for feeding once their yolk sac reserves have been absorbed. This natural productivity, critical for the success of this system, depends on a variety of factors such as the abundance and right type of available microalgae and zooplankton pumped from the adjacent farm environment into the pond, the environmental conditions and availability of nutrients for the maintenance of adequate plankton blooms.

Poor water or suboptimal weather can be debilitating, if not lethal, for fish in their most delicate and fragile life stage.

Another critical impact of larval rearing problems, which are passed on to the growers, are the life-long consequences of nutritional deficiencies and environmental stressors encountered by survivors during their early development.

Larval rearing is a period when the animalâ&#x20AC;&#x2122;s sensory, digestive, musculoskeletal, nervous, endocrine and immune systems are forming. At this stage, both the expected performance of the surviving offspring may be compromised and, as documented in other vertebrates such as humans, damage may also be inflicted to progenitor germ cells of offspring (PGCs, or cells that

17

FEATURES will undergo meiosis and become gametes). Damage caused to PGCs will unintentionally affect the performance of the next generation due to transgenerational epigenetic effects. This is because epigenetic reprograming, a critical phenomenon for normal development which occurs during gametogenesis and embryogenesis, is easily disrupted by nutritional deficiencies and environmental stressors, such as a few degrees of temperature change, or exposure to trace amounts of endocrine disrupting chemicals.

The major weak link in open systems in general, and for broodstock, eggs and larvae in particular, is the constant exposure to all kinds of parasites and pathogens. Viral nervous necrosis (VNN) is probably one of the most harmful pathogens for marine fish larvae. Recent studies powered by next generation sequencing technologies now estimate that seawater contains about one million to ten million viral particles per millilitre. Thus, it is not surprising among aquaculturists that new deadly viruses appear, and will continue to appear, frequently. In seacages and open ponds, it is impossible to maintain basic levels of biosecurity and larval quality becomes a lottery. The odds of winning – and making money out of fish farming at industrial scales using juveniles produced from these systems – are slim.

The sustainable development and expansion of tropical marine food fish aquaculture relies on the prompt availability of high quality seed stock. This implies full control over both the environment where seed stock is produced and the genetics of the broodstock. For tropical marine finfish, very few selective breeding programs exist, and for those that do, improved stocks have not yet been fully disseminated to farmers. A few tropical marine species have been domesticated to date, with life cycles fully closed in captivity and where broodstock are sourced from grow-out systems. However, the reality is that these species are likely to be highly inbred. This is due to the incredible fecundity of marine finfish, where it is common for a single female to spawn a few million eggs at a time. In practice, most hatcheries would normally stock just a few hundred thousand larvae per run. In turn, low numbers of broodstock are used and even lower numbers of broodstock actually contribute to a spawn.

In these cases, wild sourced fish must be used to recover the lost genetic diversity in inbred lines. This is because inbreeding depression takes just a few generations to manifest, usually as a loss of reproductive performance, growth rate and overall poor production performance. As such, the term “domesticated” is

18

still under debate for most tropical species. Selective breeding programs, although effective and with high return on investment of 10-15:1, are still expensive to implement and maintain for most tropical aquaculture marine finfish species because no “big players” exist (e.g. companies harvesting more than 10K tonnes p.a.). Incipient programs to date have been heavily subsidised through government funded R&D initiatives and managed by universities and research centres. However, production outcomes from business driven investments towards selective breeding programs for tropical marine fish species are still to be realised.

It is important to note that genetics does not bring value without healthy and disease-free stocks. The first and most effective step for industrial production of high quality seed stock is the investment in a fully controlled indoor production facility, with uncompromised biosecurity. Without this, the fate of any selective breeding program is at risk, because if improved broodstock become infected, with viruses in particular, there are no treatments and stocks must be culled. A hard lesson to be learned from shrimp farming is the fact that a number of breeding programs with compromised biosecurity have been terminated after years of hefty investments in genetically superior lines.

So, what are the key elements required for broodstock and hatchery facilities to deliver reliable and regular production of high quality eggs, larvae and juveniles of tropical marine food fish? Firstly, any new incoming water must be pre-filtered and effectively sterilized with ozone and/or UV and preferably stored in an intermediate tank before coming in contact with the culture species. Storage is important as it will give the facility a “buffer”, with some clean water available in times when the sterilization systems fail or need maintenance. Secondly, broodstock, hatchery and nursery facilities must be selfcontained and physically separated from one another, make use of recirculation aquaculture systems (RAS), have restricted access, individual dedicated equipment, and highly trained and attentive technical personnel. Thirdly, and this is good news, broodstock and larviculture facilities (for which biosecurity is imperative), require a very small footprint, low water usage and labour requirements, and are therefore easy to manage and control.

However, within emerging finfish industries there is a great deal of confusion about what makes a good breeding and

as fingerling production will always have commercial priority.

The following broodstock and hatchery section of this article deals with breeding for production (not for a closed nucleus), and introduces concepts that are in reach to any small scale hatchery so that they can use the best genetics possible. This article highlights some simple key technical elements for the production of high quality, disease free and non-inbred seed stock of tropical marine finfish which will have excellent performance under grow out conditions.

FEATURES

hatchery facility, and what a closed selective breeding nucleus is supposed to be. Most often, there is a misunderstanding or expectation that they can be both in one. In established industries, these are in fact separate activities and facilities with different infrastructure and different end goals. The goal of the breeding and hatchery is to produce high quality juvenile fish to be farmed, by spawning the broodstock with the very best genetics of all held in that facility, i.e. broodstock with the highest estimated breeding values (EBV). The goal of the selective breeding nucleus is to maintain genetic diversity through multiple crosses, and selecting a range of that diversity to become the next generation of breeders, for which a proportion is passed on to the breeding and hatchery facility (also known as a multiplication centre). Within the closed nucleus, no animals that have been exposed to the outside world (farmed or wild) are ever brought back to the facility, as it would be a biosecurity breach. The selection in the closed nucleus is based on the performance of relatives (siblings or offspring) tested under real-world grow out conditions, and more often also in dedicated disease challenge facilities. This requires large infrastructure and investment. When both closed nucleus selection and breeding for production activities are attempted in a commercial hatchery, they clash with each other

http://www.scienceimage.csiro.au/

BROODSTOCK FACILITY AND MANAGEMENT The broodstock facility must have an isolated and dedicated quarantine area with a tank where broodstock candidates are held until they undergo health checks as prescribed by a veterinarian knowledgeable in aquatic animal health, who will advise on effective parasite treatments. Broodstock candidates preferably with known age and superior size within their group should be selected from different batches and different origins, weighed and measured, tagged with passive integrated transponder (PIT tag), sexed with disposable cannulation tubes, and have the contents of a gonadal biopsy PCR scanned for viruses of importance, such as nodaviruses and iridoviruses. Cannulation tubes should be for single use only, never cleaned for re-use or sharing among fish, due to the high risk of transferring pathogens (especially viruses) from one fish to another. It is important to note that a negative PCR result does not imply virus free fish, either because the virus was not present in the sample, or was in quantities below the detectable levels of the assay; but a positive result implies that the fish is a carrier. Broodstock candidates confirmed to be infected by VNN for example should be rejected and not moved from the quarantine into the breeding tanks. Further, selected broodstock candidates should be fin clipped for DNA genotyping (individual “fingerprinting”) and genetics advice sought on inbreeding status, pairwise relatedness matrices and DNA parentage analysis (if possible), and best mating allocation combinations to avoid crossing relatives.

