Aquafeed vol 9 issue 1 2017

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VOL 9 ISSUE 1 March/April 2017

AQUAFEED A D VA N C E S I N P R O C E S S I N G & F O R M U L AT I O N An publication

Algae: a source of biological treasures Controlling your extruder Inulin shows promise as prebiotic Grape extract—the latest ingredient Optimizing the DHA levels of hatchery Artemia Stress resistance by stimulating protein chaperones




Absolute protection Mycotoxins decrease performance and interfere with the health status of your animals. Mycofix is the solution for mycotoxin risk management. ®

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Volume 9, Issue 1

A D V A N C E S I N P R O C E S S I N G & F O R M U L AT I O N

Contents 5




Interview with Dr. Albert G. J. Tacon


Intelligent extrusion process for aquafeed quality control


Specific Mechanical Energy (SME) in Aquafeed Extrusion


How to control the density of fish feed


Functional feed additives, key to an effective strategy against White Feces Syndrome in white shrimp


Preparing fish for stress by stimulating protein chaperones


* Algae of the seas providing for natural health in aquaculture species


Effects of a phytonutrient on aquatic species raised under commercial conditions


Inulin, a promising prebiotic in the prevention of diseases in modern aquaculture


Dry grape extracts, authorized as feed additives, and their use in aquaculture


A flexible approach to optimizing the DHA levels of hatchery Artemia


Aquafeed Horizons


Soy Aquaculture Alliance – Aquaculture America: teaming up to forge a successful future for U.S. aquaculture


* Cover story


4 Volume 9, Issue 1

A D V A N C E S I N P R O C E S S I N G & F O R M U L AT I O N

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37 empowering the aquafeed value chain since 1998. Copyright & Disclaimer


Dr Albert G.J. Tacon obtained his PhD in fish nutrition at University College (University of Wales), and went on to lecture and at the University of Aston in Birmingham and at the Institute of Aquaculture in Stirling. In 1984 he joined FAO as an aquafeed and nutrition expert. In 1999 he moved to Hawaii to become Director of the Aquatic Feeds and Nutrition Program at the Oceanic Institute. Since then Albert was has been working as Technical Director, Aquatic Farms Ltd., Hawai’i, as an independent consultant in aquaculture and aquaculture nutrition and feed. He has more than 200 scientific publications and one patent. He currently serves as Scientific Advisor on Aquatic Resources to the International Foundation for Science, Sweden, is Editor in Chief of Reviews in Aquaculture, serves on the editorial board of Aquaculture Nutrition and Aquaculture Research, and is on the board of directors of the charity Aquaculture without Frontiers (AwF).

Interview with Dr. Albert G. J. Tacon AQUAFEED.COM It seems almost everyone who has ever written a paper or made a presentation on aquaculture feed has your name as a citation somewhere, often in association with statistics. In your many years of collecting data on the industry, what do you see as the most significant change? AGJT I would say the most significant change has been the intensification of the farming systems used for the

production of freshwater fish species and marine shrimp, and the increased use of extruded fish feeds for freshwater fish, salmonids, and to a lesser extent marine fish species raised in open cage culture systems.

AQUAFEED.COM There is a major trend away from fishmeal inclusion in aquafeeds. What alternative protein sources do you believe hold most

promise in terms of nutrition, availability and sustainability? AGJT The largest source of animal protein available for use in aquafeeds are those derived from the terrestrial livestock production sector, including poultry by-product meals, feather meals, blood meals, porcine meals, and ruminant by-product meals. Provided that the meals are produced from healthy food-grade livestock, I see no

6 limitation to their use and availability. Apart from these products, there are the plant protein meals derived from oilseeds (such as soya, canola, and cottonseed), cereal grains (such corn gluten meals, corn protein concentrates, high-protein dried distillers grains and solubles, wheat gluten meals, rice protein concentrate), and other plant protein concentrates (such as pea protein concentrates, potato protein concentrate, lupin protein meals). Probably the most exciting and promising protein sources are those derived from the mass production of microorganisms from low-cost agricultural waste streams and substrates, including bacterial SCP, Yeast SCP, and algal SCP. Other widely talked about protein sources such as animal invertebrate meals (such as terrestrial insect meals and marine polychaete meals) are also of growing interest, but until production of these products reaches into the thousand of tonnes range their use will be limited.

AQUAFEED.COM Disease has decimated shrimp farming throughout its history; Acute Hepatopancreatic Necrosis Disease (AHPND), or Early Mortality Syndrome (EMS) as it's more popularly known, comes immediately to mind. Where do feeds fit in, in terms of risk? AGJT I feel that feeds pose a serious risk in terms of potential disease introduction, and in particular from the current almost universal use of nonsterilized live food organisms during the broodstock and hatchery/nursery production cycle (including the use of live marine polychaetes worms, Artemia nauplii and biomass) process, and to a lesser extent from the use of inadequately processed shrimp head

meals (derived from cultured shrimp) within grow-out feeds. The whole of the area of feed biosecurity has not been on the radar screen of most shrimp farmers and should be!

possible disease introduction from contaminated meals (as recommended by existing GAA, FAO, and Global GAP aquaculture feed manufacturing guidelines).

AQUAFEED.COM Aren't there already a slew of regulations in place governing the safe production of aquafeed?

AQUAFEED.COM In our interview with Dr. Dean Akiyama (Aquafeed Vol.8 Issue 3 2016), he said the major constraints on shrimp feed production are the shrimp farms and management. Would you agree?

AGJT No, I believe there is much more that needs to be done to ensure that the farmer receives a quality feed which meets all the dietary nutrient requirements of the target species for optimum growth and health. There are currently some major aquaculture and aquafeed producers where no government regulations exist for the manufacture of compound aquafeeds. Clearly this must be remedied for the benefit of the farmer and the consumer of the farmed fish and shrimp.

AQUAFEED.COM What steps can be taken along the supply chain to ensure aquafeeds do not introduce pathogens to the shrimp farm? AGJT I see two possible approaches, first, that individual potential contaminated marine feed ingredients or live food organisms be sold as either being pathogen free or sterilized/ pasteurized prior to shipment, and secondly, that feeds be sterilized/ pasteurized during the feed manufacturing process, either by extrusion processing, or by gamma irradiation. Moreover, the re-feeding of the same species back to the same species (intra-species recycling, ie feeding shrimp to shrimp) should be prohibited by law so as to prevent

AGJT Yes I would agree. In my opinion the shrimp industry must follow the example of the livestock sector in terms of biosecurity, and move away from the use of open pond-based culture systems to the development of closed fully biosecure culture systems where pathogens can be excluded and the rearing environment controlled so as to minimize shrimp stress and optimize water quality. Moreover, the success or not of a feed is dependent upon the management of the feed on the farm by the farmer; in this regard feed companies and government have a critical role to play in assisting the farmer; the bulk of shrimp production within most Asian countries still being conducted by smallscale farmers within limited financial resources and training.

AQUAFEED.COM Do you see a big move from pelleted to extruded shrimp feed? Any other production trends?

AGJT Yes, I believe the shrimp sector will eventually follow the finfish sector concerning the increased use of extruded shrimp feeds for both improved nutritional characteristics and shrimp performance, increased feed biosecurity,

7 and more importantly to reduce feed costs per unit of shrimp production. Other production trends will include the development of improved application technologies for the top-dressing of feeds with heat sensitive feed additives, including feed attractants, enzymes, vitamins, pigments, and probiotics.

AQUAFEED.COM We hear all the time that aquaculture holds the solution to feeding the world's exploding population. How important is fish and seafood to human nutrition? AGJT Very important. On a global basis fish and seafood products constitute the third major source of dietary protein consumed by humans after cereals and milk, representing 6.5% of total protein supply or 16.4% of total animal protein supply. However, it is also apparent that

fish supply through capture fisheries has not been able to keep up with population growth over the past two decades, and that aquaculture is the only real hope to increase production and global market availability in the long-term. Moreover, it is also clear that fish and seafood plays a greater role in the nutrition of lowincome countries within the African continent (primarily derived from capture fisheries) and within the Asian region in general (primarily derived from aquaculture). Within the Asian region, fish and seafood products were the third major source of dietary protein consumed after cereals and vegetables, representing 7.5% of total protein supply (21.9% of total animal protein supply), 1.7% of total fat supply, and 1.3% of total calorie supply.

AQUAFEED.COM Finally, what is the one thing you'd like to see happen in aquaculture before you retire? AGJT I have no intention of retiring (as long as my health holds out), but if I had to pick something it would be that the aquaculture industry realize the real value of the product they are producing, not only in terms of its content of DHA or EPA, but also in terms of all the other essential nutrients present within fish and seafood products (in marked contrast to most terrestrial animal livestock produce), including high quality digestible animal protein, omega-3 fatty acids, and much needed essential vitamins and minerals – a complete much needed superfood!



Intelligent extrusion process for aquafeed quality control By Ramesh Gangatharan and Joe Kearns, Wenger Manufacturing, Inc., Sabetha, KS, USA

The global aquaculture industry must grow rapidly to meet the increasing needs of quality seafood as by 2030 more than 60 per cent of this should come from farm based production systems. There is all round consensus that this needs to come from sustainable aquaculture practices. The scope of sustainable practices trickles down to traceability, quality control and food safety at the feedmill.

The main objective of extrusion processing of aquafeeds is to produce feed with desirable characteristics to meet the specific needs of the wide variety of farmed aquatic species depending on their farming practices – extensive/ intensive, indoor/ outdoor, and their various growing stages. Some of the key physical characteristics of the feed for important consideration are pellet size (diameter & length), floating/ sinking (bulk density), pellet durability, water stability and its ability to uptake fat coatings. Stringent quality control at various stages of extrusion processing will help to ensure that the aquafeed produced meet the highest standards. There are several inline and online tools now available which makes this job much easier and helps to integrate this into the production system with minimal human intervention. To begin it is essential to understand the benefits of quality control of the production system with lab confirmation of the results. As aquatic feed volumes increase from multiple production units or simply increased capacity of individual production machines, production of 100 tons per hour at a single facility or more is going to be more common. The actual time from start to finish producing an aquatic feed is about 1 hour including

mixing, grinding, conveying, extruding, drying coating and cooling. Want to wait until the end of the process to find out you are not in product specifications? It is considered important to assess the product as it progresses through the system with checks along the way to insure product quality is achieved greatly reducing rework and returns and reducing cost of continued processing of out of spec product. Quality control starts in the raw material receiving area, lab tests to verify the ingredients received are up to specifications defined to suppliers. In many cases a NIR system is used for these tests to verify the ingredient composition. Assuming all is good at this point the process starts. An initial quality test could be a NIR inline system to verify the formulation post grinding and mixing just prior to the extruder includes all ingredients as specified avoiding running a formula which is lacking. Additional in line systems offering visual determination to confirm the mix is ground. There are additional devices to determine grinding screen quality, no holes in the screen, due to amp load variations detected on the main drive of the grinder. The process continues and the material arrives at the extruder, an explainable

9 process but one that preforms many functions in a matter of seconds. The size, durability, density, cell structure, shape, appearance all happen so fast it is hard to visualize. Computer controls can greatly assist in these events all occurring at the same time. To start the use of loss in weight dry ingredient feeding insure the proper mix of the added liquids to the dry mix with the use of liquid flow meters and computer controls. Pellet size and density of the pellets are important due to the varieties of fish and their life stages. Size is control by the die opening and knife speed, can be monitored by a camera and measurements can occur to determine the correct specifications or the operator monitors and adjusts manually, with the latter being quite normal. Automatic Density Control has been developed due to availability of an accurate inline device where density is checked post extruder but before the dryer to insure the correct gram per liter density desired before expending energy removing water from an out of spec product. Tests occur about every 45 seconds. The Bulk Density System(BDS™) takes a sample mid steam checks the density and informs the computer control the results. Automatic modulation of the screw speed and or

Fig 1: Source Technology Bulk Density and Moisture In Line Monitoring Device.

flow restrictor behind the die achieve the needed changes to adjust the density into the range desired. Tested again and verification continues. The same unit can also detect moisture and thus additional control on water management into and out of the dryer. You do wish to sell water and or at least have the maximum level stated on the bags achieved? Durability, cell structure and possible oil up take qualities can be monitored with a

Fig 2: How the Automatic Density System Functions.

constant indication of specific mechanical energy (SME) inputs. SME is proven to be a good indication of the energy input for a product and it relates to product quality when studied based on a formulation used. In high oil content aquatic feeds this is essential data to be sure the cell structure is such that the oil will be absorbed and stay in the pellet until delivered to the fish. The other option is to wait to hear from the farmers which usually is not good news.


