Aquafeed Magazine Vol 15 Issue 4 2023 by Aquafeed Media

Vol 15 Issue 4 October 2023

AQUAFEED Advances in processing & formulation An Aquafeed.com publication

MOISTURE MANAGEMENT Feed additives Marine-based ingredients New generation of extrusion systems Published by: Aquafeed Media, S.L.U. www.aquafeed.com info@aquafeed.com

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AQUAFEED

VOL 15 ISSUE 4 2023

Contents

ALTERNATIVES TO MARINE INGREDIENTS 12 Two strategies through direct and indirect replacement using a series of high-value functional ingredients.

A PROTEIN HYDROLYSATE FOR INCREASED GROWTH 40

RENDERED ANIMAL PROTEINS FOR CARNIVOROUS FISH 47

A NEW GENERATION OF AQUATIC EXTRUSION SYSTEMS 55

New studies show that the protein hydrolysate from Calanus finmarchicus has growth-promoting properties.

A review of 20-year research at Zhejiang University replacing fishmeal with rendered animal proteins

The system responds to the demand for digital factories and is based on lean principles.

Aquafeed: Advances in Processing & Formulation Vol 15 Issue 4 2023

AQUAFEED CONTACT US Editorial: editor@aquafeed.com Editor/Publisher: Lucía Barreiro Consulting Editor: Suzi Dominy Technical Editors: Peter Hutchinson, Albert Tacon, Ph.D Assistant Editor: Marissa Yanaga Conferences and webinars: info@aquafeed.com Advertising enquiries/request media pack: sales@aquafeed.com Accounts & all other enquiries: info@aquafeed.com

SUBSCRIBE Digital editions are free to industry subscribers. You may also purchase print copies. Subscribe at Aquafeed.com to receive your own digital copy of our publications. https://bit.ly/AQUASUBSCRIBE

VOL 15 ISSUE 4 2023

Contents 6

Interview with Ramakanth Akula

News Review

12 Alternatives to marine ingredients in aquaculture to improve sustainability and profitability through nutrition

18 Digestive and metabolic enhancer offsets reduced cholesterol level in shrimp feed

20 SmartMoisture®: Improving cost and efficiency of aquafeed production *Cover story

24 Optimizing phosphorus nutrient management in aquaculture 28 Impact of phosphate sources on apparent P digestibility in juvenile Litopenaeus vannamei

31 Heads or tails? Both! 34 You can only improve what you measure!

We are grateful to the following companies for sponsoring this issue of the magazine. Their support allows us to make our publications available without charge. Adisseo......................................................................................... 2 Victam......................................................................................... 5 Wenger........................................................................................ 8 Bioiberica...................................................................................17 Devenish.................................................................................23 ADM ..........................................................................................27 ProChaete.................................................................................33 Symrise Aqua Feed..................................................................39 Extru-Tech..................................................................................49 Phodé........................................................................................58 Vetagro.....................................................................................61

40 Big things have small beginnings 43 Supporting allostasis with optimized fishmeal freshness, protein digestibility, and beneficial water-soluble composition

47 Carnivorous fish feed formulation practice with rendered animal proteins: A review of 20 year research at Zhejiang University

51 C orn fermented protein tested successfully in rainbow trout 55 New-generation extrusion system upgrades aquafeed production

59 Developing sustainable fish feed through labatory extrusion 62 Calendar of events

FeedTech Expo.........................................................................63 WAS ..........................................................................................64

To read previous issues in digital format or to order print copies, visit www.aquafeed.com

Aquafeed: Advances in Processing & Formulation Vol 15 Issue 4 2023

SAVE THE SAVE THE DATE DATE

MARCH 12 - 14, 2024

MARCH 12 - CENTER, 14, 2024 BITEC EXHIBITION BANGKOK, THAILAND

BITEC EXHIBITION CENTER, BANGKOK, THAILAND

JOIN ASIA’S LARGEST DEDICATED EVENT FOR THE ANIMAL FEED PROCESSING INDUSTRIES

Meet 16:10 (potential) customers Growing market opportunities in 27-06-2023 the booming Asian region Specific conferences with key topics in line with your interests Over 9.000 international visitors The platform to launch your See what is trending in your innovation industry Access to the industry exhibition Meet delegates from various trade missions Supported by:

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The event is held in halls 9 and 10 177 x 228 Asia 2024.indd 1

13-10-2023 15:27

INTERVIEW with Ramakanth Akula At the heart of our decision was the recognition of the immense potential for the aquaculture sector in India. India's long coastline and favorable tropical climate provided an ideal setting for shrimp farming. However, we observed that the sector was in a very nascent stage and needed a lot of support in building a strong ecosystem. We started off as an integrated aquaculture company with our own farms. To support our farms, we set up a feed plant, a hatchery, and a processing plant. The original plan was to grow shrimp, process, and export – all in-house. We were the first feed plant with pelleting technology in India. Till then, most of the feeds were either conventionally made or imported into the country. We saw a huge opportunity in the sector and embarked on a mission to contribute to the growth of the shrimp farming sector in India. Our entry into the shrimp feed market was marked by a commitment to innovation and quality. We invested in research and development to formulate nutritionally balanced and high-quality shrimp feeds. This commitment to excellence allowed us to gain the trust of shrimp farmers across the country. Today, The Waterbase Limited is not just a feed producer but a comprehensive aquaculture solutions provider. We remain dedicated to our mission of fostering sustainable shrimp farming practices and supporting the livelihoods of countless farmers. Our journey from 1987 to the present has been marked by continuous learning, adaptation, and a commitment to the highest standards of quality and innovation.

Ramakanth Akula is CEO of The Waterbase Limited

AQ: The Waterbase Limited was founded in 1987. How did it all start? Why did the company enter the shrimp feed market? RA: In 1987, The Waterbase Limited embarked on a journey that has led us to become a prominent player in the aquaculture industry today. Our foray into shrimp aquaculture was driven by a combination of factors and a vision for sustainable aquaculture that could provide gainful employment to millions of Indians and also contribute significantly to the country's GDP.

AQ: Focusing on shrimp feeds, where is the company today in terms of size, markets served and aquafeed volume? RA: We are a mid-sized player in India today. In terms of size, our installed capacities are over 100,000 MT per annum. Our state-of-the-art production facilities are equipped with advanced pelleting technology to ensure the highest quality. Our feed mills are strategically located in the province of Andhra Pradesh, which contributes to over 70% of the country’s shrimp production. Speaking of markets served, our footprint has extended beyond Andhra Pradesh. We serve aqua farmers in other states like Gujarat, West Bengal, Tamil Nadu, Maharashtra, and Goa. Additionally, we recently started exporting

Aquafeed: Advances in Processing & Formulation Vol 15 Issue 4 2023

INTERVIEW shrimp feed to Sri Lanka. Waterbase utilizes advanced pelleting technology to produce its shrimp feeds. AQ: Have the feed mills incorporated recent technology developments such as automation and energy-eﬃcient processing machines? Do you plan to change to extrusion technologies? RA: Indeed, technology plays a pivotal role in ensuring the quality and efficiency of our shrimp feed production. We have been proactive in embracing technological advancements to enhance the operations of our feed mills. Automation is a key aspect of our feed production process. We have incorporated state-of-the-art automation systems to streamline various stages of production. This not only ensures precision and consistency in our feed formulations but also improves overall efficiency. Energy-efficient processing machines are another area where we have made significant investments. Sustainability is a core value for us, and we are continually exploring ways to reduce our carbon footprint. Energyefficient machines not only help minimize our energy consumption but also contribute to cost savings, which can be passed on to our customers whenever possible. Regarding the transition to extrusion technologies, while we have been utilizing pelleting technology effectively, we are open to exploring and adopting extrusion techniques when they align with our commitment to delivering high-quality feeds. Extrusion technology offers certain advantages, such as improved digestibility and reduced feed wastage. We continuously assess market trends and innovations in aquafeed production to determine if and when it would be beneficial to incorporate extrusion into our processes. AQ: With the increase in raw materials prices, what is Waterbase’s strategy to reduce the impact on the ﬁnal feed price? Is the company currently utilizing or plans to use novel ingredients or more domestic raw materials? RA: The availability of fishmeal has become increasingly scarce, leading to upward pressure on raw material prices across the aquafeed industry. At The Waterbase Limited, we have a comprehensive strategy in place to mitigate the impact of these challenges on the final feed price and ensure the continued affordability and quality of our feeds.

We are actively working on utilizing novel ingredients, although it may take a while to see the benefits as many of the vendors have not yet ramped up production. The current costs of these raw materials are prohibitively expensive at the moment, and in these tough times when costs are high, we don’t want to pass on any additional burden to the farmer. We are aware that these ingredients, such as insect meal and SCPs, will help reduce our reliance on scarce ingredients and also contribute to sustainability. We continue to invest in research and development to enhance performance and optimize costs through the inclusion of these alternative ingredients. Moreover, we are committed to sourcing more domestic raw materials wherever feasible. This not only supports local suppliers but also reduces our dependence on expensive imported raw materials, which can be susceptible to price fluctuations and supply chain disruptions. Additionally, we have implemented efficient supply chain management practices to optimize the sourcing and procurement of raw materials. This includes establishing strategic partnerships with suppliers and closely monitoring market trends to make informed decisions. While these measures help us manage raw material challenges, our primary focus remains on maintaining the highest standards of feed quality and nutritional value. We understand that the well-being and productivity of shrimp farmers depend on the quality of feed, and we are dedicated to ensuring they receive the best possible products. AQ: Does the company also provide aquatic health care products for farmers? RA: Yes, Waterbase does provide aquatic healthcare products for farmers as part of its comprehensive aquaculture solutions. We understand that maintaining the health of shrimp is crucial for a successful crop. Our wide range of aquatic health care products includes products for managing water and soil quality parameters, immunity boosters, growth promoters, stress busters, disease management products, and other inputs like minerals that contribute to the well-being and health of the shrimp. We have developed a wide range of products that promote sustainability and farmer profitability. Our commitment to providing a holistic solution to shrimp farmers goes beyond just feed production. We aim to support farmers at every stage of farming, from the initial stages of seeding to grow-out and beyond.

Aquafeed: Advances in Processing & Formulation Vol 15 Issue 4 2023

INTERVIEW AQ: As a vertically integrated company, Waterbase operates its own shrimp grow-out farms and shrimp processing facilities. What are the main markets for processed shrimp? How is the company diving into the current low-price context? RA: Our shrimp are sold in all major markets, including Europe, the US, and China. We also export to various countries in Asia. The entire sector is grappling with the impact of low pricing, and we are no exception. We are primarily focusing on markets and customers who value sustainability, traceability, and quality consciousness. AQ: How do you see the shrimp market and what needs to be done in India? RA: The current situation is expected to persist for a while. Unless global production adjusts itself to demand, we will not see an uptick in international prices. India needs to focus on the 4Ds to tackle the current challenges posed mainly by increased global supply, contracting demand, and rising production costs:

Drive domestic consumption which is critical to have a strong domestic market to insulate us from the vagaries of international prices and demand. Concerted efforts are needed from all stakeholders to boost shrimp consumption. Diversify species since depending solely on vannamei may be too risky. The adoption of other species like SPF black tiger shrimp and indicus will bode well for the country. Disease management and diagnostics since production in India has increased primarily due to expanding the area under cultivation rather than through increasing productivity. The adoption of technology and good farm management practices, supported by disease management and diagnostics, will help boost the profitability of farmers by increasing productivity and survival rates. Defend cost structure since supply has brought down prices and the market is becoming increasingly competitive. It is very important that we defend our cost structure through building efficiencies and policy interventions.

Wenger.com

The Future Awaits Built on partnership and innovation, Wenger is providing more opportunities for client success. For almost a century, Wenger has delivered extrusionbased innovations to our partners. We’ve worked alongside you to develop new processing solutions and better products, providing our industry-leading expertise and ongoing support every step of the way. We don’t plan on stopping any time soon. Wenger’s global food processing family is growing, and we look forward to the exciting opportunities that lie ahead. We will continue to deliver even more innovations and technologies to benefit companies that share our vision of tomorrow.

Aquafeed: Advances in Processing & Formulation Vol 15 Issue 4 2023

NEWS

NEWS REVIEW Highlights of recent news from Aquafeed.com

De Heus opens aquafeed mill in Vietnam

The company officially opened its new aquafeed mill at Tra Noc Industrial Zone, Can Tho, Vietnam. The feed mill has a production capacity of 240,000 tons per year and is dedicated to producing pangasius feed. The strategic location near the river port provides outstanding advantages for transporting raw materials to the plant and finished products to farmers.

Phodé reinforces its range of aquafeed additives After launching its Olpheel range dedicated to shrimp and fish farming in 2022, Phodé has presented three new aquaculture products this year, aiming to manage different issues. Olpheel Zen is a sensory solution mainly composed of plant extracts allowing a state of well-being in fish. Olpheel Anti-Ox is an antioxidant of 100% natural origin made from grape extracts. Olpheel Eat is a neurosensory solution composed of selected attractive and botanical compounds to stimulate feed intake.

Tietjen develops machine for primary crushing The company unveiled a new crusher suitable for crushing agglomerates or pre-crushing coarse material pieces for further grinding. Crusher CR is a versatile machine for the coarse crushing of bulk materials that saves costs by pre-crushing. By homogenizing the raw material in the crusher, energy is saved in the further grinding in the hammer mill since the desired grain spectrum is achieved more quickly. At the same time, the screens of the hammer mill are protected against screen breakage. In fish feed production, where fine screens are used, the use of the crusher is particularly worthwhile. The crusher is designed for continuous industrial operation.

dsm-firmenich enters the single-cell proteins arena dsm-firmenich has developed new single-cell proteins (SCPs) for animal feeds. “dsm-firmenich has a vast library of microbial strains of algae, fungi, bacteria and yeasts. For smart protein, we are working with our library of single-cell proteins from bacteria and yeast," Karim Kurmaly, director Smart Protein at dsm-firmenich, told Aquafeed.com. Trials on rainbow trout grown for 12 weeks on different inclusions of SCP ranging from 0%, 5%, 10% to 20% on extruded feeds, with single cell protein replacing a combination of fishmeal and soy protein concentrate, showed that the inclusion of SCPs has no negative impact on fish performance measured by final body weight.

Aquafeed: Advances in Processing & Formulation Vol 15 Issue 4 2023

NEWS ViAqua Therapeutics completes $8.25 million investment to scale RNA-based solutions in aquaculture The company completed a USD 8.25 million round led by S2G Ventures with participation from Rabo Ventures, The Trendlines Group Ltd., Agriline Limited, Nutreco, I-Lab Angels and Circle Investments LLC. ViAqua has developed a biotechnology-based oral delivery platform for the targeted administration of RNA-based solutions to improve disease resistance in aquaculture. The company's first product is a feed supplement to enhance resistance to viral infections in shrimp, with the initial application targeting White Spot Virus (WSSV).