The broodstock facility needs to be within a fully enclosed building and there is no need for more than three tanks, fitted with transparent side windows for fish observation, each tank independently operated in RAS including a heater-chiller for absolute control and stability of water temperature (± 1oC), and

covered for control of lighting schedule. A simple and effective strategy for year round spawnings and commercial production is to manage these three tanks as: a) winter or recovering tank, b) summer or conditioning tank, and c) spawning tank, with broodstock in best reproductive condition (readiness), with low relatedness and/or highest genetic merit for traits of importance. Broodstock should be rotated among tanks depending on the reproductive condition of spawners.

Broodstock should be fed ad libitum three to four times per week with a high quality commercial marine finfish broodstock diet. It seems counter intuitive, but feeding more often results in broodstock accumulating abdominal fat and gonadal maturation will be hampered, with fish losing the natural aggressiveness seen in normal courtship and breeding behaviour (in lay terms, fish will get fat, lazy and males do not chase the females).

Broodstock should not be fed with commercial growout diets, ‘trash’ fish, or even human grade seafood like frozen/thawed pilchards and/or squid for obvious biosecurity reasons – these fresh/frozen diets can host unwanted bacteria and viruses such as VNN. Commercial broodstock diets are indeed several times more expensive than growout diets, and for a good reason. They are scientifically formulated to meet the highly demanding nutritional requirements for marine finfish gonadal development and maturation (instead of muscle growth as targeted in growout diets) and contain the absolute best raw materials for optimal reproductive performance.

Reliable spawnings depend on precise environmental, social and nutritional factors which influence gonadal maturation, whereas the use of hormones to induce maturation has proven unsuccessful and/or unreliable. Dependence on natural spawning, once the gonads are mature and the fish are ready,

19

FEATURES

may be possible if it fits within the hatchery schedule. However, hormone-induced spawnings can be predictable with good fertilization rates by use of hormones like Luteinizing Hormone Releasing Hormone analog (LHRHa) or Human Chorionic Gonadotropin (HCG) on females that have large and uniform oocytes (eggs) with opaque yolk (detected by ovarian biopsy), and on males with white, sticky milt and motile sperm. Fertilized eggs must be disinfected at the correct developmental stage; too soon or before gastrulation and they are too fragile, too late and bacterial/fungal load may be too high. Iodine and formalin

are the most common and safest disinfecting agents. Dedicated, well-aerated incubator tanks provided with pre-treated, flow through water should be used for hatching eggs. These tanks require siphoning several times during incubation to remove egg shells and dead eggs where bacteria, protozoa and fungus rapidly grow. Stronger larvae will often migrate (or passively float) to the surface upon turning off aeration and flow, and those should be used for stocking larval tanks on the first or second day post hatching.

LARVAL CULTURE AND MANAGEMENT Larval tanks should also be housed in a fully enclosed and biosecure building where ambient temperature and photoperiod can be fully controlled. This is important because larvae in tanks exposed to natural light will utilise energy searching for prey from dawn until the time hatchery staff are able to harvest live feeds and offer the first meal. Live feeds, in particular rotifers and Artemia, remain essential for tropical marine fish hatchery production and should be cultured in an annex adjacent and connected to the larval room.

When rearing fish species with larger mouths, which do not require copepod nauplii and accept rotifers at first feeding, microalgae culture in the hatchery facility is discouraged. Live algae culture requires unnecessary additional labour and usually open and exposed space, it does not support high rotifer culture densities (thus more live feed space and large tank volumes), is a potential source of contamination and does not provide the same nutritional profile as commercially available concentrated algal and enrichment products. Some commercial products offer a single solution for rotifer maintenance and enrichment in semi-continuous culture with high rotifer culture densities, thus requiring fewer small tanks, e.g. two 500L rotifer tanks may be sufficient for feeding and rearing 500k Asian seabass larvae.

For species in which the biology and culture requirements are well known, such as the Asian seabass, high stocking densities are preferred to optimize space, provided both water quality and high availability of live feeds can be maintained. While for some species it may be feasible to produce juveniles without highly unsaturated fatty acid (HUFA)-enriched Artemia, this practice is not advised as it will likely compromise larval quality, fish development and grow-out performance in the long run.

The use of RAS in hatcheries is critical to provide stable water quality and allows for a gradual decrease in salinity, if that is tolerated by the species and beneficial to prevent certain seawater-related bacterial and viral outbreaks (e.g. management of big belly disease in Asian seabass). Grading should be performed as often as required and before any cannibalistic behaviour takes place. Daily cleaning of the bottom of the larval tank (mortalities, organic matter, dead rotifers/Artemia, faeces, uneaten feed) is critical, starting from a few days post stocking.

In addition, standpipe screens should be cleaned, together with daily observation of water quality (temperature, DO, pH and ammonia as a minimum) and colour (turbidity).

Activity and behaviour of larvae must always be checked closely by experienced personnel, including stress indicators (e.g. feeding response; swimming pattern; cannibalism), preferably several times a day during the first month of hatchery production. Some simple strategies, such as a general microscopic examination, can help to determine the fingerling health status. The most important features to observe in fish larvae are evidence of feeding (gut fullness), the swimbladder condition (inflated, non-inflated, hyperinflated; depending on fish species), skin pigmentation/colouration, presence of parasites or abnormalities on gills and presence of external body damage (damage may be a first â&#x20AC;&#x153;doorâ&#x20AC;? for parasite/pathogen infections).

Body damage in fingerling stages can be a sign of cannibalism which may indicate low feed quantity or quality, or that grading is overdue. A regular health check of larvae and juveniles under the microscope, at least once daily, is necessary as changes on health status can happen quickly during early stages (e.g. epitheliocystis in cobia larvae culture) and allows managers to respond immediately. Examining fingerling health status is a necessary task to be implemented in fish hatcheries to guarantee the start of successful production. Partnering with molecular laboratories that can return diagnostic results in few days is also advised to assist in surveillance of pathogens in water and fish, because prevention is always better than cure.

Ultimately, there is an increasing consumer demand for tropical farmed finfish. Fingerlings of high quality, produced in biosecure facilities and with better genetics are the foundation to improve farm productivity. These fingerlings will allow aquaculture enterprises in the tropics to achieve profitability and sustainability to feed our growing population.

