Aquafeed vol 11 issue 3 2019

Vol 11 Issue 3 2019

AQUAFEED Advances in processing & formulation

PLANT-BASED INGREDIENTS Corn protein and yeast Nannochloropsis algae meal Yeast from wood Microalgae-based omega-3s Functional feeds in sea bream Soy Aquaculture Alliance new column Heat-stable probiotics Linseed as a source of fatty acids Aquafeed Innovation Awards Published by: Aquafeed.com LLC. Kailua, Hawaii 96734, USA www.aquafeed.com info@aquafeed.com

AQUAFEED

VOL 11 ISSUE 3 2019

Contents

PLANT-BASED INGREDIENTS Replacing fishmeal is closer due to new plant-based ingredients, such as the combination of corn protein and yeast.

INTERVIEW – Kevin Fitzsimmons

ALGAE-BASED OMEGA-3S

HEAT-STABLE PROBIOTICS

Kevin Fitzsimmons, Chair and judge of the F3 Challenge, talks about novel ingredients and sustainability.

Microalgae-based omega-3s are now produced at scale and offer a sustainable resource to improve omega-3 levels in farmed salmon.

How the inclusion of a new microorganism extract in extruded feed resists the mechanical and thermal exposure.

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AQUAFEED

VOL 11 ISSUE 3 2019

Contents

17

6

Interview

9

News Review

Product Focus

17 A botanical in-feed solution for growing healthier shrimp

26 Use of a high-quality corn protein product as an alternative protein source in aquaculture feeds

18

Aquafeed Innovation Awards

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Functional feed improves sea bream pigmentation

Plant-Based Ingredients

31 P ositive results from Thai Union Feedmill’s shrimp feeding trial with a sustainable Nannochloropsis algae meal

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34 A ssessment of the nutritional performance and physicochemical properties of novel yeast protein ingredient for aquaculture feed

36 M eeting the new US FDA omega-3 qualified health claim for salmon: algae-based omega-3s as a sustainable feed source

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rchaea extract: a new heat stable micro-ingredient A to improve disease resistance in aquaculture

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Ingredient Profile

Linseed, an ignored plant source of essential fatty acids

Columns

22 Greg Lutz – Trends & developments

34

46

24 Soy Aquaculture Alliance

38 Peter Hutchinson – Ask the expert

40 Albert Tacon – Recent publications

48

Calendar of Events

To read previous issues in digital format or to order print copies, visit: http://www.aquafeed.com/ publications/aquafeed-magazine/

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Kevin Fitzsimmons, Ph.D., Professor and Director of International Programs at The University of Arizona, is Chair and judge of the F3 Challenge, a global competition that aims to accelerate the replacement of fishmeal and fish oil in aquaculture feeds with innovative alternatives. He served as the President of the U.S. and World Aquaculture Societies and was the second Chairman of the global charity, Aquaculture Without Frontiers.

He received a Fulbright Fellowship in 2004 to teach and conduct research in Thailand and completed a second Fulbright Fellowship in 2017 working in Vietnam and Myanmar, where he had a 3-year USAID project supporting aquaculture. He is currently working as Team Leader for the EU supported Myanmar Sustainable Aquaculture Project.

INTERVIEW AQUAFEED: Would you tell us about your work at The University of Arizona, and how it has led you to become an advocate for fishmeal and oil alternatives?

KF: Yes, my work for the last 40 years has always focused on how we can make aquaculture more sustainable in all aspects; environmental, financial, and social. Some of the work has been with integrated farming systems in the desert, some with seaweeds, and a lot of focus on tilapia as a model crop. But one of the overarching and most global of all aquaculture sustainability issues has been the heavy reliance on fishmeal and fish oil.

with Kevin Fitzsimmons AQUAFEED: I think everyone is aware that forage fish are not going to be able to supply the protein and lipid needs of aquaculture at its current rate of growth; what is the status of fishmeal supply now and how long do you see it being able to keep up with aquafeed demand?

KF: The limited supplies of wild caught forage fishes are getting tighter all the time. Declining populations of sea birds, large predatory fishes, and several marine mammals are attributed to overfishing of forage fishes and fisheries managers are beginning to restrict harvests to below the economic maximum sustainable yield in favor of environmental sustainable yields. As aquaculture

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grows and available forage fisheries contract, we are placing a lot of hope on processing waste as a source of fish meal and fish oil. However, this is only a temporary stop gap. Living in Asia, I observe that there is no such thing as processing “waste”. Skins are used as snack foods, scraps of flesh are converted to fish balls, fish paste, and fish sauce and anything left on the frame or skeleton is converted to high phosphorus (bone), low protein, low quality fish meal. It is only a matter of time until Western processing follows the same path of higher edible recovery and economic efficiency.

AQUAFEED: So what exactly is the F3 (Future of Fish Feed) Challenge?

KF: The F3 Challenge is part of a larger F3 (Future of Fish Feed) effort to accelerate the adoption of alternative nutrients in aquafeeds. The Challenges (first, alternative replacements to fish meal and now, fish oil) are designed to educate the industry and the consumers to the fact that proper nutrients are necessary for good aquatic animal growth and health, not “proper” ingredients. Just as in human nutrition, it really does not matter where we get the amino acids, carbohydrates, lipids, and vitamins.

Just so long as we get all the macro and micronutrients in reasonable proportions. We believe that a large cash prize which generates interest and attention, has generated more buzz than the hundreds of excellent research projects which demonstrated this with literally hundreds of ingredients.

scaling-up what in the past were methods for making beer, sauerkraut, and kimchi. The insect meals are a bit more novel, but essentially using common insects to thrive on food wastes and then converting the larvae to meal and oil. One of the great side-effects is keeping large amounts of food processing by-products and spoiled foods out of landfills.

AQUAFEED: In our September 2017 issue of Aquafeed, we reported on the outcome of the first F3 Challenge.

The winner of that competition was Evergreen Feed Industry. How far has this company and the other competitors come since then, in terms of alternative feed inclusion?

KF: Our first winner, Evergreen, has continued to be a star in the industry. The farmers in China who used their F3 formulation have continued to request the feed, reporting that growth is better, waste in the pond is less and the price was the same as a fishmeal containing diet. The farmers even have petitioned us to use the F3 logo on their fish for marketing purposes. Evergreen has recently completed what is probably the largest single tilapia farm in the world in Egypt and signed a new contract to build a second farm in Saudi Arabia. Both are planning to use F3 feeds. Feed companies and farms in the US, Myanmar, and Japan are also continuing to use the contest diets.

AQUAFEED: Can you tell us about some of the novel proteins being developed?

AQUAFEED: Why are these companies embracing alternative protein sources? Is it simply because they see fishmeal as a finite resource or is it also end-user driven?

KF: First, I want to make clear that those of us involved with the F3 are not trying to promote any particular products or even classes of products. We believe that there will be demand for virtually all of them and that the well-informed free market will sort out the winners and other competitors. So, in answer to your question, in the first competition we had several teams that incorporated single cell proteins in their formulations, some who used insect meals, some included algae, and a couple used a mix of terrestrial animal by-products and plant proteins. The algae and single cell protein companies are using bioreactor and fermentation technologies that are essentially biotechnologies

KF: The most simple answer is that the companies recognize that fishmeal prices will continue to rise as supplies tighten and that the alternative protein sources will drop in cost as production scale increases. I think most feed companies, and increasingly the farmers, also recognize that the end users are very aware of the forage fish controversies and that more sustainable protein sources will gain consumer acceptance. Feed companies, who for decades have told their farmer customers that the best feeds had plenty of fish meal and fish oil have been reducing levels for a long time and now are explaining to customers that the nutrients are critical not the ingredients. The F3 folks are also promoting the

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Methane and CO2 consuming microbes, CO2 consuming and oxygen producing algae, insects devouring food wastes, are brilliant ways to address environmental problems while producing valuable feed ingredients. concept of F3 seafood as the equivalent to grass fed beef or free-range chicken to develop more demand from the consumer.

AQUAFEED: Does F3 facilitate bringing together suppliers and users of alternative proteins, besides through the competition?

KF: Yes, F3 has hosted two events in the San Francisco area bringing together major feed producers, large and small farmers, investors, NGO’s, chefs, and press to hear presentations from the alternative protein and oil entrepreneurs. These meetings have been universally acclaimed by the participants as productive and enlightening. We believe that the meetings have accelerated the process of connecting the innovators and their products with the most critical buyers. Presence of venture capital investors has further sped deal making and roll-out of larger volumes and additional products.

AQUAFEED: How are the challenges of scalability and competitive pricing in producing alternative ingredients being addressed?

KF: Several of more conventional alternatives (soy products, animal-byproducts) have long been competitive and in large supply. The more novel alternatives are rapidly increasing volumes and dropping prices. Some of the fermentation and algae products are already competitive and selling tens of thousands of tons. But others including insect meals and bacterial based proteins are still limited in volume and/or not

price competitive yet. However, the scale of investment in these companies has been impressive to say the least.

Hundreds of millions of dollars have been invested and the scale of automation means that labor costs are not likely to be a significant fraction of costs going forward.

This is in stark contrast to the overall forage fish industry where labor issues have been a constant issue and likely to increase significantly as the industry tries to improve their performance.

AQUAFEED: The latest F3 Competition is for the replacement of fish oil. This seems to be a tougher challenge. How do you see this developing?

KF: The fish oil challenge has certainly been a more difficult technology to master and judge. We thought that there would be an array of contestants representing several ingredient sources of lipid nutrients. But frankly only the algal-based companies have turned out to be competitive. That being said, we are impressed how the contestants have been ramping up sales since the start date. The new salmon products on the market touting the inclusion of omega-3 from algae leads us to believe that this will be a mainstream product in the near future.

AQUAFEED: “Sustainability” has been a big part of your career over the past 40 years; what is your definition – and goal – of sustainable fish production?

KF: My definition of sustainable aquaculture pays close attention to financial, environmental, and social aspects.

Without all three being addressed there is sure to be a breakdown in time. My goal is to see a plethora of alternative ingredients available to provide the nutrients required for healthy production of seafood containing all the healthy characteristics that we have come to expect.

I firmly believe that we will need multiple ingredients to match the mix of nutrients that come so conveniently in fish meal and fish oil. The great thing about many of these new ingredients is that they address many of our environmental problems. Methane and CO2 consuming microbes, CO2 consuming and oxygen producing algae, insects devouring food wastes, are brilliant ways to address environmental problems while producing valuable feed ingredients. Contrast these to subsidized fishing fleets, using enormous amounts of fuel to hunt and gather a limited public resource and you can see why I am so excited about the future of fish feeds.

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NEWS REVIEW Highlights of recent news from Aquafeed.com Sign up at Aquafeed.com for our free weekly newsletter for up-to-the-minute industry news

Skretting commits to insect meal volume at Protix grand opening His Majesty King WillemAlexander of the Netherlands officially opened the world's largest insect farm at Protix in Bergen op Zoom, the Netherlands on June 11, 2019.

The occasion was marked by a deal with Skretting that could see up to 5.5 million salmon servings per year brought to market with insect meal incorporated into the feed.

“We are very proud to work with Protix to unlock this new protein source into the aquaculture

feed industry to help bridge the future protein gap,” said Dr. Jenna Bowyer, Skretting Project Procurement Manager.

The guests experienced a tour of the new Protix facility, housing insects that convert vegetable residual flows into sustainable protein, contributing to a futureproof, circular bioeconomy.

The cultivation process takes place in a controlled environment and is highly automated with sensor and data systems, robots and climate control. Bühler Insect Technology Solutions has played a major part in this project.

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Cargill and Innovafeed partnership Cargill and InnovaFeed have entered into a strategic partnership to bring sustainable and innovative feed options to the animal nutrition industry. Through the partnership, Cargill and InnovaFeed will

collaborate to jointly market fish feed which includes insect protein, enabling both partners to support the growth of sustainable aquaculture. “We’ve seen a strong acceptance of our products

by consumers looking for natural and sustainable products,” said Maye Walraven, head of business development for InnovaFeed. Insectfed trout has been commercialized in France since June of last year.