Fig 3: Computer Screen with SME Indication, as seen in the dry feed Totals box gives Physical Characteristic Verification, relates to durability and cell structure of a formulation. Controls allow for predictably of results.

In high oil feeds, oil is absorbed differently in pellets of varying moisture levels. The Back-Pressure Valve, BPV, is designed to be adjusted while running allowing the cooking effect or energy inputs to be controlled without stopping the extruder to adjust the barrel configuration. Great reduction in out of spec product continuing through the process.

Automatic Control packages can also include data logging coupled with visual and photographic devices to create reports for files and future reference and traceability. Mentioned earlier is the BDS™ In line system also measures moisture, a critical ingredient and one that is monitored for obviously shelf life quality characteristics. In the aquatic arena, moisture variations in the pellets do effect profitability as well as downstream potential coatings. In high oil feeds oil is absorbed differently in pellets of varying moisture levels.

Dryers also utilizing in line system can achieve correct post dryer moisture levels at or near a 0.5% moisture variance. Quite accurate for maximizing water in the feed as well as the needed quality control for further downstream processes. These devices and others also allow for energy and exhaust air humidity readings to monitor and adjust the process for climatic conditions which vary from day to night as well as the seasons throughout the year. Advantages can also be gained in this area when changing from one product characteristic to another.


Fig 4: Computer controls in conjunction with Source Technology allows for Data Logging including photos and Data on products produced.

Current real world quality control starts with production systems that allow for product correction via computer controls coupled with In -Line systems to take critical measurements for rapid process corrections. This technology greatly assists in improved operation efficiency, achieving consistent product quality, minimizing out of spec product production, which is vital in modern day large production systems where 15 to 20MT/ hr of feed is being produced per extrusion system. Plant efficiencies show that waiting to learn the results of production by manual testing at the end of the process is not cost effective and also results in loss of production time as well as expensive ingredients, greatly impacting the profitability of the feedmill at the end of the year. Quality control is now incorporated into the production process, greatly improving the desired final product characteristic results at

extremely profitable, high tonnage per hour production rates.


More information Ramesh Gangatharan, Technical Sales Advisor, AquaFeed Division, Wenger Manufacturing. Email:

Plant efficiencies show that waiting to learn the results of production by manual testing at the end of the process is not cost effective and also results in loss of production time as well as expensive ingredients, greatly impacting the profitability of the feedmill at the end of the year.

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Specific Mechanical Energy (SME) in aquafeed extrusion By Gordon Young, Food Industry Engineering, Australia and Dennis Forte, Dennis Forte & Associates, Australia

In the most simple of terms, the transformation of raw materials into a product, involves the input of energy. The means by

which the energy is imparted, is defined by the physical design of the equipment and operational parameters of the process. The type of energy has a direct influence upon the product characteristics, for example, color, texture and flavor. Let us consider a generic cooking process, shown schematically in Figure 1. There are three mechanisms for energy transfer. These are:

Mechanical Energy (also referred to as Viscous Dissipation)

Convection Energy (associated with latent heat transfer – steam injection)

Conduction Energy (associated with

Figure 1 – The Generic Cooking Process In large-scale extrusion, conduction energy is relatively insignificant – as size of the extruder increases, the reduced surface area to volume ratio in the barrel reduces heat transfer from the barrel wall. Some of the energy in extrusion is commonly added through steam, especially via the Preconditioner, so Convection Energy can be significant. But the nature of the process means that a lot of the heating during extrusion is due to Mechanical Energy.

indirect heat transfer – conduction from the vessel wall)

SME (Specific Mechanical Energy) and Feed Extrusion This mechanical energy input, often specified as Specific Mechanical Energy (SME) is critical to the quality characteristics of the extruded feed. The stresses developed in the material (“Shear

Stress”) not only convey the product along the barrel, but cause mixing, and under the correct conditions, can break down Starches and re-align proteins and develop structure within the dough or melt. But intense shear stress can also cause breakdown at a molecular level, as will be discussed later. The shear stress applied to the product is dissipated through the viscous fluid movement of the melt (“Viscous


This temperature increase directly drives the cooking reactions that occur. But SME also affects the feed being manufactured at the molecular level. Figure 2 shows measured effect of SME on Mean Molecular Weight of the starch fraction in an extrusion operation. The shear generated in the process breaks down molecules – the higher the SME, the smaller is the average molecule size. Break-down of proteins results in the generation of peptides and changes to fatty acids. When starches are broken down, dextrins (long-chain sugars) are produced.

MMW (x106 g mol -1)

Dissipation”) leading to an increase in the temperature of the material. The bulk effect of SME is noted as a temperature rise (additional to temperature rise due to added steam).

SME ( / kg) Figure 2: Effect of SME on Mean Molecular Weight in starch extrusion.

(1) Flavor and nutritional profile – changed amino acids due to break-down of proteins, bitter notes from peptides, sweeter taste from dextrins. The Digestibility of certain components can be enhanced (for certain species). (2) Pellet durability – sufficient “cook” is essential to the development of Structure via the starches (present in very limited quantities in many aquafeeds) and any functional proteins. However, excessive breakdown of the molecules destroys binding. Therefore with respect to pellet durability, there must be sufficient SME to promote the correct Degree of Cook, but too much SME will weaken the pellet structure. (3) Water stability - The formation of dextrins lead to a reduction in the gel forming capability of any starch present;

WAI (%)

This break-down of molecules affects product characteristics in a number of ways:

SME ( / kg) Figure 3: Effect of SME on WAI (Water Absorption Index) in a starch-based extrusion process.

hence, a reduction in the Water Absorption Index (WAI). WAI is often used as a means of quantifying the water stability of a product. Some data for an extrusion process is shown in Figure 3. Note that the most significant changes to WAI occur within the same range of

SMEs that cause a major change in molecular weight (Figure 2). Smaller molecules also tend to be more water-soluble, the Water Solubility of the product increases (as measured by the Water Solubility Index WSI). The data for

WSI (%)

Starch Dry Matter Digestibility (%)


SME ( / kg) SME ( / kg)

Figure 4: Effect of SME on WSI (Water Solubility Index) in a starch-based extrusion process.

the test samples is presented in Figure 4. An increase in water solubility in aquafeed corresponds to a reduction in water stability. (4)

Digestibility – Change in level of

Figure 5: Example of the effect of SME on digestibility (starch-based cat food).

“cook” and in the molecular weight also affects digestibility. Typically, uncooked material has low digestibility. As level of cook increases, digestibility increases – up to a maximum. However, at higher SME levels, digestibility again decreases –

over-sheared product is not readily absorbed by the animal. By way of example, Figure 5 shows the effect of SME on digestibility of an extruded starched-based product (in this case in cats).

16 How is SME Measured/ Calculated?

Table 1 – Specific Energy Inputs for the manufacture of Aquatic Feeds

Remember that SME is about the mechanical energy going into the product, not Total Energy – it is important to subtract the No-Load Energy.

If the extruder is instrumented to provide a direct measure of Screw Torque, then note the torque when no product is passing through, and Net Power Input is given by: Pnet = 2 π (τ – τ0) N / 60000


where, τ = Torque (Nm) τ0 = Torque under no load (Nm) N = Screw speed (rpm)

If a torque measurement is not available, power can be calculated from amps drawn by the drive motor. For AC drives the Net Power Input (kW) can be estimated via:

Specific Convective Energy, SCE

0.040 to 0.060 kWhr/kg

Specific Thermal Energy, STE

-0.002 to 0.007 kWhr/kg

Specific Mechanical Energy, SME

0.025 to 0.045 kWhr/kg

Total Specific Energy, TSE

0.063 to 0.112 kWhr/kg

Some Guidelines for SME in Aquafeed Extrusion A very broad range of aquatic feeds, with vastly different characteristics (both floating and sinking) for a large number of species are prepared via extrusion cooking using a diverse raw material base. It is therefore impossible to define a single target value for the magnitude of the SME (or any of the other energy inputs). A typical range for the Specific Energy Inputs can be presented, however (see Table 1 above)

Pnet = √3 V [ (f η I ) - (f0 η0 I0 ) ] / 1000 (kW)

More information

For DC drives the Net Energy Input (kW) can be estimated via: Pnet = V [ (η I ) - (η0 I0) ] / 1000

“Food and Feed Extrusion Technology: An Applied Approach to Extrusion Theory”. ISBN 978-0-9945433-0-1. Visit:


where, f = Power Factor ( - ) η = Efficiency (%) V = Supply Voltage (V) I = Current (Amps) (Subscript 0 refers to no-load conditions)

This paper draws from a book by the same authors:

The authors are presenting short courses on “Aquafeed Extrusion Technology” and “Drying Technology” at Centre for Feed Technology, FôrTek, Norway, In April 2017. Also an Extrusion Course in Thailand in July 2017 Details:

(kW) Contact:

The SME is then readily calculated as: SME = Pnet / MTotal

Gordon Young, Food Industry Engineering, Australia


where, MTotal = Total Throughput Rate (kg/hr). Note: MTotal is the Total Melt Flow through the extruder – not after drying – and is the flow rate of powder, water, steam, oil and any other additives.

E: Gordon Young

Dennis Forte

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How to control the density of fish feed By Knut Szemjonneck, Key Account Manager Food/Feed, TEWS Elektronik, Germany When fish feed is put into water, it should act in accordance with the feeding behavior of the respective fish. This means that it should either float on the surface or sink to the bottom. These characteristics largely depend on the fish feed’s density, which in turn defines the different types of fish feed: • Feed that floats on the surface of the water • Feed that sinks slowly • Feed that sinks and is water resistant • Feed that sinks and is extremely water resistant Consequently, the density of the fish

flakes, granules, pellets or chips is an important aspect for the quality of the product. The moisture content of the feed is also an important parameter for the quality of the product throughout the production process. Both a high and a low moisture content often lead to loss of quality and can complicate further processing or even prevent it entirely. An ideal moisture content leads to a longer shelf life and prevents the onset of rotting. Regular checks of the moisture and density are therefore necessary to produce high-quality fish feed. The

common method of quality control is through laboratory measurements. Operators in the fish feed process are used to manually taking samples from production and analyzing the moisture and density content in a laboratory. This provides the respective moisture and density values that can, depending on the method, take between 5 minutes and a few hours to obtain. Sample taking is both time-consuming and laborintensive, and the measured values are generally available too late to be able to control the production process adequately. TEWS Elektronik has developed a measurement technique with which the


The MW 4260/70 enables the continuous in-line measurement in processing lines. Up to 3,500 measurements per second are possible.

moisture content as well as the density of the fish feed can be continuously measured during the production process. With the help of bypass sensors that are integrated into the process, the values are measured and visualized reliably and accurately in a matter of seconds. A quick and targeted intervention in the production process is thus possible. Improved control of the production processes leads to energy savings as well as less wastage and better quality of fish feed. In short, moisture and density measurements during the process increase the efficiency of the entire production chain. The patented measuring method is based on microwave resonance technology. As soon as the material passes through the field of the microwave sensor, its resonance properties change depending on the water content. A single configuration of the systems is all that’s needed for it to work continuously and reliably. The special unique feature of this technology is the independence of color,

density, grain size or surface influences of the product being measured. Contrary to the so-called nearinfrared procedure, which measures the moisture content only on the surface, the microwave measurement equipment also determines the core moisture in the inside of the product, for example in pellets or granules.

The Microwave Resonance Method Water molecules are among the smallest molecules that are positively and negatively charged. The moisture sensor generates a low-energy electro-magnetic field, which interacts with these water molecules inside the product sample. Since the microwave field penetrates a few centimeters into the sample, water is not only detected at the surface but also in the product’s core.

referred to as the dual parameter method. A moisture value is then calculated on the basis of the resonance parameters. For every product, the moisture meter is calibrated with the aid of a laboratory method (e.g. LOD method or Karl Fischer Titration), so that the product’s moisture content can be displayed, stored or processed as a percentage value. The microwave resonance method can also be used to measure bulk density, sample mass or a sample’s weight per area.

TEWS Elektronik has been developing and manufacturing its patented microwave moisture and density measurement systems, which are used for process control and laboratory QA in the feed industry, for over 30 years. Based in Hamburg, Germany, the company is also active in the USA, with its local subsidiary TEWS of America, and has established service points in Hong Kong and Bangkok.