20

ISSUE HIGHLIGHT

Evaluation of Functional Feed Additives Against Shrimp Pathogens with an emphasis on Vibrio parahaemolyticus and White Spot Syndrome Virus (WSSV) in Litopenaeus vannamei Farshad Shishehchian

Ph.D., Terrestrial and Aquatic Ecology, President & CEO; Blue Aqua International Group of Companies

Krit Khemayan

Aquaculture Technician, Blue Aqua International Group of Companies

Zahra Javidi

Technical and Registration Manager Blue Aqua International Group of Companies

S

ABSTRACT

hrimp aquaculture has been dramatically affected by many pathogenic diseases, mainly caused by V. parahaemolyticus and White Spot Syndrome Virus (WSSV). The aim of this study was to evaluate the potential use of 2 functional feed additives of AlphaGuard*L Plus (Liquid) and AlphaGuard*P (Powder) composed of Essential Oils (Eucalyptus, Thyme, Oregano belonging to Myrtaceae and Lamiaceae family respectively), Medium Chain Triglycerides (MCTs) and natural bioactive compound in shrimp against disease caused pathogens especially V. parahaemolyticus and WSSV. No significant differences in growth performance were found at 0.5% of AlphaGuard*P and AlphaGuard*L PLUS. The results indicated that AlphaGuard*L Plus effectively delayed disease progress in shrimp. These results suggest that the functional feed additive of AlphaGuard*P and AlphaGuard*L PLUS could be used in order to promote shrimpâ&#x20AC;&#x2122;s defense against pathogens.

Keywords: Functional Feed Additive, AlphaGuard, Litopenaeus vannamei, Vibrio, White spot disease

Introduction

T

he practice of aquaculture intensification is impeded by health and nutrition - affecting growth performance. To untangle these consequences, functional feed additives have been used to stimulate shrimp immune and improve shrimp performance specially to control viral and bacterial pathogens in recent treat shrimp diseases (Thimadee et al., 2016) such as V. parahaemolyticus that caused Acute Hepatopancreatic Necrosis Disease (AHPND) and White spot syndrome virus (WSSV) that caused White spot disease (WSD) combined with the implementation of biosecurity measures.

The composition of AlphaGuard composing MCT and essential oil are being recognized as GRAS practice. In this study, the efficiency of AlphaGuard product was obtained by a combination of the data from Disc-diffusion test result, disease challenge, histology and growth performance together with an evaluation of AlphaGuard product effect on the shrimp diseases.

21

ISSUE HIGHLIGHT

MATERIALS AND METHODS Pathogenic bacteria and virus Two virulent pathogens in shrimp, V. parahaemolyticus and V. harveyi in glycerol stock, have been sub-cultured in Tryptic Soy Broth (TSB) + 1% NaCl, incubated at 37oC for 24 hours before streaked onto Thiosulfate-citrate-bile salts-sucrose agar to obtain its pure culture.

WSSV suspension was prepared from the muscle of WSSV infected shrimp. Briefly, the WSSV-containing in shrimp the muscle was removed from storage at -80°C and cut into uniform pieces under cold sterile conditions then homogenized in TN buffer (20 mM Tris-HCl, 400 mM NaCl, pH 7.4) at 0.1 g/ ml. After centrifugation at 2,000 rpm for 10 min at 4°C, the supernatant was diluted to 1:100 with 0.9% NaCl and filtered through 0.45 micron. The resulting supernatant was stored at -80°C until used as a source of WSSV injection for the experiments.

The antimicrobial activity

l

Disc-diffusion test

5 µl of the of AlphaGuard*L Plus and AlphaGuard*P, concentration ranging from 0, 0.25, 0.5, 1.0, 10% dilution with NaCl 0.85%, were absorbed into 5 mm diameter, 0.9 mm thick paper disc then air drying before placed to 105 cells of bacteria culture in a Petri dish size 100 x 15 mm. 0.85% NaCl solution was used as negative control. Three replicate plates for each treatment were used. Observations were recorded after 24 hours.

Artificial infection with V. parahaemolyticus and determining the number of Vibrio in shrimp hemolymph (HL) and hepatopancreas (HP) V. parahaemolyticus was reactivated from storage at -80°C and cultured in TSB + 1% NaCl at 37°C overnight. The culture was centrifuged at 5,000 rpm for 10 min to remove the supernatant, and the pellet was re-suspended in sterile 0.85% NaCl to a density of 8.4×106 CFU/ml. The shrimps at 5th week were infected by injecting 100 μL of the bacterial suspension into the second abdominal segment of healthy shrimp at 5th week of culture and mortality was recorded for 7 days. l

Shrimps were collected randomly at 3rd day after infection for each treatment and washed three times with sterile water.

HL was withdrawn from the pericardial cavity using a 1-mL sterile disposable syringe under sterile conditions and added to an equal volume of sterile anticoagulant (adding 10 mM EDTA-Na2 to 450 mM NaCl, 10 mM KCl, 10 mM HEPES, pH 7.3, 850 mOsm.kg-1 or 10 mM Tris-HCl, 250 mM Sucrose, 100 mM Sodium citrate, pH 7.6). The HL and ground HP were serially diluted 10-fold with cold sterile PBS. Each dilution was spread on TCBS agar plates placed upside down in a 37°C incubator and cultured for 16–20 hours. Plates containing 30– 300 bacterial colonies were counted, then the numbers were recorded as CFU/ml or CFU/g.

22

Artificial infection with WSSV 0.1 ml of the filtrate was injected intramuscularly into healthy shrimp at 5th week of culture. Shrimps were collected randomly at 6 hours after infection for Histopathology examination (Bell T.A. and Lightner D.V. 1988) and recorded mortality for 7 days. l

Shrimp Growth Conditions and

experimental groups

Shrimps were purchased from local Thai farm. There were negative from V. parahaemolyticus caused AHNPD and WSSV by PCR analysis. After acclimatized for one week in aquarium prior the experiment, apparently healthy shrimp with uniform body length was divided randomly into 3 groups with 6 replicates per group, 20 shrimps per replicate. Aquarium capacity was 100 liters containing 60 liters of sea water. Saline water property was maintained at 15 ppt, pH 7.7-8.0 and DO>4.0 mg/L. Each treatment contains juvenile shrimps with an average initial weight of 2.6 g randomly stocked in each tank. Shrimps were fed to satiation around 2.5-3% of body weight, three times per day throughout 5 weeks. Feeding was adjusted daily according to ingested rate to make sure that feed was totally consumed.

Before feeding, molts, feces, and dead shrimps were removed.

20% of the water in each tank was exchanged every 3 days by new seawater.

Experimental diets for growth

performance and data collection

All groups were kept during the experiment under the same conditions as acclimation. Control group was fed by commercial shrimp pellet coated with 1% chitin-chitosan. Treatment groups were fed by AlphaGuard*L Plus or AlphaGuard*P sprayed on commercial shrimp pellet and coated by 1% chitin-chitosan at dosage 5.0 ml or g/Kg feed Feed intake, mortality and water quality parameters were recorded daily. At the end of the trial, the survival rate was evaluated. Shrimp weights were determined at the beginning (Initial Weight), third and fifth (Final Weight) week of the experiment. Weight gain and feed consumption were used to calculate several parameters; Feed conversion ratio (FCR) = feed consumed (dry weight)/live weight gain (wet weight).

Daily Weight Gain (DWG; g/day) was calculated as: (Final weight (g) − Initial weight (g))/days.