Triott Group is officially consolidated In response to increasing demand from a fastevolving global feed and food production industry, the Dutch-headquartered Triott Group has been consolidated with the launch of www.triottgroup.com. The Triott Group comprises eight complementary companies serving the global feed and food industry – including milling engineers Ottevanger and the process control and automation specialist Inteqnion. By working together even more closely and seamlessly under the Triott umbrella, the group plans to better meet both the current and future needs of the food and feed industry, but also emerging

new industries in the burgeoning bio-economy.

Triott is a 110-year-old, family-run (privately held) business that is comprised of: Ottevanger (milling engineers); Inteqnion (process improvers); PTN (Pelleting Technology Netherlands); PCE (Pelleting Consumables Europe); ALMEX (extrusion technologies); Wynveen (milling and mixing specialist); IVS (dosing technology specialists) and TSC (Top Silo Constructions). Together they have the capability to provide all (or any) elements of a turnkey milling solution, from storage, handling, dosing, milling & mixing solution to pelleting extrusion and complete plant management.

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Geelen Counterflow develops a new generation of counterflow coolers Geelen Counterflow has developed a new generation of counterflow coolers that improves product quality, operational cost and efficiency. Counterflow Cooler Plus™ allows an improved product distribution, easy inspection and optional evaporation control.

The new counterflow cooler uses Evaporation Control™ to select the ideal air volume and cooling time to maximize or minimize evaporation in the cooler when feed producers are keen to avoid insufficient or excessive product moisture after cooling due to fluctuations in raw materials or ambient circumstances. Multiple sensors for air and product temperature help to control the process. Even a 0,5% improvement in the average moisture content of product can have a very significant effect on the processor’s bottom line.

The company said that the new Topdrive product distributor has its gear motor mounted outside of the cooler, lowering the risk of pollution and overheating. Inspection of all moving parts of the product distributor is possible from the

outside. The short vertical drive shaft in solid stainless steel minimizes the need for horizontal surfaces inside of the cooler, keeping the process as clean as possible.

The drive of the product distributor is controlled by a frequency converter to allow the corners of the cooler to be reached, leading to a very even product distribution.

The new stainless steel +10% Hood design has a 10% bigger air exhaust opening which reduces airspeed to avoid product being sucked into the air exhaust system.

The round flange avoids the need for a transition piece from rectangular to round and the diagonal exhaust orientation keeps dust from accumulating on any horizontal surface.

The company also said that the big access doors in the side walls

and collecting hopper enable good access for inspection and cleaning.

The side walls in stainless steel contain multiple product sensors to allow automatic selection of bed depth, depending on product and ambient conditions.

The discharger of the cooler is a versatile swivel valve discharger, available in mild steel or stainless steel, that can handle any size, shape and condition of food, pet food, aquafeed and feed. Its hydraulic drive with angle encoder enables your control system to automatically set product specific open and close positions.

Le Gouessant Aquaculture and Innovafeed succeed in 100% fishmeal substitution with insect in rainbow trout feed Following the authorization in July 2017 to integrate insect meal in the diet of fish and shellfish, Le Gouessant, one of the major player in aquaculture nutrition in France and InnovaFeed, a French biotech company that produces a new source of protein from insect rearing for animal feed, decided to launch a

joint trial in experimental conditions on rainbow trout (Oncorhynchus mykiss). While there is an increasing number of trials using insect meal, this was the first time that 100% of the fishmeal has been substituted by high quality insect protein derived from black soldier fly larvae (Hermetia illucens).

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Bühler’s new extruder With the growing number of contamination incidents in pet food, food safety measures are being increased and continuous improvement of processes and machines is expected from pet food and aquafeed manufacturers.

That was one of the reasons why Bühler developed a new singlescrew extruder, which was launched at the Victam trade show.

High sanitation standards, excellent workmanship and an outstanding price-performance ratio make the new extruder a great solution in the field of pet food and aquafeed. But most importantly, the extruder can help minimizing the risk of non-safe pet food and aquafeed and with it the risk of costly product recalls.

The new single-screw extruder completely fulfills process requirements of the market, as capacity, screw speed, torque and pressure. Thanks to its modular design, this machine is extremely flexible in application and can be adapted to the specific needs.

What’s more, it allows for flexibility and customization from process to process and yields consistent, high-quality finished product.

The innovative design of the housing according to the “shell principle” offers special advantages: the housing excels through its mechanical stability, resistance to wear and highly efficient tempering, with spiral or conical liners.

Maintenance costs for this innovation are minimized: the

interior casing is extremely resistant to wear. The company offers a barrel wear measurement service to detect wear and tear in good time.

In this way, parts can be replaced at an early stage to ensure stable product quality.

In order to guarantee an endproduct of consistent quality, the single-screw extruder is equipped with an intelligent recipe memory.

This offers two significant benefits: susceptibility to process errors decreases, and production can be quickly adapted to other recipes.

Predefined start-up and shutdown sequences optimize the starting and shutting down of the machine.

Thus, a minimum of raw material is wasted, and the production process is considerably accelerated.

The single-screw extruder has been designed in concordance with the latest megatrends in the pet food industry; Fresh meat and Food Safety. Hence why the preconditioner is placed next –

More information: E: extrusion@buhlergroup.com

Aquafeed: Advances in Processing & Formulation Vol 11 Issue 3 2019

rather than traditionally above – to the extruder making it extremely easy to clean without spoiling the extruder with the dirty rinsing water. It also has a screw remover and handling device in order to guarantee a full cleaning accessibility to the extruder processing section, reducing the cleaning time to less than one hour.

The extruder is equipped with a user-friendly and intuitive touchscreen control system.

Safety circuits prevent damage due to improper use. Emergencies trigger self-stopping of the extruder fully automatically.

For quality and food safety reasons, the control system contains important functions such as logging and trending of actual and historical data. It also features some of the required functionalities of FDA21CFR part 11 such as user management, event logging of operator interventions, and logging of quality-related data.

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Tietjen and Neuman & Esser agreement for new solutions in fine grinding Neuman & Esser Process Technology GmbH (NEA) and Tietjen Verfahrenstechnik GmbH signed an agreement that will take over the worldwide distribution of the NEA impact classifier mill under the name "TICM" (Tietjen Impact-Classifier Mill).

The agreement refers to the aquafeed and pet food applications.

Neuman & Esser brings its proven impact classifier mill (ICM) into the partnership. The most common applications of the ICM are chemicals, pigments, powder coatings, minerals, coal, food, confectionery, cosmetics and pharmaceuticals. As a grinding specialist, Tietjen uses its mills and plants in a wide variety of areas, including the production of fish and pet food. With the "TICM", Tietjen has broadened its product portfolio and offers solutions for the high demands of the pet and fish feed industry.

The TICM fine grinding concept is specially designed for the requirements of fish feed and pet food production and thus masters the high demands in terms of fineness. The TICM combines efficient grinding with precise classification in one machine. Therefore, the companies said that it is possible to produce particularly homogeneous end products with high fineness ranges from 100 μm to 400 μm and a precise separation cut of up to 99% at high throughput rates. Another special feature is the optimized grinding chamber, designed for gentle processing with a low rise in temperature in order to preserve vitamins and microcomponents in the ground material. In addition, the single-stage grinding enables a compact system design and can thus be excellently integrated into existing systems as a dedicated fine grinding line.

PEOPLE IN THE NEWS Dhanin Chearavanont Thai family-owned conglomerate, Charoen Pokphand Group, announced the resignation of Dhanin Chearavanont, the 80-year old billionaire, who has headed Charoen Pokphand Foods since 1969. He tendered his resignation due to "other businesses refraining him from performing his duty."

Olivier Rigaud Corbion’s Supervisory Board has nominated Olivier Rigaud to succeed Tjerk de Ruiter as CEO of Corbion. Upon his appointment, Olivier Rigaud will form the Board of Management of Corbion together with Eddy van Rhede van der Kloot who will continue as CFO.

Constance Cullman The American Feed Industry Association’s Board of Directors voted on a new president and chief executive officer to head the association upon the retirement of Joel G. Newman at the end of this year. Constance Cullman, current president of the Farm Foundation, will serve as AFIA’s future president and CEO and president of the Institute for Feed Education and Research.

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BioMar’s new solutions BioMar has developed a new Symbio recipe to solve the challenges associated with dietrelated cataracts in lumpfish.

Past studies of cataracts in lumpfish have identified nutritional imbalances with high levels of specific amino acids in certain tissues. The company conducted a study on three alternative fish

Aquafeed: Advances in Processing & Formulation Vol 11 Issue 3 2019

feed diets and no traces of cataracts were found in any of these diets. The common feature of these recipes was a reduction in the content of both protein and fat when compared with a control feed.

BioMar has also completed the testing of insect meal in feeds for aquaculture and believe it shows promise as an alternative protein source. The company has been investigating insect meal since 2015 at its research and development centers, and from 2017 the company has been undertaking tests with customers who have been feeding their fish on diets containing insect meal. These fish have already made their way to supermarkets in Europe among retailers eager to implement future-oriented food solutions that focus on natural foods. Moreover, the company and Minze Long Yang Xia, the largest trout farmer in China, signed a cooperation agreement to supply high performance fish feed with almost half the environmental footprint compared to local Chinese feed. Over the last two years the BioMar BioFarm teams of China and Denmark have been collaborating closely with Long Yang Xia on technical onsite trials that consider the daily conditions of the water and the fish. The ideal recipe solution that was discovered considered the parameters of growth performance and fish welfare while limiting the discharge of nitrogen and phosphorous into the local ecosystem.

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Alltech’s energy concept to ensure consistent performance The company offers a feed formulation service to boost the consistent performance of feed and support farmers and producers to take control of their profitability.

Most aquafeed companies formulate their feed based on fixed digestible protein and energy levels, which can in part impact the variation in quality of raw materials when fish feeds are formulated in a flexible way.

Recent research from the Alltech Coppens Aqua Centre and associated universities have shown that not all forms of the digestible energy are available to use for growth with the same efficiency. By formulating feeds based on net energy, Alltech Coppens not only looks at digestible energy, but also takes into account the differences in efficiency of the different forms of digestible energy. This ensures consistent performance between batches of feed, Alltech stated.

Alltech Coppens has already started to implement this net energy approach in feed evaluation for some trout feeds where the utilization efficiency of the different macronutrients is accounted for as well.

PEOPLE IN THE NEWS Erik Visser The Board of Directors of Hamlet Protein, a world leader in soy-based specialty proteins for young animals, has appointed Erik Visser as Chief Executive Officer. Erik has extensive experience in the animal feed industry through senior management roles at Provimi, Nutriad and Adisseo.

Shauna McNeill Aker BioMarine named a new Executive Vice President with responsibility for the company’s ongoing programs to research, develop and commercialize new krill derived products and applications.

AQUACULTURE FEED FORUM Feeding for Profit

September 10, 2019 l Rennes, France.

A free half-day international conference (in English).

Part of Aquaculture SPACE, a new focus at SPACE, the world's largest livestock event.

An Aquafeed.com conference at SPACE 2019, sponsored by:

Details: http://uk.space.fr/EN/Aquaculture.aspx

Aquafeed: Advances in Processing & Formulation Vol 11 Issue 3 2019

PRODUCT FOCUS

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A botanical in-feed solution for growing healthier shrimp The complete replacement of chemical treatments with botanical-based solutions that effectively reduce disease, infection and mortality in shrimp, while at the same time safeguarding natural habitats is the mission of Israeli-based STK Aqua.

The company develops and commercializes solutions that help to increase survival rate and enable safer, healthier and more abundant production from transportation and stress reduction to staving off the onslaught of parasites and bacterial diseases.