The product sample reduces the propagation speed of the microwave field, and the water molecules in the sample perpetually align to the electromagnetic field, thus depriving it of energy. The moisture meter continuously measures these two effects, which are

More information Knut Szemjonneck, Key Account Manager Food/Feed, TEWS Elektronik E:


Page 21


Functional feed additives, key to an effective strategy against White Feces Syndrome in white shrimp By Ho Gim Chong, Maria Mercè Isern Subich, and Allen (Ming-Hsun) Wu, Nutriad International, Belgium

White Feces Syndrome (WFS) is the cause of significant crop failures of white shrimp

in Indonesia, Thailand, Vietnam and India. Although the causative agent is not yet clearly identified, WFS is easily diagnosed by the presence of white strings in the surface of the water and edges of the pond. Affected shrimp exhibit a white hepatopancreas and gut, which results in mortality and persisting stunted growth. The present series of field trials in collaboration with shrimp producers in Malaysia and Indonesia, aimed at evaluating the potential role of a health promoting feed additive in WFS prevention and treatment.

WFS is significantly affecting shrimp production in Asia In 2016, White Feces Syndrome (WFS) has been reported to be a disease problem affecting significantly shrimp farmers in Indonesia, Malaysia, Thailand, Vietnam, and India. WFS has been reported since 2010 in Penaeus monodon. From early times, the development of WFS has been related to the presence of high loads of organic matter in the culture pond caused by high stocking densities and high water

temperatures, resulting in higher feeding rates. The improvement of hygiene and biosecurity measures are helping to reduce the severity but in some cases are not enough to stop the condition and its effects. WFS is easily identified by the presence of white strings in the surface of the water and edges of the pond, and affected shrimp show white hepatopancreas and gut. The disease is mostly associated with the described initial acute symptoms, followed by chronic mortalities and stunted growth that will

22 heavily affect the yield of the culture. Different potential pathogens have been associated with the appearance of WFS, including microsporidian parasites, bacterial pathogens and gregarine-like inclusions, but till date WFS could not be induced under controlled conditions by a specific pathogen.

The affection of the digestive system (hepatopancreas and gut) leads to malnutrition. After a strong WFS outbreak, growth is never recovered to the pre-outbreak levels, typically around 0.25g/day in South East Asian intensive shrimp farms. Although some farm strategies manage to restore the macroscopic condition of the hepatopancreas and gut, growth continues to be stunted. As a result, WFS outbreaks cause many production cycles to fail since farmers prefer to harvest instead of growing WFS affected animals which show slow growth and poor feed conversion.

Functional feed additives to prevent shrimp disease Functional feeds containing gut health promotors deliver with every meal an adequate concentration of natural antimicrobial activities into the digestive system. These feeds are a key component of any strategy to prevent diseases in aquaculture, particularly where opportunistic bacteria are suspected to be a major cause of mortality. However, the success of this approach will depend on the efficacy of the gut health promotor. The gut modulating feed additive ideally is heat stable and can therefore be easily incorporated into industrial aquafeeds and be present in every meal from the starter feed

onwards, without requiring major adaptations of the production protocols. Natural feed additives combining different action mechanisms such as direct bactericide/bacteriostatic properties as well as Quorum Sensing Inhibition (QSI) properties at concentrations below MIC, are most promising to control bacterial diseases caused by opportunistic bacteria such as Vibrio spp. The inclusion of such botanical feed additive under standard industrial conditions at the feed mill improved survival under production conditions in a semi-intensive shrimp farm in Panama and Ecuador with 18 to 24% compared to the control group during two independent production cycles (Cuellar-Anjel et al., 2011; Valle and Coutteau, 2015). In these production trials, the main disease challenge at the farm consisted of White Spot Disease (WSSV) and Vibriosis. The gut health promotor was applied from the first starter feed onwards and continued throughout the entire cycle till harvest. Loc et al. (2015) was able to confirm the effect of a synergistic phytobiotic product in a controlled challenge trial with Vibrio parahaemolyticus (Early Mortality Syndrome, Acute Hepatopancreatic Necrosis Disease, EMS/ AHPND strain) under laboratory conditions; showing 62-107% increased survival in shrimp that had received the additive during 3 weeks prior to the experimental infection, compared to unsupplemented control groups. Furthermore, the addition of the phytobiotic product in the diet resulted in consistently lower Vibrio spp counts in the shrimp’s digestive system compared to the control, illustrating the capability of gut modulating additives to protect the shrimp’s gut microbiota throughout a Vibrio spp challenge. The inclusion of

natural products with bactericidal and quorum quenching activities is an important factor in current bio-security protocols to prevent the vertical spread of opportunistic bacterial pathogens such as Vibrio spp, Photobacterium spp, Flavobacterium spp, Tenacibaculum spp. Furthermore, a healthy gut microbiota enhances the overall health status and immune defenses, which may explain the positive effect of gut modulators on reducing the impact of certain endo as well as ectoparasites on productivity in fish and shrimp. The three field studies reported in this study explored the preventive and/or curative effect against WFS of a health promoting, functional feed additive with broad spectrum activity (SANACORE® GM, Nutriad), added to the feed either at the farm by top-dressing or at the feed factory, or a combination of both strategies. Field trials were executed in close collaboration with shrimp producers following standard operational procedures in Malaysia and Indonesia.

Trial 1 in Malaysia The first trial was conducted in a shrimp farm located in the Penang region, Northern Malaysia. The farm was historically affected by EMS and WSSV outbreaks, as well as WFS, usually around day of culture (DOC) 60. The trial followed standard operation procedures of the farm and was run in 8 ponds in total, 6 as treatment and 2 as control. For the treatment ponds, SANACORE® GM was top-coated in the farm and applied to the ponds since the first day of culture in all feedings until harvest. Due to mortality and stunted growth, the control ponds were harvested at DOC 42,

23 Table 1: final results on WFS - SANACOREÂŽ GM trial in Malaysia

DOC at Harvest

Number of Ponds

Survival Rate (%)

Final MBW (g)


Total Crop Yield (Kg/Ha)















Fig. 1: Survival rate at the end of culture (TRIAL 1)

while treatment ponds were continued and harvested as planned at DOC 129. Production data for final mean body weight (MBW), survival (SR), and feed conversion ratio (FCR) are shown in Figs 1 & 2 and Table 1. The average harvest yield in the treatment ponds was 13.7 MT per ha, whereas the control ponds resulted only in 1.6 MT per ha due to the emergency harvest. All treatment ponds were affected by WFS at DOC 60 (control ponds were already harvested at DOC 42). However, the WFS outbreak was mild in severity and the farmer was able to maintain culture and harvest as planned, avoiding the emergency crop. Following the WFS outbreak, the treatment ponds recovered the average daily growth within 7 to 14 days.

Fig. 2: FCR at harvest (TRIAL 1)

Trials in Indonesia The trials in Indonesia were conducted in 2 different farms located in the SubangKarawang regions in Northern Jakarta, an area that is heavily affected by WSSV and WFS. In both farms, the previous two crops were aborted with an emergency harvest due to stunted growth and mortality of the shrimp caused by WFS.

Trial 2, farm 1 in Indonesia The second farm trial aimed at evaluating a preventive protocol for the application of the functional feed additive through the addition of a basal level of SANACOREÂŽ GM via the feed, complemented during specific WFS episodes by additional dosage via top-coating application at the farm (Fig. 3). However, the protocol was adapted during the trial due to late arrival of the treatment feed

and the additive. As a result, the first exposure to the feed additive in the treatment group started only at DOC 20.

24 DOC DOC DOC DO DOC DOC DOC DOC DOC DOC DOC DOC DOC DOC DOC DO DOC DOC DOC DOC DOC DOC 1 2 3 C4 5 6 7 8 9 10 11 12 13 14 15 C 16 17 18 19 20 21 22 Proposed control Actual control Proposed preventive


Actual treatment


DOC DOC DOC DO DOC DOC DOC DOC DOC DOC DOC DOC DOC DOC DOC DO DOC DOC DOC DOC DOC DOC 23 24 25 C 26 27 28 29 30 31 32 33 34 35 36 37 C 38 39 40 41 42 43 44 Proposed control Actual control


Proposed preventive


Actual treatment



Preventive dose


Clean up preventive dose


Curative dose

Until harvest B


Until harvest

Fig. 3. Overview of the exposure of treatment and control ponds to the additive in reality during trial 2, compared with the initially proposed protocol.

Fig 4: Evolution of ADG after the WFS outbreak

Fig 5: Evolution of MBW.

* indicates statistically significant differences (P< 0.05)

* indicates statistically significant differences (P< 0.05)

Furthermore, the incidence of a WFS outbreak around days 23-26 in the treatment ponds and its subsequent successful curative treatment with

boosted levels of the feed additive during one week, resulted in the decision of the farmer to treat also the control ponds when they exhibited WFS on DOC 28.

The final exposure of the treatment and control ponds to the additive are summarized in Fig. 3.

25 The application of the curative protocol to the control ponds avoided their flushing and maintained them in the trial. All ponds, control and treatment, received 7 days of boosted levels of the feed additive as curative treatment, followed by standard diet (control ponds) or standard feed supplemented with prevention dosage of the feed additive (treatment). Recovery from WFS happened in both control and treatment. However, subsequent daily growth was significantly different for treatment and control groups. Whereas the control group did not recover growth, the treatment group recovered average daily

growth to acceptable levels within 20 days following the curative treatment (Fig. 4). During DOC 60-80 approximately, a reduction of growth was detected in both groups which could be ascribed to a reduction of feeding rates in the farm due to operational issues. Similar to the trial in Malaysia, the harvest was advanced to DOC 85 in control ponds due to the stunted growth, while treatment ponds were partially harvested on DOC 85 and finally harvested at DOC 100. Comparing results, final survival was improved in 10% (Fig. 6). Shrimp size of the treatment group was increased with

25% compared to the control group at DOC 85, and 42% at the final harvest of treatment group at DOC 100 (Fig7 and Fig 8). The treatment ponds yielded over three times more biomass than the control ponds at final harvest (Fig. 9).

Trial 3, farm 2 in Indonesia The third trial was run as a confirmation trial without control in the Northern Jakarta area. The farm had suffered crop failures during two subsequent stockings prior to the trial caused by WFS in combination with WSSV and other diseases. Due to the high probability of

Fig. 6: Final Survival of the second trial

Fig 7: Mean Body Weight at harvest (trial 2) * indicates statistically significant differences (P< 0.05)

Fig 8: Size Class harvested in the different groups (trial 2). * indicates statistically significant differences (P< 0.05)

Fig 9. Total Kg harvested per treatment (DOC 100) indicates statistically significant differences (P< 0.05)


26 DOC at harvest

Number of ponds

Survival Rate (%)

Final MBW(g)


Total crop yield (Kg/Ha)

Second trial control







Second trial treatment







Third trial treatment







Table 2: final results on WFS - SANACORE® GM trials in Indonesia

failure, the farmer did not accept to maintain control ponds and decided to run four ponds with the treatment proposal. In this trial, the preventive protocol was fully followed as proposed in Fig. 3 from the arrival of the shrimp in the grow-out ponds. In trial 3, a first, mild outbreak of WFS occurred rather late at DOC 62-66. No additional curative treatment was applied and the shrimp recovered without subsequent stunted growth. This promising outlook made the farmer decide to continue the culture in all four ponds. During DOC 83-88, a second outbreak of WFS appeared, which induced the farmer to harvest. Final harvest results of both farm trials in Indonesia are compared in Table 2. The preventive approach using the feed additive (trial 3) showed a very interesting performance with survival amounting to 68% in average for the 4 ponds, despite the absence of any curative treatments during two outbreaks of WFS, in an area where previous crops failed repetitively. The curative approach using the feed additive (trial 2) showed a clear impact on survival and productivity, with a 10% improvement of survival at harvest despite the 15 days longer period of culture. However, comparison is difficult since the control shrimp also received the curative

treatment during 7 days when WFS outbreak occurred before DOC 30.

Conclusion The current farm trials were performed in “black” areas, i.e. culture areas where previous cycles failed repetitively before the trial. This was confirmed by the need to execute emergency harvests on the control ponds in two trials due to high mortalities and / or stunted growth. These trials demonstrated the capacity of the functional feed additive treatment to delay or control the severity of WFS outbreak, and - most importantly for the farmer - recover the shrimp growth to normal levels following the WFS event. The inclusion of the additive showed the

improvement of the production yield, especially when applied as a preventive strategy since the first day of stocking, thanks to improved growth as well as survival. Furthermore, the curative treatment showed very positive effects, even when applied only during the WFS outbreak.