Statistical analysis Statistical analysis experimental units, tanks, and aquariums were distributed in a completely randomized way. Quantitative data were checked for normality and homoscedasticity. Data were analyzed using one-way analysis of variance (ANOVA) to search for significant (p < 0.05) differences among treatment means.

ISSUE HIGHLIGHT

RESULTS AND DISCUSSIONS The antibacterial activity by Disc-diffusion method and inclusion in shrimp diet on disease resistance of AlphaGuard*L Plus or AlphaGuard*P Disc-diffusion test The concentrations of AlphaGuard*L Plus and AlphaGuard*P at least 1% inhibit both V. harveyi and V. parahaemolyticus, however, there was higher inhibition efficacy for V. parahaemolyticus compare than V. harveyi. The effective concentration of AlphaGuard might be lowered if we diluted in lipid soluble such as Ethanol for testing. As the lipid compounds in AlphaGuard may not disperse appropriately across the culture medium. Essential oil compositions, assay techniques and the mode of action have been intensively reviewed (Chouhan et. al. 2017, Citarasu 2010, Faleiro 2011 Nieto 2017, Padalia et. al. 2015 and Zhang et. al. 2010). this study did not investigate at molecular details for its mechanisms, however, the main effect of bacteriocidal properties might be related to their interference at membrane integrity and permeability as the lipophilic compound from essential oil and MCTs. Different fatty acids in MCTs have a different minimum inhibitory concentration (MIC), depending on the type of fatty acid, microorganism, and environmental pH. The synergistic and antagonistic aspect between ingredients in AlphaGuard is not elucidated in this study. The essential oil in AlphaGuard also acts as an antioxidant to prevent or slow down oxidation of unsaturated MCTs to prolong its shelf life besides anti-stress in shrimp. l

Fig 2: Cumulative mortality rate of V. parahaemolyticus challenge test.

Means with different superscripts are significantly different (p<0.05).

Fig 3: Mortality and survival rate of V. parahaemolyticus challenge test.

Means with different superscripts are significantly different (p<0.05).

The ability of shrimp clearance from Vibrio spp. after 3 days of infection was plotted graph as in Fig. 4. The results indicated that the Vibrio spp. count in hemolymph of shrimp fed by AlphaGuard*P and AlphaGuard*L Plus were lower than control significantly whereas the ability of shrimp to defense against bacteria in HP found that group of shrimp fed by AlphaGuard*L Plus had better ability significantly than shrimp fed by AlphaGuard*P and control.

Fig 1: Antibacterial activity from Disc-diffusion test. Abbreviation L or P with number represented liquid or powder form with percentage of *L Plus *P AlphaGuard or AlphaGuard .

V. parahaemolyticus challenge test At the end of feed trial experiment after 5th week, shrimps were challenged by V. parahaemolyticus. Cumulative mortality rate is plotted in graph as Fig. 2. The results of mortality rate after V. parahaemolyticus challenge for 7 daysâ&#x20AC;&#x2122; interval showed that groups of shrimp fed by AlphaGuard*P and AlphaGuard*L Plus had lower mortality rate significantly than control group after day 4th of infection. Both AlphaGuard*P and AlphaGuard*L Plus had the same statistical mortality rate after day 5th of infection.

At the end of experiment as Fig. 3, in contrast, survival rate not statistically correlated to mortality rate for AlphaGuard*P l

23

ISSUE HIGHLIGHT

Log CFU/ml Vibrio count after 3 days of infection Log CFU/ml Vibrio count after 3 days of infection

5 5 4 4 3 3 2 2 1 1 0 0

5.63a 5.63a 4.22a 4.22a

0 0 0.85%NaCl 0.85%NaCl

4.16b 4.16b

4.19a 4.19a 4.02c 4.02c

4.12b 4.12b

0 0

V parahemolyticus V parahemolyticus

AlphaGuard*P AlphaGuard*P

AlphaGuard*LPlus AlphaGuard*LPlus Heamolymp Heamolymp

Hepatopancrease Hepatopancrease

Fig 4: Vibrio spp. count of V. parahaemolyticus challenge test reported as CFU per ml or gram.

Means with different superscripts are significantly different (p<0.05).

WSSV challenge test WSSV challenge test was conducted after 5th weeks of the feeding trial. The cumulative mortality was plotted as graph in Fig. 5. The group of shrimp fed by AlphaGuard*L Plus is shown significantly delay in the mortality compare to the groups of control and AlphaGuard*P during 2nd to 4th day as shown in Fig.6. This is coincided with no histopathological sign of H&E staining WSSV infection tissue at the area of injection except the generalization of muscle necrosis that is likely related to the shrimp defense mechanism after 3 days of infection.

Nevertheless, all groups ended up with 100% mortality. The group of AlphaGuard*P exhibited some sign of fungi infection by fibrous invading to sub-epithelial tissues in Fig 6 (3) and (4). l

*P

Fig 6: Histology after challenged by WSSV in shrimp fed by AlphaGuard *L Plus and AlphaGuard

(1) Control and (2) AlphaGuard*P shown the late stage of intranuclear inclusion body of WSSV infection to HP as indicated by black arrow. (3) AlphaGuard*P exhibited some signs of fungi infection by fibrous invading to sub-epithelial tissues. (4) Zoomed-in area from black rectangle from black arrow indicated in (3). (5) and (6) AlphaGuard*L Plus shown no evidence of WSSV infection but it demonstrated the generalized muscle necrosis.

Fig 5: Cumulative mortality rate of WSSV challenge test. Means with different superscripts are significantly different (p<0.05).

24

Alphaguard might have triggered or modulated a series of nonspecific immune responses against shrimp pathogens, however this needs to be confirmed via haemocyte count, pro-phenoloxidase activity or immune-related gene expression as the example indicators.

T

Conclusion

the gastrointestinal morphology such as increased intestinal microvillus or crypt depth as found in piglet or fish, nevertheless, we need to confirm this by performing more research on section of hepatopancrease lumen in shrimp. AlphaGuard could also induce better resorption of liposoluble vitamins due to its lipophilic properties. The overall of the growth performance should be elucidated further in the real pond.

ISSUE HIGHLIGHT

Effects of AlphaGuard*L Plus and AlphaGuard*P on L. vannamei growth performance Growth performances of AlphaGuard*L Plus and AlphaGuard*P at 0.5% applications starting from 1st week until 3rd week were no significant differences for all parameters from control in Fig.7. Weight gain increased and FCR decreased than control but there was no statistical difference. MCTs can be modified

ACKNOWLEDGMENTS

he results of this trial suggested that the application of AlphaGuard*P and AlphaGuard*L Plus feed additive at 0.5% with commercial feed proved the efficacy on promoting the shrimp defense against pathogen. Especially with AlphaGuard*L Plus showing a better performance.

All experiments were conducted at the Nutrition and Aquafeed Laboratory, Department of Aquaculture, Faculty of Fisheries, Kasetsart University, Bangkok, Thailand. Bacteria and virus were provided from the Department of Fisheries, Kasetsart University in Thailand.

CONFLICTS OF INTEREST The authors declare no conflict of interest.