Adisseo to build powder methionine plant in China Adisseo plans to build a new powder methionine plant with an annual production capacity of 150,000 tons in Fujian Province in China. The total investment amount is expected to be around RMB 4.9 billion (USD 680 million). The plant is expected to operate in 2027. The 150,000 tons/year powder methionine project will be located at Quanhui Petrochemical Industrial Zone, Quanzhou City, Fujian Province, China. Being adjacent to SinoChem sister company, the project is expected to benefit from strong synergies in the supply chain, receiving operational support and further gaining cost advantages by optimizing capital investments and operating costs as well as enhanced whole value-chain sustainability.

Anton Paar acquires Brabender Anton Paar acquired the German company Brabender, which will be integrated into the Anton Paar Group as Anton Paar TorqueTec GmbH. Brabender, based in Duisburg, Germany, offers measurement and process engineering solutions for the testing of various raw materials and for recipe and process development. It covers a wide range of applications – from food and feed to plastics and rubber, and even batteries and other special applications. A smooth integration of Brabender into the Anton Paar Group is planned and products and services can be purchased directly via the Brabender website and sales organization.

40 companies registered to compete in the F3 Krill Replacement Challenge Forty companies from across the world have registered for the F3 - Future of Fish Feed’s latest aquaculture feed contest — the F3 Krill Replacement Challenge. This current contest from the F3 initiative is designed to spark innovation for alternatives to krill in aquaculture feed. The companies that registered for the F3 Krill Replacement Challenge have a variety of high-quality products ranging from black soldier fly and algae to single-cell proteins. The next step is for the F3 judges to select the ten companies that will compete in the 12-week comparative feeding trial in which each competitor’s marine-animal-free product will be incorporated into an F3-designed, plant-based feed for Atlantic salmon. Aquafeed.com has been interviewing some registrants of the F3 Krill Replacement Challenge. Have a look at the Editor’s picks section on our website.

Aquafeed: Advances in Processing & Formulation Vol 15 Issue 4 2023

NEWS New trial results show PEKILO mycoprotein beneficial to salmon growth and health Finnish biotechnology company Enifer shared new trial results that detail the positive effects of their mycoprotein product PEKILO-Aqua, produced on side streams from Nordic wood pulping, on salmon health and growth. In a first trial, it was used in varying amounts to replace other common protein ingredients in salmon feed including soy protein concentrate (SPC), vital wheat gluten and fishmeal, while maintaining the total nitrogen, lipid and energy contents of the feeds equal. The growth trial was carried out with juvenile salmon in freshwater. The studies showed that not only could the mycoprotein replace conventional protein ingredients, but the feed conversion ratio in fact improved in a linear fashion with higher mycoprotein inclusion in the feed. Improvement was also seen in the nutrient utilization efficiency.

Besides a crude protein content of ~65%, the PEKILO-Aqua ingredient contains high levels of nucleotides (>10%) and β-glucan (~15%). Researchers wanted to know if they can bring benefits over conventional protein ingredients. And this is exactly what the researchers uncovered. The PEKILO-Aqua ingredient induced a systemic immune response in the fish. Even low inclusion (5%) of the ingredient led to a strong T-cell response, innate immunity responses and enhanced antimicrobial activity. With higher inclusion levels (10-20%), activation of T1 helper cells and specific responses to β-glucan were also evident. As more direct evidence of increased immune activity, the studies showed that the salmon fed PEKILO-Aqua had higher levels of antibodies against the common bacterial pathogen, Vibrio anguillarum, which can wreak havoc on farms. The trials were carried out by a group of Norwegian researchers headed by Professor Margareth Øverland at NMBU University and results were presented at Aquaculture Europe 2023 in Vienna.

Flylab, Sanyo Trading Asia to develop insect-based feeds

Extru-Tech conducts Pakistan roadshow

Sanyo Trading Asia, a subsidiary of Sanyo Trading Co., Ltd. in Japan, and Flylab have been collaborating for over a year after closing a USD 1 million in SEED and Pre-Series A round in 2022 led by Sanyo Trading. The new agreement marks a significant turning point in their partnership. Highlights of the agreement encompass ongoing negotiations aimed at securing a 6,000 metric tonnes supply contract for Flylab’s signature product, FLMeal, over the next three years. To bolster the venture, a state-of-the-art factory will be established in the Bangkok area, slated to be fully operational by the second quarter of 2024.

On August 22, 2023, a seminar was convened on the Principles of Extrusion and Technology for Feed and Food Production, sponsored by Extru-Tech, Inc., in conjunction with the U.S. Department of Commerce. The Pakistan Extru-Tech Extruder Technology Seminar and Forum, the first-of-its-kind event was held in Lahore, Pakistan and focused primarily on aquafeed and pet food. Developing countries like Pakistan are particularly interested in acquiring the flexibility to change aquafeed products as their business evolves, without the need for significant capital expenditures.

Aquafeed: Advances in Processing & Formulation Vol 15 Issue 4 2023

SUPPORTING LOW FM/FO

Alternatives to marine ingredients in aquaculture to improve sustainability and profitability through nutrition Samuel Correa, Bioiberica Animal Nutrition

The production of food to nourish the current population is highly complex and with great challenges: the continuous growth of the world population is certainly accompanied by a growing demand for quality proteins, a key element to ensure adequate food and nutrition throughout the world. Given this situation, animal protein of marine origin, becomes a product of great relevance thanks to its nutritional value, the contribution of quality protein and its healthy profile of fats and other important nutrients. In this sense, the aquaculture industry has a key role: it is a necessary industry to produce quality protein and be able to nourish the population, and in turn, it can help counteract the use of ingredients of marine origin in the feed of farmed

species. The ingredients obtained by means of direct fishing and their derived processes are not sustainable in the long term and it makes sense to look for alternative products more respectful to the environment.

Raw materials in aquaculture Nutrition plays a key role in aquaculture production. Traditionally, it has been based on a few raw materials that worked very well, such as fishmeal or fish oil. Today, with the paradigm shift that we have experienced in the last decade, there are four factors that are increasingly important and that raw materials must meet: sustainability, supply chain stability, biosecurity and profitability.

Aquafeed: Advances in Processing & Formulation Vol 15 Issue 4 2023

SUPPORTING LOW FM/FO Sustainable ingredients. The planet lives immersed in a process of climate change and global warming, promoted mainly by industrial activities, pollution and the release of greenhouse gases into the environment. This serious problem, which affects animal species, ecosystems and human populations, threatens to change the world as we know it. The raw materials of the linear economy are not products that meet sustainability requirements. Stability in supply. Basic commodities are constantly suffering from price fluctuations and disruptions in supply chains as a result of climate change and market speculation. These situations, very common nowadays, generate a lot of instability in users and considerably decrease profitability. It is important to try to work with stable values and products with little variation, since in the long run they will be more profitable. Biosafety. In the current landscape of change and uncertainty, diseases that affect both humans and animals are increasingly likely and common, affecting people's health, protein production, and the profitability of animal production businesses. Feed raw materials, especially those of animal origin such as PAPs (processed animal proteins), are the most at risk of generating global health problems. It is important to use ingredients that, regardless of their origin, have the highest quality certificates and the most robust biosecurity schemes. Efficient ingredients. The most important thing in any business, even in aquaculture, is to have good profitability. It is often assumed that profitability is achieved by lowering production costs, although the opposite effect is often achieved: cheap is expensive. It is worth working with raw materials and ingredients that, if a priori can cost more, in the long run, will have a greater economic profitability thanks to a productive improvement and a reduction of risk and problems. Through a correct nutritional strategy, we can help achieve these objectives in aquaculture. For example, fishmeal is a classic source of protein that has been used in large quantities in aquaculture nutrition and other animal species. Its convenience, qualities and characteristics make it a good product, especially for fish and shrimp. Even so, this raw material does not meet sustainability requirements, its biosecurity may be compromised and, now more than ever, price volatility and supply chain stability due to climate change are factors that can greatly condition aquaculture

production systems and their profitability. Therefore, it becomes increasingly important to look for alternatives and solutions that allow replacing these types of ingredients and reducing the dependence we have on them. In this article, we propose two replacement strategies: direct replacement and indirect replacement, using a series of high-value functional ingredients that meet all the mentioned requirements.

Replacement of fishmeal with land animal proteins Hydrolyzed porcine intestinal mucosal proteins are a protein source with a very high potential for this purpose. They are a co-product of the manufacture of heparin (which is extracted from the intestinal mucosa of pigs), one of the most widely used active pharmaceutical ingredients in human health. These porcine protein hydrolysates meet all requirements. They are sustainable, as they come from the circular economy as co-products of the pharmaceutical industry. They are totally biosafe since they share industrial processes and specific safety measures with heparin, a drug that is injected into humans. They are stable in supply and production, since, being a pharmaceutical co-product, their supply chain is secured and traceable at all steps. And finally, they are highly profitable: their origin and nutritional qualities (high digestibility, palatability and bioactive peptides that provide the nutrients necessary for the development of intestinal health) make their use as a substitute for fishmeal to increase profitability in aquaculture production. In a study on salmon (Segarra et al., 2020), conducted in Chile, fishmeal was partially replaced by two levels of porcine intestinal mucosal protein hydrolysate with 50% protein (Palbio 50) (Fig. 1). The objective of this study was to achieve nutritionally equivalent diets, but using less fishmeal, without losing productive performance and with greater profitability.

Palbio 50

Fishmeal

Diet 1

GROUP 1

20%

Diet 2

GROUP 2

2.5%

17.5%

Diet 3

GROUP 3

15%

Figure 1. Design of study diets

Aquafeed: Advances in Processing & Formulation Vol 15 Issue 4 2023

SUPPORTING LOW FM/FO The results of the study did not show significant differences between groups (Fig. 2), suggesting that the inclusion of 2.5% and 5% of hydrolysate as a replacement for fishmeal is equally efficient at the productive level (weight gain, conversion ratio, etc.). and that, therefore, it is a valid source to replace fishmeal. In addition, the inclusion of this product made it possible to reformulate the diets with a reduction in costs, which was about 2%.

profitability of aquaculture businesses. In order to make an effective substitution with vegetable meals, it is advisable to incorporate functional ingredients and products. This small investment will allow us to contribute to the development of intestinal health and immunity, and be able to take much better advantage of vegetable meals to get the most out of them and be able to obtain greater profitability. An excellent candidate for this purpose is free nucleotides. Free nucleotides are active ingredients of high nutritional interest in aquaculture, and in fact, their positive effect on fish and shrimp nutrition is well known (Fig. 3). As the main unit of DNA and RNA, nucleotides participate in the processes of tissue regeneration, growth and development of systems, having a main role in accelerating the growth and strengthening of the immune system, generating greater resistance and survival in the face of challenges and diseases. They also have a positive impact on the intestinal system: the greater the development, the greater the capacity for digestion and absorption of nutrients, as well as the greater the capacity for regeneration of damaged or injured tissues. This opens the door to the use of cheaper protein sources of lower nutritional quality than

Figure 2. Productive results in salmon: Weight, SGR, FCR, GF3 and SFR.

Indirect replacement increasing the quality of the feed As we have seen, replacing fishmeal with other protein sources is a common strategy. Plant-based proteins have been widely used for this purpose, due to their generally high availability and low cost. Even so, their nutritional qualities, digestibility, protein concentration and anti-nutritional factors will never make them a raw material to compete directly against fishmeal. Therefore, substitution with vegetable meals can greatly reduce the production cost, but also the yield, and worsen the

Figure 3. Chemical structure of nucleotides

fishmeal, the main ingredient of this type of diet. As we have already mentioned, the use of vegetable proteins such as soy, as a substitute for fishmeal, is a priori negative at the nutritional level, but positively impacts the cost of diets. The use of nucleotides, according to different studies in fish and also recently in shrimp, allows greater use of protein and softens the negative impact that anti-nutritional factors can

Aquafeed: Advances in Processing & Formulation Vol 15 Issue 4 2023

SUPPORTING LOW FM/FO Table 1. Design of study diets

Control

Diet 1

Diet 2

Diet 3

Diet 4

Diet 5

Diet 6

Diet 7

Diet 8

Diet 9

Fishmeal (%)

Soybean meal (%)

45.85

45.84

44.8

47.5

51.85

Nucleotides (%)

0.05

0.1

0.05

0.1

0.05

0.1

0.05

0.1

generate, thanks to the improvement of the digestive and intestinal systems and a greater regeneration and recovery of tissues. This makes it possible to achieve productivity levels not lower than those achieved with fishmeal but to improve the sustainability and profitability of the production. In addition, these studies also confirm that beyond nutrition, these bioactive molecules contribute to the development of the immune system and generate greater resistance and survival against diseases and biological challenges. This is also a great advantage at the productive and economic level and gives peace of mind to the producer and the industry in general. In a recently published shrimp study (Novriadi et al., 2021), two nucleotide levels (500ppm and 1,000ppm) were used with different levels of fishmeal reduction and inclusion of soybean meal (Table 1). The results showed how the inclusion of nucleotides showed the same weight after 70 days of study, even when the fishmeal was replaced almost entirely (Table 2). This is explained by the capacity of development and regeneration of the intestinal and digestive systems of the nucleotides, which allowed a greater use of the

nutrients of soybean meal. In addition, in the same study, certain immune parameters were evaluated and shrimp survival was measured 7 days after an infection with Vibrio harveyi at the end of the experiment. Shrimp fed the nucleotides significantly improved (p<0.05) the count of hemocytes, the immune cells of the shrimp, as well as the activity of lysozyme, a protein with a high antimicrobial capacity (Fig. 4, 5). Nucleotides help the maturation and tuning of the immune system, activating the responsible cells and promoting the production of defense proteins in shrimp. Thanks to this, shrimp-fed nucleotides had a significantly higher survival (p<0.05) than the control groups 7 days after infection (Fig. 6), regardless of the degree of replacement of fishmeal by soybean meal. With these results, nucleotides are presented as excellent candidates to contribute to the maintenance of the overall health of shrimp, and thanks to their addition to the diet, use of high levels of soybean meal without losing productivity while contributing to immunity and smoothing losses to any challenge or period of stress that may exist in the production system.

Table 2. Production parameters at the end of the experiment (70 days). The initial mean weight was 4.24 ± 0.3g.