For the treatment and prevention of bacterial diseases in shrimp, the company has developed a botanical-based solution that is bio-filter friendly and suitable for all growing environments, including land-based ponds and raceway systems. Eshed® is a ready-touse, non-toxic, eco-friendly formulation that can be applied in hatcheries and as a pre-mix for feed in subsequent growing stages to increase survival rate of shrimp throughout all growing stages.

When a shrimp is attacked by microbes it is unable to protect itself due to its rudimentary immune-system, in addition, a "bio-shield” is created by the microbial colony, which protects it from attacks by external molecules such as: antibiotics, formaldehyde, etc.

Yaki Keinan, STK Aqua General Manager, explains that Eshed® helps the shrimp to prevent the bio-film creation, leaving the microbes exposed to antibacterial products, such as organic acids. It has been shown that by using Eshed® synergistically with products such as organic acid, growers in all stages, has acceded up to 20-30% more in their yield. Keinan states that it saves time and money by increasing the survival rate and reducing the use of antibiotics.

Figure 1. Mode of action.

Table 1. Feeding trial results.

More information: Yaki Keinan E: Yaki@stk-ag.com

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Sustainable solutions win Aquafeed Innovation Awards 2019 A plant-based protein and an electric drier ran away with this year’s Aquafeed Innovation Awards presented during Victam International in Cologne, Germany, following Aquafeed Horizons technical conference.

The Aquafeed Technology award was given to Sander Geelen, Managing Director Geelen Counterflow and the Aquafeed Ingredients award to Mark Luecke, CEO, Prairie Aquatech by Lucía Barreiro, editor Aquafeed.com and sales manager, Michael Van den Dries.

Today’s aquafeed producers demand sustainable ingredients and methods of production, and their suppliers are not just keeping up, but often helping drive the trend. The products of two such companies impressed the Aquafeed.com panel of independent judges, who selected them from a strong line up of contenders as winners of this year’s awards.

A microbially enhanced plant-based protein Fishmeal is an ideal protein for fish feed, but finite fishery resources and a booming industry means alternatives must be developed. There has been a rush to fill the protein gap from insects, algae and single cell protein producers among others, but most of these are struggling to scale up to the production levels required by feed manufacturers. The approach taken by Prairie Aquatech is to develop a microbially enhanced plant-based protein ingredient for aquafeed, ME-PRO; it is already being incorporated into aquafeed diets in the United States and has been trialed with multiple species worldwide.

ME-PRO is produced at Prairie AquaTech’s AgTech Center in Brookings, South Dakota, a facility that focuses on research, testing and trials of innovative plant-based technologies. The center comprises fermentation units for protein production, a recirculating Aquaculture System (RAS) for multi-species testing, a feed plant for testing feed formulation and production, and a laboratory. It has recently completed construction of its first commercial-scale production facility in Volga, South Dakota. At full production capacity, the facility will produce 30,000 metric tons of ME-PRO from nonGMO soybeans per year.

Non-GMO soybean meal is treated with a natural, non-GMO, food-grade microbe (Aureobasidium pullulans), an all-natural process created at South Dakota State University, to create a high value (70% as fed, 100% digestible) protein to extend,

Left to right: Mark Luecke, CEO, and Sue Lancaster, VP of development, Prairie Aquatech.

or even replace fish meal. However, ME-PRO provides another benefit to aquaculture in that it is proven to significantly lower phosphorus discharge while providing high P availability to the animal. In one case, a pond-based trout hatchery was able to reduce their discharge by 69%, allowing the hatchery to survive and thrive, sustaining high-value customers.

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Cost and environment saving dryer The dryer is by far the single biggest energy consumer of any extrusion line, typically consuming 50-60 percent of the line’s energy. The judges were most impressed with the cost savings and environmental advantages afforded by this dryer, noting also that there are many locations around the world where gas and electricity are alternately unreliable, with huge price spikes.

The Electric Dryer uses heat-exchangers to recover up to 65 percent of all energy and water from the exhaust of the dryer and re-uses that heat after boosting the temperature of the recovered heat by industrial, high temperature heat pumps. This way the dryer can run 100 percent on electricity instead of natural gas, while ‘scope 1 CO2 emissions’ can be eliminated. By also installing gas burners, producers can switch back to natural gas in periods when electricity prices are temporarily too high, or in case technical redundancy is required. By upgrading to the new ZAS MkIV air unit, producers can choose to install electric heating capacity in small modular steps. As soon as prices for natural gas or CO2 emissions go up/and or electricity prices come down, additional heat pump capacity can be installed to optimize operational costs. By installing Counterflow Remote Diagnostics, Geelen can remotely monitor all mass and energy flows in the dryer 24/7, enabling it to provide the dryer owner or operator with detailed recommendations on how to optimize drying effectiveness and efficiency.

Left to right: Michael van den Dries, sales manager, Aquafeed.com, and Sander Geelen, managing director, Geelen Counterflow.

Reducing energy consumption by up to 65 percent and eliminating CO2 emissions, has a potentially major impact on the operation cost and sustainability of the plant. It can also have a very significant impact on the sustainability of the industry overall, with a reduction in CO2 emissions per dryer of thousands of tons of CO2.

The condensation of humidity in the exhaust air that is triggered in the heat exchanger leads to a significant reduction in odor emissions which will either reduce the risk of odor issues or reduce the cost of odor treatment – an advantage that impressed the judges.

Aquaculture is evolving rapidly. Its continued success rests to a great extent on the ingenuity and dedication of the feed industry in meeting the challenging and exacting needs of this rapidly growing and diverse sector.

Aquafeed.com has presented these prestigious awards since 2012, to recognize and honor the achievements and contribution of the allied industries to the advancement of aquafeed development.

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Functional feed improves sea bream pigmentation Roberto Anedda and Riccardo Melis, Porto Conte Ricerche

Gilthead sea bream (Sparus aurata) are one of the most widely consumed European fish species and consequently an important resource for aquaculture and coastal fisheries in Mediterranean countries.

The external appearance of sea bream, including skin coloration, is undoubtedly the first marketable index driving buyer's and consumer's preference.

While wild sea bream exhibit a more vivid pigmentation, in particular a characteristic yellow band along the eye inter-orbital line and a yellow or orange mark near the operculum, farmed specimens generally present either a pale grayish or a darker skin. The lack of the typical "wild" coloration is perceived as an objectively negative characteristic by consumers and, consequently, negatively influences their acceptance.

Cultured sea bream are generally deprived of natural sources of dyes (pigments) when fed standard

commercial diets. Therefore, it is of considerable interest developing and optimizing specific functional diets containing either natural pigments or alternative additives that would finally improve skin coloration.

Sea bream trials Aller Aqua, with Italian Palma d’Oro (Gloria Maris group) and Sardinian Porto Conte Ricerche, has recently concluded a study to test the effect of Aller Aqua Blue Organic Ex feed on sea bream pigmentation. Two groups of sea bream (190 – 260 g) were cultured in different rearing conditions, RAS system at the Porto Conte Ricerche and offshore cage at the Palma d’Oro farm.

The team analyzed the pigmentation of five different anatomical regions: IOR, inter-orbital; SOP, sub-opercolar; D1, dorsal apical; D2, dorsal medial

Aquafeed: Advances in Processing & Formulation Vol 11 Issue 3 2019

21

and D2, dorsal caudal (Fig.1). Color measurements were performed by using a hand-held Minolta CR-400 Chroma Meter (Minolta Camera Co., Japan), previously calibrated with a standard white plate (D65,Y = 93.9, x = 0.3155, y = 0.3319) and according to the CIELab system (CIE, 1976). Lightness (L*), redness (a*) and yellowness (b*) were determined as color parameters. In addition, Chroma (C*) = [(a*)2+(b*)2]1/2 was calculated as a representative index of color saturation.

Skin color improved Results showed a significant increase of Chroma C* (P<0.01) in all the examined regions from both sea bream groups regardless the rearing system (Fig. 2).

L* changes in the interorbital line (IOR) appear slightly affected (P<0.05) depending on rearing conditions.

Apical (D1) and median (D2) dorsal region are slightly affected (P<0.05) by diet in both conditions, while no difference was detected (P> 0.05) in the caudal dorsal region (D3) and sub-opercula regions (SOP).

Aller Aqua Blue Organic Ex functional diet improved skin color in sea bream raised in different farming conditions, both in offshore sea cages and in a recirculating aquaculture indoor system, over two months feeding trial. Researchers said that this effect can be mainly ascribed to the feed since no correlations were found to the rearing system and genetic breed of the two fish populations.

The optimization of a novel application of image analysis is under way at Porto Conte Ricerche which

Figure 2. Skin color changes on the five target regions in both sea bream cultured groups in response of the Aller Aqua Blue Organic Omega Ex diet.

aims at identifying quality-related features (size, morphology, color) from pictures of fish that could be potentially taken in-line before packaging.

More information: Roberto Anedda Researcher Porto Conte Ricerche Srl E: anedda@portocontericerche.it

Riccardo Melis Researcher Porto Conte Ricerche Srl Figure 1. Skin color assessment of sea bream by examining five target anatomical regions.

Aquafeed: Advances in Processing & Formulation Vol 11 Issue 3 2019

COLUMN

Trends and developments C. Greg Lutz, Ph.D. Change, alteration, revolution, upheaval, transformation, restructuring Sounds like a recipe for political and social strife, but… these are all synonyms for the term “innovation.”

Aquaculture, and the aquafeed industry in particular, has been a hotbed of innovation in recent years. In fact, this column is tasked with identifying and discussing innovations in aquaculture from time to time. But it is worth examining the concept of innovation so that each of us can consider how it relates to our role in this industry.

Most people would currently define an innovation as a new method, idea or product. Many would probably confuse or equate innovations with inventions, but this is not necessarily the case in most instances. An invention can be seen as a discovery, a new work or a novel idea. In contrast, an innovation is more like a new or modified device, method or process. Like… applying an invention under real-world conditions.

The implication is that an innovation provides a solution or improvement.

Seen as catalysts for economic growth, innovations involve how new things and ideas are put into practice.

The scholarly study of innovation involves entire schools of philosophy, and many theories

and definitions overlap each other to some degree. When considering the broad range of aquaculture species, production methods and policy frameworks, several basic innovation concepts seem most applicable for our industry.

Disruptive innovations are perhaps the most familiar type of innovation, where businesses and researchers continuously seek to discard the old and embrace the new in an effort to improve competitiveness.

In this scenario producers of similar products are competing in the same marketplace for a finite number of customers. Consumers in many societies tend to tire of traditional products and perceive newer products as superior, even when the differences are imperceptible (we’ve all read the phrase “new and improved!” on the labels of countless products).

This type of innovation involves not only products but also processes and business models.

Many examples from aquaculture production come to mind.

Innovative filtration configurations for RAS have made the production concept much more feasible compared to what was available 15 years ago. The development of submersible cages for offshore

Aquafeed: Advances in Processing & Formulation Vol 11 Issue 3 2019

Dr. Greg Lutz is a Professor with Louisiana State University Agricultural Center. He is also an author and consultant, and serves as the Editor in Chief of Aquaculture Magazine.

E: lutzaqua@att.net

production has reduced operating risks and allowed for more management options. Even something as simple as a crawfish trap that stands upright on its own serves as an example of innovation translating into economic expansion (seriously – the latest figures indicate it’s a $211 million industry in Louisiana).

Foundational innovation is characterized by slower, yet equally innovative, changes in business models and competitive environments. Foundational innovation is often the result of an industry’s incorporation of and adaptation to disruptive innovation.

In turn, foundational innovation can serve to promote institutional and policy changes. It’s an incremental process.

Clearly, diffusion of innovations can be far more rapid in the digital age than was the case 30 years ago. But this tends to put

23

an overemphasis on disruptive innovation such that foundational and institutional structures have difficulty responding in a timely manner. This can result in sector-wide dysfunction due to the relative inflexibility of the pervasive bureaucratic mentality infesting many regulatory organizations and agencies.