REFERENCES Bell TA and Lightner DV. 1988. A Handbook of Normal Penaeid Shrimp Histologyâ&#x20AC;? The World Aquaculture Society.

Chouhan S, Sharma K, Guleria S. 2017. Antimicrobial activity of some essential oils-Present status and future perpectives. Medicines 4, 58.

Citarasu T. 2010. Herbal biomedicines: a new opportunity for aquaculture industry. Aquacult Int 18:403-414.

Faleiro ML. 2011. The mode of antibacterial action of essential oils. Science against microbial pathogens: communicating current research and technological advances.

Nieto G. 2017. Biological activities of three essential oils of the Lamiaceae family. Medicines 4, 63.

Padalia H, Moteriya P, Baravalia Y, Chanda S. 2015.

Antimicrobial and synergistic effects of some essential oils to fight against microbial pathogen. The battle against microbial pathogens: Basic science, Technological advances and educational programs.

Thitamadee S, Prachumwat A, Srisala J, Jaroenlak P, Salajan PV, Sritunjalucksana K, Flegel TW. 2016. Review of current disease threats for cultivated penaeid shrimp in Asia. Aquaculture 452:69-87

Zhang J, An M, Wu H, Stanton R, Lemerle D. 2010. Chemistry and bioactivity of eucalyptus essential oils. Allelopathy Journal 25 (2): 313-330

25

INTERVIEW ISSUE HIGH LIGHT

7Questions With

Ravi Kumar Yellanki Mr. Ravi Kumar Yellanki CEO

Vaisakhi Bio-Marine Pvt. Ltd

1.

Tell us a little bit about yourself!

I did my graduation in civil engineering and Master's in Management. I have been working in shrimp aquaculture for the last 20 years. So today, I call myself an aqua culturist than anything else.

2.

How did Vaisakhi Bio-Marine start its operations?

I have been in shrimp aquaculture for the last 20 years. I started my career as civil engineer working for the leading consultants in India. Then I started a consultancy firm with group of friends to completely man the hatcheries and produce seed. That was the time we were looking at the business very closely. Soon we started Vaisakhi Bio-Marine (P) Ltd hatchery operations and carved a niche by producing disease free seed using wild monodon broodstock. During the vannamei boom, we have acquired three more companies. One of the companies we acquired has a large farm and a hatchery. That paved way for our entry into farming. Today, we produce about 3.5 billion fry and about 1500 tons of shrimp per annum. We have plans to make our company vertically integrated by coming in to exports.

26

3.

What are the current challenges in Indian aquaculture?

In the beginning of this year, the Indian industry struggled because of low global shrimp prices. India produced a record high 700,000 tons of shrimp this year and was trying to sell most of it to US and China. India could not sell much to Europe as the EU came up with a new regulation of testing 50% of the shipments. Coming up with a change in processes throughout the value chain and convincing the EU was one of the challenges before the industry. The industry is very much up to the task and trying to achieve zero tolerance for antibiotic residual loads. The other major issue is disease threat. On one side we have good old WSSV which never gives any respite to the farming fraternity; on the other side we have the emerging issues like EHP, white feces, and running mortality syndromes.

In India, we have more small scale farms than medium and large scale farms. The farmers operating small scale farms are less aware of measures to be taken to avoid these new issues and by losing big time money. Low prices in tandem with disease problems could bring the production down substantially in the coming years.

4.

What about the local market trends and opportunities in the industry?

Right now there are no organized domestic markets for farmed shrimps in India. The small shrimps that are caught because of distress harvest are finding their way to wet markets. The intermediaries in between the consumer and the farmer are making profits as it is neither planned harvest nor sale. The major impediment for shrimps to be sold in dry markets is lack of cold chain infrastructure. Right now the organized retail

5.

There are lots of talk on 'sustainable practicesâ&#x20AC;&#x2122; in aquaculture. In your opinion, what kind of strategy should businesses focus on?

Farmers should work on efficiencies rather than cutting corners in the cost. Today the problems faced by the industry are different from what it used face earlier. So farmers should move away from their conventional approach and come up with new methods and tools to operate efficiently. In the process they may have to also incur some capital cost. In India, there is a school of thought that going for lower densities is more sustainable. But I donâ&#x20AC;&#x2122;t think that is going to solve the problems. When you are affected with disease, it does not really matter whether you have gone for lower densities or higher densities. Pond preparation, water treatment, seed selection, farm management in terms of feed and microbes and so forth have to be revisited and changed to counter todayâ&#x20AC;&#x2122;s problems. Hatcheries have to suppress bacterial loads to minimum possible levels and give seed that is free of EHP. Health care companies should come up with new age strains of microbes to competitively exclude or suppress the fast changing pathogens in the culture systems. Feed companies should work on functional feeds that reduce FCRs and thereby organic loads in the pond and also enhance the immunity of the shrimps. The other solution could be from genetics: coming up with tolerant stocks for bacteria and WSSV.

6.

7.

Where do you see Indian aquaculture in the next 5 years?

With new problems emerging by the day, shrimp farming is becoming more of a silence than an art. So I think, slowly small farmers could be phased out in the coming years as they are not ready to understand new methods fast nor they are interested in investing. Sooner than later domestic market for farmed shrimp is going to be established. That could change the entire landscape and give opportunities for further growth. The other change I foresee is that companies might go in for closed/semi closed farms with smaller ponds and higher densities and aim for higher productivity.

INTERVIEW

sector is only about 7%. Further liberalization on multi-brand retail could remove the barriers for global retail giants to peg their tents in India. This could auger well for shrimp industry as retail giants would set up cold chains with in no time.

What are the most recent advances in Hatchery management in India?

India is the only country in the world that is not using any pond raised broodstock or for that matter broodstock from any loose breeding programs. Indian hatcheries have come of age in terms of the way they disinfect their facilities and feed the animals in maturation. Hatcheries have started washing the eggs thoroughly to reduce the pathogens. Most of the hatcheries either pasteurize or freeze their feeds before feeding. Hatcheries mostly do not operate their facilities for more than one cycle; after through disinfection only they go for the second cycle. Many are completely doing away with antibiotics and using probiotics right throughout the cycle. Some of the hatcheries test their seed in house for the pathogens before they release to the market

27

NEWSHIGH & PRESS ISSUE LIGHT

One industry, One Source, One IntraFish The one-stop-shop for seafood news

IntraFishâ&#x20AC;&#x2122;s web-platform brings all our industry-leading titles underneath the IntraFish.com umbrella.

We call it One IntraFish - the one-stop-shop for seafood news.

Your live news coverage is enhanced with more exclusive news, in-depth analysis, opinion and insight than ever before.

The responsive design allows readers to enjoy an improved reading experience on desktop, laptop, tablet or smart phone.

Visit IntraFish.com now for an easier and even better way of accessing your seafood news.