The results were achieved in different farms and countries, were production strategies, daily routines and farming conditions are different. This further confirms the broad spectrum action radius of the feed additive and its capability to reduce the impact of WFS on economics of shrimp farming in SE Asia.


More information Ho Gim Chong, MSc, Product manager Aquaculture Farm Products SE Asia, Nutriad International E:

Maria Mercè Isern Subich, DVM, Business development manager Aquaculture Health , Nutriad International E:

Allen (Ming Hsun) Wu, MSc., Regional manager Aquaculture Asia Pacific, Nutriad International E:

Ho Gim Chong and Mercè Isern at work in the field




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13 – 14



FVG Select 2017 is a new event which will be organized by Victam International BV. The event will focus on a series of high quality industry conferences and matchmaking with colleagues and clients in the animal feed processing, grain processing, ingredients & additives, aquafeed, petfood and biomass pelleting industries. The event will be complemented by an industry expo for a select group of companies. THE CONFERENCES AND DELEGATE PROFILES Tuesday 13th June

Wednesday 14th June

1. PETFOOD FORUM EUROPE Production technology and specialist ingredients for dry petfoods. Organizer: WATT Global Media Delegate profile: CEOs, directors, mill and plant managers, head and senior nutritionists, formulators from dry petfood production plants.

1. FIAAP ANIMAL NUTRITION The use of specialist additives and ingredients used within the production of animal feeds. Organizer: WATT Global Media Delegate profile: Head and senior nutritionists, feed formulators, CEOs, directors, mill and plant managers from commercial feed production plants, integrators, specialist feed producers, etc.

2. THE GLOBAL MILLING CONFERENCE WITH GRAPAS Processing technology and additives used within flour milling and grain processing. Organizer: Perendale Publishing Delegate profile: CEOs, directors, mill and plant managers, nutritionists from flour mill, rice mills and grain processing plants. 3. VICTAM FEED PROCESSING CONFERENCE Production technology and equipment used within the animal feed production processes. Organizer: Wageningen University and IFF Delegate profile: CEOs, directors, mill and plant managers, head and senior nutritionists, feed formulators from commercial feed production plants, integrators, etc. G Evening network reception for exhibitors, delegates and visitors.

2. AQUAFEED HORIZONS Production technology and specialist ingredients for aquaculture feeds. Organizer: Delegate profile: CEOs, directors, mill and plant managers, head and senior nutritionists, feed formulators, integrators, specialist aquaculture feed producers, etc. 3. BIOMASS PELLETING Production technology for the pelleting of bio-degradable materials for biomass pellets. Organizer: AEBIOM (The European Biomass Association) Delegate profile: CEOs, directors, mill and plant managers from biomass pelleting plants.

A fee of â‚Ź95 per person (valid for two days) will be charged to each individual attending the event, both exhibitors and visitors, and includes lunch, two coffee and tea breaks a day, the network reception and access to our matchmaking service. A separate fee will be charged for each conference. More details can be found on our website.



There will be 9m2 and 18m2 standard shell scheme booths available during the two day event. The expo will take place in the adjoining hall to the conference rooms and will also be the venue for all the refreshments.

Victam International BV P.O. Box 197, 3860 AD Nijkerk, The Netherlands T: +31 33 246 4404 F: +31 33 246 4706 E:


Preparing fish for stress by stimulating protein chaperones By Eva BequĂŠ1, Geert Rombaut1, Patrick Lavens1, Tania De Wolf2, Mathieu Wille3, Peter Bossier3 1. INVE Technologies, Belgium. 2. INVE Aquaculture Research Center, Italy. 3. Laboratory of Aquaculture & Artemia Reference Center, Ghent University, Belgium

Introduction During standard aquaculture practices, fish are affected by stresses such as being handled, transported or vaccinated. These stresses affect fish physiology and immunity and can also cause minor physical damage (Portz et al 2006). This can then result in loss of appetite, poor growth, deformities, infections and in the worst case mortality. Managing stress in fish is therefore an essential part to create more robust animals. Attaining robust animals throughout the entire production cycle is an essential part in the holistic approach to combat diseases for all aquaculture organisms. For fish, investing in the quality of larvae and juveniles results in a significant return on investment for subsequent cage or pond farming. Nutrition and health protocols have a large impact on fish quality that lasts beyond the early life stages. The third factor that affects production and returns in pond and cage culture is the quality of fry, as transport to the on-growing site and stocking into cages or ponds with less controlled culture conditions is very stressful and may affect the vulnerable fry. Stocking high-quality, robust fish will in turn result

in a more reliable production and a reduced disease risk in grow-out facilities.

Aim and Rationale INVE Aquaculture, through its Innovations Department and in collaboration with recognized research institutes, is continuously striving to bring innovative and cost-beneficial concepts to the market. To document the proof of concept of innovative methods of improving the host’s defense system, a 3year joint research project with the Laboratory of Aquaculture & Artemia Reference Center (ARC) at Ghent University was set up. In a previous article (Aquafeed Vol 8 Issue 4 2016) we described the mechanism of Heat Shock Protein induction (HSPi) which results in having more chaperones pro-actively produced in the animal so that they can better cope with stressors and thus having a spectacularly better performance in abiotic stress tests and bacterial challenges. A promising way to improve the host’s defense is thus by stimulating its endogenous protective pathways, such as increased heat shock

protein synthesis (Fig.1). And the validation of HSPi in the model organism Artemia and in larval shrimp culture was demonstrated.

In this second article we cover our research results with fish fry, showing our approach to prepare fish for anticipated stressful events. When fish are exposed to stressful conditions, the hypothalamic-pituitary-interrenal axis (HPI) is activated and the release of corticosteroid hormones such as cortisol is induced and at the cellular level, fish can respond to these stressors by an increased synthesis of heat shock proteins (Poltronieri et al 2007). By using formulations of selected natural components in the culture water of the fish we aim to trigger the Heat Shock Protein (HSP) chaperones in the cells of the fish and thereby prevent protein damage caused by oxidative stress from challenging events such as transport and grading, rather than attempting to recover from an existing insult. These treatments with compounds from plant origin will enable higher survival and more robust fry during and after these stressful conditions.



Fig.1: Mechanism of Heat Shock Protein 70 (HSP70) induction.

Results in Seabass (Dicentrarchus labrax) A variety of compounds from plant origin were selected and tested on HSP inducing properties. Seabass juveniles (DAH30) were exposed for 2 hours to the test compounds at different concentrations. After different recovery times the production of HSP70 was detected and quantified by Western blot analyses. Results showed that the relative amount of HSP70 increased in the seabass treated with some of the tested compounds from plant origin. Figure 2 shows the amount of HSP70 analyzed by Western blot in seabass juveniles. Exposure to a HSPi concentrate significantly increased HSP70 levels after 2 and

2h recovery

12h recovery


2 mg/l

5 mg/l


2 mg/l

5 mg/l







Fig.2: Western Blot results showing the amount of HSP70 proteins in seabass juveniles after 2 hours and 12 hours of recovery from a 2 hour exposure to different concentrations of a natural HSPi compound in the culture water.

12 hours recovery. In a subsequent starvation challenge trial, where the fish weren’t fed for 6 days, pretreated seabass juveniles could resist starvation

conditions much better than the control animals (Fig.3). This could be very beneficial in events such as transportation when fish aren’t fed for 2 to 3 days.

30 As described in the FAO review of Berka (1986), the fish to be transported should be left to starve for at least a day; fish with full digestive tracts need more oxygen, are more susceptible to stress, and produce excrements which take up much of the oxygen of the water. However, when fish larvae are transported, their time of survival without food should also be taken into consideration.

Fig.4: Survival of juvenile sea bream (10-15g) subjected to an air exposure stress test after being pretreated with different concentrations of a HSPi concentrate (10-100mg/l) for 24 hours in the culture water.

Fig.3: Survival of Seabass juveniles in a 6 day starvation challenge trial. Seabass were pretreated for 2 hours with different concentrations of a natural HSPi concentrate (10-20 mg/ l) in the culture water before starvation challenge.

Results in Sea bream (Sparus aurata) Optimal doses for application and different modes of application of natural HSPi compounds in the culture water of sea bream fry were determined. Trials were carried out during the nursery stage of sea bream, from 60 days post hatching (dph) up to the pre-ongrowing stage (<15g). Selected plant compounds were applied shortly before critical moments such as grading and transports when stress related problems could occur. Different mixtures of HSPi compounds were tested for optimal stress resistance

Fig.5: Survival of juvenile sea bream (70 dph) subjected to an air exposure stress test 48 hours after being pretreated with different HSPi compounds (P1, P2 & P5) for 2 hours in the culture water.

improvement, as evaluated by air exposure stress tests (Fig 4 & 5). Bath treatments with different HSPi concentrates resulted in significantly increased survival of the fish during air exposure stress tests up to 3 days after treatment without affecting overall survival or growth. These findings were reinforced by the relative amount of HSP70 proteins present in the fish 24 hours after a 2 hour bath treatment for different ages of fish, showing a consistently higher HSP70

content (12-33%) in fish that were pretreated with different HSPi compounds for 2 hours (Fig.6). A 48 hours transport simulation trial in sea bream of 160dph with different doses of HSPi compounds and different exposure times showed the beneficial effect on stress resistance of the treatments as evaluated by air exposure stress tests (Fig 7).


Fig.6: Amount of HSP70 relative to control in seabream juveniles of different ages 24 hours after a 2 hour bath treatment with a HSPi compound

Results in Tilapia As for seabream, a transport simulation trial was conducted for Tilapia of 40g

Fig.7: Survival of juvenile sea bream (160 dph) subjected to air exposure for 8 minutes after a 48 hour transport simulation and different treatments with HSPi compound 1 (P1). TR2 was exposed to 100mg/l P1 for 2 hours before transport; TR3 was exposed to 100mg/l P1 for 3 hours before transport; TR4 was exposed to 50mg/l P1 for 48 hours during transport.

with different doses of different HSPi concentrates applied in the water during transport (Fig 8). Results showed the

beneficial effect on stress resistance of the treatments as evaluated by salinity stress tests after transport (Fig 9).

32 References: Berka, R. (1986). The transport of live fish: a review (No. 48). Rome: Food and Agriculture Organization of the United Nations. Poltronieri, C., Maccatrozzo, L., Simontacchi, C., Bertotto, D., Funkenstein, B., Patruno, M., & Radaelli, G. (2007). Quantitative RT-PCR analysis and immunohistochemical localization of HSP70 in sea bass Dicentrarchus labraxexposed to transport stress. European journal of histochemistry: EJH, 51(2), 125.

Fig.8: Transport simulation set-up in 100L-tanks.

Portz, D. E., Woodley, C. M., & Cech, J. J. (2006). Stress-associated impacts of short-term holding on fishes. Reviews in Fish Biology and Fisheries, 16(2), 125170.


Fig.9: Cumulative stress index of Tilapia (40g.) subjected to 45ppt salinity after a 48 hour transport simulation with different treatments of HSPi concentrates during transport.

Conclusion Through extensive scientific research and development it was found that natural HSPi components enhance the stress resistance of fish by inducing the production of Heat Shock Proteins. These documented effects proved to significantly improve stress management for different fish species in different age groups. We showed that resistance to air exposure during events such as grading and vaccination procedures can be measurably improved as also resistance to starvation conditions and other

stresses during transport operations. Overall the robustness of the fry was visibly improved, which is an essential part in the holistic approach to combat diseases.

More information

Acknowledgements: Part of this research was funded by the Belgian Agency for Innovation by Science and Technology (IWT), project number: 120404.

Eva BequĂŠ, INVE Technologies, Belgium E:


Algae of the seas Providing for natural health in aquaculture species By Maarten Jay van Schoonhoven, Aqua Care manager, Olmix, France

Macro algae, or seaweeds, play their role in shaping the marine underwater world, from the seashore to depths, where there is enough light for seaweeds to grow. Seaweeds were mainly used as a food source by coastal communities or used in folklore and medicinal purposes. Seaweeds were and are still being used as fertilizers, benefiting among others from the rich mineral content. They are also widely used in the food industry for various purposes such as gelling, thickening and stabilizing agents. More recent screening has revealed a whole new range of biological activities, such as anti-bacterial, anti-viral, anti-hyperlipidemic, mucin inducing, etc.