Final biomass (g)

Final mean weight (g)

Survival (%)

WG1 (%)

FCR2

TGC3

Control diet

217.43

16.50

87.78

290.99

2.80

0.0463

Diet 1

222.97

15.74

96.67

272.78

2.95

0.0443

Diet 2

219.02

16.21

90.00

283.59

2.85

0.0455

Diet 3

215.41

16.65

86.67

294.89

2.79

0.0467

Diet 4

232.42

16.58

93.33

292.00

2.80

0.0464

Diet 5

235.40

16.76

93.33

298.48

2.78

0.0469

Diet 6

231.73

16.55

93.33

291.52

2.79

0.0464

Diet 7

215.37

16.38

87.78

286.60

2.83

0.0458

Diet code

Diet 8

230.98

16.33

94.43

286.09

2.83

0.0472

Diet 9

225.55

16.86

88.89

300.38

2.75

0.0472

P-value

0.5834

0.6238

0.1194

0.5738

0.5358

0.5855

PSE

6.8429

0.2924

2.1505

7.0733

0.0469

0.0007

WG1 = Weight gain; FCR2 = Feed conversion ratio; TGC3 = Thermal growth coefficient; PSE4 = Pooled standard error

Aquafeed: Advances in Processing & Formulation Vol 15 Issue 4 2023

SUPPORTING LOW FM/FO

Figure 4. Hemocytes count. Adapted from Novriadi et al., 2021.

Figure 5. Activity of lysozyme. Adapted from Novriadi et al., 2021.

Figure 6. Survival 7 days after infection with Vibrio harveyi. Adapted from Novriadi et al., 2021.

Aquafeed: Advances in Processing & Formulation Vol 15 Issue 4 2023

SUPPORTING LOW FM/FO improve their sustainability. Conclusion Faced with current uncertainties and the great challenges of the world, aquaculture can provide solutions, specifically, through aquaculture precise and sustainable nutrition. Working efficiently with tailor-made strategies and ingredients as specific as nucleotides or hydrolyzed proteins of the porcine intestinal mucosa, big gains can be achieved and do our bit to make production more efficient, more economical, more profitable, and more sustainable. Studies and references are available upon request.

More information:

Using nucleotides intelligently, we can improve the competitiveness of our production by minimizing the cost of diets and increasing profitability, while we can BIO_2023_AnimalNutrition_NucleoForceAqua_anunci 177x114_AF.pdf

Samuel Correa Technical Manager Bioiberica Animal Nutrition E: scorrea@bioiberica.com

5/10/23

11:36

NATURAL GROWING INNER PROTECTION

The real value is in free nucleotides Nucleoforce is a functional ingredient for formulating precision aquaculture diets and supplementing the animal’s needs during early stages, stress conditions and tissue recovery periods.

Follow Bioiberica on:

Aquafeed: Advances in Processing & Formulation Vol 15 Issue 4 2023

For more information: animalnutritioncontact@bioiberica.com

SUPPORTING LOW FM/FO

Digestive and metabolic enhancer offsets reduced cholesterol level in shrimp feed Marleen Dehasque, Waldo G. Nuez-Ortín, Martin Guerin, Adisseo SAS

Global fishmeal and fish oil prices are, to a large extent, linked to the supply situation in South America (Peru and Chile) and demand from Asia (primarily China). Currently, Peru is the largest source of fishmeal (FM) and fish oil (FO) output and also the world’s largest producer and exporter of FM and FO. An unsuccessful season in Peru might cause as much as a 20% decrease in global output. Impacted by El Niño, Peru has recently canceled the first fishing season for anchovy in the north-central zone, creating new challenges for the global market of FM and FO.

Impact on essential lipids Cholesterol is an essential nutrient for shrimp. It is a key compound of membranes and a precursor for the synthesis of ecdysteroid molting hormones. Reducing FM and FO in shrimp feeds will directly affect the level and availability of essential lipids such as n-3 HUFA, phospholipids and cholesterol. Cholesterol requirement studies show a wide range of cholesterol requirements from 0.5 to 5 g/kg for L. vannamei (Chen, 1993; Duerr & Walsh, 1996; Gong et al., 2000). Practical shrimp diets are usually formulated with cholesterol levels starting from 0.07-0.1%. Lower levels than 0.07% are considered insufficient. Purified cholesterol can also be added to reach a targeted level. These sources are, however, becoming very expensive, therefore, requiring more cost-effective alternative options. In times of high FM and FO prices, it is important for nutritionists to search for strategies to fulfill the cholesterol requirement in shrimp feeds. One of the solutions is LIPOGEST®, a digestibility and metabolic enhancer based on bile salts. Bile salts constitute an

Table 1. Experimental feed formulation and chemical composition

Ingredients (%)

CHOL8

CHOL6

Peruvian fishmeal

Tuna fishmeal

Soybean meal

34.38

34.11

Rapeseed meal

Corn gluten

Wheat bran

Whole wheat

19.09

20.39

Rice bran

Squid meal

Vit/min premix

Lecithin (liquid, 60% PL)

1.5

Fish oil

0.5

Cholesterol (92%)

0.025

Total

100

Crude protein

37.35

37.31

Crude fat after hydrolysis

6.35

5.61

Crude ash

7.56

7.66

Crude fiber

3.2

3.3

Analysis (as % product)

HUFA

0.89

0.75

Cholesterol

0.080

0.064

Moisture

11.46

10.62

alternative source for the steroid ring and, therefore, can maintain the synthesis of molting hormones in situations where cholesterol levels are deficient in the diet (Lin et al., 2017). Furthermore, bile salts improve the digestive capacity for lipids in the digestive system of shrimp by improving lipid emulsification

Aquafeed: Advances in Processing & Formulation Vol 15 Issue 4 2023

SUPPORTING LOW FM/FO Table 2. Effect of LIPOGEST® on the performance of white shrimp L. vannamei fed a diet containing a low level of cholesterol. Different letters show significant differences (p<0.05). CHOL8: positive control with a cholesterol level of 0.08%; CHOL6: negative control with a cholesterol level of 0.06%; LIPOGEST: 1.25 kg/MT added to the CHOL6 feed.

CHOL8

CHOL6

LIPOGEST

Survival (%)

90 ± 0.00

91.67± 2.89

90 ± 5.00

Initial weight (g)

1.05 ± 0.01

1.06 ± 0.00

1.05 ± 0.02

Final weight (g)

12.44 ± 0.84ab

10.83 ± 0.32b

12.21 ± 0.49ab

SGR (%/d)

4.42 ± 0.11a

4.15 ± 0.05b

4.38 ± 0.1ab

Growth (g/wk)

1.42 ± 0.1ab

1.22 ± 0.04b

1.39 ± 0.06ab

Feed intake (g)

28.58 ± 1.82

28.47 ± 2.06

27.16 ± 0.72

FCR

2.51 ± 0.03

2.91 ± 0.13

2.44 ± 0.11a

PER

1.07 ± 0.01a

0.92 ± 0.04b

1.10 ± 0.05a

and micelle formation, resulting in faster absorption of lipids in the hepatopancreas.

Shrimp trial A feeding trial with white shrimp (Litopenaeus vannamei) demonstrated the effective replacement of FO and purified cholesterol with LIPOGEST®. As shown in Table 1, the positive control feed (CHOL8) was formulated to reach an optimal cholesterol level of 0.08%. The negative control feed (CHOL6) was formulated at a cholesterol level of 0.06% by lowering the fish oil and by omitting the purified cholesterol compared to the positive control diet. To this diet, 0.125% bile salt-based LIPOGEST® was added and compared to CHOL8 in an eight-week feeding trial. Reducing the level of cholesterol significantly affected growth and feed conversion in white shrimp (Table 2). However, the addition of LIPOGEST® improved the growth compared to the negative control and restored the performance of the shrimp to the same level as that of the feed with higher cholesterol levels. FCR of the LIPOGEST® treatment was similar to that of the feed with higher cholesterol levels. Such improvements can be attributed to a more optimal digestion efficiency and regulation of cholesterol metabolism. In this line, Lin et al. (2017) demonstrated the significant effects of adding bile salts in cholesterol-deficient soybean mealbased diets for white shrimp. Furthermore, this study showed that several ecdysteroid signaling response genes, such as the expression of ecdysteroid receptor, chitin synthetase

and muscle growth markers, were improved by adding bile salts in cholesterol-deficient diets. In a more recent trial, the supplementation of bile acids at 0.025-0.1% in 14% FM shrimp feeds reduced the FCR and increased weight gain (Wang et al. 2023). Altogether, these results show that bile salt supplementation can mitigate the adverse effects of cholesterol deficiency caused by low FM inclusion.

Conclusion In conclusion, within the current context of high prices of FM, FO and cholesterol, aquafeed formulators can count on LIPOGEST® to make aquafeeds perform with less marine lipids and cholesterol and, therefore, achieve significant cost-reductions without losing the performance and metabolic activities related to dietary cholesterol. References available on request

More information: Marleen Dehasque Global Product Manager Aqua Nutrition Adisseo SAS E: marleen.dehasque@adisseo.com

Aquafeed: Advances in Processing & Formulation Vol 15 Issue 4 2023

ADDITIVES

SmartMoisture®: Improving cost and efficiency of aquafeed production Dr. Antonios Chalaris, Devenish

The global challenge of driving efficiencies Feed represents over 50% of the production cost in intensive aquaculture. Soaring ingredient and energy costs have resulted in significant feed price increases in recent years, and it seems highly unlikely that these will drop significantly anytime soon. Managing costs in aquafeed production is becoming increasingly important. Any strategy to reduce cost has to find a balance between optimal formulation cost and impact on production efficiency, while also maintaining feed quality, and fish and shrimp performance. A common but misunderstood component and one

that has a direct impact on the cost and efficiency of aquaculture production is moisture management.

The role of moisture in aquafeeds Aquafeed requires high moisture addition during processing. Moisture is added at various stages of production to ensure optimal cooking and starch gelatinization for pellet binding and expansion which is important for post-extrusion oil addition and consistent pellet quality. Moisture extends its role to the safety of aquafeeds. Hot temperatures cause water to migrate during storage,

Aquafeed: Advances in Processing & Formulation Vol 15 Issue 4 2023

ADDITIVES while humid conditions will result in condensation, both resulting in higher activities of free water and facilitating microbial growth. By managing water activity (aW) during processing, storage, and transportation, the proliferation of microorganisms can be reduced and ensure aquafeed safety. It is not just about optimizing and safeguarding the current levels of moisture but also increasing them. Elevated levels of moisture in aquafeeds have been well documented to result in much better absorption of nutrients by the fish and shrimp. This creates a lot of downstream performance benefits, such as improved feed quality and durability, and increased feed intake and performance.

How SmartMoisture® improves cost efficiency, quality, and safety The patented technology of SmartMoisture® is based on a unique blend of glycerol esters of propionic acid, and surfactants. Glycerol acts as a humectant and emulsifier and can hold up to 1,000 times its own weight. The surfactant reduces water’s surface tension and consequently improves moisture distribution. Glycerol and the surfactant improve water absorption into the aquafeed particles, facilitating steam uptake during extrusion. Aquafeed raw materials need to absorb moisture in a very short period of time, and this is becoming more of a challenge with the increasing inclusion of vegetable raw materials. Vegetable ingredients are generally hydrophobic and do not easily absorb moisture. This results in adding additional moisture which has the effect of increasing drying times, energy consumption, and reducing production throughput. More efficient starch gelatinization and superior pellet quality are effects of increased steam uptake. Specifically, improvements have been recorded in feed durability and reduced levels of dust and fines. Additionally, allowing for a reduction of starch in formulations can drive down recipe costs by creating more space in the formulation thus widening the potential raw material basket. It has been shown that SmartMoisture® improves the absorption and retention of moisture through the aquafeed production process. This efficient use of moisture allows improvements in throughput of up to 18% (Fig. 1), drier capacity, and energy savings of up to EUR 11/t (Fig. 2).

Figure 1. Consistent throughput improvements across feed trials.

Figure 2. Energy savings across the trials will contribute to reducing feed costs.

SmartMoisture® is more concentrated than any other product on the market due to the stability of glycerol esters. The propionic acid esterified to glycerol is a patented process used to buffer and stabilize propionic acid. It increases the retention time of propionic acid in feed, ensuring a longer-lasting microbial inhibition. The rate at which mold and bacteria grow after feed processing depends on factors such as pH, temperature, oxygen availability, and most importantly, water activity of the feed. The decision on what the final moisture content will be is based on risk. Lower moisture content in the final feed is targeted considering the volume of moisture used in the extrusion process and the hydrophobic nature of aquafeed raw materials to avoid free water or high water activity. SmartMoisture® technology is very effective in reducing water activity in aquafeed thereby reducing risk and allowing manufacturers to produce feed with higher moisture, improving margins.

Aquafeed: Advances in Processing & Formulation Vol 15 Issue 4 2023

ADDITIVES

In summary, SmartMoisture® provides a holistic solution to driving down costs in aquaculture production. It provides opportunities to reduce feed formulation, energy, and production costs, whilst driving improvements in feed quality. The benefits provided are not only to the feed producer but also to the farmer. Trials with customers have identified possible savings of up to €70 per tonne of feed.

About Devenish Founded in 1952 in Belfast, Devenish is a sciencebased company, headquartered in Northern Ireland, with sites across Ireland, the UK, USA, Mexico, Turkey, and Uganda focusing on developing sustainable food solutions. Today, Devenish manufactures premix, starter feeds and speciality feed additives across eight production sites, which are sold and delivered to over 40 countries worldwide. The company’s strategy is to grow substantially across multiple species and geographical boundaries, with major investments in plant, machinery, and human resource. A major part of this expansion strategy has been aquaculture, where a team of nutritionists work with aquaculture customers around the world, providing bespoke technical solutions and nutritional formulations that help them exceed their targets.

More information: Dr. Antonios Chalaris Aqua Business Development Manager Devenish E: antonios.chalaris@devenish.com

Aquafeed: Advances in Processing & Formulation Vol 15 Issue 4 2023

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MICRO-INGREDIENTS

Optimizing phosphorus nutrient management in aquaculture Frederic Baron, Delphine Weissman, ADM Animal Nutrition

As aquaculture production increases with the rising global population and growing consumer demands for food security, optimizing nutrient management in aquaculture systems is essential to ensure the industry is both efficient and sustainable. Like any industry, aquaculture has its environmental challenges, and one of the most pressing concerns is the management of nutrients found in feed and in the ecosystem, such as phosphorus. Phosphorus is an essential mineral mainly brought through the feed for aquatic organisms, playing a crucial role in their growth and development.

However, excessive phosphorus in aquaculture systems can have detrimental environmental effects, including water pollution, algal blooms, eutrophication and more.

The role of phosphorus As a primary mineral, phosphorus contributes to bone and scale formation. Adequate levels of phosphorous, along with the right balance of other minerals like calcium in the diet, are needed to prevent skeletal deformities and promote proper growth. Phosphorus is also fundamental to deoxyribonucleic acid (DNA),

Aquafeed: Advances in Processing & Formulation Vol 15 Issue 4 2023

MICRO-INGREDIENTS ribonucleic acid (RNA) and adenosine triphosphate (ATP), which are essential primary molecules found in all live organisms. Phosphorus is a key element of phospholipids, which are integral to cell membranes and an important precursor for a range of highly biologically active mediators of the metabolism and physiology of aquatic species (i.e., eicosanoids, inositol phosphate). Phosphorus is involved in many metabolic and physiologic processes needed to guarantee optimal growth, health, reproduction and overall development in fish species.

Challenges of phosphorus management While phosphorus is an important nutrient, excessive phosphorus in bodies of water can trigger eutrophication, leading to algal blooms, oxygen depletion and habitat degradation, which can negatively impact overall water quality. Eutrophication can also disrupt the balance of aquatic ecosystems, leading to shifts in species composition and potential loss of biodiversity. Excessive excretion of phosphorous in the environment can happen if phosphorous is used in excess in feed if its digestibility is low or if the mineral balance of the feed is not correct. One source of phosphorus used in fish feed is collected from open-pit in the form of phosphate, which is a limited resource. Therefore, the aim is to reduce or even eliminate this form of inorganic phosphate added to diets by utilizing phosphorus present in other ingredients. This will improve the cost-effectiveness of the feed and conserve the limited phosphate resource. At the same time, regulatory compliance requirements vary from country to country, as do fines and penalties that vary. Many regions around the globe have established water quality regulations that aquaculture operations are required to follow. Phosphorus management strategies Phosphorus is primarily supplied to fish species through their diets. To mitigate its environmental impact, researchers and feed manufacturers like ADM are continually investing in new nutritional strategies to optimize phosphorus levels and digestibility in feed to match the specific needs of different species at varying life stages. The development of effective nutritional strategies to manage phosphorus waste outputs

without impacting zootechnical performances requires a detailed understanding of phosphorus utilization by the animal according to its source and form (phosphate, phytic acid, hydroxyapatite, phospholipids). Proactive approaches include the use of phosphorus-efficient feeds formulated under dedicated digestible nutrients (precise nutrition), well-balanced mineral input and potential utilization of feed additives like enzymes. For several years, ADM has developed tools to better measure and characterize the digestibility of ingredients for diverse aquatic species. This approach has allowed us to define new digestible nutrients enabling more precise nutrition. Among nutrients like protein and amino acids, the development of a specific “digestible phosphorus” nutrient has been established and evaluated through in vivo zootechnical trials. There was a clear improvement in the prediction of growth performance using this new nutrient versus total or standard ones. The inclusion of a digestible phosphorus nutrient leads to improved animal performance and reduced feed costs and environmental impact by enhancing calcium and phosphorous balance. To complete this nutritional approach, adding exogenous phytase to aquafeeds can enhance phosphorus availability and reduce the need for inorganic phosphates by breaking down phytic acid, which is an indigestible form of phosphorus found in plant-based ingredients. What’s more, phytic acid can represent up to 30% of the phosphorus found in tilapia feeds, for instance. Several ADM studies demonstrated that using exogenous phytase is an effective strategy for improving phosphorus availability in feed without increasing total phosphorus levels and with decreasing phosphorus excretion. Including phytase in aquafeeds can enhance phosphorus availability, especially in diets that rely on plant-based protein sources – ultimately improving phosphorus utilization, increased feed efficiency and overall health and performance, while reducing the need for phosphorus supplementation. In addition to this purely nutritional approach, the aquaculture industry also invests in better feeding management practices and technologies that better filter the waste output of fish species, such as Recirculating Aquaculture Systems (RAS). RAS technology, advanced management and monitoring tools in aquaculture operations can offer improved control over water quality, feeding strategies,

Aquafeed: Advances in Processing & Formulation Vol 15 Issue 4 2023

MICRO-INGREDIENTS

Figure 1. Phytase: an efficient strategy to increase phosphorous availability, without increasing total P and with decreasing P excretion

disease management and overall production efficiency. RAS systems continuously recirculate and filter water within the farm, reducing water consumption and waste release while allowing for better control of the water quality. The control of mineral excretion, like phosphorous, in such a system is important to ensure good water quality. Automatic feeders and feed monitoring systems are used to regulate feeding rates and can be adjusted accordingly based on the growth and behavior of the fish. Efficient phosphorus nutrient management is crucial for ensuring a sustainable and environmentally responsible aquaculture industry. To achieve this, it is important to balance the nutritional requirements of fish species with the aim of reducing phosphorus waste. This requires careful planning, efficient feeding practices and the use of advanced technologies. By adopting these strategies, the aquaculture industry

can increase productivity while minimizing the potential environmental impact of excess phosphorus. In doing so, they contribute to the long-term sustainability of the industry and help protect aquatic ecosystems for future generations. More information: Frederic Baron Aquaculture Nutritionist, Creation Design & Development ADM Animal Nutrition E: Frederic.Baron@adm.com

Delphine Weissman Services Growth Director ADM Animal Nutrition E: Delphine.Weissman@adm.com

Aquafeed: Advances in Processing & Formulation Vol 15 Issue 4 2023

MICRO-INGREDIENTS

Impact of phosphate sources on apparent P digestibility in juvenile Litopenaeus vannamei Jean-Gabriel Reynaud, Phosphea

Phosphorus (P) is an essential micromineral for fish as it contributes to skeleton and exoskeleton maintenance and formation. It is also involved in cellular energy production or in muscle tissue synthesis (NRC, 2011; Milián-Sorribes et al., 2021; Truong et al., 2022). P is mainly supplied by feeding as P concentration in water and phosphorus absorption through gills are low (Lall & Kaushik, 2021). Fishmeal is the main source of protein and phosphorus in aquafeed, but it is step-bystep replaced by plant protein ingredients due to the increase in its price and its lower availability (Morales et al., 2018). However, phosphorus coming from these raw materials is mainly in phytate form, a P-form less available for fish due to their low level of intestinal phytase (Milián-Sorribes et al., 2021; Truong et al., 2022). Dietary deficiency of P leads to several problems such as reduced growth, bone mineralization and fecundity or an increase FCR and fat flesh (Lall, 2002; Mabroke et al., 2013). Supplementation with inorganic feed phosphate (IFP), a common and stable source of phosphorus, is

Table 1. Water parameters, mean (s.d.)

Salinity (ppt)

34.7 (0.26)

Temperature (°C)

30.1 (0.27)

Dissolved oxygen (mg/L)

5.80 (0.22)

Dissolved oxygen (%)

86.9 (2.16)

8.13 (0.09)

Alcalinity (mg CaCO3/L)

118.7 (5.75)

Amminia-N (mg/L)

0.15 (0.05)

Nitrite (mg/L)

0.15 (0.05)

nowadays a good solution to meet P requirements for fish (Morales et al., 2018). Several IFP exist on the market with different characteristics (P content, P water solubility, calcium content or pH). Thus, P digestibility and retention and P level released in the water can vary depending on IFP source (Lall, 2002; Hua & Bureau, 2006; Hossain & Yoshimatsu, 2014; Morales et al., 2018; Hossain et al., 2020). Today, there is still a lack of data on P requirements and digestibility data in fish in general and especially in crustaceans (Truong et al., 2022).

Table 2. Inorganic feed phospate inclusion and nutrient profile, mean (s.d.)

Negative CTRL

MCP

MAP

MSP

Inclusion rate (%)

1.20

1.00

1.10

Moisture (g/100g)

6.53 (0.19)

5.88 (0.09)

5.59 (0.24)

5.81 (0.25)

Crude protein (g/100g)

38.6 (0.31)

38.0 (0.21)

38.6 (0.34)

37.9 (0.25)

Lipid (g/100g)

7.29 (0.20)

7.48 (0.13)

7.41 (0.05)

7.57 (0.10)

Ash (g/100g)

3.55 (0.01)

4.23 (0.02)

4.00 (0.06)

4.15 (0.08)

P (mg/kg)

4,720 (14.1)

7,900 (113.1)

7,860 (14.1)

7,915 (49.5)

Ca (mg/kg)

1,805 (63.6)

3,675 (106.0)

1,755 (7.07)

1,820 (14.1)

Cr2O3 (g/100g)

0.40 (0.00)

0.40 (0.02)

0.42 (0.02)

0.42 (0.00)

Aquafeed: Advances in Processing & Formulation Vol 15 Issue 4 2023

MICRO-INGREDIENTS Table 3. Inorganic feed phosphate specifications

MCP

MAP

MSP

P (%)

22.7

23.5

Ca (%)

N (%)

Na (%)

P 2% citric acid solubility (%)

>95

P water solubility (%)

>85

>95

4-5

4.6-5

In shrimp, one of the main farmed species in the world (FAO, 2020), just three publications are available on this subject (Davis & Arnold, 1994; Zwart, 2018; Lemos et al., 2021). The aim of the present study is to compare the phosphorus digestibility and growth performance of monocalcium phosphate (MCP), monoammonium phosphate (MAP) and monosodium phosphate (MSP) supplemented in low P plant-based diets for juvenile whiteleg shrimp (Litopenaeus vannamei).

Materials and methods Trial system and water parameters The study was conducted at USP Aquaculture Laboratory in Ubatuba (São Paulo, Brazil) for 52 days. The feeding trial was performed in a saltwater recirculated 500L tanks stocked with 39 individuals (100 ind/m3; 4g/ind), with continuous feed delivery (2022h/day, belt feeders), heating, physical and biological water treatment (Table 1). Four replicate tanks were used per treatment. Diets Feed started with 5% biomass and followed daily adjustments according to observation on the presence or absence of pellet leftovers. The basal

diet provides rich nutrients (according to estimated nutrient requirements and recommendations) and palatable pellets for juvenile shrimp to have similar performance to a commercial diet (survival, growth and FCR). Experimental diets were formulated to be isoprotein, isolipidic and isophosphorous with Cr2O3 as a marker. The (negative) control diet was tested with a similar formulation devoid of IFP supplementation (0.50% total P) (Table 2). Added IFP have been characterized in Table 3. They were grinded at 250 μm before mixing with the other ingredients. Pellets were dried and stored in plastic bags at -15°C until use. Analysis of ingredients, diets and feces followed AOAC (AOAC, 2005) for proximate composition (crude protein, lipid, and ash contents) and ICP-OES for mineral determination (phosphorus, magnesium, calcium, and potassium) (EPA, 2014). Parameters and statistics Fecal sample collection was done 5 days/week and 4-6 times/day over the whole trial. Feces stood in water for one hour maximum before being collected. At the end of the experiment, shrimp were weighed to evaluate their weight gain (WG). During the trial, survival and feed intake were recorded to evaluate the survival rate (SR) and food conversion rate (FCR). Apparent digestibility coefficients for dry matter and nutrients (%) of diets and apparent digestibility coefficients for phosphorus (%) of phosphate sources were calculated as follows in Table 4. Data was submitted to normality and homoscedasticity test prior to the application of analysis of variance (ANOVA) or Kruskall-Wallis ANOVA to compare diet performance. Differences between means were analyzed by post hoc Tukey HSD or Dunn tests and considered significant at P<0.05 (Zar, 1984).

Table 4. Determination of Apparent Digestibility Coefficient (ADC)

( [(

( ((

ADC (dry matter, %) = 100 – 100 x

% Cr203 in diet % Cr203 in feces

ADC (nutrients, %) = 100 – 100 x

% Cr203 in diet x nutrient in feces % Cr203 in feces nutrient in diet

([

ADC (phosphate source, %) = (A + B) x C – (A x D) B

Aquafeed: Advances in Processing & Formulation Vol 15 Issue 4 2023

A: contribution of phosphorus in control diet to nutrient content of experimental diet. B: c ontribution of phosphorus in test ingredient to phosphorus content of experimental diet. C: ADC of phosphorus in experimental diet. D: ADC of phosphorus in control diet.

MICRO-INGREDIENTS Table 5. Shrimp performance, mean (s.d.) (P < 0.05)

Negative CTRL

MCP

MAP

MSP

Initial body wt (g/ind)

4.17 (0.17)

4.06 (0.04)

4.11 (0.19)

4.07 (0.04)

Final body wt (g/ind)

17.7b (0.77)

22.8a (0.40)

23.1a (1.32)

23.3a (1.96)

Growth (g/week)

1.82b (0.11)

2.53a (0.05)

2.56a (0.16)

2.59a (0.26)

Weight gain (%)

324.7b (25.7)

461.7a (8.37)

460.9a (27.1)

471.9a (44.7)

FCR

2.07a (0.40)

1.33b (0.11)

1.28b (0.09)

1.34b (0.16)

Survival (%)

76.3b (6.61)

88.1a (6.57)

90.6a (3.75)

86.9a (13.1)

Negative CTRL

MCP

MAP

MSP

Dry matter

70.7 (1.93)

75.7 (1.44)

74.5 (1.00)

74.8bc (1.68)

Crude protein

77.5a (2.22)

81.4bc (1.15)

80.6abc (0.96)

80.2abc (1.51)

Lipid

87.9 (3.10)

92.3 (2.58)

92.6 (1.93)

92.8 (1.33)

Ash

-33.2 (5.03)

-6.68 (8.92)

-20.4 (5.10)

-11.4b (2.81)

53.0a (3.82)

63.3bc (2.05)

65.3cd (1.81)

67.0d (2.37)

79.0ab (5.15)

84.2b (4.59)

88.2b (5.94)

Table 6. Apparent digestibility, mean (s.d.) (P < 0.05)

P sources

Results and discussion Performance parameters Shrimp growth performed well for diets supplemented in IFP: growth and FCR values were comparable to practical farming (Table 5). Survival was also increased. Negative CTRL has shown significant negative effects with at least 15% lower survival, lower growth and thus higher FCR. Overall, there were no significant statistical results in the growth performance of shrimp among all P source treatments.

studies (Davis & Arnold, 1994; Zwart, 2018; Lemos et al., 2021). In this study, P digestibility was the highest for MSP (88.2%). To our knowledge, these results constitute the first comparative reference in the literature between MCP, MSP and MAP in shrimp trials. In the manner of Lemos et al., 2021, who worked on MAP, MCP and DCP, further studies may be conducted to check the ideal supplementation level of these valuable P sources in shrimp diets.

Apparent digestibility coefficient (ADC) Dietary treatments supplemented with each of the three P sources showed no significant difference in ADC of dry matter, crude protein, lipid, ash and P. In addition, ADC nutrients of the test diet were overall higher than those of other trials that used diets with the same P supplements from other suppliers (MCP and MAP) (Lemos et al., 2021). In terms of ADC PIFP sources , the data showed the following trend: MSP (88.2%) > MAP (84.2%) > MCP (79%) (Table 6).

Acknowledgment We would like to thank Professor Daniel Lemos and his team from the University of Sao Paolo (Brazil) for their great contribution.

Conclusion Inorganic feed phosphate sources tested are adapted ingredients to support growth in shrimp. They showed higher P digestibility values compared to previous

References available on request.

More information: Jean-Gabriel Reynaud Product Manager Phosphea E: jeangabriel.reynaud@phosphea.com

Aquafeed: Advances in Processing & Formulation Vol 15 Issue 4 2023

ADDITIVES

Heads or tails? Both! Oddgeir Oddsen, Prochaete

“When it comes to Gain PRO, I really do think that exclamation marks are called for,” OddGeir Oddsen, managing director of ProChaete, can hardly contain his enthusiasm for the feed additive Gain PRO. “With its unique blend of ingredients and nutritional benefits, Gain PRO is not only a high-quality protein source for fish and animal feed but also a great example of the kind of product the world needs right now to provide a growing population with healthy food.”

Full circle As the world’s population grows and climate change challenges us, the need for more efficient use of our resources is more important than ever. “When you eat shrimp, you only eat about half of it. The rest (head and shell) is usually discarded. Byproducts of shrimp farming are regarded as waste – which means

you’re throwing away half of your product. That’s just bad business, both for the company and the planet.” Gain PRO is the result of ProChaete’s philosophy. “I am proud to work at a company whose core values are innovation and sustainability. This is very inspiring and motivates me to focus on finding ever better ways to develop a healthy and sustainable business that’s also profitable.”

Fresh, functional and future-proof Gain PRO is a powder made from byproducts of shrimp production that can be added to fish and animal feed. There are many varieties in the feed market that use byproducts from food production, but what sets Gain PRO apart is that the raw material is fresh. Shrimp deteriorate quickly, especially in hot climates. Fermentation and rot start almost immediately,

Aquafeed: Advances in Processing & Formulation Vol 15 Issue 4 2023

ADDITIVES causing a loss of nutrients. But if you collect the heads and shells while they’re still fresh, you have a raw material that is a valuable and versatile resource for feed manufacturers. Gain PRO has many functional qualities that other feed additives lack. Minerals and vitamins are essential of course, but Gain PRO also contains high levels of antioxidants and natural pigments. It also has a probiotic effect that helps digestion and nutrient intake, providing a healthier feed for the fish. “I hope that feed manufacturers will discover the positive effects of Gain PRO and that it will inspire them to use byproducts more effectively. While Gain PRO is a great product, the solution to more sustainable food production is something that requires a collective effort across the entire business.” “With its unique blend of ingredients and nutritional benefits, Gain PRO is not only a high-quality protein source for fish but also a great example of the kind of product the world needs right now to provide a growing population with healthy food.”

Gain more! The University of Texas A&M and Sea Farms Nutrition Ltd have conducted a study that evaluates the use of shrimp byproducts as a feed ingredient in growth trials.

Figure 1. Feed conversion ratio (FCR) in an 8-week trial with Gain PRO10%. Feed conversion ratio is the amount fed in grams weight gained, the lower the number the more efficient feed is utilized.

The method Super fresh shrimp byproducts such as heads and shells have been processed in a gentle manner and added to tilapia feed. The dose and response have been measured. The results The use of Gain PRO showed a clear boost to the animal’s performance. Gain Pro has a functional and nutritional input. Using Gain PRO up to 5% gives a good boost in growth and feed efficiency.

More information: Figure 2. Specific growth rate (SGR) in an 8-week trial with Gain PRO. Final weight increased with the use of 2.5% Gain PRO. SGR is the Specific Growth Rate, this is % weight gain per day from the start of the trial to end of trial, the higher the number indicates better performance.

Oddgeir Oddsen Managing Director Prochaete E: oddgeir@prochaete.com

Aquafeed: Advances in Processing & Formulation Vol 15 Issue 4 2023

Holt Paulsen

FINAL WEIGHT ENCREASED WITH USE OF 2,5% GAIN PRO

Gain PRO Functional proteins for fish

Gain health Gain taste Gain pigmentation

Make your feed more valuable Meet Gain PRO, our innovative feed ingredient crafted from residual shrimp raw material. Not only does it deliver essential minerals, vitamins, antioxidants, and natural pigments, but it also boasts a pro-biotic effect for improved digestion and nutrient absorption. The science is clear: Gain PRO significantly boosts animal performance, growth, and feed efficiency, making it a game-changer in sustainable food production. Gain PRO — where sustainability meets profitability.

RESEARCH

You can only improve what you measure! Mikael Herault, Clément Martineau, Dachawat Poonnual, Freddy Balbi, Symrise Aqua Feed

In the aquafeed industry, more and more stakeholders own their testing facilities to control and improve the performance of their products and customize them to various species, growth stages and rearing environments. Historically, most big aquafeed manufacturers were owning in-house testing facilities to develop and assess dietary tailor-made formulations meeting the nutritional requirements of various aquaculture species, and their specific growth stages. More recently, they started to develop feed ranges dedicated to rearing environments (RAS farms) or environmental challenges (summertime or wintering). Today, even medium-sized and small

aquafeed manufacturers have set up their own testing facilities to optimize their testing capacities, operational costs and control over the methods and results. Regarding the feed additive suppliers, in many cases, they will rely on studies implemented on terrestrial mammals to promote their products to the aquaculture industry. Most committed ones will contract public institutions, such as universities or private CROs, to assess the value of their technical solutions in some very specific conditions. However, very few of them have the dedicated budgets to invest in several aquaculture-dedicated studies and/or set up aquaculture testing facilities.

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RESEARCH team of passionate researchers and technicians relies on internal and external testing facilities, to assess the performance of the technical solutions developed internally or by industry partners. In that way, aquafeed manufacturers are ensured that the proposed solutions will meet their needs.

Figure 1. Aqualis EMEA technician filling up automatic feeders

At Symrise Aqua Feed, a company developing and producing palatability enhancers and functional protein hydrolysates for shrimp and fish feed, a dedicated

Aqualis EMEA, a pioneer in fish testing among aquafeed ingredient suppliers Since the beginning of Symrise Aqua Feed in 2006 (formerly known as Aquativ), Aqualis EMEA has been a mandatory step before releasing a new dietary solution on the aquafeed market. Aqualis EMEA facilities consist of the permanent rental of two independent testing units belonging to a French research institute dedicated to marine sciences and consisting of 24 tanks of 100L capacity (Fig. 1). Both units are supplied with sand filtrated seawater thermoregulated to 20°C all year round. The minimum water exchange rate is 200% per hour, which guarantees the best water quality to the European seabass juveniles used as model fish species for all commercially farmed marine fish species because of its high requirements in feed palatability and nutrition. Experimental conditions are standardized as much as possible to reduce the variability of the different KPI measured during the palatability, nutritional and health studies. Fish are mostly fed ad libitum using online automatic feeders to maximize dietary palatability performance, which can be specifically measured with short-term feeding trials of 2 to 3 weeks.

Figure 2. Ranking of palatability performance of different aquafeed solutions as measured at Aqualis EMEA (% of relative growth gain between positive control diet at 100% and negative control diet at 0%)

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RESEARCH

Figure 3. View of RAS and testing facilities at Aqualis APAC

In this case, palatability performance is defined as the percentage of the maximum growth gain observed when top-coating an internal reference palatability enhancer (PE) onto an internal reference basal diet (extruded plant-based feed, nutritionally balanced). This strategy allows the ranking of palatability performance resulting from all the products compared to these 2 control diets while reducing fish response variability (Fig. 2). Due to its long tracking record, palatability performance assessment is also used in routine to control the batch quality of the proposed solutions or to determine the product shelf-life. For nutritional studies, an on-site feed processing lab allows the pelleting process of experimental feeds where dietary variables can be graded fishmeal and/ or feed additive qualities or levels. In-house NIR device will confirm that feed specifications meet formulation expectations, before starting the feeding trials. While it is not possible to run disease challenges within these testing facilities, environmental (temperature, anoxia) or husbandry (net chasing, crowding, grading, vaccination, swim tunnel) challenges remain possible, along with the measurements of some immune or stress-related parameters such as fish serum lysozyme activity or cortisol levels. In this case, procedures are required to be submitted, and approved, by a local Ethical Committee and the 3R rule will apply (replace, reduce, refine). In respect of local regulations, technicians and researchers are regularly trained in animal experimentations and their legal requirements.

Figure 4. View of the aquaponic system at Aqualis APAC

Aqualis APAC as a testing hub for main APAC species and rearing practices Asia is the 1st regional producer of aquaculture products in the world with the widest diversity of farmed species and rearing practices. It was therefore obviously necessary to work locally and target new model fish species such as tilapia for omnivorous fish species and Asian seabass, or snakehead for carnivorous ones. In this context, Aqualis APAC opened in 2020 and has been upgraded till now. Today, the facilities consist of two RAS units of 32 tanks of 500L capacity, possibly running both with brackish and fresh waters (Fig. 3). As white shrimp is another widely farmed species in Asia, two RAS sets of 32 aquaria were set up to allow specific palatability assessments with this species. A feed processing lab also permits the production of sinking feed by the pelleting process. As at Aqualis EMEA,

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RESEARCH

Figure 5. View of Aqualis LATAM shrimp testing facilities

experimental diets can be checked by in-house NIR or external lab analysis. Fish and shrimp are currently fed manually ad libitum while waste feed and feces are removed one hour after feeding either by siphoning or using individual tank, or aquarium, pump aspiration. In the four mentioned species, palatability, nutritional and health studies are feasible, thanks to partnerships with local universities for analysis of immune or physiological parameters. CSR is also an important component of Aqualis sites, and Symrise Aqua Feed DNA. Tested individuals are purchased from qualified local hatcheries and given to local farmers at the end of feeding studies, while RAS waste waters are reused for their nutritional value within an in-house aquaponic system (Fig. 4).

Aqualis LATAM as a regional shrimp dedicated testing platform Ecuador is now the largest exporter of farmed shrimp. Farming practices are completely different in LATAM compared to Asia, not to mention the significant differences in post-larvae genetic (SPR vs. SPF) or access to feed raw materials. This is why Symrise Aqua

Feed has set up Aqualis LATAM operations, relying on two local partners, one university in Ecuador and one private CRO in Peru. Their facilities consist of a set of 30 to 40 tanks of 1m² area, where shrimp are tested in nutritional and health studies at a density of 40 shrimp/ m² in clear water conditions. Experimental diets are pelleted within a Symrise Aqua Feed feed processing lab using local feed manufacturers’ raw materials, with in-house NIR controlling feed specifications.

Conclusion The aquafeed industry is still growing at a high pace facing new challenges, in addition to the ones listed at the beginning of this article. The need for testing more sustainable and functional diets is increasing as fishmeal and fish oil prices soar while various new ingredients are proposed on the market at an industrial scale. Another illustration of sustainability is water eutrophication, which is now more and more controlled, mostly to ensure required water quality but also to not compromise biodiversity. Dietary formulations, and resulting dietary digestibility, play an important role in this important topic.

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RESEARCH

It is therefore mandatory for the main aquafeed stakeholders to rely on their own testing facilities to ensure they have enough testing capacities while making sure the implemented experimental conditions fully meet their specifications. External CROs, or public institutions, will remain essential for specific needs or more fundamental projects. Thus, at Symrise Aqua Feed, more than 60 feeding trials have been implemented in 2022 with 75% of them focusing on dietary palatability while 25% of them were investigating deeper proposed solutions at nutritional and health levels. These testings are essential at all levels of Symrise Aqua Feed organization reflecting the different steps of a product life cycle. As an illustration, 35% of these assays aim at improving proposed solutions and 50% at demonstrating their performance within the local aquaculture environment and economics, including collaborative trials with aquafeed manufacturers. Last but not least, 20% of Symrise Aqua Feed feeding trials are implemented to control the batch-to-batch product quality and most of all to improve testing methods, especially on more challenging species such as white shrimp, because you can only improve what you measure accurately!

More information: Mikael Herault Performance Measurement Global Symrise Aqua Feed E: mikael.herault@symrise.com

Clément Martineau Performance Measurement Aqualis EMEA Symrise Aqua Feed

Dachawat Poonnual Performance Measurement Aqualis ASPAC Symrise Aqua Feed

Freddy Balbi Performance Measurement Aqualis LATAM Symrise Aqua Feed

Aquafeed: Advances in Processing & Formulation Vol 15 Issue 4 2023

always inspiring more …

Aqualis, performance measurement centers aquafeed.symrise.com

MARINE-BASED INGREDIENTS

Big things have small beginnings Isak Bøgwald, Hogne Abrahamsen, Zooca The zooplankton company Zooca is set for growth, both in production volumes and nutritional terms. New studies show that the protein hydrolysate from Calanus finmarchicus has growthpromoting properties.

Figure 1. Zooca is looking to grow beyond the snowy mountains of northern Norway following the completion of the new factory in Sortland, Vesterålen. Credits: István Kiss.

Small beginnings with big potential The bold idea of using the tiny zooplankton Calanus finmarchicus for humans and animals was conceived by Prof. Kurt Tande in the early 2000s. Its big potential was the combination of extraordinary nutrients and an enormous annual production estimated at 300 million tonnes in the Norwegian Sea. Calanus AS was founded in 2002 and named after the resource it was centered around. It has since rebranded to Zooca (The Calanus Company) and currently employs over 40 people. Through developments of harvesting, processing, novel products, regulatory approvals, marketing, and sales, the efforts to commercialize the resource have now culminated in the world’s first factory specifically designed to process C. finmarchicus

(Fig. 1). With an increased production capacity from 600 to 10,000 tonnes of raw material every year, the company is looking to grow beyond the Arctic Circle and offers aquaculture ingredients for various fish and shrimp species.

A protein hydrolysate for increased growth The company's main protein product is Zooca® Hydrolysate, a pure protein hydrolysate from C. finmarchicus. New studies reveal its potential to improve growth even when compared to other protein hydrolysates of marine origin. A feeding trial was performed by SPAROS in Portugal, where European seabass juveniles were fed diets with low fishmeal contents and 4% inclusion of the various hydrolysates.

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MARINE-BASED INGREDIENTS

Figure 2. Body weight growth and feed conversion ratio (FCR) from a feeding trial with European seabass juveniles fed diets with 4% marine hydrolysate inclusion. Lowercase letters denote statistically significant differences (P<0.05).

Results showed that the diet with Zooca® Hydrolysate inclusion was associated with increased body weight growth throughout the trial combined with a lower feed conversion ratio (Fig. 2). Motivated by the good performance of the Zooca® Hydrolysate in the feeding trial, follow-up studies by the Norwegian research institution Nofima were initiated with the aim of investigating the growthpromoting properties in skeletal muscle cells. The Zooca® Hydrolysate was split into six fractions of different molecular weights to determine if the degree of hydrolysis of the proteins influenced muscle growth. Primary skeletal muscle cells were isolated from Atlantic salmon and treated with each of the six Zooca®

Hydrolysate fractions, using standard growth medium as control. The results indicated that the fraction of the lowest molecular weight (<260 Da) was associated with increased viability and proliferative activity in the skeletal muscle cells. A gene expression study found that the low-molecular fraction upregulated genes coding for structural muscle proteins, suggesting increased differentiation of muscle cells into muscle fibers. The low-molecular fraction was characterized to allow a deeper look into the contents of the fraction, possibly to identify its bioactive constituents for muscle growth. The average molecular weight of the peptides in the fraction was calculated to be 260 Da, which approximately corresponds to an average length of

Figure 3. Dr. Sileshi Wubshet and colleagues at Nofima found that a low-molecular fraction of a hydrolysate from C. finmarchicus positively affected muscle cell differentiation in studies with salmon skeletal muscle cells. Credits: Jon-Are Berg-Jakobsen, Nofima.

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MARINE-BASED INGREDIENTS

Figure 4. Characterization of the growth-promoting fraction of Zooca Hydrolysate by NMR spectroscopy.

2.3 amino acid residues. Short peptides are known to have efficient uptake by peptide transporters in the intestines and to improve growth performance. Further characterization by NMR spectroscopy (Fig. 4) revealed that the bioactive fraction contained amino acids and marine metabolites known for their growth improvement. Branched-chain amino acids (isoleucine, leucine, and valine), glutamate, arginine, dimethyl glycine (DMG), and lactate have all been described as improving growth when supplemented in feeds.

The future could bring big things With increased production capacity and a patent pending for a growth-promoting hydrolysate fraction, the new Zooca factory is ready to produce big things in the future. The company still focuses its efforts on R&D activities, and several new studies on the functional properties of the Zooca® Hydrolysate in feeds are right around the corner. Future studies include its health implications for European seabass, and growth, health, and attractivity for whiteleg shrimp (Litopenaeus vannamei). While the quote “big things have small beginnings” is borrowed from a classic movie, it could hold true for the expanding zooplankton company from northern Norway.

More information: Hogne Abrahamsen CSO Animal Health & Nutrition Zooca E: hogne.abrahamsen@zooca.no

Isak Bøgwald Researcher Animal Health & Nutrition Zooca E: isak.bogwald@zooca.no

References available on request.

Aquafeed: Advances in Processing & Formulation Vol 15 Issue 4 2023

MARINE-BASED INGREDIENTS

Supporting allostasis with optimized fishmeal freshness, protein digestibility, and beneficial water-soluble composition Ola Flesland, Dag Gjerde, Pelagia As part of its science-based innovation pipeline, Pelagia has developed a new fishmeal, Pro-Allo, which creates optimal conditions in fish to support allostasis – the biological process of fighting stress caused by factors such as environmental changes and handling. Pro-Allo is produced in Norway from sustainably sourced fish and to the same strict standards as high-quality Group 1 fishmeal. However, tests conducted by Pelagia in seabass demonstrate that, depending on the levels of water-soluble protein present, Pro-Allo reduces the number of vulnerable fish by 27-86% compared with standard Group 1 fishmeal (Fig. 1). It has also been shown to improve the specific growth rate by 4-19% (Fig. 2) and feed efficiency by 6-30% (Fig. 3).

Developed in close collaboration with fishmeal specialist Dr. Anders Aksnes, Pro-Allo supports allostasis and delivers improved feed efficiency and more robust fish by focusing on three key areas: raw material freshness, protein digestibility, and levels of watersoluble components. In this article, we will look at why these three factors are so important, and how Pro-Allo is formulated to align with them.

Raw material freshness A substantial body of research shows that the freshness of the raw materials in fishmeal is directly correlated to growth rates in fish. In a study (Fig. 4), after 11 weeks, salmon given feed made from stored fish had grown at

Figure 1. Impact of Pelagia Pro-Allo on the number of vulnerable fish. Pelagia’s own trial.

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MARINE-BASED INGREDIENTS

Figure 2. Impact of Pelagia Pro-Allo on specific growth rate in seabass. (SGR). Pelagia’s own trail.

half the rate of those consuming feed produced from fresh fish (Opstvedt et al., 2000). This supports previous research (Aksnes & Mundheim, 1997) indicating that spoilage of fish raw material before fishmeal production, measured by levels of biogenic amines such as cadaverine, led to reduced growth and reduced feed efficiency ratios in halibut (Fig. 5). To ensure the freshest raw material for Pro-Allo, Pelagia is working closely with fishing fleets through a dedicated supply chain. This is centered on specific procedures from catch to arrival at the factory for processing into fishmeal. Pelagia has incorporated new quality parameters to maximize levels of freshness above and beyond existing industry standards. This results in fishmeal that exceeds the levels of freshness

currently found in aquafeed products.

Protein digestibility The extent of protein digestibility in fishmeal is influenced by the processing method. In particular, digestibility is reduced when raw materials are dried at higher temperatures. Furthermore, research indicates a link between protein digestibility in fishmeal and growth rates achieved in fish. In a study on halibut, researchers found that processing raw fishmeal materials at 100°C resulted in a 4% reduction in protein digestibility, compared with processing at 70°C (Aksnes & Mundheim, 1997). This reduction in protein digestibility led to a 7% decline in specific growth rate (Fig. 6). In addition, higher

Figure 3. Impact of Pelagia Pro-Allo on feed efficiency in seabass. Pelagia’s own trial. *Average of the 15 most recently published data (2017-2022)

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MARINE-BASED INGREDIENTS

Figure 4. Impact of freshness in salmon feed (Opstvedt et al., 2000)

digestibility was associated with a significantly improved feed-conversion ratio. Gentle drying of the raw materials in Pro-Allo preserves a higher level of protein digestibility to optimize specific growth rates, feed efficiency and feedconversion ratios. These benefits translate into bigger, healthier fish and greater operational efficiency.

Levels of water-soluble components The water-soluble fraction contains several molecules, of which the major part comprises small proteins and free amino acids – the building blocks of protein that exist in isolation or in short chains (peptides). These amino acids and peptides are important for growth, well-being and survival in fish. Existing aquaculture quality parameters state that Group 1 fishmeal should comprise no less than

18% and no more than 32% water-soluble protein. Anything higher than 32% has historically been seen as a quality concern because it might indicate adverse microbial growth and the presence of growthretarding components. However, research shows that water-soluble proteins are critical for fish growth and well-being. Figure 7 shows results from a study where increasing levels of stickwater (SW) – a water-soluble protein solution – were added to fishmeal, and fish weight was evaluated (Kousoulaki et al., 2009). A direct correlation was found between SW levels and weight, with greater quantities of SW in the fishmeal leading to bigger fish. These findings indicate that levels of water-soluble protein higher than 32% could be desirable in fishmeal – provided that the molecules in question are beneficial in nature to fish growth and wellbeing. In practice, this

Figure 5. Impact of freshness on specific growth rate (SGR) and feed conversion ratio (FCR) in halibut (Aksnes & Mundheim, 1997)

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MARINE-BASED INGREDIENTS Table 1. Nutrient composition of Pro-Allo

Figure 6. The impact of true protein digestibility on specific growth rate (SGR) and feed conversion ratio (FCR) in halibut (Aksnes & Mundheim, 1997)

Specification

Values

Raw protein

69-72%

Lipids

11% maximum

Crude ash

18% maximum

Water

6-10%

Cadaverine

< 0.6g/kg

Histamine

< 0.2g/kg

Water soluble protein

32% of crude protein minimum

Microbials

Values

Salmonella

Negative

Enterobacteriaceae

< 10 CFU/kg

Pro-Allo contains more than 32% water-soluble protein (Table 1) with a guarantee of increased levels of beneficial components and reduced levels of undesirable growth-retarding components. As a result, it delivers superior fish well-being, growth and robustness by promoting allostasis phase in fish during challenging periods in their lifecycle, particularly in fish with specific vulnerabilities. Incorporating Pro-Allo in commercial feed may therefore be especially useful when fish are facing the challenges of the larvae and smolt stage, disease, unfamiliar temperatures, physical handling, or a feed regime with high levels of plant protein. Figure 7. The effect of water-soluble protein on fish weight (Kousoulaki et al., 2009)

suggests it would be a positive step to incorporate free amino acids and peptide fractions into fishmeal. A later study supports this hypothesis (Kousoulaki et al., 2012), with the results indicating that the inclusion of the smallest molecules in SW resulted in bigger fish. There are several possible reasons why water-soluble protein fractions result in bigger and more robust fish. Free amino acids have a specific smell and taste and therefore might stimulate increased feed uptake. This is especially important in fish struggling to achieve adequate uptake. The digestive tract contains sensors for free amino acids and peptides, indicating that some free amino acids and peptides may have specific neurological effects that could affect fish physiology and well-being.

References available on request

More information: Ola Flesland Product & Process Developer Pelagia

Dag Gjerde Senior Sales Manager Pelagia E: feed@pelagia.com

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PROTEINS

Carnivorous fish feed formulation practice with rendered animal proteins: A review of 20 year research at Zhejiang University Yan Wang, Ocean College, Zhejiang University, Peng Li, North American Renderers Association

Aquaculture is one of the fastest growing sectors in agriculture. Feed technologies have been recognized as one of the major driving forces to the expansion of aquaculture worldwide. The rapidly growing demand for digestible proteins for aquacultured species has been resulting in the depletion and deficiency of dietary protein sources, especially fishmeal, which has been used as the most ideal protein source in aquafeed. In 2023, the market price of quality fishmeal skyrocketed to RMB 18,000-20,000 (USD 2,500-2,775) per metric ton due to the dramatically declined fishmeal production. Fishmeal deficiency becomes a challenge threatening the sustainability of aquaculture of crustacean and carnivorous fish, which traditionally rely on high levels of fishmeal in formulated feeds.

Rendered animal proteins to replace fishmeal Research on fishmeal replacement in aquafeed started in the mid-1970s, following the research on fishmeal replacement in chicken diet. Before 2005, hundreds of papers were published to assess the potential replacement of fishmeal with rendered animal protein ingredients, such as poultry byproduct meal (PBM), meat and bone meal (MBM), feather meal (FEM) and blood meal (BD), in shrimp and finfish diets. During this period, fishmeal replacement level with PBM, MBM, FEM and BM singularly or in combination, was examined in diets for various carnivorous fish species, including rainbow trout, barramundi, sunshine bass, yellowtail, grouper, red drum, hybrid striped bass, gilthead seabream, Australian silver perch, red snapper, silver

seabream, chinook salmon, palmetto bass, European eels and black sea turbot. Since 2000, supported by the Fats and Protein Research Foundation (FPRF), North American Rendered Association (NARA) and other local funding sources, we have been developing cost-effective formulation technologies to replace fishmeal inclusion with PBM, MBM, FEM and BD in diets for those local carnivorous fish commercially important for mariculture in China. Previous studies by other researchers showed the inclusion of 15% FEM in chinook salmon diets and 12% FEM in Japanese flounder diets as a fishmeal alternative did not result in significantly lower growth, and the inclusion of 24% MBM and 16% FEM in rainbow trout diet containing 30% fishmeal did not negatively influence fish growth. Red drum fed the diets with fishmeal replaced by PBM grew faster than that of the fish fed the diets with the same amount of fishmeal replaced by MBM. These studies all demonstrated nutritional values of PBM, MBM and FEM in carnivorous fish diets, but did not fully elucidate the efficiency of those three promising protein ingredients. Therefore, we conducted the first feeding trial to compare the efficacy of PBM, MBM and FEM as a single fishmeal alternative in cuneate drum diet and found PBM delivered the best performance to cuneate drum, followed by MBM and FEM. In latter studies, we confirmed similar findings with Malabar grouper. Those early findings allowed local aquafeed millers of drum and grouper to include PBM, MBM and FEM in

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PROTEINS Table 1. Growth and feed utilization efficiency of some carnivorous fish fed the diets with fishmeal replaced by rendered animal protein ingredients

Fish species Cuneate drum Malabar grouper Japanese seabass Golden pompano Largemouth bass

Dietary fishmeal level

Alternative ingredients

Initial body weight (g)

35.0 17.5 PBM 24.5 MBM 31.5 FM 35.0 24.5 MBM 17.5 PBM 7.0 MM1 7.0 MM2 40A 8A MM2 40B B 8 MM2 50.0 25.0 MM2 12.5 MM2 50.0 25.0 PBM 12.5 PBM 25.0 MBM 25.0 FEM 40 8 PBM 0 PBM 35a 21a SPC 14a SPC 35b 21b SPC 14b SPC 30 24 FEM 18 FEM 24 I-FEM 18 I-FEM

Weight gain (g/d)

Feed conversion ration

Protein retention efficiency (%)

67.1 56.6 46.4 55.6 68.6 67.0 77.8 74.5 74.6 59.8 65.5 60.7 55.9 86.2 79.1 73.6 101.3 92.1 91.0 93.6 70.5 37.1 47.9 39.9 43.0 34.6 34.3 39.6 38.3 35.9 55.3 49.3 47.8 49.2 52.6

1.05 1.07 1.07 1.34 1.12 1.05 1.14 1.09 1.07 1.12 1.21 1.21 1.46 1.00 1.09 1.21 1.14 1.17 1.16 1.31 1.43 1.15 1.13 1.23 1.50 1.86 1.87 1.62 1.66 1.76 0.91 0.87 0.86 1.06 1.02

35 33 35 28 37.2 38.6 42.3 41.0 40.4 34.8 31.4 33.5 33.1 32.0 30.0 27.3 30 29 29 28 24 31.4 32.1 28.7 23.8 19.3 19.3 21.9 21.9 20.3 42.1 41.2 40.6 37.0 35.3

27.4 27.4 27.4 27.4 36.8 37.5 36.7 37.6 37.0 41.6 40.8 41.9 42.2 50.9 50.9 51.1 49.5 49.9 50.6 50.6 50.2 8.6 8.5 8.5 16.7 16.7 16.5 16.8 16.7 16.6 14.4 14.7 14.7 14.2 14.0

Reference

Wang et al., 2006, Aquaculture

Guo et al., 2007, Aquaculture Nutrition Wang et al., 2010, Aquaculture Nutrition Wang et al., 2008, Aquaculture Li et al., 2009, Journal of the World Aquaculture Society Wang et al., 2015, Aquaculture Research Wang et al., 2017, Aquaculture Research

Ren et al., 2020, Animal Feed Science and Technology

PBM, poultry byproduct meal; MBM, meat and bone meal; FEM, feather meal; MM1, a blend of poultry byproduct meal, meat and bone meal and blood meal; MM2, a blend of poultry byproduct meal, meat and bone meal, feather meal and blood meal; SPC, soy protein concentrate; I-FEM, gamma ray irradiated feather meal. Duration of the feeding trials was eight weeks for cuneate drum, Japanese seabass, golden pompano and largemouth bass were lasted for, and ten weeks for Malabar grouper. Dietary protein level was 47% for 40A and 8A, and 41% for 40B and 8B; Dietary PBM level was 15% for 35a, 21a and 14a, and was 10% for 35b, 21b and 14b.

their formulation based on the pricing and availability of those three rendered proteins. At present, PBM is recognized as an excellent fishmeal alternative and is widely used at high levels in carnivorous fish diets worldwide. MBM, FEM and BM are also well recognized as valuable aquafeed ingredients used at lower inclusion levels in diets of various carnivorous/omnivorous fish and diets of herbivorous fish at early stages or cultured at cold water temperatures.

Combining ingredients We carefully reviewed research results from our own lab and other fish nutrition laboratories and we hypothesized certain nutrients in individual rendered

proteins might be limiting, after comparing their nutrient profile with fishmeal. We started to explore combinations of various rendered proteins to further reduce fishmeal inclusion in various marine fish. We compared the performance of cuneate drum fed diets containing fishmeal and various fishmeal replacement levels with PBM, MBM and two combinations of PBM, MBM, FEM and BM (60% PBM + 30% MBM + 10% BM; 60% PBM + 20% MBM + 10% FEM + 10% BM). Our results showed that the growth of cuneate drum fed the diets containing fishmeal partially replaced by the blend of PBM, MBM and BM or a blend of PBM, MBM, FEM and BM were comparable to that of the fish fed the diet with fishmeal replaced by PBM. This

Aquafeed: Advances in Processing & Formulation Vol 15 Issue 4 2023

PROTEINS finding was then confirmed in other studies on Malabar grouper and largemouth bass in our lab. The proper combination could increase fishmeal replacement levels with MBM and FEM. Combination technologies based on the consideration of balancing amino acids and other nutrients are proven to enhance the nutritional value of MBM and FEM in feed formulations for those high-value aquatic animal species and reduce the total ingredient cost in aquafeeds without compromising the performance of those carnivorous species. In the meantime, we conducted a series of feeding trials to determine the minimum fishmeal inclusion in diets for cuneate drum, Malabar grouper, golden pompano, Japanese seabass, giant croaker, largemouth bass and large yellow croaker as well as factors that could affect the performance of rendered proteins in diets of those fish to make fishmeal replacement by rendered proteins more predictable and replicable. We found that fishmeal replacement level with a blend of PBM, MBM, FEM and BM in cuneate drum diet could be increased by elevating protein level, but dietary lipid level did not significantly influence the efficacy of blended rendered proteins in fishmeal replacement. We also found that fishmeal replacement level with soy protein concentrate in golden pompano diet could be increased by either increasing PBM content in basal diet formulation or exogenous selenium supplementation. These results revealed that fishmeal replacement level with alternative ingredients is closely dependent on the composition of basal diet formulation and probably micronutrient availability. When we estimated how gamma-ray-irradiation could affect amino acid availability of rendered proteins after gamma-ray-irradiation was widely

used to control Salmonella at ports, we found that gamma-ray-irradiated FEM could further enhance the nutritional value of FEM and replace more fishmeal in the diets of largemouth bass and golden pompano without compromising production. We also found supplementation of protease could increase fishmeal replacement level with FEM in large yellow croaker diet. Those studies indicated the new potential of feather meal processing towards a more desirable high-protein ingredient in aquafeeds.

Re-establishing the balance We reviewed scientific research on fishmeal replacement technology in our lab and other labs and we concluded that fishmeal replacement is a systemic work to re-establish nutrient balance in the diet formulation with reduced fishmeal content and increased alternative ingredients, rather than a simple project to search for an ideal and novel alternative ingredient that can replace fishmeal in a control diet at high levels. We proposed the technique diagram for fishmeal replacement, which include (1) screening the suitable alternative ingredients by assessing the potential of the ingredients singularly to substitute fishmeal in a basal diet formulated at the optimal protein level to the tested fish; (2) assessing the potential of several alternative ingredients in combination to substitute fishmeal in a basal diet formulated at the optimal protein level; (3) improving nutrient bioavailability of alternative ingredients with poor protein digestibility or rich in antinutritional factors; (4) establishing micro-nutrient balance by supplementation of functional constitutes, including amino acids, micro minerals and proteases, in low-fishmeal diets; (5) establishing nutrient-balance at

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Aquafeed: Advances in Processing & Formulation Vol 15 Issue 4 2023

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PROTEINS

Figure 1. Diagram to formulate low fishmeal diets

multiple dimensions, including proximate composition, digestible protein or amino acid, bioavailable microminerals, as well as inactivation of antinutritional factors and supplementation of growth-promotion-factor and palatants, and other functional supplements. According to the technique diagram, we developed an innovative fishmeal analog based on rendered animal protein ingredients and functional additives and demonstrated this fishmeal analog could significantly improve the growth of carnivorous fish fed with lowfishmeal diets. We currently created a series of lowest dietary fishmeal meal records for major carnivorous fish cultured in south China (16% for large yellow croaker; 8% for largemouth bass; 14% for golden pompano, 8% for Japanese seabass, 25% for Malabar grouper and 8% for cuneate drum) in the past 15 years and published those studies in peer-reviewed journals. We assessed production performance, feed cost, fishmeal reliance and waste outputs of these fish species fed with the low-fishmeal diets. The low-fishmeal diets for largemouth bass, large yellow croaker, Japanese seabass and golden pompano, with rendered animal protein ingredients as the main fishmeal alternatives have been successfully used in commercial farming practices.

The challenge of replacing fishmeal The learning and practice in the past two decades have demonstrated that complete replacement of fishmeal in diets for carnivorous fish species

appears an unconquerable challenge despite the fishmeal content in diets for marine fish has been significantly reduced to an average of 14%. Our objective is to reduce dietary fishmeal levels for carnivorous fish to 5%. A 5% fishmeal inclusion will allow 70% of the current global fishmeal production to produce 70 million MT formulated feeds, which can afford to produce about 46 million tons of carnivorous fish (feed conversion ratio is 1.5). Since 2020, we have attempted to reduce fishmeal levels from 8% to 4% in largemouth bass diet, however, fish fed the diets containing 4% fishmeal exhibited retarded growth compared to fish fed the diet containing 8% fishmeal. Another research group recently reported that largemouth bass and red drum could grow well when fed with fishmeal-free diets, highlighting the potential for further improvement in the use of rendered proteins for global finfish aquaculture. Those findings on fishmeal replacement in various marine fish were provided to aquafeed millers and fish farmers as technical support to improved uses of rendered proteins and repeatedly confirmed by various aquaculture entities on commercial scales. Research in our lab and aquaculture nutrition labs worldwide have contributed to affordable nutritional solutions in support of the development of aquaculture in China, Vietnam and other countries in Asia and in other continents in the past two decades and served as foundations for potential innovations and improvements of fishmeal replacement technology for the future aquaculture. More information: Yan Wang Professor Ocean College, Zhejiang University E: ywang@zju.edu.cn Peng Li Asia Regional Director North American Renderers Association E: hi-peng@hotmail.com

Aquafeed: Advances in Processing & Formulation Vol 15 Issue 4 2023

PROTEINS

Corn fermented protein tested successfully in rainbow trout Dr. Steve R. Craig, Louis Rens, Green Plains

A 12-week feeding trial with Green Plains’ corn fermented protein (CFP) has aligned with previous research in showing the ingredient is suitable for certain fish diets. This most recent study was conducted with rainbow trout at the Optimal Aquafeed AquaLab in Shenandoah, Iowa, in the United States. Findings show that CFP60 (60% crude protein, as fed) inclusion rates ranging from 12-21% of the total diet are suitable for commercial diet formulations for rainbow trout and have significantly lower coloration impacts than corn gluten meal, even at much higher inclusion rates.

Methodology Rainbow trout (Oncorhynchus mykiss) were obtained from the same cohort of eggs (Steelhead strain, Riverence Brood, Rochester, U.S.), hatched in the AquaLab hatchery and reared using standard hatchery protocols. After reaching an average weight of 200 grams, 30 fish were weighed and placed into each of the 21 individual experimental tanks (630 fish total), as part of a recirculating aquaculture system equipped with a moving bed biological filter, mechanical filtration, UV sterilization, temperature control and pure oxygen

Table 1. Composition of the evaluated diets for rainbow trout during the 12-week feeding trial. See text for details.

Ingredient

No Corn

12CFP

15CFP

18CFP

21CFP

8CGM

Commercial

Wheat flour

27.10

29.75

28.95

27.95

27.05

31.40

Poultry byproduct meal

18.00

Fishmeal

12.00

Soybean meal

13.00

----

L O

CFP60

----

12.00

15.00

18.00

21.00

----

Corn gluten meal

----

8.00

Fish oil

12.00

Feather meal

7.50

6.50

5.20

3.70

2.00

7.25

Blood meal

4.50

3.50

2.50

2.00

1.50

5.00

Vitamin premix

1.25

Mineral premix

0.75

Choline chloride

0.60

Lysine

1.75

2.10

2.20

2.10

Methionine

0.50

0.45

0.40

Threonine

0.30

0.40

0.45

0.50

Stay-C

0.25

Taurine

0.50

No Corn

12CFP

15CFP

18CFP

21CFP

8CGM

Analyzed composition

Commercial

Crude protein

43.3

42.8

43.5

42.8

42.5

41.8

45.8

Crude lipid

14.0

15.4

15.2

14.4

13.0

14.1

13.4

Crude fiber

4.0

4.8

4.7

6.0

5.4

3.9

3.2

Ash

6.9

5.9

6.1

6.0

6.3

6.4

8.0

Aquafeed: Advances in Processing & Formulation Vol 15 Issue 4 2023

PROTEINS supplementation. Water quality parameters were all within acceptable ranges for rainbow trout under culture conditions. Each tank was randomly assigned to a dietary treatment with three replicate tanks per treatment (n=90 fish/treatment). A control diet was formulated that contained no corn products (No Corn) and a commercially available diet commonly used in the North American trout industry was also used as a control (Commercial). The subsequent experimental diets consisted of CFP60 at 12%, 15%, 18%, and 21% inclusion levels, with an additional diet containing corn gluten meal (CGM) at an 8% inclusion level. The six formulated diets provided 42% crude protein and 16% crude lipid, while the Commercial control diet provided 45% crude protein and 14% crude lipid (Table 1). All feeding was conducted by hand and fish were fed to apparent satiation from measured bins twice daily. Fish were monitored for average individual fish weight, specific growth rate, percent increase from initial weight, and average daily feed intake. Additionally, somatic indices, hematocrit, weight and length distributions, fillet

Hunter Color score, and fillet carotenoid analysis were conducted at the conclusion of the trial.

Results Generally, fish fed the CFP60 diets performed equally well across all the inclusion levels evaluated in this trial, returning growth metrics statistically similar to those observed in the No Corn and Commercial control diets. The final average weights of fish are presented in Figure 1 and cumulative feed conversion ratio (FCR) values are provided in Figure 2. Due to the previous successes with the CFP50 product, these results are not surprising, with inclusion levels of CFP60 up to 21% of the dry diet performing equally to all other evaluated diets. Fish fed all the CFP-based diets returned positive, equivalent responses as related to the No Corn control and Commercial control diets, indicating the CFP60 inclusion rates ranging from 12-21% of the total diet are suitable for commercial diet formulations for rainbow trout. These findings are similar to those previously obtained in rainbow trout with the CFP50 product.

Figure 1. Final average fish weights of rainbow trout fed varying levels of CFP60 at the end of the 12-week feeding trial.

Figure 2. Mean feed conversion ratio (FCR) values rainbow trout fed varying levels of CFP60 at the end of the 12-week feeding trial.

Aquafeed: Advances in Processing & Formulation Vol 15 Issue 4 2023

PROTEINS The 8CGM diet was included in this trial as this protein source is widely utilized as a complementary protein source in commercial diets for a range of cultured fish species. Typically, corn gluten meal inclusion levels are limited to a high of 4-8% of the total diet in aquafeed formulations for white-fleshed final products due to the high total carotenoid levels found in this product and the concern of flesh coloration. Fish fed the 8CGM diet did not perform well in the current trial, returning significantly (P = 0.0087) lowest growth parameters (i.e. average fish weight), as well as other parameters not presented in this article (percent increase from initial weight, specific growth rate), as compared to fish fed the Commercial control diet and fish fed the 15% CFP60 diet. Coloration of the fillets from fish at the end of the trial was evaluated through total carotenoid analysis of the experimental feeds as well as in homogenized fillet samples (Fig. 3, 4). The total carotenoid analyses of the feeds followed what was to be expected, with increasing inclusion levels producing increasing dietary carotenoid levels. Tissue carotenoid levels followed a similar trend to the

dietary levels, with increasing carotenoid deposition with increasing CFP inclusion rates, although these levels were substantially lower than those observed in the Commercial control and 8CGM diets, which contained corn gluten meal, 8% in the 8CGM diet and an unknown inclusion level in the closed formula Commercial control. Additionally, fillets from fish fed their respective diets were subjected to Hunter Color analysis which provides three metrics of coloration and data in a threedimensional format (Fig. 5) that illustrates the coloration impacts of dietary manipulation of feeds fed to rainbow trout. To simplify the graphics, we present the threedimensional graph with data from selected treatments: Commercial control, No Corn Control, 12CFP, 21CFP and the 8CGM diets. The three-dimensional graphing of the L, a and b scores from the Hunter Color analysis clearly shows the impacts of carotenoid levels on flesh color, with the No Corn control with no corn products grouping in the upper left quadrant—the whiter, less yellow and less red area of the graph. Fillets from the fish fed the 12CFP diet group are in this area as well. Fillets from

Figure 3. Analyzed total carotenoids (ppm) for the feeds evaluated in rainbow trout during the 12-week feeding trial.

Figure 4. Analyzed total carotenoids (ppm) in the final fillet (homogenized sample of five fillets per treatment) of rainbow trout after the 12-week feeding trial.

Aquafeed: Advances in Processing & Formulation Vol 15 Issue 4 2023

PROTEINS in terms of coloration. This despite an inclusion level of nearly 3 times the corn gluten meal in the 8CGM diet. These three analyses of color clearly show that CFP60 has significantly lower impacts on fillet coloration than corn gluten meal, and at much higher dietary inclusion levels. Green Plains has revolutionized the ethanol industry with its introduction of CFP feed ingredients, which provide 50% and 60% crude protein levels for the aquaculture industry. The 50% product has been widely investigated with tremendous success in over 25 feeding trials with many fish species, as well as in marine shrimp. This trial clearly demonstrated the efficacy of CFP60 as a major protein source for rainbow trout at inclusion levels ranging from 12-21% of the total diet, as fish performance Figure 5. Three-dimensional illustration of Hunter Color scores for selected fillets from Rainbow trout fed the feeds evaluated during the 12-week feeding trial. at all inclusion levels was equivalent to that of the two control diets the 8CGM diet group are in the lower right quadrant utilized in this trial. This trial also demonstrated the - darker, more yellow and more red. Fillets from the coloration impacts of corn gluten meal, even at low 21CFP diet group are along the plane separating the inclusion levels. Conversely, fillet coloration was not graph in half, meaning that these fillets were in between impacted by CFP60 inclusion levels of up to 18% of the the No Corn control and 12CFP and the 8CGM diets total diet. Table 2. Carotenoid analysis of CFP50 and 60 compared to corn gluten meal (CGM). Values in µg/g (mg/kg) as received. Analysis was performed utilizing high performance liquid chromatography (HPLC) in duplicate from samples sent to Eurofins Analytical in Wilson, NC.