For advertising and subscription enquiries, please contact John Dogma at +65 65570047 or email john.dogma@intrafish.com

NEWS & PRESS

Ecuadorean

News Around the World EU to fund Ecuadorean shrimp sustainability initiative Organized by

https://www.undercurrentnews.com/2018/07/25/eu-to-fund-ecuadorian-shrimp-sustainability-initiative/

T

he EU will fund the Sustainable Shrimp Partnership (SSP) to boost Ecuadorean shrimp exports to European countries, through "EXPORT-DES", a program run by the Corporation for Exports and Investments Promotion (CORPEI).

SSP is an initiative launched by some of the largest Ecuadorean shrimp farmers last March, with the objective "to transform the future of the industry and turn it in a new direction – a race to the top".

The EU is supporting SSP with the aim of increasing the number of actors, especially small and middle-size producers, to participate in the initiative, it said.

A total of $148,000 will be invested in the initiative, of which half will be financed by the EU and the other by the Ecuadorean national chamber of aquaculture (CNA), according to a cooperation agreement confirmed last week by by Eduardo Egas, executive president of CORPEI, and Jose Antonio Camposano, CNA executive president.

"The European Union, within the framework of the free trade agreement with Ecuador, created the fund to boost the competitiveness of the national product, and increase the exportable supply to that destination. EU resources, which are managed by CORPEI, seek to invest in projects such as SSP that promote sustainable production," Egas said.

“Ecuador has always demonstrated its leadership in sustainability, but we must continue to evolve. Sustainability criteria are not static -- they are always changing as circumstances change and as technology advances. What is sustainable today, will likely not be done tomorrow," said Camposano.

“Via the SSP platform, we are looking to the future and are recognizing where we can evolve and how we can ensure we are continually providing a product to meet changing consumer demands. This investment fund will support us in getting more Ecuadorian production into the program and increasing the supply of sustainable, traceable and antibiotic-free shrimp into the EU marketplace," Camposano added.

29

NEWS & PRESS

Investments in Russian aquaculture on the rise

R

ussia is experiencing a boom in aquaculture and is seeking to increase its farmed seafood almost threefold, to 700,000 metric tons (MT) through 2030. Investment activity in the sector during the last three years suggests the projected figure can be exceeded provided some cornerstone problems be solved and international experience be used.

This is the second article in a two-part series exploring Russia's aquaculture industry. Part one, “Russia’s aquaculture industry brimming with potential,” appeared on Thursday, 23 August.

Systemic efforts taken by the government coupled with growing demand on domestic market due to the import food embargo have encouraged massive investments into Russian aquaculture, with a few large-scale projects launched every year.

In 2017-2018, more than a dozen new initiatives to build infrastructure for fish farming were announced.

Most projects are strongly supported by local authorities as aquaculture is the best tool to get relatively cheap fish for regions located far from coastlines and to create more jobs.

Some regions in Russia are even forcing the development of the sector, making it one of the top priorities.

One example of this is the government of Chelyabinsk region, which has established a special fishery council, headed by local governor Boris Dubrovsky, to advance projects in aquaculture, initially by helping in the organization of sites for farms. The region has adopted an aquaculture development program and has achieved an increase in production from 3,200 MT in 2013 to 4,700 MT in 2016, with a desire to further increase that tonnage to 7,000 MT through 2020.

“Aquaculture is as important for the region as cattle breeding and crop growing,” said Boris Dubrovsky of the initiative in one of his public speeches.

In Vologda region, Aquaproduct company is establishing a newly-branded facility for farming char, with a capacity of 2,000 to 2,500 MT a year. The investment is projected to amount to RUB 1.7 billion (USD 25.5 million, EUR 22.14 million) through 2021.

The Siberian Investment Group is also diving into the aquaculture sector, and has started the first phase of a farm in the Siberian region of Kemerovo, intent on producing 2,500 MT of rainbow trout a year. The total investment will be RUB 1.6 billion (USD 24 million, EUR 20.83 million). Next year, the company will start building a capacity for manufacturing aquaculture feed.

In the republic of Dagestan, more than 20 aquaculture projects were launched, with total investments of RUB 560 million (USD 8.4 million, EUR 7.3 million).

Primorsky Krai region in the Russian Far East attracts attention from Asian companies due to its promising potential and geographical proximity. China’s Yantai Tunsyan Foods company and Weng Lyan want to establish their farms in the territory.

30

NEWS & PRESS Meanwhile, The Institute of Agroecology and Biotechnology is going to build a farm to grow 1,500 MT of sharptooth catfish, with investments amounting for RUB 591 million (USD 8.9 million, EUR 7.7 million), in Yaroslavl region.

In Kaluga region, The White Shrimp, the biggest farm of Pacific cleaner shrimp in Russia, was launched in 2017, with annual projected capacity of 35 tons and the possibility to increase output to 100 tons, which will make it possible to entirely substitute the current import of the species into the country.

Being on the rise and still having enough space for new projects, the aquaculture sector in Russia seems to be favored by investors. Russian Aquaculture company, the biggest player in the industry, conducted a secondary public offering on the Moscow stock exchange in late 2017. Other players are likely to post their public offerings in a few years.

Some barriers to overcome Investment into the sector is sure to increase, as long as regulators and businesses have managed to solve some fundamental problems for the industry.

Vadim Likhachev, chairperson of the Association of Mariculture Organizations of Primorksy Krai, points out serious flaws in current legislation, which doesn’t clearly regulate how water areas can be jointly used for multiple purposes – for example, aquaculture and tourism. This flaw made authorities in the Primorsky Krai region, in the Russian Far East, deny granting 91 sites to aquaculture farms.

Some federal laws contradict each other as well. While the Ministry of Agriculture put standards for minimal harvest from a site auctioned to a private business, the Ministry of Natural Resources has recently enacted requirements for aquaculture companies to get an environmental seal of approval, which in turn is linked to the size of harvest. More harvest may lead to more environmental impact, thus making getting the seal that much more difficult. This doesn’t mention fact that the

environmental impact audit to be renewed every three years costs about RUB 2 million (USD 29,400, EUR 25,355).

“Today, the investor rents a site for aquaculture for 25 years, but cannot be sure that it can be really used for aquaculture,” explained Likhachev to SeafoodSource.

Evgeny Podyapolsky, a representative of the Siberian Investment Group, highlighted the problem of getting state-funded subsidies during a meeting with regulators, Russian media reported.

Podyapolsky complained that subsidies are given to a farm, which generates at least 76 percent of its profit from selling agriculture product. But aquaculture farms operate differently – it takes up to 16 months for fish to grow and, consequently, for an aquaculture company to start generating revenue after setting up a new operation. An additional year is then needed to have a proper annual account, which means that state support is in fact only given to farms working for no less than two years, while startups are forced to go without financial aid, Podyapolsky said.

Among other problems not linked to state policy and regulation approaches, is a lack of smolt and fish feed, which are nearly 100 percent imported. Storage capacities for these items are also limited. Such issues even inspired Russian Aquaculture to purchase smolt production facilities abroad. These pains are sure to alleviate fast, however, given the boom the industry is seeing now.