Algae, a source of biological treasures Macroalgae, are divided in three different groups: green, red and brown. They are made up of a variable part of carbohydrates (mainly polysaccharides), proteins, minerals, lipids and vitamins. Nutritional studies on marine algae indicate that green, brown and red seaweeds possess good nutritional characteristics and could be used as an alternative source of dietary fiber, protein, vitamins and minerals (Chojnacka et al., 2012; Raposo et al.,

2013). In addition, detailed screening of macroalgae functions have revealed a new range of biological activities including anticoagulant, antiviral, antibacterial, anti-tumoral and immunomodulatory activities. All of them could be of relevance in nutraceutical functional foods (Wijesekara et al., 2011a). Cell walls of green, brown and red macroalgae contain large amounts of sulfated polysaccharides, named ulvans, fucoidans and carrageenans respectively, which can range from 4 up to 76% of seaweed dry weight (Holdt et al., 2011). The high content of these sulfated polysaccharides, their unusual structure, and their biological properties reveal these compounds as promising natural products for medicinal and dietary applications (Rioux et al., 2007; Laurienzo et al., 2010). The biological specificity of these marine algal polysaccharides stem from the complexity of their structure. Marine algal polysaccharides are branched polysaccharides, they are composed of various sugars (including rare sugar units like xylose, rhamnose) and these sugars can be sulfated, conferring them a

special reactivity. Together these parameters show a phylogenic similarity with polysaccharides from the animal kingdom such as heparin, which explains their unique biological properties. Marine algal polysaccharides’ reactivity, and biological properties, vary a lot according to the type of sugars and linkage they contain, their level of sulfation and also their molecular weight. Therefore, several marine algal polysaccharides, each with distinct biological activities, can be found in algae. Their specific extraction is a key to ensure a targeted effect on animals.

34 Algae extracts to support natural defenses Polysaccharides can be extracted and purified and one of these purified groups is recognized as having immunomodulating properties. Studies evaluating the immunological properties of algaederived polysaccharides were mainly carried out using primary culture of macrophage like cells lines (Chen et al., 2008; Jaswir et al., 2011). However, although macrophages are essential effector cells of innate immunity, intestinal epithelial cells are also of interest since they express patternrecognition receptors (PRRs) that enable them to act as dynamic sensors of microbial environment and foreign antigens. They are therefore active participants in coordinating the mucosal immunity by producing a broad range of mediators involved in adjacent immune cells activation (Peterson et al., 2014).

Assessing the immunomodulatory effects of macroalgae extract in vitro

Assessing the immunomodulatory effects of algae extract in aquaculture

A recent study, from Berri et al. (2016), conducted in collaboration with INRA (National Institute for Agronomy Research) and published in the Journal of Applied Phycology, demonstrated that the sulfated polysaccharides with specific immunomodulating properties, prepared from the green algae Ulva armoricana (Olmix MSPIMMUNITY), are able to activate immune receptors of intestinal cells, and to induce the expression of some immune mediators (cytokines and chemokines), thus modulating both innate immunity (macrophages, neutrophils) and adaptive immunity (T and B lymphocytes). This suggests that this extract could be used as a new prophylactic strategy to stimulate the immune response of animals, and thus reinforce their natural defenses.

To evaluate the capacity to stimulate the immune response in aquaculture, MSPIMMUNITY in the form of an end product named Searup was tested, on shrimp. Stress and bacterial challenge tests were set up in a University trial. Stress tests were set up because this is an often used method to estimate PL quality in a hatchery and a challenge test to evaluate the shrimp response to a pathogenic bacteria which has a serious impact on health.

Stress test Shrimp post larvae (PL5), having tested negative to WSSV, YHV, TSV, MBV and AHPND by PCR method, were reared in 4 tanks of 500L each, with an average

35 stocking density of 50pcs/L until stage PL12. Water in each rearing tank had a salinity of 25ppt. Tanks were randomly allotted to one of four treatments during rearing:

Searup 3d: Searup application (0.1%) during 3 consecutive days prior to stress tests

At stage PL12, shrimp in each treatment tank were allotted into twelve 30L tanks (500pcs/tank). Each treatment received 3 different stress tests (2 levels of salinity (10ppt and 30ppt) and formalin (100ppm)) which were conducted with 4 replicates for each stress test. Mortalities of PLs were recorded at 3h, 6h, 9h and 24h after exposure to salinity and formalin stress.

Searup 5d: Searup application (0.1%) during 5 consequtive days prior to stress tests



Control: no Searup application

Searup 7d: Searup application (0.1%) during 7 consequtive days prior to stress tests

24h after exposure, PLs from Searup groups had significantly lower cumulative mortality than the control group (P<0.05) for all stress tests.

Figure 1. Cummulative mortality after 10ppt salinity challenge. The best performance was obtained when Searup was administered for 5 and 7 consecutive days.

The use of Searup showed a dose dependent increase in survival rates after the stress tests, suggesting an increased tolerance to stress such as during handling, transport and stocking of growout ponds.

EMS challenge Shrimp post larvae were reared from PL5 to PL25 in 4 tanks of 500L, with an average density of 50pcs/L. All PLs tested negative to WSSV, YHV, TSV, MBV and V. parahaemolyticus causing EMS/AHPND by PCR method. Tanks were randomly allotted to one of the following 5 treatments:

Figure 2. Cummulative mortality after 30ppt salinity challenge. All Searup treatments showed similar improvements against the control (P <0.05).

Figure 3. Cummulative mortality after 100ppm formalin challenge. Application of Searup during 3,5 and 7 consecutive days prior to the stress test showed significantly improved survival rates (P<0.05).

36 • Negative control: no application of Searup – no challenge • Positive control: no application of Searup – EMS challenge • Searup 3d: Searup application (0.1%) during 3 consecutive days from PL5 to PL8 - EMS challenge

• Searup 5d: Searup application (0.1%) during 5 consecutive days from PL5 to PL10 – EMS challenge • Searup 7d: Searup application (0.1%) during 7 consecutive days from PL5 to PL12 - EMS challenge Searup was coated onto the diets with water, after which the diets were allowed to air-dry prior to feeding. At stage PL25, each tank was allotted to 30L tanks with an average stocking density of 500pcs/tank. Shrimp were then submitted to an immersion challenge with EMS causing bacteria (Vibrio parahaemolyticus). There were 4 replicates per treatment. PL mortalities were recorded daily over a 15-day period.

Results In the positive control group, first mortalities were recorded 9h after the EMS challenge. In the Searup treated groups, first mortalities were recorded 24h after the challenge test. A dose dependent effect was observed, with 5 and 7-day administrations showing the best survival rates after challenge (P<0.05). EMS has a major impact in the shrimp industry worldwide. Using Searup, which contains immunomodulating algal polysaccharides (MSPIMMUNITY), has shown in this trial to be an effective product to

Figure 4. Survival rate post EMS-challenge.

improve survival rates after a challenge with EMS causing bacteria, even when the challenge takes place two weeks after the last Searup application. Searup has a long lasting protective effect.

Conclusion The use of MSPIMMUNITY was able to support the natural defenses of the shrimp. This suggests that specific algae extracts are promising candidates to help farms achieve a good performance and improving the overall health of shrimp using natural products from the seas.


More information Maarten Jay van Schoonhoven, Aqua Care manager, Olmix E:

World Aquaculture 2017

Sustainable Aquaculture – New Frontiers For Economic Growth

June 26-30, 2017

Cape Town International Convention Centre Cape Town, South Africa The Annual International Conference & Exposition of World Aquaculture Society Hosted by Aquaculture Association of Southern Africa Department of Agriculture, Forestry and Fisheries, Republic of South Africa Associate Sponsors Aquaculture Engineering Society International Association of Aquaculture Economics & Management WorldFish


Effects of a phytonutrient on aquatic species raised under commercial conditions By Josselin le Cour Grandmaison, Product Manager – XTRACT® range, Pancosma SA, Switzerland Tilapia and shrimp farming are currently booming: Food and Agriculture Organization predicts that 12 million tons of tilapia feed will be produced by 2020. Shrimp farming is following the same dynamic from 2001 to 2014 shrimp aquaculture production has been multiplied by 13. These sharp rises are not without consequences and numerous challenges have to be overcome by industry and farmers. Enhancement of growth performance and improvement of the animal health status are currently the two main issues for these aquatic species. As a matter of fact, intestinal health, integrity and functioning are key elements in animal production and aquaculture. Physiological research has demonstrated that digestive tract is not only a place where digestion is occurring, but other systems as important as immune defenses are also taking a crucial place. Some products are able to increase digestive secretions, protect gut epithelium and possess immune

modulating properties. However their effects are often tested in vitro or in research-like conditions precluding their real assessment at farm level. The Swiss based company Pancosma, a global leader in feed additives, has developed concrete solutions for aquaculture farming. XTRACT® 6930 is a product composed of standardized protected particles containing carefully selected combination of bioactive substances found naturally in aromatic plants and spices. Focusing on a few, well known ingredients, a set of physiological reactions are triggered inside aquatic species digestive tracts: boosting feed utilization and modulating natural defense of aquatic species. For instance, XTRACT® allows to better utilize feed via an enhancement of digestive enzyme secretions, the improvement of nutrients absorption and a reduction of maintenance requirements via its immune modulation action. It also permits to better endure external challenges without any need for antibiotics. New trial results demonstrating XTRACT®

positive action on tilapia and shrimp have been set up in commercial-like conditions. We present the results in this article.

Evaluation of XTRACT® on tilapia growth performance Juvenile tilapia fish with a body weight of 2.56 g were transferred to 24 tanks: 200 fish were allocated per tank. Water in tanks was mimicking commercial situation. Tilapia were fed with 3 different diets: negative control, 200 g/ton and 300 g/ ton of XTRACT®. Each treatment was replicated eight times. The negative control consisted of a basal diet made of soy bean meal, fishmeal, wheat flour, rice, blood meal and a premix. The XTRACT® 200 g/ton and XTRACT® 300 g/ ton diets consisted in the basal diet supplemented with of XTRACT® at 200 g/ ton and 300 g/ton respectively. XTRACT® was previously mixed with fish oil and top coated at 2% onto the pellets in order to reach additive desired concentration.

39 Final fish weight (g/tank)

Negative control

XTRACT® 200 g/ton

Feed intake (g/tank)

XTRACT® 300 g/ton

Negative control

XTRACT® 200 g/ton

Economic FCR (g/g)

XTRACT® 300 g/ton

Negative control

XTRACT® 200 g/ton

XTRACT® 300 g/ton

Figure 1, 2 and 3: Final fish weight, fish feed intake and Economic Feed Conversion Ratio (FCR) in function of treatment groups (a, b, P< 0.05). Economic FCR = Feed Intake / (Biomass Final – Initial Biomass)

Table 1: Histological evaluation of tilapia hindgut and midgut in function of two dietary treatments.

Negative control

Tilapia fed with XTRACT® at 200 g/ton

A dose response was noticed on tilapia growth performance: when XTRACT® dose was increased from 200 g/ton to 300 g/ton tilapia performance was also improved in a linear manner.


These results corroborate previews research performed on young tilapia in Thailand in 2015. Histological observation suggested that XTRACT® supplemented at 200 g/ton had a positive effect on absorption surface and intestinal integrity of tilapia (see Table1). Intense and numerous well shaped villi were observed in compassion to control group.



Compared to fish administered with negative control, tilapia fed with XTRACT® at two levels displayed better growth performance: higher final body weight (p<0.05), higher feed intake and improved economical feed conversion ratio (Figures 1, 2 and 3 ).

Disrupted and heterogeneous groups of villi

By courtesy of Dr Thomas Wilson

Intense and numerous well shaped villi

Improvement of tilapia survival rate was also noticed when XTRACT® was supplemented at 200 g/ton and 300 g/ ton of feed (Figure 4). This lower mortality could be explained by the ability of XTRACT® to modulate immune system and help aquatic animals to endure external challenges.

40 Survival rate (%)

Negative control

XTRACT® 200 g/ton

XTRACT® 300 g/ton

Figure 4: Tilapia survival rate in function of treatment groups

Evaluation of XTRACT® on shrimp growth performance Pacific white shrimps (L. vannamei) with a body weight of 3.06 g were allocated into 36 cages and raised during 63 days: 225 shrimps were allocated per cage. Cages were allocated in commercial shrimp pond. Shrimp were fed with 3 different diets: negative control, XTRACT® 100 g/ton and XTRACT® 150 g/ton. Each treatment was replicated twelve times. The negative control consisted of a basal commercial shrimp diet with 35% of Crude Protein. The XTRACT® 100 g/ton and XTRACT® 150 g/ton diets consisted in the basal diet supplemented with of XTRACT® at 100 g/ton and 150 g/ton respectively. XTRACT® was added directly in to feed Final shrimp biomass (g/cage)

Negative control

XTRACT® 100 g/ton

XTRACT® 150 g/ton

via the premix.