Governments often dedicate significant funding to foster technical and technological innovation, and this is certainly the case in aquaculture. Examples come to mind from countries across the globe, such as Mexico, the European Union, South Africa, India and China (just to name a few). Those same governments, however, usually fail to invest in policy structures that promote the adoption of such innovations.

Sadly, this is also the case in aquaculture. So â&#x20AC;Ś how would a government normally structure and implement policy-making in a way that fosters innovation and its diffusion? There are several major policy categories.

Mission-oriented policies, as the name suggests, pull together resources and expertise across a broad range of stakeholders to get something done. And itâ&#x20AC;&#x2122;s usually something big, like food security or bolstering foreign exchange earnings. Aquaculture is frequently a component (on paper, at least) of many of these policies, especially in developing countries.

Invention-oriented policies are for innovations focused on technical research and development that usually involve traditional science and technology policy-making agencies. The

emphasis here is more disruptive, leaving foundational innovations to take care of themselves.

System-oriented policies that in the case of aquaculture, are more likely to promote innovations in supply chains, value chains, trade, cooperatives, technology transfer and infrastructure needed for production and/or export.

Having access to information is not necessarily enough to allow for diffusion of technical innovations. Potential users must also have the means and facilities to put the information to use. Research has repeatedly demonstrated that factors such as expertise, financial resources, market demand and R&D facilities are all required for successful innovation. The extent to which these requirements are met by public or private entities will vary, as will philosophies regarding this division of effort. In any event, if they are not met innovation simply does not occur. Orâ&#x20AC;Ś it will occur elsewhere. Innovation policy literature suggests that if governments want to truly foster innovation and the economic growth that comes with it, all of these systemic factors must be addressed.

To do so, however, requires the involvement of many traditional governmental agencies with varied areas of focus (financial policy, science & technology, education, labor and environment, to name a few). And so, instead of technical innovation, policy innovation is rapidly becoming the big sticking point for the growth of aquaculture (and aquafeeds) in the coming decades.

Aquafeed: Advances in Processing & Formulation Vol 11 Issue 3 2019

COLUMN

Combined ultrafiltration and reverse osmosis technologies to increase soy protein use in aquaculture feeds Andy Tauer, Soy Aquaculture Alliance, USA The Soy Aquaculture Alliance starts a new column providing the latest results of their broad research on the utilization of U.S. soybeans in the diets of fish and shrimp.

The Soy Aquaculture Alliance (SAA) conducted research with Brookings, South Dakota biotech company, Prairie AquaTech in 2018 to better understand how adding ultrafiltration (small particle water filtering) and reverse osmosis (membrane technology for water filtration) in recirculating systems (RAS) could impact body weight, mortality and ultimately, cost to production. Two diets, with and without soymeal, and various stocking densities replicated possible commercial production practices.

Purpose and goals The central goal of this project was to quantify the benefits of commercially available ultrafiltration (UF) and reverse osmosis (RO) water treatment technologies in rainbow trout fed either fishmeal or soy-based diets in recirculating aquaculture systems (RAS). The study evaluated whether combined UF and RO technologies increase soy protein use in aquaculture feeds.

Study design Two separate feeding trials were conducted using two matched tank-based RAS. One RAS system was used to rear fish at increasing stocking densities utilizing conventional water filtration technology, while the other system utilized an identical rearing configuration but possessed a UF/RO system to treat rearing water.

In trial #1, juvenile rainbow trout with an average weight of 40 +/- 0.5 gm, were stocked in tanks at increasing densities, 25-125 fish, in replicates of 4 tanks/stocking densities for 49 days on an extruded diet which contained 13% fishmeal and no soybean meal.

In trial #2, juvenile rainbow trout with an average weight of 20 +/- 1.2 gm, were stocked in the same two RAS system tanks and fed a 40% soybean meal diet for 42 days.

Fish characteristics (growth, survival, feed utilization), water quality and biological parameters were periodically measured.

Figure 1. Average body weight of rainbow trout reared under Trial #1 and Trial #2 conditions.

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25

Results Trial #1 UF/RO technology increased production of RAS systems by 25 kg/m3 of biomass as compared to rearing of fish in the same system under control conditions.

Productivity corresponds to 333 fish/m3: £ 333 fish/m3 = $1.50/fish = $500/m3 £ $500/m3 = return of $38,879 or $151,515/year The UF/RO system used 35-40% less makeup water, decreasing operation costs. l

l

l

Trial #2 UF/RO technology showed benefits when applied to trout reared on high soybean meal inclusion diets.

Trout fed a very high (40%) soybean meal diet grew less than fish fed a fishmeal diet under the same control condition.

Benefits of UF/RO technology were most apparent at higher stocking densities.

Trout reared under UF/RO conditions achieved larger

l

l

l

Figure 2. Mortality rate of rainbow trout reared on a diet which contained 13% fishmeal and no soybean meal and under UF/RO conditions.

body weight when compared with the control trout.

Trial #1: Trout stocked at higher densities (75-125 fish/ tank) in control tanks displayed higher mortality rates, especially at 125 fish density. The average biomass mortalities at 125 density is 1,300 gm/tank. Whereas, the UF/RO tanks had zero mortalities until the 125 density.

As a result, the percentage of biomass gain was 10-15% greater for UF/RO trout compared to the control group.

Trial #2: Final stock densities for control trout declined significantly when fish were stocked over 100 fish/ tank and both control and UF/RO tanks had trout

Figure 3. Mortality rate of rainbow trout fed on a 40% soybean meal diet and under UF/RO conditions.

achieve similar final average body weight of about 50 gm. Compared to the data in trial one, control group mortality had about 50% cumulative mortality and UF/ RO fish had 33.4% mortality rate.

The study showed that utilizing ultrafiltration (UF) and reverse osmosis (OS) in farm-raised systems increases body weight and reduces mortalities, even at high stocking densities. Adding these modifications to commercial systems can increase overall yield and decrease operation costs.

Based on data collected from both trials, when paired with UF/RO systems, soybean inclusion diets increased trialed fish body weight and reduced mortalities. In future trials, diets containing 20% soybean meal are hypothesized to yield more consistent average body weights compared to fishmeal.

More information Combined Ultrafiltration and Reverse Osmosis Technologies to Increase Soy Protein Use in Aquaculture Feeds. Prairie AquaTech LLC. Full research brief and technical bulletin on file at www.soyaquaalliance.com.

(Research funded with soybean checkoff dollars).

The Soy Aquaculture Alliance (SAA) works to create new opportunities for soybean farmers within a growing domestic market: aquaculture.

SAA funds programs and research that increases the utilization of U.S. soybeans in the diets of fish and shrimp through affiliations with academic and private researchers and industry leaders.

SAA is governed by a volunteer board representing U.S. soybean farmers and industry members. Membership is open to Qualified State Soybean Boards and public and private entities in the soy, aquaculture and seafood industries. To learn more about SAA-funded research projects or becoming a member, visit www.soyaquaalliance.com.

More information: Andy Tauer, Executive Director, Soy Aquaculture Alliance, USA E: atauer@soyaquaalliance.com

Aquafeed: Advances in Processing & Formulation Vol 11 Issue 3 2019

PRODUCT FOCUS

Use of a high-quality corn protein product as an alternative protein source in aquaculture feeds Dr. Scott Tilton, Flint Hills Resources As consumer demand for dietary protein continues to grow on a global basis, aquaculture producers are finding receptive consumer markets for their products. But as producers look to optimize and grow their operations to meet consumer demand, one of the constraints they face is the availability of cost-effective, highly digestible protein sources for use in their feed operations.

Flint Hills Resources, one of the largest ethanol manufacturers in the United States, recently introduced a high-quality corn protein product to the aquaculture feed market. NexPro is a lower cost alternative to ingredients like fishmeal and protein concentrates commonly used in feed formulations, providing nutritionists in the aquaculture industry with a steady supply of a high quality, cost-effective protein ingredient.

University researchers working with Flint Hills Resources have evaluated NexPro in multiple aquaculture feeding studies. This article summarizes some of their findings in warm water species, including shrimp and tilapia.

Historically, corn protein ingredients including corn gluten meal and dried distillersâ&#x20AC;&#x2122; grains with solubles (DDGS) have been used as protein sources for a variety of aquaculture species, including carp, shrimp, tilapia, salmon, and trout. Also, the use of yeast and yeast products in aquaculture have long been shown to promote growth. Since NexPro is a combination of both corn protein and yeast, it works as a single product with the characteristics of both to provide a unique nutritional profile (Table 1).

NexPro is a 50% protein product derived from the whole stillage that remains after the fermentation process in ethanol manufacturing. Flint Hills uses a

Ingredient (%)

NexPro

Corn Gluten Meal

93

90

90

Crude Protein

50.1

60

47.7

Crude Fat

3.1

4.74

1.5

Crude Fiber

5.5

0.7

3.9

Ash

4.0

1.7

6.3

Phosphorus

0.40

0.49

0.71

Lysine

2.01

0.93

2.96

Methionine

1.01

1.21

0.66

TSAA

1.88

2.22

1.36

Threonine

2.00

3.52

1.86

Tryptophan

0.43

1.81

0.66

Phenylalanine

2.57

3.52

2.4

Valine

2.87

2.42

2.23

Leucine

5.57

9.82

3.62

Isoleucine

2.19

2.23

2.14

Histidine

1.33

1.32

1.28

Arginine

2.30

1.66

3.45

Dry Matter

Soybean Meal

Table 1. Typical nutrient content of the NexPro protein product compared to corn gluten meal and soybean meal.

patented physical separation technology to separate both yeast and corn protein from the remaining fiber.

Traditionally, ethanol plants process their whole stillage into distillers dried grains with solubles (DDGS) and distillersâ&#x20AC;&#x2122; corn oil (DCO). The mechanical process Flint Hills uses allows the creation of a higher-protein product that is valuable to aquaculture producers.

The process uses a series of screens and centrifuges to separate fiber from protein and remove solubles.

Yeast from the fermentation process becomes concentrated in the remaining protein, with the yeast content estimated at 25% of the product by weight. This allows for production of a consistent, high-quality ingredient for use in aquafeed applications.

In addition to containing 50% protein, the amino acid profile provides significant advantages to aquaculture producers; it contains nearly twice as much lysine as corn gluten meal, and over 50% more methionine than soybean meal.

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Tilapia performance Tilapia is a popular warm water omnivorous fish produced for the consumer market. With an estimated 5.9 million metric tons produced in 2016 (Tacon, 2018), it is the second-most important farmed fish species after carp. Tilapia are well suited for aquaculture-based production systems as they are relatively disease resistant and able to utilize a wide variety of feedstuffs.

Corn-based products including corn gluten meal and corn protein concentrate are used in tilapia feeding programs with considerable success.

Flint Hills wanted to demonstrate to its customers that replacing some of the more-expensive protein products with NexPro was both economically and nutritionally viable. The studies created by our research partners examined the effects of the ingredient in tilapia diets at a variety of inclusion rates.

First, preliminary research was conducted to attempt to determine maximum inclusion rates in tilapia diets.

Diets were formulated to be equivalent in protein and lipid content, with lysine and methionine requirements met through the addition of crystalline amino acids as needed.

The initial 56-day study indicated that with increasing inclusion, tilapia growth performance was numerically but not statistically improved as NexPro inclusion was increased (Fig. 1). Overall feed conversion ratios also did not differ among the different dietary treatments in this

experiment. The digestible energy content of NexPro was estimated at 98.3%, while the digestible protein content was estimated at 88.9% for tilapia.

Subsequent research using NexPro as an ingredient in the diet of juvenile tilapia was presented by Amorocho et al., 2018. This nine-week feeding study involved replacing corn protein concentrate in low fishmeal diets. Corn protein concentrate, which contains 75% crude protein, was included in the basal diet at 8%. Treatment diets replaced the corn protein concentrate at 2% increments on an equal protein basis, resulting in 8%, 6%, 4%, 2%, and 0% c orn protein concentrate in the test diets. NexPro inclusion rates in the same treatment diets were increased from 0% to 3.15%, 6.30%, 9.45%, and 12.6% as corn protein concentrate was reduced.