Skyrocketing prices for aquaculture sites have also given stakeholders pause in their efforts. For instance, one 9.6-hectare site in the Republic of Karelia (Russian North West) was auctioned for RUB 1.5 million (USD 22,536, EUR 19,550), 500 times the asking price [RUB 3,072 (USD 46.2, EUR 40)]. Another site in the same region went for RUB 1.1 million (USD 16,540, EUR 14,334), or 2,644 times more than the initial price of RUB 416 (USD 6.3, EUR 5.4).

https://www.seafoodsource.com/news/aquaculture/investments-in-russian-aquaculture-on-the-rise

31

NEWS & PRESS

Vietnam poised to become top player in ocean aquaculture https://www.seafoodsource.com/news/aquaculture/vietnam-poised-to-become-top-player-in-ocean-aquaculture

V

ietnam has set an ambitious goal of becoming a leading country in aquaculture – specifically in the productive development of its coastal marine environment.

Currently ranked as the fourth-largest producer of seafood from aquaculture, behind China, Indonesia, and India, Vietnam produced 3.84 million metric tons (MT) of farmed seafood in 2017. That was more than 53 percent of Vietnam’s total seafood production of 7.23 MT, which itself represented an increase of 5.2 percent year-on-year over Vietnam’s total from 2016.

Vietnam’s government and industry stakeholders have recently taken a more serious interest in the development of Vietnam’s aquaculture sector, Tran Dinh Luan, the deputy director of Vietnam’s Fisheries General Department, told a workshop in Hanoi in early July.

The workshop, co-organized by the Vietnam Seaculture Association, Vietnam’s Fisheries General Department, and the U.S. Soybean Export Council (USSEC), centered around Vietnam’s draft national strategy for marine aquaculture development through 2030. The strategy, with an addendum that proposes a vision through 2050, was prepared by the Ministry of Agriculture and Rural Development and will be submitted to Vietnam Prime Minister Nguyen Xuan Phuc for final approval.

The plan calls for the country to implement – on a trial basis – several policies designed to encourage industrial sea farming, particularly in offshore areas, by 2020. The plan aims to double the farmed output from the sea by 2020 to 750,000 MT total, comprising 200,000 MT of fish, 400,000 MT of mollusks and 150,000 MT of seaweed, according to the draft strategy.

Luan said while sea farming in Vietnam is still at its early stages of development, the strategy is designed to develop the whole production chain of the sector at larger and more advanced levels. In its latter stages of execution, the plan calls for production to rise to 1.75 million MT by 2030, and to three million MT by 2050.

As a result, Vietnam hopes to gain USD 1.5 billion (EUR 1.29 billion) from exports of its farmed marine products by 2020. That total is estimated to grow by between USD 5 billion and 8 billion (EUR 4.3 billion and 6.9 billion) by 2030 and more than USD 10 billion (EUR 8.6 billion) by 2050. The country aims to become the leading player in the Southeast Asia and Asia in marine aquaculture sector, with the eventual goal of ranking in the top five in the world in terms of output and value of farmed marine

32

products exports by 2050, according to the draft strategy.

The plan also calls for a more intensive focus on developing trading relationships and technology and training partnerships.

The country aims to deepen ties with top countries in terms of sea farming, including Norway, Denmark, Japan, the U.S., and Australia. Investors from these countries will be welcomed to transfer modern aquaculture technologies to Vietnam. Vietnam will also look to import high-quality fingerlings from Japan, South Korea, Taiwan and Australia, the draft strategy showed.

Last year, Vietnamese aquaculture produced 2.69 million MT of fish and 723,800 MT of shrimp, data released by Vietnam’s Ministry of Agriculture and Rural Development shows. However, output from sea farming remain modest, producing a total of 377,000 MT of farmed marine products, including fish, mollusk, lobster, crab, and seaweed in 2017.

Nguyen Huu Dung, the president of the Vietnam Seaculture Association, told SeafoodSource sea farming has not developed much yet in Vietnam, but “it is very promising.”

With a coastline of more than 3,260 kilometers (2,026 miles) and numerous islands and bays, Vietnam has huge geographic potential for aquaculture, Luan said. Its exclusive economic zone accounts for nearly 30 percent of the South China Sea. In particular, the waters in the country’s west, with fewer storms, and the deep waters in the central region are ideal for largescale farming of marine fish species.

Pangasius and shrimp are Vietnam’s two major aquaculture products. They are mainly raised in Mekong Delta, though other areas across the country are also home to aquaculture development. Developing ocean aquaculture will bring greater diversity and balance to the country’s seafood production, especially if Vietnam can apply advanced sea farming technologies in deeper waters, Dung said.

Currently, there are about 50,000 households farming marine products across the country. Most are independent, small-scale farmers using old equipment and outdated practices. But a number of companies – particularly those operating near Phu Quoc Island in the south and Van Phong Bay in the central region – have scaled up industrial farming practices using modern technologies from Norway.

Initial results showed that these sea farming models can be applied in other sea areas of the country, Dung said.

NEWS & PRESS

â&#x20AC;&#x2DC;Omni-channelâ&#x20AC;&#x2122; shrimp tech platform looks to modernize Indian sector "Omni-channel" shrimp platform Aquaconnect is ready to take the next step in its ambitious plans towards making the Indian aquaculture sector simpler and more modern for rural farmers.

T

he Chennai, Tamil Nadu-headquartered project is now hiring business analysts to "develop strategies for our omni-channel marketplace", as well as "digital assistants" to help farmers with technology adoption, CEO Rajamanohar told Undercurrent News.

Its stated mission is to simplify the shrimp farming business experience for farmers in rural India, according to an investor brief shown to Undercurrent.

"Our omni-channel marketplace provides quality and affordable inputs to the shrimp farmers. Also we provide them market access to sell their harvested shrimp."

Aquaconnect also locates good quality postlarvae from the hatcheries; procures other inputs such as feed, chemicals and probiotics at optimum prices; and creates a transparent and accessible export market for their harvest, it claims.

"Increasingly, hatcheries, input providers and processors prefer to work with Aquaconnect as we help them reduce the dedicated sales force and associated operational cost," said Rajamanohar.

Aquaconnect has identified a number of challenges in the Indian shrimp sector, including supply chain inefficiency; a lack of transparency in the marketplace; exploitative middlemen; and a lack of formal financial access.

As well as supplying farmers, it hopes to help educate them using its website "for tech-savvy farmers"; a multi-lingual toll-free number, and digital assistants "on the ground".

It plans to take service fees on aquaculture inputs, and sees an overall market potential for Aquaconnect of $558 million, based on a three-crop cycle per year. It believes it can serve some 2,200 farmers in Tamil Nadu; 5,00 in Gujarat, on the west coast; and a huge 42,000 farmers in Andhra Pradesh.

Ultimately, it sees opportunity beyond shrimp farming too.

For now, though, it is on track with its timeline, having launched a pilot of the digital farm management tool in July 2018. It secured its required $29,000 pre-seed funding via aquaculture accelerator HATCH, and next it hopes to raise $2m for market expansion in the fall of 2018.

Currently Aquaconnect works with around 3,000 farmers in Tamil Nadu and parts of Andhra Padesh, and has completed 100 paid deals, he said. 65 hatcheries and 15 exporters are also on board.