Discussion and Conclusion

Cages containing shrimps fed with diets supplemented with XTRACT® displayed higher final biomass and higher feed intake. When XTRACT® was supplemented at 150 g/ton of feed, large improvement of shrimp Feed Conversion Ratio was observed (-4.8%).

Dietary supplementation of this phytogenic feed additive successfully improves performance of juvenile tilapia and pacific white shrimps. For these two aquatic species final body weight, feed intake, economic feed conversion ratio and survival rate were improved. Using comparative physiology it is possible to get insights of the product’s true mode of action (Bravo. 2015).

More importantly, shrimp fed with XTRACT® had a strongly higher survival rate in comparison with shrimp fed negative control diet. Again this better shrimp survival rate can be explained by Shrimp survival rate (%)

Negative control

XTRACT® 100 g/ton

XTRACT® 150 g/ton

Figure 8: Shrimp survival rate in function of treatment groups

the ability of XTRACT® to help aquatic species to support external challenges and stresses.

Cumulative Feed Intake (g/cage)

Negative control

XTRACT® 200 g/ton

XTRACT® 300 g/ton

Higher bile acid secretion and enhance lipase activity leading to a higher fat digestibility are effects mainly coming from capsaicin, a component of XTRACT® (Bravo et al., 2014; Ganesh Bhat et al., 1984; Jamroz et al., 2005). In addition anti-oxidant and immunomodulation properties exert a protective effect on gut membrane improving intestinal integrity and gut absorption surface (Awaad et al., 2014; Karadas, Pirgozliev et al, 2014). Thus phytonutrient XTRACT® answers one of the main demands of industrials and farmers regarding tilapia and shrimp farming: better feed utilization and resistance to external challenges and stresses. This solution has the potential to generate greater financial profit to Economic Feed Conversion Ratio (g/g)

Negative control

XTRACT® 200 g/ton

XTRACT® 300 g/ton

Figure 5, 6 & 7: Final shrimp biomass, Cumulative Feed Intake and Economic Feed Conversion Ratio (FCR) in function of treatment groups. Economic FCR = Cumulative Feed Intake / (Biomass Final – Initial Biomass)

41 Dimitrov, O. Oduguwa, and D. Bravo. 2014. Dietary essential oils improve the hepatic antioxidative status of broiler chickens. British Poultry Science 55 (3): 329-334. AFΩ

Trials at experimental farm in Vietnam aquaculture stakeholders. In addition to phytonutrients, Pancosma has designed additional feed additives based on their expertise. This portfolio for aquatic species includes organic trace minerals and health enhancers.

References Awaad, M. H. H., M. Elmenawey, and A. A. Kawkab. 2014. Effect of a specific combination of carvacrol, cinnamaldehyde, and Capsicum oleoresin on the growth integrity of broiler chickens performance, carcass quality and gut integrity of broiler chickens. Veterinary World, EISSN: 2231-0916. Bravo, D. M. 2015. Comparative Gut Physiology Symposium. J. Ani. Sci. 93. Ganesh Bhat B, Srinivasan M. R., and Chandrasekhara N. 1984. Influence of curcumin and capsaicin on the composi-

tion and secretion of bile in rats. J. Food. Sci. Technol. 21, 225-227. Bravo, D., V. Pirgozliev, and S. P. Rose. 2014. A mixture of carvacrol, cinnamaldehyde and capsicum oleoresin improves energy utilization and growth performance of broiler chickens fed maizebased diet. Journal of Animal Science 92:1531–1536.

Thus the phytonutrient answers one of the main demands of industrials and farmers regarding tilapia and shrimp farming: better feed utilization and resistance to external challenges and stresses. This solution has the potential to generate greater financial profit to aquaculture stakeholders.

Jamroz D, Wiliczkiewicz A, Wertelecki T, Orda J, and J. Skorupinska. 2005. Use of active substances of plant origin in chicken diets based on maize and locally grown cereals. British Poultry Science 46 [4]: 485-493. Karadas, F., V. Pirgozliev, S.P. Rose, D.

More information Josselin le Cour Grandmaison, Product Manager – XTRACT® range, Pancosma, Switzerland E:


Inulin, a promising prebiotic in the prevention of diseases in modern aquaculture

Introduction Application of prebiotics is a common practice in functional feeds for fish farming today to improve fish or shrimp health status, promote gut integrity and improve sustainable productive animal performance in general. The most recent definition describes prebiotics as follows

(Gibson , 2010): “A dietary prebiotic is a selectively fermented ingredient that results in specific changes in the composition and/or activity of the gastrointestinal microbiota, thus conferring benefit(s) upon host health.� Typically prebiotics resist gastric acidity and are not hydrolyzed by digestive enzymes nor absorbed in the digestive

system. Prebiotics are fermented by the intestinal microbial community and selectively stimulate the growth and/or activity of intestinal favorable bacteria that are associated with health (Gibson, 2010; Robertfroid, 2010). Hoseinifar (2015) published a comprehensive overview on the role of the most studied and applied prebiotics and the fish

43 immune response. Mannanoligosacharides (MOS), derived from yeast cell walls are probably the most evaluated in fish. Oligofructose (or fructooligosaccharides, FOS) and inulin, both β-D-fructans differing in their degree of polymerization, have also been tested extensively. Next to these there are plenty of candidate ingredients with potentially prebiotic properties in fish such as galactooligosaccharides (GOS), arabinoxylanoligosaccharides (AXOS), etc. Next to the specific proven prebiotic ingredients there are several yeast extracts in the market as commercial prebiotics.

Inulin and oligofructose as prebiotics in aquaculture species Beneo is a producer of inulin and oligofructose, soluble indigestible polysaccharides consisting of β(2,1) bonded fructosyl molecules derived from chicory root. Inulin is already successfully used in food and feed applications. Whereas oligofructose is fermented in the midgut, inulin reaches the hindgut. Inulin and oligofructose are selectively fermented and induce a shift in microbial composition. Addition to feed stimulates selectively the growth of “friendly bacteria” as illustrated in the study of Mahious (2006) in weaning turbot in

which an increase from 0% to 14% of Bacillus was observed after feeding of oligofructose. A higher bacterial diversity as well as positive effects on growth have been observed as well. Changes in intestinal microbiota after addition of inulin, oligofructose or shortchain fructooligosaccharides (scFOS) were also observed in Pacific white shrimp (Li, 2007), Stellate sturgeon (Akrami, 2013) and rainbow trout (Ortiz, 2013). The short-chain fatty acids (SCFA) produced by the microbiota after fermentation of chicory root fiber lead to a stimulation of the villi growth. The associated reduction of the pH in the gut

Fig. 1: Effect of dietary oligofructose as prebiotic for weaning turbot on microbiota composition (data taken from Mahious, 2006).

Figure 2: Production levels and relative amounts of SCFA during in vitro fermentation of inulin (IN) and oligofructose (OF) (internal research, unpublished results).


Fig. 3: In vitro reduction of intestinal pH due to fermentation of inulin and oligofructose (internal research, unpublished results).

creates a less favorable environment for pathogens. Investigations by Beneo on in vitro fermentation of prebiotics in sturgeon showed an increased production of SCFA and a shift towards higher butyrate production (unpublished results).

Fig. 4: Serum total immunoglobulin (Ig) levels of Caspian roach fry fed different levels of oligofructose (data taken from Soleimani, 2011).

In the trial the shift in SCFA lead to a reduced pH in the intestinal tract as illustrated in the below graph. Prebiotics sometimes referred to as “immunosaccharides� exert a direct stimulating effect on the innate immune

systems (Akhter, 2015). In Caspian roach fry Soleimani (2012) discovered a doseresponse effect on total serum immunoglobulin levels when fed different levels of oligofructose. The results are illustrated in figure 4.

45 Inulin in diets for Atlantic salmon smolts (internal research, unpublished results). In the remaining part of this article a promising challenge study on the effect of inulin on productive performance, gut health and immune response in Atlantic salmon is presented. Atlantic salmon smolts of 71 grams average weight were fed during 60 days a control diet or test diets containing two different levels of inulin (0.2% and 0.5%). After 60 days growth performance and villi heights were compared. The remaining fish were challenged with different dosages of Piscirickettsia salmonis after which fish mortality was monitored during 30 days post challenge.

Figure 5: Microvilli height (Âľm) at 0 and 60 days after feeding 0%, 0.2% and 0.5% inulin to Atlantic salmon smolts.

Results After 60 days the growth of the tested groups was not significantly different however the specific growth rate (SGR) was highest in the 0.5% inulin group and higher in all the inulin fed groups compared to the control group. The most favorable feed conversion ratio (FCR) was observed in the 0.2% and 0.5% inulin groups. Table 1: Growth performance of Atlantic salmon smolts after 60 days feeding with different levels.

Inulin in feed

Initial weight (g)

Final weight (g)





1.93 1.12




2.03 1.08




2.11 1.09

Figure 6: Effect of inulin supplementation on accumulated mortality 30 days after P. salmonis challenge in Atlantic salmon smolts.

Histological evaluation revealed a marked increase of the intestinal villi height in the inulin fed animals after 60 days as illustrated in figure 5. The accumulated mortality 30 days after the challenge with Piscirickettsia salmonis is illustrated in figure 6. It ranged from 55.6% in the control group

to 43.8% in the 0.2% inulin fed group and 32.5% in the 0.5% inulin fed animals.

Conclusion From the presented study it may be concluded that addition of chicory derived prebiotics in casu addition of

46 0.2% and especially 0.5% inulin to feeds of Atlantic salmon smolts, results in a better intestinal health, an improvement in productive performance and a higher resistance to infectious diseases in this case Piscirickettsia salmonis.

“A dietary prebiotic is a selectively fermented ingredient that results in specific changes in the composition and/or activity of the gastrointestinal microbiota, thus conferring benefit(s) upon host health.” — Gibson, 2010

Based on internal Beneo research and published literature, the addition of prebiotic inulin and oligofructose in aquafeed is an excellent solution to contribute to fish farmers’ profitability by improving the health condition of the farmed fish and productive performance. Beneo-Animal nutrition is dedicated to a more sustainable aquaculture industry by offering scientifically proven innovative nutritional concepts.

Acknowledgement: SGS Aquatic Health, Chile Aquaculture Experience, The Netherlands

References Akhter, N. (2015). Probiotics and prebiotics associated with aquaculture: A review. Fish and Shellfish Immunology, 45(2):733-741. Akrami, R. (2013). Effect of dietary supplementation of fructooligosaccharide (FOS) on growth performance, survival, lactobacillus bacterial population and hemato-immunological parameters of stellate sturgeon Acipenser stellatus juvenile. Fish Shellfish Immunology, 35(4):1235-9. Gibson, G. (2010). Dietary prebiotics: current status and new definition. Food Science & Technology Bulletin, 7:1-19. Hoseinifar, S. (2015). Prebiotics and fish immune response: A review of current

knowledge and future perspectives. Reviews in Fisheries Science & Aquaculture, 23: 315-328. Li, P. (2007). Dietary supplementation of short-chain fructooligosacharides influences gastrointestinal microbiota composition and immunity characteristics of pacific white shrimp Littopenaeus vannamei cultures in a recirculation system. Journal of Nutrition, 137, 27632768. Mahious, A. (2006). Effects of dietary inulin and oligosacharides as prebiotics in weaning turbot. Aquaculture International, 14, 219-229. Ortiz, L. (2013). Effects of inulin and fructooligosaccharides on growth performance, body chemical composition and intestinal microbiota of farmed rainbow trout (Oncorhynchus mykiss). Aquaculture Nutrition, Volume 19, 4, 475 –482. Roberfroid, M. (2010). Prebiotic effects: metabolic and health benefits. . British Journal of Nutrition, 104(S2):S1-S63. Soleimani, N. (2012). Dietary supplementation of fructooligosaccharide FOS improves the innate immune response, stress resistance, digestive enzyme activities and growth performance of Caspian roach Rutilus rutilus fry. Fish Shellfish Immunol. 2012 Feb;32(2):31621, 32(2): 316-21. AFΩ

More information Beneo Technical Staff E:


Dry grape extracts authorized as feed additives and for their use in aquaculture A few weeks ago, the Dry Grape Extract (Nor-Grape® 80) developed and commercialized

by Nor-Feed, became the first botanical sensory additive authorized for animal nutrition by the European Commission.