Tilapia were fed for nine-weeks starting at an average weight of 7.49 g. No differences were seen in any of the response variables, including final weight, weight gain percent, or feed conversion ratio. Therefore, we conclude that NexPro can replace other, more expensive high-quality protein ingredients in tilapia feed formulations.

Shrimp performance NexPro has been used in several Pacific white shrimp feeding studies. Most of these experiments have been aquarium based, with between ten and 30 post-larval

Figure 1. Growth performance of tilapia fed increasing levels of NexPro in the diet for 56 days.

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Figure 2. Effect of replacing fishmeal with NexPro in the diet of juvenile shrimp (initial weight 0.25 g) on final weight and feed efficiency.

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Figure 3. Replacement of corn protein concentrate or fishmeal with NexPro in a green water environment for Pacific white shrimp.

shrimp per tank. Initial research focused on maximizing inclusion rates with levels up to 30% of the diet being tested. In these studies, researchers found slightly higher final biomass per tank for the NexPro-fed treatments compared to the control (Qui et al., 2017).

Subsequent research examined the use of NexPro as an alternative to fishmeal, with NexPro replacing a combination of fishmeal and soybean meal. Diets contained 12%, 9%, 6%, 3%, and 0% fishmeal, and 0%, 6%, 12%, 18%, and 24% NexPro, respectively. Final weights were greatest for shrimp that were fed the 6% NexPro diet, followed by the 12% diet, but were only significantly increased compared to the 24% NexPro diet (Qui et al., 2017; experiment 3). Note that the control diet was not statistically different from any of the treatment diets for final weight.

More recent research conducted with NexPro in shrimp diets have used an outdoor green water recirculating aquaculture system with 30 shrimp per tank and four replications per treatment. This research evaluated the ability to use NexPro to replace either fishmeal or corn protein concentrate. The result of this research showed that a shrimp diet completely replacing corn protein concentrate at a 12.6% inclusion with a 20% inclusion of NexPro (Fig. 2) produced no difference in final weights. The same result was observed when reducing fishmeal from 17.4% to 6% inclusion, while replacing it with a 15% NexPro inclusion.

Note that performance could not be maintained at the lowest fishmeal inclusion level (2.21% fishmeal; Fig. 3).

Overall, this research demonstrated that in shrimp

diets, fishmeal inclusions could be reduced to just 6% of the diet while simultaneously adding up to 15% NexPro, and growth performance would not be affected.

Conclusions We conclude that if formulated properly, NexPro can replace higher-cost protein ingredients without a reduction in growth performance in common aquaculture production operations. Multiple studies have independently demonstrated that the product can be used as a substitute to high cost ingredients like fishmeal and protein concentrates. In the most recent shrimp study, feed cost savings per metric ton of $43, to $85 were obtained for diets replacing corn protein concentrate with NexPro. Diets replacing fishmeal with NexPro showed a savings of $77 to $153 per metric ton. Given both animal growth performance and opportunities for substantial feed cost savings, Flint Hills Resources recommends using up to 15% NexPro in shrimp and tilapia feed formulations.

References available on request.

More information: Scott Tilton, Ph.D., provides research and technical support for Flint Hillsâ&#x20AC;&#x2122; Ingredients business.

Manager of Nutrition Services Flint Hills Resources, USA E: NexProSales@fhr.com

Aquafeed: Advances in Processing & Formulation Vol 11 Issue 3 2019

PLANT-BASED INGREDIENTS

31

Positive results from Thai Union Feedmill’s shrimp feeding trial with a sustainable Nannochloropsis algae meal Martin Sabarsky, Barry Raleigh and Avery Kramer, Cellana LLC Increasingly, seafood consumers prefer sustainable products over their conventional counterparts.

For aquafeeds, this involves replacing fishmeal and other feed inputs with more sustainable ingredients.

Photosynthetic algae farmed on non-arable land using seawater and recycled CO2 represent one of the most promising new ingredients for sustainable aquafeeds given the low land, fresh water, and carbon footprints.

While economic production of photosynthetic algae has proven challenging for aquafeeds, utilizing algae meal (the residual algae biomass remaining after extraction of lipids) as part of a multi-product algae “biorefinery” is a more viable option that companies like Cellana LLC have successfully pursued.

Cellana’s strategy has been (1) to select and grow EPArich, photosynthetic algae, with EPA representing the most valuable omega-3 oil in the world, (2) to provide a nutraceutical-quality oil product using food-grade

Parameter Commercial Control (%) RD-A10 (10% Nannochloropsis) (%) Protein Fat Fiber Ash Moisture

37.86 6.41 2.73 10.48 10.60

Table 1. Chemical composition of the two feed formulations tested.

extraction technologies, and (3) to utilize the remaining high-protein algae meal containing valuable residual EPA omega-3s and pigments in aquafeeds and other food applications.

This advantageous strategy doesn’t require a price premium or government subsidy. Economically sustainable returns can be generated from multi-product algae biorefineries while supplying increasing quantities of competitively-priced commodity inputs to food and feed customers.

Treatment Parameter Commercial Control RD-A10 (10% Nannochloropsis) P-value Initial Weight (g/shrimp)

3.60±0.09

3.60±0.07

1

Final Weight (g/shrimp)

14.36±2.12

14.00±0.71

0.763

Weight gain (g/shrimp)

10.75±2.13

10.40±0.68

0.763

298.92±60.35

288.81±17.35

0.758

0.17±0.03

0.17±0.01

0.791

Survival Rate (%)

62.86±14.75

55.00±8.21

0.388

Feed intake (g/shrimp)

14.20±2.30

14.52±0.77

0.804

Feed Conversion Ratio (FCR)

1.33±0.14

1.40±0.02

0.441

Protein Efficiency Ratio (PER)

2.00±0.20

1.80±0.03

0.136

Weight gain (%) ADG (g/days)

39.87 7.90 1.94 10.49 9.51

Table 2. Growth performance of Litopenaeus vannamei fed control diet and Nannochloropsis containing diet (10%) at week 9 (63 days). All values given as mean±sd (n=4).

The recent feed trials conducted by Thai Union Feedmill with Cellana algae meal A large feed trial involving Nannochloropsis algae meal was recently conducted

by Thai Union Feedmill Co., Ltd. over 9 weeks, from late 2018 through February 2019, on the commercial shrimp species Litopenaeus vannamei. This trial

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32

Figure 1. Average weight, weight gain and average daily growth of Litopenaeus vannamei fed control diet and Nannochloropsis containing diet (10%) at week 9 (63 days). All values given as meanÂąsd (n=4). All values are means of four replicates per treatment. Each bar represents the mean value from four determinations with SD.

represents the second shrimp feeding trial performed

by Thai Union Feedmill with Cellana algae meal and builds on the successful results from the first joint Cellana-Thai Union Feedmill trial conducted in 2015.

The purpose of this second trial was to compare the effects of feeding commercial shrimp food (commercial control) versus an experimental diet consisting of 90% commercial control and 10% Nannochloropsis algae meal provided by Cellana LLC. The Nannochloropsis algae meal was derived from whole algae originally containing approximately 29% lipids at harvest, from which approximately 88% of its lipids were extracted via a conventional food-grade solvent extraction method.

The algae meal incorporated by Thai Union Feedmill

Figure 2. Feed conversion ratio and protein efficiency ratio of Litopenaeus vannamei fed control diet and Nannochloropsis containing diet (10%) at week 9.

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33

Figure 3. Survival rate of Litopenaeus vannamei fed control diet and Nannochloropsis containing diet (10%) at week 3, week 6 and week 9 (63 days). All values given as meanÂąsd (n=4). All values are means of four replicates per treatment.

into the experimental feed mix, designated RD-A10, was provided from the same lot of algae meal produced

by Cellana in 2015 for the first trial. In this second trial, shrimp were cultivated for 9 weeks in a 500-liter tank with recirculating water prior to harvest. After harvest, Thai Union Feedmill analyzed relevant growth, feed conversion, protein conversion and survival rate factors.

In Table 1, the composition of the experimental feed RD-A10 differed primarily in a 5% increase in protein and a 23% increase in fat over the commercial control used by Thai Union Feedmill.

The data from the trials collected from four replicates per treatment (Table 2, Fig. 1-3) showed that, within one standard deviation, the shrimp fed Nannochloropsis algae meal gave equivalent results to those fed the commercial control diet.

Conclusion Given the successful performance of the algae meal at the 10% inclusion rate tested, even if pricing of the Cellana algae meal were to be at a 10% to 20% discount to the current price of fishmeal, or approximately $1.16/kg to $1.35/kg based on fishmeal prices of $1.45/kg to $1.50/kg, this would be an important revenue contributor as part of an economically sustainable, multi-product biorefinery business model. This, together with the superior

environmental sustainability of Cellanaâ&#x20AC;&#x2122;s algae meal, makes for a very exciting combination in the universe of emerging aquafeed innovations.

More information: Martin Sabarsky CEO Cellana LLC, USA E: martin.sabarsky@cellana.com

Barry Raleigh, Ph.D. Chairman and Co-Founder Cellana LLC, USA E: cbarryraleigh@gmail.com

Avery Kramer Vice President of Operations Cellana LLC, USA E: avery.kramer@cellana.com

Aquafeed: Advances in Processing & Formulation Vol 11 Issue 3 2019

PLANT-BASED INGREDIENTS

Assessment of the nutritional performance and physicochemical properties of novel yeast protein ingredient for aquaculture feed Ricardo Ekmay and Elimy Glenn, Arbiom

Sustainability challenges for proteins in aquaculture The aquaculture industry has been at the forefront in the use of novel proteins and particularly those that contribute towards a circular economy. In one regard, fishmeal supply shortages and price volatility have contributed toward this movement to adopt new, more sustainable protein sources. In another regard, alternatives to fishmeal have largely resulted in the adoption of plant proteins such as soy, wheat and maize. However, the adoption of these conventional vegetable proteins has not necessarily resolved the issues of sustainability and has, in some instances, introduced new issues surrounding gastrointestinal health and nutritional performance relative to fishmeal. There have been extensive efforts by the industry to mitigate these issues, but the drive toward alternative proteins that can deliver nutritional health performance and contribute to the overall sustainability of the aquaculture is strong.

One of the under-scrutinized attributes of novel proteins is the material handling properties of the ingredient. Materials that are not free flowing, are

significantly hygroscopic or produce high levels of dust, may pose significant logistic and safety issues in a feed mill. Further, the ingredientsâ&#x20AC;&#x2122; functional properties in extrusion applications may differ from the ingredient(s) they are replacing and, therefore, may not be suitable for the existing feed formulations. It is imperative to consider the physicochemical characteristics along with the nutritional attributes of an ingredient when evaluating novel proteins.

In this article, weâ&#x20AC;&#x2122;ll look at an alternative protein on the horizon that recently underwent material handling testing with promising results.

High-quality alternative protein Arbiom offers a next-generation protein source for aquaculture, which delivers science-backed performance across several dimensions of a novel protein ingredient: nutritional, economic, traceability and sustainability value. This first commercial product is a high-protein ingredient, SylProÂŽ, that is a strong alternative to fishmeal or vegetable proteins for sustainable animal nutrition. The product is an enhanced strain of yeast produced from wood through a process Arbiom has developed to produce food/ feed and other intermediates from wood. The protein ingredient is composed of dried inactive microorganisms, a proprietary strain of yeast, which are grown on woody substrate material in fermentation.

The microorganism has a history of safe use and is already approved by major regulatory agencies. The enhanced amino acid content (particularly methionine, lysine and threonine), high digestibility and rich beta

Aquafeed: Advances in Processing & Formulation Vol 11 Issue 3 2019

35

glucan/mannan content helps address some of the critical issues facing aquafeed formulators.

In a pilot-trial conducted by Arbiom, SylPro bulk density, Hausner ratio and angle of repose were assessed to determine the flowability of the ingredient.

Further, the product underwent a variety of extrusion processes to assess the impact of the product’s inclusion rates and barrel temperature on the quality characteristics of a resulting fish feed pellet. Using a single screw extruder, 12 hybrid striped bass diets were produced that consisted of no yeast, 5, 10, or 20% inclusion, respectively, and three barrel temperatures: 100, 120 and 140°C, respectively. SylPro replaced a combination of fishmeal and soybean meal as a protein source in these trials, with the target pellet size being four millimeters.

The angle of repose and Hausner ratio suggest that torula yeast had good flowability and did not require external agitation for movement. Inclusion of SylPro linearly increased the water absorption index (WAI) of extruded pellets, but had no effect on water solubility index (WSI). Further, inclusion rate had no effect on expansion ratio up to 10%. No significant barrel temperature effects were observed for WAI or WSI. However, a significant interaction effect was observed for expansion ratio where a significant inclusion by temperature interaction was observed, driven primarily by the 20% inclusion level.

These results support the initial assessment that SylPro is flowable and its incorporation into extruded aquaculture feeds will be similar or superior to existing conventional protein ingredients utilized by feed manufacturers.

Nutritional performance analysis Arbiom followed up its physicochemical analysis with an in-depth nutritional evaluation of SylPro in hybrid striped bass and Atlantic salmon. In hybrid striped bass, diets were formulated using SylPro as a replacement for vegetable proteins at various inclusion rates up to 30%. The study measured indicators including growth, body composition, nutrient digestibility and general gastrointestinal health over a period of 60 days.

The results show no differences in mortality or feed intake across all diets, and there was no statistical difference in body weight gain, feed intake or feed efficiency up to 20% inclusion. Further, apparent

SylPro

Bulk density (g/cm3) Hausner ratio Angle of repose (°)

0.76 ± 0.02 1.06 ± 0.03 30

Soy protein concentrate Fishmeal 0.50 1.11 51.9

0.56 1.15 53.3

Material Handling Study of SylPro conducted by Texas A&M University, 2018.

Jiang, X. 2016. Properties of feed ingredients and extruded products. MS Thesis. Iowa State University, Ames, Iowa.

Table 1. Comparison of material handling properties: SylPro and common protein ingredients.

crude protein and lipid digestibility were 97% and 98%, respectively.

A similar experimental design was used for evaluation of Atlantic salmon. However, with salmon, the trial was terminated at 35 days and inclusion rates went up to 20%. In addition to replacement of solely vegetable proteins, a set of diets were prepared that replaced fishmeal instead of vegetable protein. In both diet sets, where vegetable proteins or fishmeal were replaced, performance was equivalent up to 20% inclusion.

Results indicate that SylPro can be used to replace fishmeal or vegetable proteins in feed up to 20% inclusion and deliver the same nutritional performance.

Arbiom is committed to enabling the safe and sustainable growth of aquaculture and animal protein production as a leading alternative protein production platform. The company is partnering with aquaculture feed producers and forest products companies to bridge the wood-to-food chain, bringing new protein source to market at commercial-scale.

More information: Ricardo Ekmay Vice President of Nutrition Arbiom, USA E: rekmay@arbiom.com

Emily Glenn Director of Business Development Arbiom, USA

Aquafeed: Advances in Processing & Formulation Vol 11 Issue 3 2019

PLANT-BASED INGREDIENTS

Meeting the new US FDA omega-3 qualified health claim for salmon: algae-based omega-3s as a sustainable feed source Jill Kauffman Johnson and Katie Compton Yahna, Corbion Algae Ingredients Strong research has supported the vital role that long chain omega-3s play when it comes to supporting normal brain development and function, as well as reducing the risk of coronary heart disease.1,2 One of the primary ways that long chain omega-3s – specifically docosahexaenoic acid (DHA) and eicosapentaenoic acid (EPA) – make their way into the human diet is via seafood, especially salmon and other oily fish.3 The importance of long chain omega-3s recently got a new boost from the U.S. Food and Drug Administration (U.S. FDA). On June 19, 2019, the FDA announced that it “does not intend to object to the use of certain qualified health claims stating that consuming omega-3 EPA and DHA fatty acids in food or dietary supplements may reduce the risk of hypertension and coronary heart disease.”4 This is a significant step for the U.S. FDA, as it builds on a Qualified Health Claim (QHC) issued by the agency in 2004 relating to the role EPA and DHA omega-3 fatty acids may play in helping to reduce the risk of coronary heart disease. This new QHC discusses the role of EPA and DHA in helping to lower blood pressure – but in order for foods to be labeled with a QHC, there needs

to be a combined total of at least 0.8 grams of EPA and DHA (combined total) per serving.5 What does this mean for the salmon aquaculture industry? The US FDA has specified that the QHC can be used if the fish contains at least 0.8 grams of EPA and DHA in an 85 g serving of uncooked fish (or 55g of smoked salmon), setting a new minimum omega-3 threshold that U.S. consumers should expect from their salmon products making this claim.6 This poses a challenge for feed producers and salmon farmers. The major source of long chain omega-3s today is fish oil – salmon farmers understand the importance of omega-3s for fish health and highquality seafood, but there is consistent pressure to reduce the amount of marine ingredients used in feed to move closer to being a “net protein producer.”

Fish oil is a constrained resource Approximately 1 million metric tons of fish oil are produced each year, primarily for use in aquaculture, terrestrial animal feed, and human nutrition.7 About 70% of the available crude fish oil is consumed by the aquaculture industry, with use in farmed

Source: US FDA website: https://www.fda.gov/food/cfsan-constituent-updates/fda-announcesnew-qualified-health-claims-epa-and-dha-omega-3-consumption-and-risk-hypertension-and

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Source: Salmon Farming Industry Handbook 2019, MOWI, pg. 26.

salmon feeds accounting for over 50% of this amount (400,000 MT in 2014).8, 9 Aquaculture is the fastest growing animal-based food producing sector in the world.10 While the aquaculture industry is growing, the availability of long chain omega-3s from wild caught fish is essentially flat. Over the last 10 years, the global Atlantic salmon industry had a 6% compound annual growth rate (CAGR) continuously driving demand for long chain omega-3s.11 Continued availability of long chain omega-3s depends on sustainable fishery management policies, productivity of natural systems (which have been impacted by climate change), compliance with country quotas, and other factors.

Microalgae as an alternative to meet rising demand for long chain omega-3s Recognizing this looming resource constraint, the development of alternative sources of long chain omega-3s have been underway for decades.

Genetically engineered crops, engineered yeast, and natural marine microalgae have all been contenders. Today, microalgae have emerged as the first commercially viable new source of long chain omega-3s at scale and already incorporated into salmon feed. The advantage is that microalgae are the original source of marine omega-3s (EPA & DHA) and are at the base of the food chain – salmon and other fin fish eat forage fish and accumulate high levels of omega-3s. By growing microalgae on land, feed suppliers can go straight to the source, microalgae, for a traceable, high quality and sustainable alternative for long chain omega-3s.

Source: Corbion Algae Ingredients

One example of a commercial product on the market today is AlgaPrime™ DHA12, produced by the global ingredients company, Corbion. They announced recently that in Norway alone, over 350,000 tons of fish feed with AlgaPrime™ DHA has been delivered to Norwegian salmon farmers since 2016 to improve the omega-3 content and reduce the fish-in, fish-out ratio of farmed salmon.13 When salmon are fed algae-based omega-3s, the fish-in, fish-out ratio often drops below 1:1 – and in some cases as low as 0.5:1.14 As salmon producers look to meet consumers’ expectations for good levels of omega-3s in their salmon and other seafood, additional sources of long chain omega-3s are needed for the growing aquaculture sector. Microalgae-based omega-3s are now produced at scale and offer a sustainable resource to improve omega-3 levels in farmed salmon and beyond.

References available on request.

More information: Jill Kauffman Johnson Head of Global Market Development Corbion Algae Ingredients, USA E: jill.kauffmanjohnson@corbion.com

Katie Compton Yahna Marketing & Communications Manager Corbion Algae Ingredients, USA E: katie.compton@corbion.com

Aquafeed: Advances in Processing & Formulation Vol 11 Issue 3 2019

COLUMN

Ask the Expert Your aquafeed processing questions answered

Q: We are getting a lot of pellet variation and wedging.

See attached pics. Any ideas to help? Do we need to change formulation?

Without having pellets in my hand it is hard to tell, however looking at the pictures you sent, there is some variation in length and I’m not seeing signs of curling, I’m going to discount a slow knife speed/wide blade problem for now. Don't go changing formulation at this stage.

My instinct tells me any one of or a combination of the following are the cause of your problem; water is too low, die temp is too high and/or die open area is too high.

The reason I suggest this is I can see quite a range in product size, which suggests poor die fill, poor flow distribution and potentially low die viscosity. I also note that the back end cuts don’t appear to have concave cupping normally associated with a well-cooked high protein/low starch pellet achieved with appropriate water addition (conversely, high starch will tend to have convex cut surface under ideal process conditions).

Try increasing water until pellets become too tacky to handle, then back water up until pellets free flow.

The level of water required will be very much formulation dependent.

Run significant cooling on final head (try setting temp low enough that cooling controllers never close off if necessary). You may also

require cooling further back down the barrel (depending on the type of extruder, profile and other process conditions). Bear in mind that if you are not currently cooling the final head, when you first introduce cooling the viscosity of the melt will increase. This will increase pressure at the die, extruder load, SME and potentially lead to increased expansion. It is often only through significant cooling, sometimes required on more than one section/ head, that you will see an increase in density.

If you are running a lot of steam injection directly through the barrel into product, you may want to try backing this off a bit, as this could be reducing your viscosity leading to “run out” at the die. At the same time, try to get a bit more cook out of your conditioner if you can.

Finally, reduce the die open area and if open area allows, reduce the hole pattern to one row only with symmetrical pattern. Also check to see that flow behind the die is not being disrupted by any pre-die configuration. A cone on the back of die would likely help as well if you are not already using.

There are other considerations such as die land length and screw/ head wear; however you can start with the above before looking too closely at hardware.

The next issue you face will be balancing density. Some pointers here:

Aquafeed: Advances in Processing & Formulation Vol 11 Issue 3 2019

Peter Hutchinson is a Technical Editor at Aquafeed.com, the owner – director of ENH Ltd., New Zealand, and an aquaculture feed consultant.

Send your questions to: pete@aquafeed.com

Max expansion will occur when starch reaches optimum moisture for cook at given process conditions, leading to peak viscosity. Below the required level of water to achieve this cook, pellets will often be dense with poor structure and tend to have square cuts. Above the water level required for optimum expansion (if you can get there without pellets becoming too sticky), density increases again.

Pellet quality and water stability will be better at higher water levels, provided cook (particularly through SME) is maintained.

Injection of oil into the barrel will have the biggest impact on reducing expansion (increasing density), however consideration of SME,

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cook and ability to coat oil afterwards will need to be taken into account.

Restriction plates behind the die will increase SME and expansion, so adjust accordingly if you have these. Even better a variable restriction valve will make it easier to adjust SME and expansion during operation.

Adjusting rates of cooling and

screw speed can also be used to alter density.

Q: What is the difference between an electric and steam powered extruder?

I believe there may be some misunderstanding here and think what you may be referring to is the difference in delivery of Specific Thermal Energy (STE) in different

makes of extruders. STE is the energy delivered through external inputs rather than through the friction or shear created by the turning of the extruder screw called Specific Mechanical Energy (SME).

Some extruders utilize jackets around the barrel to allow steam or water for additional heating and cooling as required. Others use induction elements surrounding the barrel. Some older style extruders have no means of temperature regulation on the barrel at all.

You may also be referring to steam preconditioning, which is used to increase the level of cook and improve pellet quality, by injecting steam through the mash in a mixing device immediately prior to extrusion. Some older style extruders (more commonly referred to as dry expanders) donâ&#x20AC;&#x2122;t have preconditioning systems fitted and thus do not utilize steam.

By far the majority of extruder screws are powered by electric motors, although not necessarily all, and I have heard of machines historically which have been driven by things such as the PTO shaft from a tractor. There may well have been instances of steam engine driven extruders, however I am confident the barrel temperature control described above likely explains the question you have raised.

Aquafeed: Advances in Processing & Formulation Vol 11 Issue 3 2019

COLUMN

Recent publications Albert G. Tacon, Ph.D. Aloha. Continuing the theme from the last issue, I have collected some of the most recent publications in 2019 that I think will be of interest.

Juvenile Procambarus clarkii farmed using biofloc technology or commercial feed in zero‐ water exchange indoor tanks: A comparison of growth performance, enzyme activity and proximate composition.

Li, Jinghao; Li, Jiayao; Li, Wei; Sun, Ying; Liu, Xiaofei; Liu, Mingming; Cheng, Yongxu. (2019). Aquaculture Research, 50(7):1834-1843.

A 60‐day study comparing the growth performance; proximate composition and the digestive, immune and antioxidant enzyme activities of juvenile red swamp crayfish (Procambarus clarkii) reared using biofloc technology (the biofloc group) or a commercial diet (the control group) in zero‐ water exchange indoor tanks was performed. A commercial diet was fed to both groups, and the daily feeding amount in the biofloc group was 40% less than that in the control group. Wheat bran and glucose were used in the biofloc group to ensure a C/N ratio >15 and the same feeding energy as that given to the control group.

After 60 days of feeding, the final weight, weight gain and specific growth rate in the biofloc group were significantly higher than those in the control group. Furthermore,

proximate composition analysis showed that the total hepatopancreatic lipid and ash contents of the biofloc group were significantly higher than those of the control group, and the intestinal and hepatopancreatic pepsin activities in crayfish in the biofloc group were higher than those in crayfish in the control group. Moreover, higher hepatopancreatic lipase and cellulase activities were observed for the biofloc group than for the control group. Biofloc technology promoted the health of the crayfish in terms of their immune and antioxidant enzyme activities.

Based on the results of this study, biofloc technology is more effective than traditional commercial diets for the farming of juvenile crayfish.

Nutritional regulation of pyruvate kinase and phosphoenolpyruvate carboxykinase at the enzymatic and molecular levels in cobia Rachycentron canadum. Li, Ruixin; Liu, Hongyu; Li, Shuyun; Tan, Beiping; Dong, Xiaohui; Chi, Shuyan; Yang, Qihui; Zhang, Shuang; Chen, Liqiao. (2019). Fish Physiology and Biochemistry; Dordrecht, 45(3): (Jun 2019): 1015-1028.

Despite being a carnivorous fish species, cobia (Rachycentron

Aquafeed: Advances in Processing & Formulation Vol 11 Issue 3 2019

Dr. Albert Tacon is a Technical Editor at Aquafeed.com and an independent aquaculture feed consultant.

E: agjtacon@aquahana.com

canadum) can utilize high levels of dietary carbohydrate (up to 360 g kg−1). By contrast, rainbow trout (also carnivorous) cannot, due to the absence of molecular induction of glycolytic enzyme and inhibition of gluconeogenic enzyme gene expressions such as pyruvate kinase (PK) and phosphoenolpyruvate carboxykinase (PEPCK). We hypothesized that this phenomenon is species-specific and will not be observed in cobia. Our results show that, at the molecular level, the mRNA abundance of the important glycolytic (PK) and gluconeogenic (PEPCK) enzymes in cobia liver are regulated by dietary carbohydrate-to-lipid (CHO:L) ratios and nutritional status (fed, unfed, and refed). Significantly upregulated hepatic PK and depressed PEPCK gene expressions were observed when the fish were fed with an

41

increasing CHO/L-ratio diet or were refed. However, in contrast to gene expression, there was no significant effect of dietary CHO/L ratios on PK enzyme activity. The decrease in PEPCK activity was significantly found between low CHO/L ratio and high CHO/L ratio diets, whereas the moderate CHO/L ratio group showed intermediate values. But PEPCK activity appeared to be independent of nutritional status. These results suggest that nutritional regulation is obvious, at least at the molecular level, in the key hepatic enzymes (PK and PEPCK) of the glucose metabolism pathway, in response to different dietary CHO/L ratios and to the transition from being starved to fed. Determining whether other key enzymes involved in hepatic glucose metabolism contribute to glucose tolerance in cobia is necessary for further investigation of this phenomenon at the enzymatic and molecular levels.

Astaxanthin enhances hematology, antioxidant and immunological parameters, immune-related gene expression, and disease resistance against in Channa argus. Mu-Yang, Li; Xin-Yu, Liu; Chang-Ge, Xia; GuiQin, Wang; Dong-Ming, Zhang.

(2019). Aquaculture International; Dordrecht,27(3): (Jun 2019): 735-746.

The present study was aimed to add 0, 50, or 100 mg/kg of astaxanthin (AST) to the basal diet of Channa argus and determine the effects on hematology, antioxidant and immunological parameters, immune-related genes expression, and disease resistance. Adding 50 or 100 mg/kg AST significantly

enhanced (P < 0.05) Hb content at 14, 21, and 28 days and also the levels of Ht and RBC at 21 and 28 days. Supplementing 50 or 100 mg/kg of AST for 21 and 28 days significantly increased (P < 0.05) the level of serum antioxidant enzymes (SOD, CAT, and GSH-Px) or decreased (P < 0.05) serum MDA content. The level of serum lysozyme significantly enhanced (P < 0.05) at 28 days and also IgM and C3 at 21 days in 100 mg/kg AST treatment and at 28 days in 50 or 100 mg/kg AST treatment.

Meanwhile, 50 and 100 mg/kg AST treatment significantly upregulated (P < 0.05) the expression of HSP70, HSP90, IκB-α, and GR or downregulated NF-κB p65 and inflammatory cytokines (TNF-α, IL-1β, and IL-8) transcription at 21 and 28 days. Fish fed 50 and 100 mg/kg AST enhanced (P < 0.05) disease resistance against Aeromonas hydrophila (survival rate 51.0% and 58.0%, respectively). The present results suggested that AST enhanced hematology and serum antioxidant and immune response, regulated immune-related gene expression, and increased disease resistance against A. hydrophila in C. argus.

Spirulina platensis in rainbow trout (Oncorhynchus mykiss) feed: effects on growth, fillet composition, and tissue antioxidant mechanisms. Sheikhzadeh, Najmeh; Mousavi, Shalaleh; Ali Khani Oushani; Firouzamandi, Masoumeh; Mardani, Karim. (2019). Aquaculture International; Dordrecht (Jun 2019): 1-11. An experiment was conducted to investigate the effects of Spirulina

platensis (SP) on growth, fillet composition, and intestinal, skin, and gill mucosal antioxidants of rainbow trout, Oncorhynchus mykiss (initial weight 17.18 ± 0.59 g). One-hundred and thirty-five fish were randomly distributed among nine cement tanks (1.8 m × 022 m × 0.35 m) with 15 fish per tank.

Three isonitrogenous (37.8% crude protein) feeds containing either 0, 2.5, or 5% SP were prepared, randomly assigned to triplicate tanks of fish, and given at 2% body weight per day for 7 weeks. At the end of the trial, fish in each tank were assessed for growth (final weight, thermal growth coefficient), condition factor, fillet composition, mucosal antioxidant activity, and gene expression analysis related to the antioxidant enzymes catalase, glutathione peroxidase 1, and glutathione S-transferase. Neither growth nor fillet composition were influenced by inclusion of SP in feed. Total antioxidant activity in three mucosal tissues including the intestine, skin, and gill was significantly increased by 2.5% SP whereas administration of 5% SP only increased total antioxidant activity in the intestine.

Feeding fish with 2.5% and 5% SP could upregulate the expression of catalase in the intestinal tissue whereas 5% SP enhanced the expression of glutathione peroxidase 1 in this tissue.

Glutathione S-transferase gene expression was also increased in the intestinal and skin tissues of fish administrated with 2.5% SP while in the fish that received 5% SP-supplemented diet, an upregulation of this gene was

Aquafeed: Advances in Processing & Formulation Vol 11 Issue 3 2019

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only noted in the intestinal tissue.

It was concluded that 2.5% SP had a potential to enhance some antioxidant parameters mostly in the intestine, followed by the skin and gill.

Soybean glycinin decreased growth performance, impaired intestinal health, and amino acid absorption capacity of juvenile grass carp (Ctenopharyngodon idella). Ya-Lin, Zhang; Xu-Dong, Duan; Wei-Dan, Jiang; Lin, Feng; Wu, Pei; Liu, Yang; Jiang, Jun; Sheng-Yao Kuang; Tang, Ling; Wu-Neng Tang; Xiao-Qiu, Zhou. (2019). Fish Physiology and Biochemistry; Dordrecht (Jun 2019): 1-14. The present study evaluated the influence of dietary soybean glycinin on growth performance, intestinal morphology, free intestinal amino

acid (AA) content, and intestinal AA transporter (AAT) mRNA levels in juvenile grass carp (Ctenopharyngodon idella).

Results were displayed as follows: (1) 8% dietary glycinin decreased growth performance, inhibited intestinal growth, and caused intestinal histology damage of grass carp; (2) dietary glycinin decreased the content of free neutral AAs including Val, Ser, Tyr, Ala, Pro, and Gln in all intestinal segments, and Thr, Ile, Leu, Phe, and Gly in the MI and DI while downregulated the mRNA levels of corresponding transporters including SLC38A2, SLC6A19b, and SLC6A14 in all intestinal segments, and SLC7A5, SLC7A8, and SLC1A5 in the MI and DI. Dietary glycinin decreased the content of free basic AAs

Aquafeed: Advances in Processing & Formulation Vol 11 Issue 3 2019

including Arg in the MI and DI and His in all intestinal segments while downregulated cationic AAT SLC7A1 mRNA levels in the MI and DI. Dietary glycinin decreased the content of free acidic AAs including Glu in all intestinal segments and Asp in the MI and DI while decreased mRNA levels of corresponding transporters including SLC1A2a in all intestinal segments and SLC1A3 in the MI and DI; (3) the digestion trial showed that basic subunits of glycinin was hard to digest in the intestine of grass carp; (4) co-administration of glutamine with glycinin partially alleviated the negative effects.

Overall, glycinin decreased intestinal AA absorption capacity partly contributed by decreased AATsâ&#x20AC;&#x2122; mRNA levels and the indigestibility of glycinin.

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Archaea extract: a new heat stable micro-ingredient to improve disease resistance in aquaculture Duncan Sutherland and Tray Ghidossi, TwentyGreen Group Disease burden and antibiotics dependence remain a major challenge for the aquaculture industry. Probiotics and microbe derived extracts are useful strategies to improve survival rates and FCR in aquaculture, although these products are typically not heat-stable and therefore difficult to integrate in commercial granulate and extruded feeds.

How conventional probiotics work The idea of integrating microbes into granulated feed is aimed to help create a natural barrier against pathogens preventing cell invasion and disease onset.

Probiotics and microbe-based extracts are a much more preferred alternative to antibiotic treatments.

Antibiotics overuse in agriculture has been linked to the emergence of unprecedented levels of antibiotics resistance worldwide.

Conventional probiotics incompatible with industrial processes Unfortunately, routine industrial heat processes destroy microbe-based feed additives diminishing the potency of such products significantly.

Hatchery feeds are typically produced using granulation, microencapsulation, spray-drying and extrusion processes. Commercially available microbe-based products are insufficiently heat-stable to endure heat-exposure. It is cumbersome to apply probiotics and microbial extract type products separately â&#x20AC;&#x201C; e.g. by top-coating etc.

High level resistance to heat Pioneer research performed at the Institute of Technology in Lausanne (EPFL) in Switzerland, identified archaea as a high potential microingredient for the animal feed industry.

Figure 1. Archaea cell wall structure versus bacterial cell wall structure.

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Archaea are anaerobes forming a separate domain to bacteria. Some strains are adapted to thrive in extreme conditions such as thermal vents above 100°C – so called thermophiles and extremophiles.

Here, we report that TwentyGreen archaea strains remain viable after 80°C exposure. In vivo studies in shrimp and tilapia indicate the bioactive fragments remain functional after 120°C exposure. The heatstability of archaea extract facilitates the integration into the feed.

Heat-resistance comparative analysis: archaea versus yeast A study investigating the tolerance level of TwentyGreen archaea to heat stress was carried out and compared to a yeast, Saccharomyces cerevisiae.

S. cerevisiae was chosen as a probiotic model because it is widely used in probiotic preparation, it is low-cost, fast-growth and efficient against gastrointestinal tract disorders, both in humans and animals. Heat treatments (5, 30 minutes at 65°C and 72°C and 5, 10 minutes at 80°C) were performed on freeze-dried biomass of TwentyGreen archaea and S. cerevisiae. The positive control (CP) was kept at 37°C for 30 minutes and the negative control (CN) was autoclaved at 121°C for 20 minutes. To study the cell viability, archaea cells were cultivated anaerobically according to the standard cultivation method defined by TwentyGreen. The optical density of the samples was measured at 600 nm (OD600nm) with a spectrophotometer (6320D, Jenway, UK) to monitor the cell growth. S. cerevisiae is an aerobic microorganism and the cell viability study was performed by estimation of colony forming units (CFU), through serial dilutions plating on TSA agar

plates (Tryptic Soy Agar, MP Biomedicals, France).

Each experiment was repeated in triplicate (n=3) for statistical analysis.

Results The cell growth of archaea and S. cerevisiae after heat treatment is represented and compared in Figure 3.

After 30 minutes at 72°C and after 5,10 minutes at 80°C, no growth of S. cerevisiae was observed in the agar plates. Under the same conditions Archaea not only showed growth, but cells achieved the same concentration as the positive control (OD600nm 0.8 1.0) during the same time of cultivation. Such results show a high potential of archaea to withstand high temperature exposure compared to a normal probiotic.

Existing studies claim that strains of Archaea similar to those used in this study are extremely sensitive to oxygen. Here cells were in contact with oxygen during the whole downstream processing part and also during the storage of the final product. It is important to mention that heat resistance trials were performed on freeze dried biomass (Fig. 2). Freeze drying is an excellent method for long-term preservation but undesirable effects such as protein denaturation, might lead to a decrease in the cell viability. Results shows that TwentyGreen Archaea were able to survive after a combination of unfavourable environmental conditions. The rapid activation of Archaea in the anaerobic environment of gastrointestinal tract of fishes and crustaceans is of great interest.

Biological activity of vegetative cells Bacteria, such as Bacillus sp., with the ability to form endospores – which are rigid structures that can

Figure 2. Flow diagram. Dry heat inactivation of TwentyGreen archaea versus Saccharomyces cerevisiae.

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Figure 3. Growth study of TwentyGreen archaea and S. cerevisiae, after being subject to different conditions (5, 30 min at 65°C and 72°C and 5, 10 min at 80°C). Results of positive control (CP, 30 min at 37°C) and negative control (CN, 20 min at 121°C) are showed. The cell growth of archaea is represented as the difference in the initial OD 600nm (t=0h) and the beginning of the stationary growth phase (t=168h), whereas the growth of S. cerevisiae is represented as colony-forming units (CFU) per mL. The experiments were repeated in triplicate (n=3), the standard deviation is represented.

survive under harsh conditions – are attractive for commercial application as they can endure feed processing steps and have long shelf life. However, the disadvantage is that endospores are metabolically inert and that germination in gastrointestinal tract might be reluctant or require longer time than the intestinal transit time. Archaea do not form spores and for this reason are expected to be more efficient and potent in aquaculture applications compared to sporebased products.

Archaea extract suitable for extruded aquafeed Research trials suggest bioactive archaea fragments withstand the harsh extrusion process. Studies performed in collaboration with TwentyGreen Solutions R&D facility demonstrated an improvement in Japonicus shrimp survival rates (85% survival with TwentyGreen versus 45% survival in control) when archaea were included in shrimp pellets prepared with a BCTM10 Twin-screw extruder (at 110°C exposure). Similarly, tilapia grew 20% faster at a commercial tilapia hatchery when TwentyGreen was included in the extruded fish

pellets. The inclusion of archaea extract in extruded feed (at micro-doses) demonstrates potent bioactivity despite the mechanical and thermal exposure.

Summary TwentyGreen archaea extract has excellent potential for granulated, microencapsulated and extruded aquaculture feeds. Archaea extract has only recently been registered in Europe and registration is planned in South-East Asia territories in 2020. TwentyGreen’s pioneer premix will be available exclusively from select distributors.

References available on request

More information: Duncan Sutherland CEO TwentyGreen Group, Switzerland E: duncan@twentygreen.com

Aquafeed: Advances in Processing & Formulation Vol 11 Issue 3 2019

INGREDIENTS PROFILE

Linseed, an ignored plant source of essential fatty acids Humaira Amman, Shafaq Fatima, RSN Janjua, Lahore College for Women University

A massive investment in scientific research has taken place to address the challenges of using plant proteins and oils in fish feed formulation. In developing countries where fish oil is not an ingredient of fish feed due to its high cost, the use of economical plant oils rich in n-3 PUFAs can remarkably improve feed quality.

Soybean has been widely used in aquafeeds and supports healthy and efficient fish growth.

However, its polyunsaturated alpha-linolenic acid (ALA) content is low (6.8%) when compared with other vegetable oils such as linseed oil (LO), with a 53.3% of ALA.

Production of linseed Flax (Linum usitatissimum) belongs to Linaceae family.

It is an annual herb that produces small flat and oval seeds. Flaxseed, commonly known as linseed, is a source of fiber and ALA. The main product obtained from linseed is oil and a residual paste that is used as an animal feed ingredient. It is produced in 34 countries, mainly in Russia, India, China, United States, Canada and Ethiopia (Fig. 1). The remaining world production of linseed are 2,793,344 tons (Goyal et al., 2014).

Composition of linseed Linseed is rich in fat, protein and dietary fiber and it is valued by its good flavor and nutritional properties. It contains 40% oil, 30% fiber, 20% protein, 4% ash and 6% moisture. Out of this 40% of oil proportion, LO fatty acids levels are 9% of saturated, 18% of monounsaturated and 73% of polyunsaturated fatty acids (PUFAs).

The PUFAs content comprises 16% omega-6 fatty acids, primarily as linoleic acid (LA), and 57% alpha-linolenic acid (Zuket et al., 2015; Goyal et al., 2017).

Linseed in fish feed Linseed has attained the attention of scientists, researchers and industry due to high proportion of omega-3 fatty acids. Even though flaxseed oil, unlike fish oil, does not contain eicosapentaenoic acid (EPA) and docosahexaenoic acid (DHA), it is gaining popularity in India and Western countries due to its high ALA content that can be converted into EPA and DHA with the help of enzymes elongase and desaturase in fresh water fish (Nayak et al., 2017).

Figure 1. Production of linseed at global level.

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Figure 2. Weekly (mean ± SE) variations in condition factor in control and treatment groups of grass carp (Ctenopharyngodon idella). Diet of treatment group A, B and C contained 5% of linseed flour, 5 % linseed oil, combination (2.5 % linseed flour + 2.5 % linseed oil), respectively. Trial continued for 36 days.

Control group was fed with soybean feed. Initial: Pre-treatment value, W1: first week post-treatment, W2: second week post-treatment, W3: third week post-treatment.

Carbon # Fatty acids C 12:0 C 14:0 C 16:0 C 16:1 C 17:0 C 18:0 C 18:1 C 18:2 C 18:3 C 18:3 C 20:0 C 20:1 C 20:2 C 20:3 C 20:5 C 22:0 C 22:6

Lauric acid Myristic acid Palmitic acid Palmitoleic acid Heptadecanoic acid Stearic acid Oleic acid Linoleic acid α-Linolenic acid β-Linolenic acid Arachidonic acid Eicosaenoic acid Eicosadienoic acid Eicosatrienoic acid Eicosapentaenoic acid Docosanoic acid Docosahexaenoic acid

Control

A

B

C

1.34 0.44 26.10 1.02 - 5.49 36.50 19.87 4.46 2.15 1.10 0.74 - - 0.79 - -

1.32 0.47 28.79 0.40 0.73 0.91 39.89 24.85 0.57 0.36 0.32 0.33 - - 0.56 0.20 0.31

1.30 0.15 38.69 0.31 1.30 1.20 39.11 12.71 1.07 - 0.35 1.13 0.24 - 1.64 0.22 0.41

0.76 33.66 0.39 0.52 0.50 39.09 17.23 1.73 2.31 0.73 1.22 1.35 0.23 0.40

Table 1. Comparison of fatty acid profile (%) in total liver lipids of grass carp (Ctenopharyngodon idella) fed with three different diets (A: 5% linseed flour, B: 5 % linseed oil, C: 2.5 % linseed flour and 2.5% linseed oil) over the period of 36 days. Control group was fed with soybean feed.

Previous studies suggested that the supplementation of linseed (meal and oil) showed higher levels of C16:0, C18:1n and n-3 LC-PUFAs, EPA and DHA, in liver and muscle than non-supplemented, soy-based feed in common carp, Cyprinus carpio (Zhelyazkov et al., 2014; Zupan et al., 2016), rainbow trout (Masiha et al., 2013), Senegalese sole (Benítez et al., 2013), Nile tilapia (Omolo et al., 2017) and juvenile turbot (Peng et al., 2017).

A new study tested on grass carp (Ctenopharyngodon idella) three different diets: A, 5% linseed flour; B, 5 % linseed oil; C, 2.5 % linseed flour and 2.5% linseed oil (Fatima et al., unpublished data). A control group was

fed with a soybean feed. Fish average weight was 75.55 g and the study lasted 36 days. It was observed that dietary total proportion of LA and ALA was higher in the control group than in the treatment groups.

High levels of DHA were observed in total liver lipids of grass carp in all three treatments groups (A: 0.31%, B: 0.41%, C: 0.40%), while no DHA was detected in the control group (Table 1.).

Similarly, high levels of EPA were observed in the three treatment groups (A: 0.56%, B: 1.64%, C: 1.35%). The high levels of EPA and DHA found in treatment groups may be due to enzymes Δ6 and Δ5 fatty acyldesaturase and elongase that are involved in the biosynthesis of LC-PUFAs from the catalysis of the C18 substrates LA/ALA. Both LA and ALA are considered essential in fish, due to fish incapability to biosynthesize these fatty acids de novo.

Despite the fact that other studies on common carp showed significant increase in growth rate by increasing the concentration of linseed oil (Zupan et al., 2016), non-significant differences in growth parameters were observed between different treatments in grass carp (Fig. 2), possibly due to the short duration of study and the fish stage.

It has been reported that high dietary lipid level (>7%) could reduce fish growth (Pei et al., 2004; El-Marakby, 2006; Zupan et al., 2016) due to the reduction in feed intake, the low ability to digest and absorb lipids and palatability issues.

The use of linseed in fish feed is increasing.

The high availability of LO and the sporadic use in human consumption makes it an interesting option for low-cost fish feed production. It can be used mixed with other vegetable oils for further enhancing its nutritive values. *References available on request

More information: Humaira Amman Ph.D. scholar Lahore College for Women University, Pakistan E: mairi.aquarius@yahoo.com

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