"We recently won a contract from [one of the] worldâ&#x20AC;&#x2122;s largest feed companies, worth about $72,000, to launch their farm management tool in India," he said. This has not yet been announced publicly, and so he could not confirm the company, he said. However, Dutch aquafeed firm Nutreco has recently invested in Indian startup Eruvaka, which aims to use the "internet-of-things" and mobile technology to help aquaculture farmers increase productivity.

https://www.undercurrentnews.com/2018/08/13/omni-channel-shrimp-tech-platform-looks-to-modernize-indian-sector/

33

www.asianaquaculturenetwork.com

VOL. 3 ISSUE 3 AquaSG'18 Special Edition

PROACTIVE PROFESSIONAL PRACTICAL

ww w.a sian

aqu acu ltur

VOL. 3 ISSUE 3

ene two rk.c

om

Aqu aSG ’18 Spe cial Edi tion

3-6 Octo

ber 20

School of Applied Scie 18 Temasek nce Pol Singapore ytechnic

"Aqua Practical" brings practical knowledge sharing on technical issues and best practices to its network of members and subscribers. Published quarterly by the Asian Aquaculture Network (AAN).

INV EST ME NT & CU RR EN T PR AC TIC ES IN AQ UA CU LTU RE TEC HNOL

Asian Aquaculture Network (AAN) was established in 2009 as a regional professional network of communication, knowleadge and sharing practical technical information about aquaculture.

The primary focus is on promoting sustainable development and profitable practices of aquaculture around the world.

ADVERTISE WITH US!! TODAY

E DITORIAL CALENDAR 2018 JAN−MAR APR−JUN JUL−SEP OCT−DEC

Sustainable Aquaculture Responsible Aquaculture AquaSG'18 Special Edition Challenges in Aquaculture 1/2 (H)

Full Page

- HIT THE RIGHT TARGET - EXPEDITE YOUR BUSINESS -

Avertising Rate: W EB BANNER RATES (SGD)

1/4 (V)

1/2 (V) 1/4 (H)

M AGAZINE ADVERTISING SIZE (mm.)

Full page:

215.9 x 279.4

Vertical

Horizontal

1/2 page:

107.95 x 279.4

215.9 x 139.7

1/4 page:

107.95 x 139.7

215.9 x 69.85

Artwork file format: .ai .pdf .epi .psd .tiff .jpg Image resolution: 300 DPI (dots per inch) with bleed lines 3 mm included artwork Color: mode CMYK in process for printing / RGB for web banner advertisment Font: Create outlines

34

OG Y

Size Position Frequency

6 Months 12 Months 212 x 140 px Right Column 2,500 4,500 520 x 140 px Middle Column 2,800 4,900

M AGAZINE RATES (SGD) Page Sizes Frequency

1x 4x Full page 1,700 1,400 x 4 1/2 (vertical/horizontal) 1,050 840 x 4 1/4 (vertical/horizontal) 650 500 x 4 Premium Placement: Inside cover Add 300 Outside back cover Add 600

What we do

Membership

• Provide updated information and emerging news about aquaculture through magazines and Facebook.

• Conduct aquaculture trainings and practical workshops by a team of industry experts.

• Hold annual conferences and seminars.

• Collaborate with research institutions and universities on research, educational and technological development information exchange and student exchange program.

• Assist its members in advocating to national goverments.

• Provide latest pioneering and new technology knowledge and other discovery information to its members.

• Provide aquaculture supplier directory to members annually.

•

• •

4 issues of "Aqua Practical" and free acces to the online version of the magazine.

Free online access for supplier directory.

Discount on annual seminar and conference and customized seminar and training.

Our Readers • Network of professionals in aquaculture industry.

• Memberships, University libraries, and trade shows.

AAN facebook

Supplier Directory • Asian Aquaculture Network (AAN) Supplier Directory contains a wide listing of aquaculture suppliers from all over the world • Members get to know suppliers from outside their home country or current location that might be able to offer lower prices for a better quality • They are categorized into specific products and services • AAN Supplier Directory is available online to our members only

Total page likes as of today:

UK 51

Egypt 206 Ecuador 124

Country

India Malaysia Thailand Indonesia Philippines Vietnam Bangladesh Egypt Brazil Singapore Mexico USA

1,984 578 503 464 351 277 231 206 186 186 136 133

Bangladesh 231 India 1,984

Taiwan 89 Thailand Hong Kong 102 503 Vietnam 277

Malaysia 578 Singapore 186 Indonesia 464

Brazil 186

AAN Fans

7,651

China 36

USA 133 Mexico 136

AAN Website

City

AAN Fans

Bangkok 315 Singapore 185 Kuala Lumpur 120 Ho Chi Minh City 109 Chennai 104 Hong Kong 100 Andhra Pradesh 98 Surat, Gujarat 86 Yangon 83 Mumbai 78 Cairo 67 Jakarta 64

Australia 58

Language

AAN Fans

English (US) 3,214 English (UK) 1,587 Spanish 384 Indonesian 359 Thai 312 Vietnamese 230 French (France) 192 Arabic 189 Portuguese 184 Chinese 98 Malay 62 Turkish 49

35

EVENT CALENDAR

Aquaculture

EVENTS 3-6

8-9

17-20

18-21

23-26

25-27

6-8

17-19

Oct

AquaSG'18 Temasek Polytechnic, Singapore www.aquasg.com

International Congress for Marine Biotechnology Hotel SENTIDO Rosa Beach, Monastir TUNISIA www.icmb2018.com

Nov

LATIN AMERICA & CARIBBEAN AQUACULTURE 2018 Bogotรก, Columbia www.was.org

SEA WORK ASIA 2018 Commercial Marine and Workboat Exhibition & Forum www.myanmar-aquafisheries.com

2018

10th Euro-Global Summit on Aquaculture & Fisheries Park Inn by Radisson London aquaculture-fisheries.aquaconferences.com

Gender in Aquaculture and Fisheries Conference 2018 Bangkok, Thailand www.gafconference.org www.myanmar-aquafisheries.com

(ICAI) 2018 INTERNATIONAL CONFERENCE OF AQUACULTURE INDONESIA Hanover Messe, Germany www.eurotier.com

HIGH ENERGY EUROPE MARINECULTURE CONFERENCE The Corfu Imperial Hotel, Corfu, Greece www.offshoremariculture.com/europe

Jun (2019)

Mar (2019)

Dec

3-6

36

Asian Aquaculture 2018 Asian Institute of Technology (AIT), Thailand www.asianaquaculture.org

13-15

VIV ASIA 2019 BITEC 88 Bangna-Trad Road, Bangna Tai, Bangna, Bangkok, Thailand www.viv.net/events/viv-asia-2019-bangkok

19-21

Asian-Pacific Aquaculture (APA) 2019 Chennai, India www.was.org www.asianaquaculture.org

22-23

12th Global Summit on Aquaculture & Fisheries Innovative and Sustainable Aquaculture Sydney, Australia aquaculture.global-summit.com

13-16

EUROTIER 2018 - GERMANY Hanover Messe, Germany www.eurotier.com

EVENT CALENDAR