The Dry Grape Extract is rich and standardized in total polyphenols, procyanidins and anthocyanins, active molecules well known for their strong antioxidant potential. It is used in many countries to fight against oxidative stress which is the disbalance of the equilibrium state between antioxidants and free radicals in favor of the latter. This phenomenon can be caused by many factors (water temperature, water quality, level of dissolved oxygen, stress…) and may have dramatic effects on the fish growth, health and the quality of its products. Its potential applications in aquaculture are numerous. In particular, grape polyphenols have been shown to have a protective effect on fish meat when added directly on it after slaughter (Pazos et al. 2005, Gai et al. 2015, Magsood et al. 2013). A trial was set up in an experimental station in the south of France (CREUFOP, Montpellier) on sea bass (Dicentrarchus labrax) to establish if such a protective effect would be observed on the meat when orally supplementing the fish (drip

loss, during storage and cooking, or oxidation state) and whether it would at the same time benefit the fish by improving its antioxidant defenses. 120 170-gram sea bass were divided in two homogeneous groups: a control group (“CTL”), fed a classic grower extruded feed (43% proteins, 20% fats), and a Dry Grape Extract group (“NG”), fed the same feed supplemented with 40ppm of Nor-Grape®80. The fish were fed for 66 days, until slaughter, when blood samples were taken and fish were fileted. Total glutathione peroxidase was measured on blood samples. 30 filets per group were kept at +4°C and individually weighed at D1 and D10. Moreover, 16 other filets were weighed before and after being cooked for 12 min at 80°C to evaluate drip loss. Finally, oxidized fats (TBARS) were analyzed on D10 on 5 filets per group. The measure of total glutathione peroxidase on blood samples evidenced a significantly higher level (p<0,05) in sea bass

48 supplemented with the Dry Grape Extract (see figure 1), demonstrating a beneficial effect of the grape extract on the stimulation of endogenous antioxidant defenses. Moreover, this improvement of the fish defenses resulted in a less oxidized meat (see figure 2) and a reduced drip loss during storage in the supplemented group (NG: 4,93% vs. CTL : 5,15%). Furthermore, the improvement of the oxidative stability of the

Registering the Dry Grape Extract A long and rigorous process The authorization process began in 2004, when Nor-Feed declared the Dry Grape Extract (DGE) in the European Register of Additives to maintain Nor-Grape 80’s right of sale. In 2010, following a request from the European Union, the company filed the EFSA (European Food Safety Authority) Registration. Then, between 2014 and 2016, Nor-Feed exchanged information with the EFSA, the EURL (European Union Reference Laboratory) and the European Commission on the DGE safety and efficacy and the analysis and characterization of its active substances (total polyphenols, proanthocyanidins and anthocyanins). In March 2016, the EURL proceeded to an evaluation of Nor-Feed methods of analysis, and in June 2016, the European Food Safety Authority Scientific Committees published a positive opinion on the DGE.

During a regulatory session, on 14th and 15th November 2016, the authorization of the Dry Grape Extract dossier submitted by Nor-Feed was voted unanimously by the Plants Committee of the Animal Nutrition & Veterinary Medicines (ANVM), and published in the official journal of February, 22nd 2017. Fig, 1. Blood GPx levels of sea bass.

Fig. 2. State of lipid oxidation in sea bass filets.

EFSA had stressed the efficacy and safety of the use of dry grape extract in animal feed. This decision is important in two ways: "The presence of the dry grape extract on the official European Union Register is for us - and our customers - the guarantee that our product is duly standardized and based on sound and validated scientific bases.", said Alexia Lepont, Head of Nor-Feed Quality and Regulatory Affairs. "To our knowledge, there is no other product based on grape extract currently being evaluated by EFSA", she added. Moreover, since the authorization was based on the results provided by Nor-Feed, it is the company methods of analysis and characterization of active substances (total polyphenols, proanthocyanidins, and anthocyanins) that will be used for the evaluation of possible future grape extracts looking for the same authorization. Many actors of the Animal Health & Nutrition industry are already using Nor-Grape 80 to "compensate for oxidative stress in animal farming" and "to bring watersoluble biological antioxidants", using Nor-Feed publications to build their own allegation dossier.

49 The supplementation of the diet with 40ppm of Nor-Grape® 80 for 2 months before slaughter therefore increased the fish antioxidant defenses. This resulted in an improved meat quality by improving the oxidative stability of the meat leading to a reduction of drip loss during the cooking process. Other studies on the use of Dry Grape Extract in aquaculture have demonstrated similar improvements on the products’ quality (meat, eggs) as well as growth performances improvement in several species (fish and crustaceans). This demonstrates the need for a combination of synergistic water- and fatsoluble antioxidants in aquaculture to better protect fish against oxidative stress. AFΩ

More information References available on request. Fig.3. Drip loss of sea bass filets during the cooking process.

filets in the NG group resulted in a significant reduction of drip loss during cooking (-25%, p<0,01, see figure 3).

Paul Engler, R&D and Product Manager, Nor-Feed, France E:


A flexible approach to optimizing the DHA levels of hatchery Artemia R&D

Although Artemia nauplii are widely used in the hatchery rearing of marine fish larvae, by themselves they do not contain a lot of the nutrition that is essential to young fish development. In particular, they are lacking in the highly unsaturated fatty acids (HUFAs) – docosahexaenoic acid (DHA) and eicosapentaenoic acid (EPA) – that are essential to marine fish growth and development. Therefore, to increase the DHA and EPA content, hatcheries typically subject Artemia to an enrichment phase before it is fed to fish, whereby it is placed in an enrichment suspension that it will proceed to ingest and incorporate into its biomass. There are, though, two fundamental problems associated

with this standard practice model, explains Eamonn O’Brien, Product Manager for Skretting Marine Hatchery Feeds (MHF). Firstly, not all Artemia are the same – as a live animal, there are significant disparities between its different strains and their ability to incorporate enrichment as well as variances in quality and strength from year-to-year, which is termed as ‘enrichment kinetics’. And yet the aquaculture industry has a strong tendency to overlook these inconsistencies and look at Artemia as a constant standard.

Secondly, there is the added complexity that the metabolism of Artemia will very efficiently break down the essential DHA into other fatty acids, in particular EPA, thereby depriving the marine larvae of their very high DHA requirement. “Hatcheries typically want a DHA-EPA ratio of 2:1 in their Artemia, which can often be difficult to achieve because of this inherent ability,” says O’Brien.

To mitigate these and other constraints associated with Artemia, a number of hatchery operations have replaced it with a formulated diet. But recognising that Artemia substitution is not an option for every marine hatchery and that it is important that the many hatcheries that continue to follow this protocol make their Artemia as nutritionally rich as possible, Skretting MHF has invested considerable R&D into the delivery of a feed expressly focused on optimising the level of DHA in the live feed. The result is its ORI-N3 Artemia enrichment, which delivers a highly efficient incorporation of DHA into the live feed while also providing far greater flexibility in this essential marine hatchery process compared to standard enrichment protocols. Developed at MHF’s Centre of Excellence in Vervins, France, ORI-N3 is an algaebased liquid that is aligned with its other ORI range concepts in that it is designed to be ingested by the animals more efficiently than traditional methods, as well as being easy to use – it is supplied in 2kg foil stand-up spout bags. The dosage is simply weighed, mixed with water and then administered to the Artemia tank. Because it is algal-based there are no superfluous oils, while the risk of oxygen

52 level challenges are much less likely than with standard enrichments. The Artemia also maintain a good protein base. A direct outcome of these benefits is a greatly reduced mortality rate and a much cleaner enrichment.

Flexibility “It was also very important to provide a solution that is adaptable to each hatchery’s specific requirements, rather than just a single general application protocol,” says O’Brien. “All hatcheries are different; each has individual needs and motivations, so it is vital that the products made available to them are sufficiently flexible to fulfil those aims.” Based on what hatchery managers want to achieve with their Artemia as well as what best suits the operations within their facility, ORI-N3 offers everything

between 12- and 24-hour enrichment. If a medium level n-3 HUFA content and highest DHA-EPA ratio is the target, for example, then a single-dose 12-hour enrichment tends to present the best option. Alternatively, if a hatchery is more interested in achieving a maximum HUFA entry, it should elect for a 24-hour enrichment and two doses, says O’Brien. “Because ORI-N3 establishes a much more efficient intake of DHA, its application can be as low as 400 ppm and still providing very good incorporation into the Artemia and an excellent DHA:EPA ratio. Such low dose application also assists in keeping the enrichment clean and is of course very cost-effective.

“In addition to providing unmatched DHA content, the flexibility and easiness-touse allows each hatchery to tailor the product to its own specific requirements.

With commercial trials imminent, our goal is to have the market fully up to speed ahead of the new season, starting in September,” says O’Brien. AFΩ

More information Eamonn O’Brien, Products Manager Skretting Marine Hatchery Feeds E:!/ SkrettingMarineHatcheryFeeds


Aquafeed Horizons 2017 Advances in Processing & Formulation Koelnmesse, Cologne, Germany. June 14, 2017.

Join us for the 10th in's series of international technical conferences for aquafeed industry professionals.

Limited places REGISTER NOW

Aquafeed Horizons conferences focus on practical information and have become the recognized event in the calendar for industry stakeholders worldwide; it has

been arming delegates with the latest processing and formulation solutions to help keep ahead of the trends and changes in the industry since 2006.

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Aquafeed Horizons 2017 PROGRAM Unravelling the mysteries of extrusion die flow using CFD

Daniel Stoffner; Bühler Nutrition, Product Management Aqua Feed & Pet Food, Switzerland Today, with the power available in standard computer workstations it is possible to realistically model the flow through elaborate extrusion dies, even in an industrial R&D environment, in particular, the effects of viscous heating and shear-thinning viscosity on aquafeed die flow. Typical models spanned the final windings of a twin-screw, the transition from a twin-screw barrel to an axisymmetric die, as well as inserts and nozzles. However, more complicated aspects such as heterogeneous material, viscoelasticity, thixotropic viscosity or gas expansion have not been addressed.

Extruded aquafeed quality management; Relations between technology and extruded aqua feed quality

Thomas Ellegaard Mohr, Segment Manager/ Sales Application Manager, ANDRITZ Feed & Biofuel, Denmark Industrially produced feed is undergoing a continuous quality improvement process. The different types of feed are subject to different demands, the feed/food’s functionality. e.g. on fish farms in terms of floatability or sink ability and pellet durability to assist mechanical handling without the generation of fines. Increased demand for feed also influences the availability of "classic" raw material, this gives new challenges to ensure that the extrusion technologies and process control are developed to adapt accordingly.

New developments in aquafeed production by extrusion

Joe Kearns, Vice President Aquafeed Division, Wenger Manufacturing Inc., USA Aquafeed production methods continue to advance in order to keep pace with changes and requirements desired by the industry. High capacity sinking and floating feed machines are now capable of making products as small as 0.5 mm with production rates measured in tons per hour. AquaFlex designs are available for making a complete, wide range of all styles of aquatic feed. There are density control packages for instantaneous corrections of density while running as well as computer and In-Line measuring devices, all controlled automatically for speed and accuracy avoiding costly rework and improving efficiency by reduced down time and higher plant yield. Capacity increases in large single screw extruder's also improve cost efficiencies by producing higher capacities per hour. Dryer improvements also reflect the industries desire to improve efficiency in production of predicable results.

Alternatives to water-plasticization in the extrusion process – reduced drying costs and improved physical pellet quality Rafiq Ahmad, Industrial PhD candidate, Cargill Aqua Nutrition, USA Possible plasticizer candidates such as free amino acids and pH have been evaluated combined with moisture in a soy protein concentrate model system. The study demonstrates that free amino acids and low molecular weight water-soluble peptides can replace moisture as a plasticizer in the extrusion process. Due to the need for higher moisture level during extrusion processing of plant-based diets these require more energy for drying compared to fishmeal based diets. The use of protein

55 based plasticizers opens up the possibility to obtain a satisfactory “cook� at reduced moisture level with a potential for significant reduction of the energy consumption. The results from this study documents the possibility to develop a new processing add for the fish feed industry serving multiple purposes as nutrient, plasticizer and binder in extruded fish feed.

system to conserve energy and improve efficiency. Water can also be condensed and reintroduced back into a preconditioner or extruder, reducing the volume of fresh water needed for the process and eliminating effluent concerns from a heat recovery system and a processor can raise discharge moisture and get more out of the production line with no additional cost or additional dry ingredients. The value of this recaptured production can be surprising.