CFP50

CFP60

CGM

Lutein

22.0

22.7

102.0

Zeaxanthin

28.9

34.2

169.0

Β-carotene

1.13

1.94

6.49

Cis-β-carotene

0.50

0.95

3.52

Trans-β-carotene

0.64

0.99

2.97

α-Cryptoxanthin

4.62

4.46

12.2

Β-Cryptoxanthin

4.79

5.25

19.3

Total carotenoids

61.5

68.6

309.0

More information: Steven R. Craig Vice President of Nutrition Green Plains E: steve.craig@gpreinc.com

Louis Rens Louis Rens Senior Vice President of Global Ingredient Sales Green Plains E: louis.rens@gpreinc.com

Aquafeed: Advances in Processing & Formulation Vol 15 Issue 4 2023

PROCESSING TECHNOLOGY

New-generation extrusion system upgrades aquafeed production Garrick Yan, David Ma, Famsun

Figure 1. Control system user interface

With the advent of Industry 4.0 and the pervasive adoption of lean manufacturing practices across various industries, digitalization has emerged as a prominent trend in recent years. In keeping with the digital transformation, Famsun has developed a new generation of aquatic extrusion systems that are based on lean principles. These cutting-edge systems are designed to provide enhanced value to customers and exemplify Famsun's resolute commitment to actively respond to the demand for digital factories. In doing so, Famsun will become a veritable "engine" that drives the high-quality development of the global agriculture industry.

Visualized production data From a lean perspective, data-driven improvement is critical. However, numerous factories either do not

collect production data or still rely on paper-based records. The value of this data is restricted due to inaccessibility, inconsistent readings, and the inability to analyze and use the data in real time. To tackle this issue, Famsun upgraded its control system. At first glance, key monitoring data such as preconditioning temperature, pre-conditioning retention time, melt temperatures and melt pressures before the die plate, main motor load, and SME are prominently displayed on the control panel, making it easier for operators to determine if production and safety requirements are being met. The upgraded system surpasses mere data display and allows digital collection, standardization, and realtime analysis to identify improvement opportunities. Automated alarms notify operators of deviations from set points to minimize waste and downtime. In

Aquafeed: Advances in Processing & Formulation Vol 15 Issue 4 2023

PROCESSING TECHNOLOGY

Figure 2. Customized report and data analysis

Figure 3. Maintenance warning

Figure 4. Comprehensive efficiency analysis interface

summary, the digital control system helps transform data into actionable insights to drive continuous lean improvements. FAMSUN Digital Extruder offers a visual data management platform containing various tools. Customized production reports automatically grab production data in real time and generate daily, monthly, or annual reports according to the established template. The data sheet also has rich functions such as classification, sorting, filtering, and comparison, which can help the manager efficiently compare and analyze the data. Moreover, in the new generation of digital extrusion systems, we have added the function of maintenance reminder. The program has set up reminders and overdue alarms to facilitate maintenance personnel to arrange their work reasonably and avoid unplanned downtime caused by negligence in maintenance. Do not underestimate this small tool since, according to a survey, the total maintenance cost with no plan is more than three times that of a periodic plan. The production efficiency analysis module monitors and analyzes equipment performance in real-time. For instance, it can categorize the running status based on a combination of production parameters, then calculate the time spent and material usage at each status. The module also identifies the reasons for and locations of downtime and provides recommendations to improve production efficiency. This module supports lean production teams by helping them analyze the root causes of inefficiencies and continuously improve key performance indicators like overall equipment efficiency (OEE) and yield efficiency (YE). Overall, the produc tion efficiency analysis module aims to transform data from extruder operations into actionable information that can guide decisions for enhancing efficiency, yield and equipment utilization through a lean lens. By presenting insights in a clear and easy-to-understand manner, it can help extruder operators and managers make more informed choices to optimize operations on an ongoing basis.

Aquafeed: Advances in Processing & Formulation Vol 15 Issue 4 2023

PROCESSING TECHNOLOGY Table 1. Example of feeding stability with different methods

Original method

New method

Setting value

Fluctuation range

Deviation

Setting value

Fluctuation range

Deviation

Feeding capacity (4,600kg/hr)

4,531-4,657kg/hr

±1.8%

Feeding capacity (4,600kg/hr)

4,591-4,600kg/hr

<1.9%

Water & steam addition

<1.8%

Water & steam addition

<1%

Fluctuation of motor load

64.2-65.6%

<1.8%

Fluctuation of motor load

64.8-65.5%

<0.7%

Drying moisture deviation

±0.71%

↓

Drying moisture deviation

±0.62%

↑

Uniform quality comes from stable feeding Stable and consistent feeding of dry and wet materials is paramount to pellet uniformity and quality. Our team of control engineers has conducted extensive research on various operating conditions in different applications and has developed an advanced automatic feeding control program. This program precisely controls the feeding rates of dry and wet materials to maintain a consistent ratio during extrusion. It can compensate for fluctuations in raw material properties and machine variables, ensuring stable material flow into the extruder barrels. By monitoring sensors in real-time and automatically adjusting feeder speeds, the control program reduces variations in pellet properties, responding quickly to maintain a uniform material mix and feeding rate under different operating conditions. Less variation in material feeding also helps improve the throughput and energy efficiency. In all, this upgrade not only satisfies the traditional requirement of automatically adding water and steam according to the proportion of dry materials to ensure constant moisture and temperature but also significantly reduces motor load fluctuations and even decreases the moisture deviation in the finished product. For a 10-ton/hr single dryer, this improvement may increase profits by more than $60,000 per year.

Reduce “startup waste” Excessive startup waste is a major issue in extrusion production management. For instance, a typical 10-tonper-hour extrusion line can generate over 600 kilograms of startup waste every time it starts from cold. This not only wastes labor but also raw materials if the startup waste cannot be recycled and reused in time. The startup waste reduction module aims to solve this problem as a unique technology in the industry. It aims to speed up the heating of the system and reduce the amount of waste generated during the initial run. For a standard aquatic feed production run, this technology can shorten the time to reach stable running conditions by more than 20%, reduce pre-conditioner startup waste by up to 70%, and reduce the total extrusion system's startup waste by over 60%. The module achieves this through optimized control of heating elements, automatic adjustments to process parameters, and active monitoring of material flow and temperatures. It works to quickly bring the extruder barrel and the melt before the die plate up to stable operating temperatures while minimizing material wastage. Therefore, the startup waste reduction module provides significant benefits in terms of material savings, production efficiency, and environmental impact. By optimizing the system's warmup process, it helps maximize yield and productivity from the very beginning of each production run.

Table 2. Amount of the startup waste with different technology

Control mode (for one startup)

New technology

Manual

Reduction (%)

Scrap from pre-conditioner (kg)

155

500

~70%

Scrap from extruder (kg) 145 90

~38%

Total 645 245

~62%

Aquafeed: Advances in Processing & Formulation Vol 15 Issue 4 2023

PROCESSING TECHNOLOGY Additionally, after reducing the total waste, it is easier to reuse these materials in time. We have several processing solutions to recycle the remaining scraps, reducing labor intensity and avoiding material loss in all aspects, especially when changing the formula frequently.

Make all operations easy In order to consistently reproduce the qualified finished product, it is important to have a recipe management system (RMS) that stores the production data from successful runs. Famsun's RMS allows users to save all relevant parameters and recall them with one click. This ensures that future productions using the same input materials and equipment status can match previous results. Users no longer need to manually re-enter settings each time. The RMS also features other integrated functions to simplify operations. These include one-click commands to flush the extruder, adjust jacket temperatures, and empty the conditioners.

Conclusion In summary, the Famsun Digital Extrusion System integrates a stable feeding system, user-friendly control interface, functional modules and data analytics platform to embody the principles of reducing waste, recycling materials and reusing resources. This helps feed producers reduce costs, improve efficiency, optimize processes and produce consistent products of high quality.

More information: David Ma Deputy Director of R&D Institute Famsun Co. Ltd. E: fengde.ma@famsungroup.com

SUPPORT OF NATURAL DEFENSES www.phode.com / phode@phode.com Follow us

Aquafeed: Advances in Processing & Formulation Vol 15 Issue 4 2023

PROCESSING TECHNOLOGY

Developing sustainable fish feed through laboratory extrusion Jens Müller, Brabender

Despite increasing demand, fish is becoming more and more difficult to obtain and is often caught unsustainably. Yet it is used not only for consumption as food fish but also as a component of fish feed. The black soldier fly is now providing a remedy. In the meantime, every second edible fish comes from aquaculture. This is because the demand for protein-rich fish is increasing – but, at the same time, the number of wild-caught fish has been stagnating for 30 years. Today, as many marine animals are raised in aquaculture as are caught in the oceans – and the trend is rising. Aquaculture fish are fed feed consisting of vegetable ingredients, minerals, fishmeal and fish oil. Fishmeal and fish oil in the feed are problematic for the environment.

These are simply processed fish, which in turn are fed to fish in aquaculture. Fish feed made from fishmeal is not sustainable and ensures that we are fishing our oceans dry. Around 16 million tonnes of small fish are caught every year to produce five tonnes of fishmeal and one million tonnes of fish oil. As long as feed from marine sources is used, the growth of aquaculture is limited despite increasing demand for fish.

Natural fish feed from the extruder To counteract the problem, Brabender has reduced the amount of fishmeal in fish feed by testing more sustainable fish feed formulations. Fishmeal was replaced with insect meal from black soldier fly and

Aquafeed: Advances in Processing & Formulation Vol 15 Issue 4 2023

PROCESSING TECHNOLOGY

produced fish feed pellets on a trial using our TwinLab-F 20/40 extruder. The insects used are not only natural, but they also have a high protein content. Different proportions were tested. In addition to fishmeal and insect larvae meal, wheat meal was also used. Step by step, the fishmeal was reduced and replaced by black soldier fly larva meal. The starting recipe was a standard recipe for carp feed. The fishmeal was reduced by half, and in a subsequent trial, the fishmeal was finally replaced completely by the insect meal. The result was good. Substituting fishmeal with black soldier fly larvae meal is possible so that products of acceptable quality can be obtained. In addition, the use of insect larvae would be more cost-effective in the future, and insects would be subject to lower price fluctuations on the market than fish.

Cost-effective and sustainable production In the animal feed industry, black soldier fly is becoming increasingly popular as an ingredient in animal feed. However, they can also be used in the production of food, oil, diesel as well as fibers. There are also companies producing the insects as an alternative to soy feed. Breeding black soldier flies is also simple. For example, an adult soldier fly can neither bite nor sting. Since the adult insects do not eat and thus do not need food, reproducing the black soldier fly is more cost-effective than breeding fish, for example.

The Duisburg-based mechanical engineering company Brabender is a global manufacturer of measurement and process engineering solutions for testing the quality of various raw materials as well as for developing and optimizing products, recipes, methods and processes. As a long-standing partner of industry and research in the field of food and feed, Brabender offers a wide range of modular and compact solutions for laboratory-scale rheology and extrusion, as well as application technology consulting and expertise. The use of the TwinLab-F 20/40 extruder is well suited for the production of feed pellets, as all operations can be implemented with one unit. First, the material is filled and conveyed in the extruder, mixed, and heated under shear. Finally, the extrudate is formed in the die. It is also advantageous that the extruder runs continuously and automatically. This manufacturing process avoids downtimes on the production line on an industrial scale but also offers advantages on a laboratory scale. For example, different formulations can be run or process parameters tested in a short period of time for the purpose of research and product development without blocking the production extruders. A laboratory extruder also consumes significantly lower quantities of raw materials. A laboratory extruder is thus suitable for the development of feed formulations where the production process can be adapted to achieve more similar and better product properties by varying, for example, the moisture of the raw material mixture, the screw speed or the temperature profile.

Fat content decisive for feed pellet quality However, there are also challenges in sustainable fish feed extrusion. For example, it can happen that fish feed based on fishmeal is preferred to that made from insect larvae in terms of the fish's eating behavior. Nutritionally, insect protein is of course not as valuable as fish protein. Therefore, fishmeal may be preferred because the nutritional value of the fish decreases when the protein of the black soldier fly larva is used.

Aquafeed: Advances in Processing & Formulation Vol 15 Issue 4 2023

PROCESSING TECHNOLOGY Since fish also need vitamins and unsaturated fatty acids, which are normally found in fish, the extruded fish feed pellets made from insect larvae have to be enriched with fats and vitamins in a separate step. Fat addition is mostly done by vacuum coating but also requires resources such as time, machinery, labor and energy. In addition, a high-fat content affects the quality of the feed pellets. They might lose stability if the fat content is too high. In addition, too high a fat content in the extruder reduces friction and thus energy input, which has a negative impact on the extrusion process and thus on the quality of the pellets.

process parameters. For example, fish feed with different sinking properties or fat contents and thus fish with different eating habits can be developed.

More information: Jens Müller Application Engineer Food & Feed Extrusion Brabender

A new process saves a step In order to optimize the process in animal feed production and avoid the additional work step of adding fat, Brabender has carried out a research project together with Fulda University of Applied Sciences and Evonik Operations GmbH. The project partners have developed a process to increase the fat content in extruded fish feed pellets without additional vacuum coating by adding silica, presented at Victam International. By omitting the work step, energy and resources are saved. This is then of course even more sustainable. Another challenge in processing is that the fish feed contains temperature-sensitive c o mp o n ent s . T h es e ar e polyunsaturated fatty acids and vitamins that fish need to live and absorb through their food. Quality determination solutions for food, feed, plastics and rubber industries The TwinLab-F 20/40 twin-screw extruder can also be used to test various raw material mixtures for fish feed pellets for their processing properties at different

Aquafeed: Advances in Processing & Formulation Vol 15 Issue 4 2023

Industry Events 2023 NOVEMBER 2 – 4: 8 – 10: 8 – 10: 13 – 15: 13 – 16: 20 – 22:

Aquaex India 2023, India 2023 Equipment Manufacturers Conference, USA X International Symposium on Fish Nutrition and Health, Brazil Food & Feed Extrusion Technology, UK Aquaculture Africa 2023, Zambia VIV MEA 2023, Abu Dhabi

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