Processing efficiencies for sustainability and profits Scott Vallette, Market Manager, Buhler Aeroglide, U.K. This presentation will introduce three opportunities for processing efficiencies in drying that can ensure sustainability and improve bottom line profits: if fines are continuously collected in the dryer, they can be carefully reintroduced upstream to eliminate waste, product loss and the need to introduce additional raw material for the same output. Removing the fines from the dryer on a continuous basis can also reduce the cost and time required for cleaning and reduce potential fire risks; heat can be captured and reused with a heat recovery

Hybrid dryer exhaust air processing; towards a sustainable solution for odor abatement with concurrent energy- and water recovery Anders F. Haubjerg, Sr. Process Engineer, Graintec A/S, Denmark Integrating the goals of reducing odor, recovering energy and reusing condensed water back into the process, in a single endpipe drying solution has been the subject of a recent collaborative development project. A technical-economical comparison with conventional odor abatement technologies will be given, alongside results from pilot-scale experimental stages: condensation based odor

56 abatement efficiency, energy recovery potential and feeding trials using feed produced with recycled water, condensed from the drying process. This new solution brings a promising alternative to the existing technology pool.

Feed and food processing – Technology transfer and novel ingredients

Dr. Olav Fjeld Kraugerud, Manager, Centre for Feed Technology (Fôrtek), NMBU, Norway The drive for new ingredients puts a timely pressure on processing evolution, with respect to both physical quality of the pellets, and nutritional quality. The talk will give some examples on processing of novel ingredients, and strategies to reach the goal. E.g., the food sector has valuable knowledge on ingredients processing, that the feed sector can benefit largely form.

Mycotoxins in aquaculture: occurrence and impact in rainbow trout (Oncorhynchus mykiss). Rui Alexandre Gonçalves, Scientist Aquaculture, BIOMIN Holding GmbH, Austria To evaluate the consequences of mycotoxin contamination in European aquaculture finished feeds, two experiments were performed to study the effect of short and long term feeding of Fusarium mycotoxins to rainbow trout, using natural contaminated raw materials and pure mycotoxins. Moreover the influence of mycotoxins against Yersinia ruckeri susceptibility was evaluated. All experimental diets revealed the presence of other mycotoxins metabolites/toxins alongside added Fusarium. The presence of these naturally-occurring metabolites/toxins, highlights the high risk of mycotoxin contamination in aquaculture finished feeds within Europe. Results confirmed that the ingestion of mycotoxins, within the levels found on finished feeds across Europe (up to 1000ppb), can lead to an overall decline in performance which ultimately results in economic losses.

Toward higher inclusion of soybean meal in salmon feeds André Dumas, Director of Fish Nutrition, The Center for Aquaculture Technologies Canada, Canada The negative effects of soybean meal on Atlantic salmon reported in the literature are variable. Growth of Atlantic salmon is not always negatively impacted by diets containing relatively high levels of soybean meal. Moreover, the inducement of enteritis differed between soybean sources and we have demonstrated that soybean tolerance/utilization in this species is also influenced by genetics in a recent study. This presentation will address and explain further these discrepancies to ensure soybean meal is used increasingly in an optimal manner in Atlantic salmon feeds.

Effect of an immuno-modulating algal extract on disease prevention in aquaculture Maarten Jay van Schoonhoven, Aqua Care Manager, Olmix SA, France Nutritional studies have shown that green, brown and red seaweeds possess good nutritional characteristics, and in addition, also have biological active compounds including proteins, polyunsaturated fatty acids, pigments, polyphenols, minerals, vitamins and polysaccharides. Biological activities related to these compounds include: antiviral antibacterial, antitumor, anti-proliferative and immunomodulatory activities. Studies in collaboration with INRA have demonstrated high immunemodulating activities. More recently University trials where shrimp were challenged with Vibrio parahaemolyticus bacteria causing EMS, showed long lasting effects that helped improving survival rates. Since these University trials, field trials in both fish and shrimp have confirmed the beneficial effects of these sulphated polysaccharides from marine algae.


57 Breaking the bottleneck: Feed-based solutions to the recurring challenges in aquaculture

Deforestation-free and sustainable vegetable fats and proteins for aquaculture feed

Kabir Chowdhury, Global Product Manager Aquaculture, Jefo Nutrition Inc., Canada Despite the significant growth in the share of three major commercially farmed species tilapia, shrimp and salmonids, issues such as poor consumer perception due to the lack of social responsibility, leadership and poor governance, and as well as environmental pollution and frequent outbreaks of old and new diseases remain major hindrances. These issues have been creating a bottleneck for the sustainable growth of the industry. The major contributing factors, which can be partially solved through dietary means, are mainly environmental and health issues. Reducing waste output for better environment, and ensuing better performance and immune response are possible through various dietary means such as better formulation and using various in-feed additive based solutions. This presentation will discuss recurring and upcoming issues that the industry is facing and highlights some of the commercially available feed-based solutions.

Dr. Norbert Schmitz, Managing Director, ISCC, Germany The use of protein and fats sources of vegetable origin have potential negative effects on the environment, as most of the fishmeal alternatives are cultivated in highly bio-diverse tropical and equatorial regions and could derive from unsustainable deforestation practices. This presentation discusses an innovative system based on remote sensing data (Global Risk Assessment Services), able to verify whether unsustainable production practices are used for the production of raw materials in aquaculture feed. The system allows the identification of agricultural land obtained through deforestation. In addition, the several stages of the feedstock preparation and processing can be mapped, offering a complete traceability of the supply chain of the feed products. Cases based on soybean meal and palm oil production are presented in this contribution.


FVG Select Opening Hours

FVG Select—the networking event Aquafeed Horizons is routinely held alongside Victam International’s tradeshows, FIAAP/Victam/Grapas (FVG) that take place in Cologne every three years and Bangkok every other year. This year is a ‘gap year’ – and Victam International has taken the opportunity to fill it with a new event, called FVG Select. It is a two-day, conference-led networking event, which will take place on June 13 and 14, 2017 and which will have an exhibition by a select group of 55 companies. Aquafeed Horizons is one of six independent conferences taking place during the event. A highlight of FVG Select will be a business matchmaking program. It will be done through an easy-to-use technology platform combined with a personalized service to help maximize your networking opportunities. A network reception will be held on Tuesday, June 13th from 17:00 - 19:00 giving exhibitors, delegates and visitors the opportunity to relax, socialize and network.

TUESDAY, JUNE 13, 2017 08:00 Registration & Badge Pick up 09:00—17:00 Conferences 17:00—19:00 Network reception

WEDNESDAY, JUNE 14, 2017 08:00 Registration & Badge Pick up 09:00—17:00 Conferences 10:00—17:00 Exhibition & Matchmaking

ALL GENERAL INFORMATION Brochure, Registration, Exhbitor List, Travel and Accommodation, Conferences


Conference Report

Soy Aquaculture Alliance – Aquaculture America: teaming up to forge a successful future for U.S. aquaculture The Soy Aquaculture Alliance (SAA) Annual Coalition Meeting, held during the Aquaculture America Conference, featured speakers with a wide range of experiences and expertise who shared exciting updates on work underway to support the U.S. Aquaculture Community By Bridget Owen, Executive Director, Soy Aquaculture Alliance, USA The Soy Aquaculture Alliance (SAA) recently held the 13th Annual Coalition Meeting during the Aquaculture America Conference. The annual event includes speakers with a wide range of aquaculture experiences and expertise who

share exciting updates on work underway to support the U.S. Aquaculture Community. This year proved to be another full slate of exciting updates and discussion topics. Nermeen Youssef Abass, 2017 U.S. Aquaculture Society and Soy Aquaculture Alliance Student Travel Award recipient, began the meeting with a presentation on her current research work at Auburn University working with channel catfish. SAA is honored to work with U.S. Aquaculture Society to support this important award. U.S. Aquaculture benefits greatly from the high quality research conducted in our nation and the great educational programs that develop high quality students to support U.S. Aquaculture well into our future. Research remains of key importance to aquaculture and to helping continue to grow in smart and efficient ways. The United Soybean Board and many State

Photo: Scott Bauer, USDA-ARS

Soybean Organizations support the Soy Aquaculture Alliance’s Soy in Aquaculture Research Program. Researchers currently working on research in this program presented updated on their work, including Dr. Tiago Hori from the Center for Aquaculture Technologies and the work they have been doing with SAA to Identify Genetic Markers Associated with Growth Performance on a Soybean Meal Based Diet for Atlantic salmon. Dr. Jesse Chappell, Auburn University, shared the work he and Dr. Terry Hanson have underway with In Pond Raceway Systems (IPRS) development including the rapid adoption of this production system in China and introduction of the system elsewhere in the world. Dr. Guillaume Salze also shared the work he and his team from Auburn University worked on regarding the Effect of Pellet Size, Extrusion Technology, and Water Temperature on Taurine Leaching in SoyBased Fish Feeds. This year we had an


Photo: United Soybean Board

important announcement to make. Dr. Salze announced the very recent news of following years of research, the Association of American Feed Control Officials (AAFCO) has approved a new definition for the use of taurine in fish feeds. Taurine is an important amino acid for fish, but it was not approved for use in aquafeeds produced in the United States – until now. “This had left feed formulators in the United States with little choice but to provide taurine through an animal-based ingredient, typically fishmeal” said Guillaume Salze, the research associate at Auburn University’s School of Fisheries Aquaculture and Aquatic Sciences who spearheaded the project. “Being allowed to add crystal taurine to fish feeds means that feeds can be formulated with less fishmeal and more plant proteins (such as soy), thereby reducing costs and improving sustainability at the same time” Salze added. The new definition includes all life stages in all fish species. Multiple groups collabo-

rated on this project, including universities, federal agencies, and non-profit organizations, and funding sources. This was a major research effort supported by SAA members, the United Soybean Board, National Oceanic and Atmospheric Administration (NOAA) and United States Department of Agriculture (USDA) partners that began a few years ago. Advanced Soy Proteins are important resources for aquafeeds and SAA Members Prairie AquaTech (ME-PRO®), the Ohio Soybean Council (Enzomeal) and Midwest Ag Enterprises (NutriVance) discussed their advanced soy protein products and work that was underway in this sector. Pentair shared an update on their work and an innovative, virtual tour of their facility at PAES W.A.T.E.R. and Insta-Pro shared information on their aquafeed production equipment and on the work they do with extrusion equipment. The aquaculture industry relies on great partners in feed ingredient innovation and in equipment innovation.

“Being allowed to add crystal taurine to fish feeds means that feeds can be formulated with less fishmeal and more plant proteins (such as soy), thereby reducing costs and improving sustainability at

the same time”

60 Aquaculture in the United States also has critical partners in the U.S. Government Agencies and Departments. SAA was honored to have Dr. Gene Kim from USDA National Institute of Food and Agriculture provide an update on their work with aquaculture and insights on a new effort underway on an Innovation Challenge to explore new approaches to increasing the consumption of U.S. Aquaculture production domestically. Dr. Caird Rexroad III provided an update on USDA Agricultural Research Service Aquaculture priorities and research programs. Dr. Michael Rubino, Director of the Office of Aquaculture at NOAA's Fisheries Service, shared with the group an update on work in the offshore efforts including new discussions for the Western Pacific and NOAA programs working at creating models utilized in exploring siting opportunities in the offshore environment.

“Offshore Aquaculture remains an important opportunity for U.S. Aquaculture to grow and to supply high quality marine products for our consumers” Offshore Aquaculture remains an important opportunity for U.S. Aquaculture to grow and to supply high quality marine products for our consumers. Don Kent from Hubbs Sea-World Research Institute provided a presentation updating the audience on their work in establishing a farm off the Coast of San Diego, California. Their work continues in the permitting process. Significant challenges remain in the area of permitting for aquaculture and are a key area for all of us in the U.S. Aquaculture

Community to focus on collaboration for solutions.

Photo: NOAA National Ocean Service

SAA shared updates on the sustainability of U.S. Soybean Production and the U.S. Soybean Sustainability Assurance Protocol. We also updated the audience on the work of the Coalition for U.S. Seafood Production (CUSP) and the efforts underway in this coalition to educate and advocate for the growth of U.S. Aquaculture. The final speaker was a new participant to Aquaculture America and proved the importance of collaboration. Harlon Pearce with the Gulf Seafood Institute (GSI) shared with the work GSI has underway to support aquaculture in the Gulf of Mexico. GSI is approaching this work in an innovative and successful way through building important partnerships and sharing important outreach on the value aquaculture can bring to the region. It takes a team for us to achieve greater success for U.S. aquaculture and this meeting highlighted the importance of all the work underway to support our U.S. aquaculture community as we grow and thrive. AFΩ

More information

Bridget Owen, Executive Director, Soy Aquaculture Alliance E: