Aquafeed Magazine Vol 16 Edition 1 2024 by Aquafeed Media

Vol 16 Issue 1 January 2024

AQUAFEED Advances in processing & formulation An Aquafeed.com publication

EXTRUSION OF PREMIUM SHRIMP FEEDS Feed additives Slow-release amino acids Updates in insect meal performance Published by: Aquafeed Media, S.L.U. www.aquafeed.com info@aquafeed.com

HEALTH IS AT THE HEART OF OUR CONCERNS

BOOST YOUR BUSINESS BY OPTIMIZING YOUR HEALTH ADDITIVE STRATEGY

RESTRICT

inappropriate use of antibiotics and chemicals

www.adisseo.com

REDUCE

losses from subclinical disease and outbreaks

INCREASE

economical and ecological sustainability

BOOST

feed perfomance and farm productivity

AQUAFEED

VOL 16 ISSUE 1 2024

Contents

LOW-FOOTPRINT INSECT-BASED FEED 53 Insect-derived meal at an inclusion rate of up to 10% has positive effects on dealing with stress, the mucosal barrier, wound healing and liver health in salmon.

ENHANCING FISH GUT HEALTH FOR OPTIMAL FEED PERFORMANCE 12

NEW NUTRIENT UTILIZATION ENHANCER 26

MOISTURE MEASUREMENT AND CONTROL IN AQUAFEEDS 42

Marine protein hydrolysates enhance gut health in marine fish species, leading to a significant improvement in feed assimilation.

A combination of active ingredients with a synergistic effect decreases the energy requirements of diets designed for aquaculture species.

In-line moisture measurement to optimize the entire process, from ingredient reception and storage to mixing, drying, and final storage.

Aquafeed: Advances in Processing & Formulation Vol 16 Issue 1 2024

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

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:

This event is held at BITEC in halls 9 and 10 Bangkok International Trade & Exhibition Center

See you at VICTAM ASIA!

4 17.78 x 22.86 Aquafeed.indd 1

15-12-2023 13:21

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 16 ISSUE 1 2024

Contents 6

Interview with Hernan Lim

News Review

12 'You are what you eat’: Enhancing fish gut health for optimal feed performance

18 A phytobiotic-based health solution to promote gut health at multiple levels

21 Spicing up aquafeeds: A novel strategy to mitigate reduced fish oil dietary inclusion, showcased in seabream fed a low fish oilhigh poultry oil diet

26 Nutrient utilization enhancer goes beyond lysophospholipids 30 Dietary potassium diformate in bacterially challenged whiteleg shrimp in Latin America

32 Dietary browning products and mineral availability in rainbow trout

36 Extrusion of premium shrimp feeds on single screw extruders: A review *Cover story

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.

42 Is water killing your fish? Benefits of moisture measurement

Adisseo......................................................................................... 2

46 Slow-release amino acids: How they work in aquafeeds

VICTAM Asia................................................................................ 4 Berg + Schmidt........................................................................... 8 Symrise Aqua Feed..................................................................17 Houdek......................................................................................20 Lucta..........................................................................................24 Nouvelles Vagues....................................................................29 ISFNF ..........................................................................................35 Extru-Tech..................................................................................41 Aquafarm..................................................................................61 WAS ..........................................................................................62

and control in aquafeed production

49 How AI and big data discovered new optimal EPA + DHA levels for salmon

53 Low-footprint insect-based feed shows positive effects on salmon health

57 Reshaping aquafeed formulation with black soldier fly larvae meal: Why the time is now

60 Calendar of events

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

Aquafeed: Advances in Processing & Formulation Vol 16 Issue 1 2024

INTERVIEW with Hernan Lim Hocpo was first founded in 1988 with partners from Taiwan who became the pioneers in the production of prawn feeds during the golden age of monodon prawn farming. In that decade, diseases were uncommon and there were relatively fewer environmental and ecological issues, making it highly profitable especially considering the East Asian market. AQ: Why did you decide to also enter the fish feed market? HL: During the mid-90s a widespread outbreak of luminous bacteria in Visayas caused the monodon industry to collapse. This was largely due to poor farming practices and during those years disease prevention technology was still in its infancy. While finding solutions for the prawn industry in the country and investing in R&D, we had an opportunity to diversify and cater to the fish feed market for both milkfish and tilapia. We took advantage of this to offset the volume loss we incurred from the outbreak. AQ: Where is the company today in terms of size, markets served and aquafeed volume produced? HL: Today the company operates integrated business units that accommodate the various demands of the aquaculture industry in the Philippines; from fry to growout farms, and aquafeeds to the trading of raw material ingredients and farmcare products for both fish and shrimp. Hocpo is primarily concerned with producing feed for shrimp, tilapia, and milkfish as well as our upcoming catfish feeds.

Hernan Lim is President at Hoc Po Feeds Corporation

AQ: Hoc Po Feeds was founded in 1988 as a shrimp feed manufacturer. How did it all start? HL: Our family originated from a province in the Philippines called Cotabato. Having resided alongside a body of water, it allowed us to catch wild milkfish fry (Chanos chanos, locally called Bangus) and trade it locally and internationally as well, Taiwan specifically. This endeavor paved the way for opportunities that opened up different business ventures in the aquaculture industry such as feed milling.

AQ: The Philippines banned the import of PAPs from certain ASF-affected countries in 2022 and reauthorized it again after complaints from the aquafeed industry. How important are these ingredients in the feed formulas in the country? HL: Processed Animal Protein (PAPs) is a key ingredient in our formulation, providing essential amino acids, vitamins, and minerals crucial for fish growth. It's a valuable alternative, reducing costs compared to traditional protein sources like fishmeal. This ensures economic sustainability in producing quality feeds without compromising nutritional content. Notably, formulations without PAP tend to be higher in cost.

Aquafeed: Advances in Processing & Formulation Vol 16 Issue 1 2024

INTERVIEW AQ: How are you diving into the current volatile market conditions for all feed ingredients? Are you prioritizing local ingredients? HL: We are living in a VUCA world, volatile, uncertain, chaotic, and ambiguous. Many factors can influence the prices of raw materials at any time. Therefore, it is crucial to maintain fundamental principles of managing our risk. When the pandemic hit us in 2020, we were forced to reduce our dependence on imported ingredients and prioritize the sourcing of local ones due to limited containers and vessels. Being able to respond to each challenge allowed us to pause, reevaluate our strategies, consistently innovate our protein sources, and pivot when necessary. This kind of positioning served us well during post-pandemic geopolitical conditions that spiked raw material costs globally. Although we are situated in a country surrounded by bodies of water with available local fishmeal, there has been a noticeable decline in the country's fish harvest over the years; therefore, we've established strategic ties with key suppliers to ensure a consistent supply of quality fishmeal. AQ: What is your strategy to reduce the impact of the current context on the final feed price? HL: To address the price volatility of raw materials, we customize feed formulations and utilize feed additives to optimize the national content of the formulation. We consistently pursue innovation as we explore alternative protein sources and innovative technologies that aim to enhance cost-effectiveness without compromising the nutritional value of our feeds. Maintaining access to quality raw materials is a priority, that is why we have long since established a business subsidiary that directly trades and hedges major raw materials. This allows us to secure our prices months in advance to gain leverage and economies of scale. AQ: Hoc Po Feeds utilizes pelleting technology to produce its shrimp feeds. Do you plan to change to extrusion technologies? Are energy costs an issue for feed mills in the Philippines? HL: Energy cost is definitely an issue for feed mills in the Philippines as it is one of the highest in Asia, second only to Japan. That being said, extruded feeds require more power thus having a higher manufacturing cost when compared to pelleted feeds. We are still evaluating the environmental and nutritional viability of adopting extrusion technology for our shrimp feeds.

AQ: Hoc Po Feeds operates two aquaculture farms and a hatchery. Why did you decide to set up your own farms and what kind of services do they provide? Do you have a specific selective breeding program for the hatchery? HL: Hocpo established its subsidiary HP Aquafarms to complement the feedmill, and engage in research and development for fry, feeds, and various farm care products such as probiotics and immune-stimulants. This approach allows us to observe and learn more about the effect the feeds have compared to all other factors and conditions that affect the growth and performance of farmed aquatic animals. This helps sustain our learning, innovation and product development. In line with Hocpo’s commitment to support the advancement of aquaculture in the Philippines, the company maintains ongoing partnerships with research institutions and universities within the country. These collaborations enable the company to conduct technical trials and provide training opportunities for students. These farm trials utilize various protocols aimed at enhancing productivity and strengthening the Philippine aquaculture industry. As of this time, we do not have any breeding programs for vannamei. We source our SPF (specific pathogen free) and SPR (specific pathogen resistant) broodstock from a facility in Hawaii. AQ: A wide variety of species are farmed in the Philippines. What are the main issues farmers are facing in the Philippines? HL: The aquaculture industry in the Philippines faces several challenges. Firstly, unsustainable aquaculture methods caused by greed have led to the deterioration of marine ecosystems. The overcrowding and overfeeding of fish in cages have led to degraded water quality, slow growth, higher feed conversion ratios, and a higher risk of disease outbreaks. Additionally, in the Philippines, there is a lack of cold storage and processing plants that affect the farmers' ability to sell their goods directly to the market. It is important as well for our government to establish policies to properly regulate the import of marine products, to give consideration to local production and provide local farmers adequate opportunities and fair pricing for their goods. Lastly, the effects of El Niño on the country's aquaculture industry are also a concern for farmers.

Aquafeed: Advances in Processing & Formulation Vol 16 Issue 1 2024

INTERVIEW

AQ: Specifically in shrimp, what are the main challenges? How Hoc Po Feeds is supporting farmers? HL: High energy costs hinder the Philippines' shrimp industry competitiveness. While we predominantly sell our shrimp locally, the sector lags in growth compared to other countries. Growth is limited due to disease outbreaks, lack of government support, and insufficient processing facilities. Hocpo addresses these challenges by developing its functional shrimp feed and innovating sustainable protocols to lower power costs and improve

production efficiency. We also assist farmers with our diagnostic laboratories to aid them in managing water quality, monitoring animal health and implementing disease risk management to enhance harvest success. Our dedicated technical team provides steadfast support for farmers to optimize their farm operations. Our objective is to approach each customer's farm with the same level of care and attention as we would our own. AQ: What are Hoc Po Feeds projections for growth going forward? HL: The Philippines, Asia's 3rd largest archipelago with 7,600 islands and the world's fifth longest coastline, faces a threat to food security due to the decline in wild-caught fish. However, this challenge presents a significant opportunity for the aquaculture industry. With the persistent decline in wild fish and the growing Philippine population, there is a rising demand for farmed aquaculture. With an annual per capita consumption of 40 kilograms in 2022, fish is still the most popular animal protein consumed in the Philippines, according to the Philippine Statistics Authority. This is where Hocpo is able to create value; by having the means to manufacture environmentally responsible aquafeed. Simultaneously, it has a huge potential to expand its manufacturing capacity to be able to meet the demands and support the industry. It's crucial to recognize that several obstacles and unknowns could hinder our progress. It would be appropriate to state that we have a cautiously optimistic outlook for the future in light of these circumstances. We must continue to be vigilant and adaptable to overcome any potential challenges that may come up in the future.

Aquafeed: Advances in Processing & Formulation Vol 16 Issue 1 2024

NEWS

NEWS REVIEW Highlights of recent news from Aquafeed.com

Cargill opens state-of-the-art premix plant in Vietnam The company recently opened a new Provimi premix plant in Giang Dien commune, in the southern province of Dong Nai, that will improve access to world-class additives and specialty ingredients for Vietnamese livestock and aquaculture farmers, as well as the nation’s feed mills. Spanning 30,000 square meters, the Giang Dien factory is more than nine times the size of its 27-year-old predecessor, and its 40,000-ton annual capacity represents a 100% increase over the plant it supersedes.

Skretting unveils new generation of hatchery feed with better resource utilization and lower emissions The company introduced Nutra Terra, a novelty in hatchery feed with documented high performance and reduced emissions. “Nutra Terra represents our largest innovation in grower feed for juvenile fish in over a decade. All available knowledge and research have been gathered to develop a new and optimized feed recipe. The result is a feed that provides better performance while reducing emissions from the fish and making better use of raw materials in the feed,” the company said.

Nicovita unveils innovative feed that takes care of shrimp pond soil At Aqua Expo Guayaquil, Nicovita unveiled a new line of products, Proterra, a mix of sporulated Bacillus sp. strains that begin their action in the digestive tract of shrimp. When the active particles come into contact with the moisture present in the shrimp's digestive system, it triggers the activation of compounds that interrupt the nitrogen cycle, preventing the formation of additional nitrogenous compounds. This process is maintained after being converted into feces, continuing to act when they reach the ground, which prevents the increase of the levels of organic matter.

Payper updates its open-mouth bagging range In a market where precision and efficiency are principal, PAYPER introduces its upgraded open-mouth bagging range. This improved series comprises six fully automatic models capable of reaching speeds of up to 1,800 bags per hour. They adapt to a diverse range of dry bulk materials, whether free-flowing or non-free-flowing. The new CSA open-mouth bagging line was redesigned and optimized to reduce energy consumption while elevating production speeds.

Aquafeed: Advances in Processing & Formulation Vol 16 Issue 1 2024

NEWS ADM to acquire PT Trouw Nutrition Indonesia

ADM has reached an agreement to acquire PT Trouw Nutrition Indonesia, a subsidiary of Nutreco and a provider of functional and nutritional solutions for livestock farming in Indonesia. Incorporated in 2007, PT Trouw Nutrition Indonesia is a premix manufacturer, providing innovative and comprehensive nutrition solutions for the animal industry. With the planned acquisition, ADM will be strengthening its premix and feed additives & ingredients (FA&I) business and strategically positioning itself to meet the anticipated market growth to sustain the rising demand for protein.

Veolia gets approval to export insect meal and oil into Europe Veolia Bioconversion Malaysia, part of Veolia Group, received approval from the Ministry of Agriculture in Malaysia to export its insect meal Entomeal™ and insect oil Entolipid into the European markets for use in pet food, aquaculture and livestock. Insect meal and oil are produced with EU industrial standards and fully compliant with EU regulations to be used in animal feed applications. TRACES ensures the safety and full traceability from feedstock to finished products of our production.

Entobel opens the largest insect protein production plant in Asia The state-of-the-art facility is currently the world’s most CAPEX-efficient BSF production facility. The Vung Tau facility marks Entobel’s second industrial-scale production facility in Vietnam and will have an annual production capacity of 10,000 MT of insect protein.

Northern Europe’s largest insect factory opens in Denmark

Enorm Biofactory opened the doors to the largest insect factory in Northern Europe, which is set to produce over 10,000 tons of insect meal per year using byproducts from the food industry. Covering 22,000 square meters, processing 100 tons of black soldier fly larvae per day, and yielding more than 10,000 tons of insect meal annually, the newly constructed factory is placed in Eastern Jutland, Denmark.

Aquafeed: Advances in Processing & Formulation Vol 16 Issue 1 2024

NEWS Yield10 Bioscience, BioMar partner to grow fish oil on land

Yield10 Bioscience and BioMar Group have signed an agreement to form a long-term partnership to commercialize a camelina crop containing enriched levels of EPA and DHA equal to fish oil. Over the next year, Yield10 expects to scale up planted acres of camelina to supply BioMar with oil for formulation and testing while also working towards securing regulatory approval for commercial production of omega-3 camelina oil and meal in the targeted production geographies.

Andritz opens Food Innovation Xperience test center The company opened a leading-edge test and research center for the food and feed industry in Waddinxveen, the Netherlands. The test center is equipped with the latest milling, extraction, dewatering, and drying technologies from Andritz, enabling customers to conduct feasibility studies, and pilot plant tests for scale-up or R&D activities under foodgrade or even ATEX conditions. Covering a total area of 850 m2, the center is available to customers in the food and feed industry as well as to research and development organizations.

Enifer secures €12 million grant to build a first-of-its-kind mycoprotein ingredient factory

Protix, Tyson Foods to build insect facility in the US

Business Finland has conditionally approved a recycling and reuse investment grant of more than €12 million for biotech startup Enifer to build its first commercialscale PEKILO® mycoprotein ingredient factory. The new facility will be built in the Uusimaa region and will have a production capacity of 3 million kilograms a year of sustainable, locally sourced protein. The factory is currently projected to cost €30 million to build and is expected to be completed by the end of 2025, with production rampup occurring in 2026.

Tyson Foods reached an agreement for a two-fold investment with insect ingredients company Protix. The strategic investment will support the growth of the emerging insect ingredient industry and expand the use of insect ingredient solutions to create more efficient sustainable proteins and lipids for use in the global food system. In addition, Tyson Foods and Protix have entered a joint venture for the operation and construction of an insect ingredient facility in the continental United States.

Aquafeed: Advances in Processing & Formulation Vol 16 Issue 1 2024

PROTEINS

'You are what you eat’: Enhancing fish gut health for optimal feed performance Mikael Herault, Symrise Aqua Feed, Dr. Kyeong-Jun Lee, Jeju National University The consolidation of 16 trials conducted in red seabream demonstrates the potential of marine protein hydrolysates to enhance gut health in marine fish species, leading to a significant improvement in feed assimilation.

The benefits of using Marine Protein Hydrolysates (MPH) in aquafeeds have been widely documented. In this way, our team has demonstrated the advantages of incorporating shrimp and tilapia powder hydrolysates (SPH* and TPH*, respectively) to restore the performance of olive flounder when fed diets with reduced levels of fishmeal (FM) (Gunathilaka et al., 2020). Numerous clinical trials were also conducted in red seabream using these two products for internal reference purposes. Some of them have been published (Khosravi et al., 2015; Gunathilaka et al., 2021) but some haven’t yet, and their consolidation provides further insights to better understand their potential for improving feed assimilation and the underlying modes of action.

A comprehensive and standardized study of MPH uses in the contexts of FM replacement or supplementation Between 2014 and 2019, 16 trials were conducted at Jeju University in South Korea (Fig 1A, 1B) to assess the nutritional and health performance of new dietary solutions in red seabream, with SPH and TPH as our internal benchmarks. Both MPH were incorporated at 5% to facilitate the substitution of approximately 50% of the initial fishmeal with plant proteins, primarily soybean protein (Table 1, depicting typical High Fishmeal (HFM) and Low Fishmeal (LFM) formulations for red seabream). These designs provided the opportunity to evaluate the benefits of MPH in both dietary fishmeal substitution

A B

Figure 1. (A) Jeju University fish testing facilities. (B) Red seabream.

and supplementation contexts (LFM + 5% MPH vs. HFM diets and LFM + 5% MPH vs. LFM diets, respectively). After 12 to 15 weeks of feeding trials, fish were sampled to collect and measure an extensive array of zootechnical, physiological, and immune parameters. For this study, we will specifically focus on the following parameters: Specific Growth Rates (SGR in %/d), FCR and its mirror parameter Feed Efficiency (FE, calculated as 1/FCR), Villi Length (VL in µm), Enterocyte Height (EH

Aquafeed: Advances in Processing & Formulation Vol 16 Issue 1 2024

PROTEINS Table 1. Feed formulations (dry basis) and nutritional composition used in the trial.

Experimental diets Ingredients HFM LFM SPH/TPH FM FAQ65

30.00

15.00

SPH/TPH

15.00 5.00

Soy protein concentrate

15.00

26.40

21.40

Corn gluten meal

12.00

Wheat flour

30.00

Fish oil

4.65

6.00

Soybean oil

4.65

4.20

3.90

Mineral/vitamin Mix

2.00

Starch

1.20

1.50

Choline chloride

0.50

L-Lysine

0.00

0.50

L-Methionine

0.00

0.20

Taurine

0.00

0.50

Di-calcium phosphate

0.00

1.50

Proximate composition (%, dry matter) Crude protein

45.4

45.7

45.4

Crude lipid

16.6

17.1

16.8

5.76

5.00

5.26

Gross energy (MJ.kg )

21.78

21.59

21.65

Moisture

5.37

7.25

6.63

Crude ash -1

in µm) and Goblet Cell density (GC, number per crosssection). Simultaneously with these feeding studies, Apparent Digestibility Coefficients for Dry Matter (ADCDM in %) and Crude Protein (ADCCP in %) were assessed with fish from the same hatch for 3 weeks. The same dietary formulation was used, and Chrome Oxide (Cr2O3) served as an inert marker for these assessments.

SPH demonstrates more versatility when substituting FM with plant proteins in carnivorous fish species Red seabream is a marine carnivorous fish species that requires a high level of dietary crude protein, which must be of good nutritional quality and palatable. SGR and FCR indicators, as illustrated by Figure 2A and 2B, confirmed that it was possible to effectively substitute 50% of fishmeal with plant proteins as long as 5% MPH were used to compensate for the missing soluble protein, associated bioactive peptides, and dietary palatability.

It is important to note the differences in observed benefits based on the context and the source of MPH inclusion. When it comes to dietary supplementation, we observed higher SGR and FCR gains compared to fishmeal replacement (p-value < 0.05). In less challenging nutritional conditions, both TPH and SPH are equally effective, with approximately 9% higher SGR and 13% lower FCR to be expected compared to the control diet (LFM). Additionally, in the case of dietary application, the use of TPH in the LFM diet successfully restored the growth and feed conversion performance observed in the HFM diet, but dietary SPH outperformed it with a 5% higher SGR and a 10% lower FCR when compared to the LFM diet. The use of MPH led to higher feed efficiency, and this improvement was in part attributed to enhanced ADC for dry matter and crude protein. In the context of fishmeal replacement, 5% MPH dietary inclusion allowed for maintaining the observed ADC values seen in the high-level FM diets (Figure 2C, 2D). When used as a dietary supplement, it resulted in a significant increase in ADC by 6% (SPH) to 8% (TPH). These improvements far exceed what would typically be expected when replacing only 5.0% of dietary dry matter, or 7.8% of dietary crude protein considering the 75% average protein content for MPH. This suggests that, in addition to promoting increased growth rates, there are other physiological changes associated with the use of MPH in diets, which contribute to improved nutrient assimilation and, consequently, feed efficiency.

Unraveling the role of MPH in enhancing fish gut physiology and nutrient assimilation: Differential actions and performance The study of fish gut histology helps us better understand how the dietary use of MPH enhances dietary nutrient assimilation. The Spearman correlations (Table 2) indicated strong and significant relationships between FE and ADCs, with ADCs themselves being correlated with VL and GC. Notably, VL was found to be highly correlated with EH. While there may be additional mechanisms of action undescribed in this study, such as modulation of intestinal amino acids and/or peptide transporters like Pept1, we can reasonably infer that MPH functionalities

Aquafeed: Advances in Processing & Formulation Vol 16 Issue 1 2024

PROTEINS

Figure 2. Normalized feed assimilation and ADC gains over controls (LFM + 5% MPH vs. HFM diet for FM replacement topic and LFM + 5% MPH vs. LFM diet for the supplementation one) (A) SGR; (B) FCR; (C) ADC for DM; (D) ADC for CP.

Aquafeed: Advances in Processing & Formulation Vol 16 Issue 1 2024

PROTEINS Table 2. Spearman correlation coefficients between fish gut morphometrics, FE and ADCs for DM and CP (n=16 individuals, p-values are mentioned below the coefficients). FE= Feed Efficiency, VL= Villi Length; EH = Enterocyte Height; GC = Goblet Cell density; ADCDM = Apparent Digestibility Coefficients for Dry Matter; ADCCP = Apparent Digestibility Coefficients for Crude Protein.

FE (1/FCR)

ADCDM

ADCCP

0.7055 p < 0.05

0.7059 p < 0.01

0.3588 p > 0.10

0.1647 p > 0.10

0.5618 p < 0.05

0.8374 p < 0.01

0.6220 p < 0.05

0.4154 p > 0.10

0.5165 p < 0.10

0.6471 p < 0.05

0.3971 p > 0.10

0.5618 p < 0.05

0.8500 p < 0.001

0.1735 p > 0.10

ADC DM

0.7055 p < 0.05

ADC CP

0.7059 p < 0.01

0.8374 p < 0.01

0.3588 p > 0.10

0.6220 p < 0.05

0.6471 p < 0.05

0.1647 p > 0.10

0.4154 p > 0.10

0.3971 p > 0.10

0.8500 0.0706 p < 0.001 p > 0.10

0.5618 p < 0.05

0.5165 p < 0.10

0.5618 p < 0.05

0.1735 p > 0.10

promote the development of enterocytes (EH), affecting the size of gut villi (VL) and the density of goblet cells (GC, Fig. 3A). After having undergone an enzymatic hydrolysis process, a large proportion of the protein amino acids from MPH is found in free form. These free amino acids are known to have a direct trophic effect on the enterocyte layer. Antioxidative and antimicrobial peptides, commonly found in MPH, also play a role in maintaining gut cell integrity and their defense against stressors like anti-nutritional factors in plant protein or disruptions in gut microbiota. This is how dietary changes at the molecular level (the change of MPH native protein form and resulting functionality) significantly impact animals at the cellular and tissue levels, with bigger enterocytes and gut villi. Numerous references demonstrate that the size and shape of vertebrate gut villi modulate nutrient absorption and feed assimilation. In parallel, goblet cells play an important role by secreting protective mucus, reducing the risks of gut inflammation, or infection, while increasing nutrient bioavailability. Enhanced fish gut physiology ultimately leads to improved ADC for both macro and micro-nutrients. It is important to note that the modes of action vary when using different MPH; SPH demonstrated higher values for villi length (VL), while TPH exhibited

0.0706 p > 0.10

higher values for goblet cell densities (GC, as shown in Figure 3B, 3C, 3D). These differences may partially explain why SPH was found as more effective than TPH in the context of FM replacement.

Conclusion and perspectives Marine protein hydrolysates are widely recognized for their palatability and high digestibility as functional ingredients. However, there is a limited number of references proving their potential to enhance fish gut health and subsequent nutrient assimilation. Through the consolidation of data from several trials conducted in red seabream with dietary MPH, we have brought to the forefront this potential and have elucidated some of the underlying modes of action. Our findings illustrate the practical applications of dietary MPH, showcasing the effective reduction of FM usage in marine fish species, and offering advantages both in terms of feed efficiency and growth performance. These benefits become more pronounced when opting for MPH dietary supplementation. Our analysis also showed that the raw material origin of MPH affected the functional benefits of the finished product. The hydrolysis of protein coming from fish or shrimp can lead to the production of different peptide profiles and AA peptide sequences carrying various biological activities and thus affecting different tissues and functions.

Aquafeed: Advances in Processing & Formulation Vol 16 Issue 1 2024

PROTEINS Taking a holistic approach to MPH dietary supplementation to enhance feed and nutrient assimilation can lead to additional advantages for the industry, such as improved water quality and reduced environmental eutrophication. Simultaneously, besides the decreased environmental impact on feed production (Life Cycle Analysis), the industry can anticipate enhanced Fish In Fish Out (FIFO) ratios when using MPH for reducing FM in diets. Further research concentrating on the influence of MPH on the assimilation of crude fat in marine fish species, ideally with detailed Apparent Digestibility Coefficients (ADC) for critical fatty acid components (PUFA), holds promise in reducing the reliance on fishmeal and oil in current dietary formulations.

References Khosravi Sanaz, Rahimnejad Samad, Herault Mikael, Fournier Vincent, Lee Cho-Rong, Bui Hien Thi Dieu, Jeong Jun-Bum, Lee Kyeong-Jun (2015). Effects of protein hydrolysates supplementation in low fish meal diets on growth performance, innate immunity and disease resistance of red sea bream Pagrus major. Fish & Shellfish Immunology, 45, 858-868. Gunathilaka Buddhi E., Khosravi Sanaz, Herault Mikael, Fournier Vincent, Lee Chorong, Jeong Joon-Bum, Lee Kyeong-Jun (2020). Evaluation of shrimp or tilapia protein hydrolysate at graded dosages in low fish meal diet for olive flounder (Paralichthys olivaceus). Aquaculture Nutrition, 26(5), 1592-1603. Gunathilaka Buddhi E., Khosravi Sanaz, Shin Jaebeom, Shin Jaehyeong, Herault Mikael, Fournier Vincent, Lee Kyeong-Jun (2021). Evaluation of shrimp protein hydrolysate and krill meal supplementation in low fish meal diet for red seabream (Pagrus major). Fish Aquatic Sciences, 24(3), 109-120. *SPH and TPH, standing for Shrimp Powder and Tilapia Powder Hydrolysates are known on the feed market as Actishrimp and Actifish respectively.

More information: Mikael Herault R&D Performance Measurement Manager Symrise Aqua Feed E: mikael.herault@symrise.com

Figure 3. Normalized red seabream gut morphometric gains over controls (HFM diet for FM replacement topic and LFM diet for the supplementation one). (A) microscopic details of (B) VL, (C) EH and (D) GC (u).

Dr Kyeong-Jun Lee Professor in the Department of Marine Life Sciences Jeju National University, South Korea E: kjlee@jejunu.ac.kr

Aquafeed: Advances in Processing & Formulation Vol 16 Issue 1 2024

always inspiring more …

ActiShrimp, functional hydrolysate for aquafeed Reach optimum feed intake Improve resistance to stress and pathogens Better yield aquafeed.symrise.com

FEED ADDITIVES

A phytobiotic-based health solution to promote gut health at multiple levels I-Tung Chen, Maria Mercè Isern-Subich, Waldo G. Nuez-Ortín, Adisseo

Gut health is fundamental for the well-being of farmed fish as it supports nutrient absorption, disease prevention, and growth performance. The complex gastrointestinal tract ecosystem, consisting of the epithelium, microbiota, and immune system, works harmoniously to promote metabolism and maintain animal health. The epithelium, a protective layer lining the gut, serves as the first line of defense against harmful microorganisms and parasites while also playing a crucial role in nutrient transport, digestive enzyme secretion, and gut microbiota balance. However, challenges in aquaculture production conditions, such as using plant ingredients in feed, high rearing densities, or suboptimal temperatures, can impair immunity and disrupt the delicate balance of the gut ecosystem. These factors can lead to chronic inflammation and compromised gut integrity, which in turn increases the risk of infections and reduces nutrient absorption. To address these negative effects, it is important to focus on nutrition strategies that emphasize gut health to prevent the entry of bacteria and parasites into extraintestinal tissues and mitigate chronic inflammation. This article describes the role of Sanacore® GM (Adisseo) as a synergistic blend of phytobiotic extracts. One of the purposes of the application of Sanacore® GM is to strengthen gut health, resulting in better fish growth, less impact of bacterial and gut parasite infections, and overall vitality.

Gut health through integrity, anti-inflammatory benefits, and immunocompetence Gut integrity is an indicator of gut health. Transepithelial electrical resistance (TER) in the intestine of gilthead

seabream was measured to determine the integrity of the intestinal barrier. The higher the resistance value, the better gut integrity. Results revealed a 30% increase in TER of fish fed a 10% fishmeal (FM) feed supplemented with Sanacore® GM, suggesting better villi development along with stronger intestinal connections. Tighter junctions prevent harmful substances and pathogens from passing through the gut lining, leading to better overall health and growth. Gut integrity enhancement by Sanacore® GM was also measured in tilapia fed a zero-FM feed. In this species, the additive increased TER by 16%. Overall, results across species validate the efficacy of the additive to support fish gut health under challenging feed formulations, at early stages of development, or during critical periods of production. Gut morphometry is crucial for optimal gut health in fish as it encompasses aspects such as mucosal surface area and the potential for optimal nutrient absorption. An indicator of the mucosal surface area is the ratio of the internal mucosal perimeter in relation to the external/section perimeter, the higher the mucosal surface area, the higher the internal perimeter, the higher the ratio. In the hindgut of gilthead seabream fed 10% FM feed, Sanacore® GM increased the ratio by 7%, providing a larger mucosal surface and suggesting more optimal nutrient absorption under supplementation. Intestinal inflammation can arise from various factors, including dietary factors and infections. Inflammation can be elicited in the lamina propria, a layer of the gut epithelium hosting immune cells and supplying irrigation to the epithelium. Histopathological analysis of the different gut sections of gilthead seabream fed 10% FM feed has proved a consistent decrease in

Aquafeed: Advances in Processing & Formulation Vol 16 Issue 1 2024

FEED ADDITIVES

Figure 1. Schematic representation of the beneficial effects of Sanacore® GM on gut health at multiple levels. (A) Production conditions such as plant-based aquafeeds, infections, or thermal stress, can impair gut health and lead to chronic inflammation and compromised immune function. Sanacore® GM promotes gut health by (B) strengthening gut barrier integrity to prevent the entry of harmful substances and pathogens; (C) enhancing nutrient absorption through well-developed villi and mucosal surface area; (D) supporting immunocompetence by optimizing antioxidant mechanisms, phagocytosis, and anti-inflammatory response. Figure created with BioRender.com.

Aquafeed: Advances in Processing & Formulation Vol 16 Issue 1 2024

FEED ADDITIVES

lamina propria infiltration and thickness by 15-55%. This observation indicates that Sanacore® GM can effectively alleviate gut damage and inflammation. The gut also plays an important role in regulating the full immune system and general health of fish. Sanacore® GM has been shown to regulate expression changes in proteins involved in intestinal antioxidant defenses as a basic pillar of optimal immunity, phagocytosis as the major innate mechanism for eliminating pathogens, and intestinal inflammatory response. These results indicate that the additive supports gut immunocompetence not only via reinforcement of physical barriers but also via active cellular and humoral responses.

Conclusion This article briefly describes some evidence regarding the benefits of Sanacore® GM to support gut health. Under preventive supplementation strategies, such gut health-promoting mechanisms can support fish in dealing with intensive production conditions and current aquafeed formulations and reduce the severity of parasite and bacterial infections.

More information: Waldo Nuez, PhD Global R&D Manager Aquaculture Adisseo E: waldo.nuezortin@adisseo.com

Aquafeed: Advances in Processing & Formulation Vol 16 Issue 1 2024

FEED ADDITIVES

Spicing up aquafeeds: A novel strategy to mitigate reduced fish oil dietary inclusion, showcased in seabream fed a low fish oil-high poultry oil diet Alberto Ruiz, Enric Gisbert, IRTA, Thiago Raggi, Sofia Morais, LUCTA S.A.

The fish oil challenge Availability and cost of fish oil has become a main bottleneck for the growth of the aquaculture sector, and for maintaining the end-quality and welfare of farmed fish. Fish oil provides energy and essential nutrients, including n-3 long-chain polyunsaturated fatty acids (LC-PUFA), mainly eicosapentaenoic acid (EPA) and docosahexaenoic acid (DHA), cholesterol and fat-soluble vitamins. A steady growth of aquaculture production, with declining or stagnant fish oil production in the last two decades, has only been possible through the gradual replacement of fish oil with other fats, mostly of vegetable origin. However, the dietary level of fish oil is reaching, or has already reached, its lower limit in most industries, and cannot be further reduced without significant negative effects on fish performance, health, and end-quality. Most of these impacts are associated with the anti-inflammatory, cardioprotective, and lipotropic role of omega-3 fatty acids (particularly of n-3 LC-PUFA). Conversely, omega-6 (highly abundant in plant oils) have opposite effects. Commercially viable alternative sources of omega-3 are emerging in the industry, but cost and volumes are still limiting for use in most species, markets, or production stages. Therefore, new strategies are urgently needed to optimize the efficiency of the use of fish oil and

alternative omega-3 sources. Formulation and supplementation strategies to aid in the strategic use of fish oil The overall composition and fatty acid class balance of fish oil have been given less attention than its level of omega-3. The principal fatty acids of most commercial fish oils used in aquafeeds are 14:0, 16:0, 16:1n-7, 18:1n9, EPA and DHA, and they typically contain 25-30% saturated fatty acids (SFA), 20-50% monounsaturated fatty acids (MUFA) and 20-45% polyunsaturated fatty acids (PUFA), of which most are omega-3. Replacing fish oil with vegetable oils greatly changes the dietary fat profile, most notably by reducing SFA and increasing omega-6 levels concurrently with omega-3 reduction. The physiological consequences of these imbalances are magnified by the fact that omega-3 and omega-6 are competitively metabolized by the same set of enzymes, and the metabolites of these enzymatic conversions perform antagonistic functions. On the other hand, SFA and MUFA are preferential substrates for metabolic energy production, sparing LC-PUFA, notably EPA and DHA, from catabolism. Several fish studies have already demonstrated that this omega-3 sparing effect can increase LC-PUFA in the edible fraction and/or reduce the dietary levels needed to satisfy essential fatty acid requirements. Hence, optimized formulation strategies should aim to reduce

Aquafeed: Advances in Processing & Formulation Vol 16 Issue 1 2024

FEED ADDITIVES omega-6 and increase SFA levels, maintaining high MUFA contents, in aquafeeds. In this respect, rendered terrestrial animal fats present an advantageous profile. Moreover, besides being affordable, they meet circularity criteria and are widely and often locally available. Spices, on the other hand, present a range of welldemonstrated nutraceutical benefits, with the potential to be integrated into dietary strategies to mitigate the negative impacts of low fish oil feeds. Many mammalian studies have established that pungent spices have hypocholesterolemic and hypotriglyceridemic roles (similarly to n-3 LC-PUFA in fish oils), stimulate digestion, and have anti-inflammatory, antioxidant, and antimicrobial effects.

Gilthead seabream fed poultry oil-rich diet supplemented with a blend of spices showed improved performance In order to demonstrate the beneficial effects of pungent spices in fish feeds containing a reduced fish oil level and poultry oil as the principal fat source, a 90-day feeding trial was performed in IRTA research facilities with gilthead seabream juveniles (start weight: 44.1 ± 4.1 g), in tetraplicate 450 L recirculation tanks, at 22.5 ± 0.5°C. Fish were fed two isoproteic, isolipidic and isoenergetic diets: a basal feed (Control; formulation and composition shown in Table 1) and an experimental feed with the same formulation but supplemented with a combination of pungent spices (Lucta, Spain), at an inclusion level of 0.1% (Spicy). At the end of the trial, BW and SGR increased significantly in fish fed the Spicy diet compared with the Control diet, while FCR was significantly decreased (Table 2). Spices changed lipase activity, lipid accumulation and lipid metabolism in gilthead seabream The concentration of bile salts, measured in the anterior intestine of fish at 2h after feeding, was 37% higher (although P<0.05; Fig. 1A), and the activity of bile saltactivated lipase was significantly increased in the Spicy group relative to the Control (Fig. 1B). Accumulation of fat in enterocytes and hepatocytes was evaluated histologically in 48h-fasted seabream. Fish fed the Spicy diet showed substantially lower lipid accumulation in the anterior intestine, with a higher percentage of specimens with very low (1) and none with moderate (3) scores (Fig. 2A). An important reduction in high (4) lipid accumulation

Table 1. Main ingredients (≥2%) and composition of the basal (Control) diet. The experimental (Spicy) diet was further supplemented with 0.1% of a combination of pungent spices.

Ingredients (%)

Control

Fishmeal Super Prime

7.50

Fishmeal 60

5.00

Fish protein concentrate

2.00

Feathermeal hydrolysate

5.00

Porcine blood meal

3.00

Poultry meal

15.00

Aminopro NT70

4.00

Corn gluten meal

8.00

Soybean meal 48

12.00

Sunflower meal

5.00

Wheat meal

10.31

Whole peas

5.00

Pea starch (raw)

2.40

Fish oil

3.02

Soybean oil

2.35

Poultry fat

8.04

Proximate composition Crude protein (%)

44.14 ± 0.05

Crude fat (%)

18.10 ± 0.04

Gross energy (MJ/kg)

21.38 ± 1.11

Fatty acid profile (% of total fatty acids) Saturated fatty acids (SFA)

27.19 ± 0.40

Monounsaturated fatty acids (MUFA)

36.61 ± 0.73

n-6 polyunsaturated fatty acids (n-6 PUFA) 26.65 ± 0.06 n-3 polyunsaturated fatty acids (n-3 PUFA)

9.55 ± 0.39

Table 2. Growth and feeding performance at the end of the 90-day trial. Different letters in each row indicate significant differences (P<0.05) between treatments. Values are mean ± SD (n=4 tanks per group).

Control

Spicy

BWf (g)

215.80 ± 1.06

221.96 ± 3.46b

SGR (%/day)

1.81 ± 0.01a

1.84 ± 0.02b

FI (g fish-1)

195.89 ± 8.70

192.29 ± 9.94

FCR

1.21 ± 0.05

1.13 ± 0.02a

PVFI (%)

3.01 ± 0.28b

2.32 ± 0.37a

Aquafeed: Advances in Processing & Formulation Vol 16 Issue 1 2024

FEED ADDITIVES

Figure 1. Concentration (µg/mg) of total bile acids (A) and enzymatic activity (mU/mg protein) of bile salt-activated lipase (B) in the anterior intestine of seabream, 2h after feeding. Values are mean ± SD (n=4 tanks per group). Different letters indicate significant differences (P<0.05) between treatments.

Figure 3. Differentially expressed lipid metabolism markers in the liver of seabream fed Control or Spicy feeds, 2 h after feeding. Values represent normalized relative gene expression mean ± SD (n=8 fish per group). ** indicates significant differences (P<0.05) and * trends (P<0.1), between groups.

Figure 2. Semi-quantitative histological scoring of fat accumulation in the anterior intestine (A) and liver (B) of 48h-fasted seabream at the end of the trial. Scores describe very low (1), low (2), moderate (3) and high (4) lipid accumulation in enterocytes and hepatocytes. Values represent the score frequency mean ± SD (n=12 fish per group).

scores in the liver was also observed in the Spicy group (Fig. 2B). Perivisceral fat index (PVFI), assessed in whole fish at the end of the trial, significantly decreased by more than 20% in the Spicy group (Table 2). These results were associated with changes in 2 h-postprandial hepatic gene expression of several biomarkers involved in the incorporation of fatty acids into tissues (lipoprotein lipase: lpl), in de novo fatty acid synthesis (fatty acid synthase: fasn, elongation of very long chain fatty acids: elovl6, stearoyl-CoA desaturase 1b: scd1b), in the regulation of fatty acid β-oxidation (peroxisome proliferator-activated receptor β: pparβ), and bile acid synthesis from cholesterol (cholesterol 7-alpha-monooxygenase: cyp7a1) (Fig. 3). The up-regulation of de novo fatty acid synthesis and down-regulation of fatty acid β-oxidation biomarkers

Aquafeed: Advances in Processing & Formulation Vol 16 Issue 1 2024

FEED ADDITIVES in fish fed the Spicy diet might seem contradictory given that spices have been reported to regulate these pathways in an inverse way, but it is likely a response to attempt to restore the low hepatic fat stores. The down-regulation of lpl is consistent with the decreased PVFI and lower hepatic fat deposits in fish fed the Spicy diet, given the key role of this enzyme in the hydrolysis of circulating lipids and their incoporation into tissues. Enhanced expression of cyp7a1, on the other hand, indicates a higher synthesis of bile acids, correlating with results shown in Figure 1.

The immunomodulatory and gut microbiota modulation potential of pungent spices Changes in immune gene markers were observed in the intestine of 48h-fasted fish, with down-regulated genes - pattern recognition receptor: cd302, cell markers: cd4-1 and cd8b, and interleukins: il-15 and il-34 - indicating an anti-inflammatory response in fish fed the Spicy diet (Fig. 4). Regarding gut microbial communities, results indicate a healthy intestinal status in

Figure 4. Differentially expressed immune markers in the anterior intestine of 48h-fasted seabream from Control or Spicy groups. Values represent normalized relative gene expression mean ± SD (n=8 fish per group). ** indicates significant differences (P<0.05) and * trends (P<0.1), between groups.

both treatments, although the Simpson’s Diversity Index was significantly higher in the posterior intestine of fish fed the Spicy diet. This was associated with an increased relative abundance of the phylum Chloroflexi and lower relative abundances of the genera

Aquafeed: Advances in Processing & Formulation Vol 16 Issue 1 2024

FEED ADDITIVES Campylobacter, Corynebacterium and Peptoniphilus (Fig. 5). Such changes might be caused by a potential microbial effect of the pungent spices on the different bacterial strains or associated to the higher levels of total bile acids in the intestine, which also have a modulatory role on the diversity and composition of gut bacterial populations.

Conclusion In conclusion, the blend of pungent spices significantly improved growth and feeding efficiency in gilthead seabream. A lower fat accumulation in the visceral cavity was also observed, with the potential to enhance consumer perception and extend the shelf-life of the end product. Reductions in intracellular lipid accumulation in the intestine and liver, coupled with changes in the expression of lipid metabolism genes, and increased activity of bile salt-activated lipase, confirm the lipotropic and lipid digestion-enhancement (likely through higher hepatic biosynthesis of bile acids) effect of the tested spices. Regulation of the intestinal immune response and subtle changes in gut microbiota were also found, suggesting an improved health status and condition of seabream fed the Spicy-supplemented feed. Overall, the results showcase that Spicy supplements, by mitigating several of the negative effects of low fish oil diets, could be used as part of a functional nutrition approach to optimize the use of fish oil as a strategic ingredient in the aquafeed industry. Figure 5. Relative abundances of gut bacterial taxa in the posterior intestine of 48h-fasted seabream (n=12 fish per group). Data shown are at the level of phylum (A) and genus (B) (excluding unassigned genera). Taxa with an abundance <1% are classified as others in the bar graph and not represented in the bubble plot. Asterisks represent significant differences between dietary treatments (P<0.05).

This article presents a summary of a more complete publication in Front. Immunol. 14:1222173 (https://doi. org/10.3389/fimmu.2023.1222173). References available on request.

More information: Sofia Morais Innovation Aqua Team Leader Lucta S.A. E: sofia.morais@lucta.com

Aquafeed: Advances in Processing & Formulation Vol 16 Issue 1 2024

FEED ADDITIVES

Nutrient utilization enhancer goes beyond lysophospholipids Elvira Alcalde, Kemin AquaScience

According to the FAO, the world's population is set to surpass 9 billion by 2050. This will inevitably increase the demand for food to meet the nutritional requirements of the growing population. The challenge is to meet the demand in a sustainable and environmentally friendly way, without any risks to the parties involved. Currently, organizations and companies are working on solutions to alleviate population growth by tackling this issue from different perspectives. In this sense, aquaculture is playing a crucial role in meeting the increasing demand for animal production to feed the growing population sustainably. Recently, we acknowledged the constraints in the procurement of raw materials typically utilized in aquafeeds, including fishmeal and fish oil, which are progressively becoming scarce for diverse reasons. At Kemin AquaScience™, a division of Kemin Industries

focusing on aquaculture solutions, we pledge to promote eco-friendly solutions specifically designed for the aquafeed sector. According to FAO (2009), “the cost of fish feed accounts from 50-70% of total production costs depending on the species.” It is crucial for farmers to monitor and manage this expense by linking it to technical parameters. Regrettably, over the past few years, the aquaculture industry has faced unpredictable increases in the cost of fish feed. This has been caused by a convergence of global events. Initiated by the effects of COVID-19, conflicts, and compounded climate disturbances, the price of raw materials and energy saw a significant increase. Additionally, the demand for certain fish species remained static, and as a result, fish prices did not follow inflation, ultimately eroding margins. The situation seems to be stabilizing, but one

Aquafeed: Advances in Processing & Formulation Vol 16 Issue 1 2024

FEED ADDITIVES

certainty remains: fish feed prices will not revert to pre-crisis levels. Therefore, it is even more important to improve the feed efficiency.

A new nutrient enhancer One of our newest innovations for the aquafeed industry is Aquatria™. It is the result of extensive research into lipid metabolism with various species, combined with advancements in application systems. Aquatria™ is a combination of active ingredients with a synergistic effect that can decrease the energy requirements of diets designed for aquaculture species, fish and crustaceans. This results in maintaining production rates while enhancing the animal's health, due to the positive impact of Aquatria™ on their liver and intestine. The most recent tests of this product were conducted in two different aquatic species, Atlantic salmon presmolt juveniles at Temuco Catholic University, Chile, for 81 days, and in juvenile European seabass at the Hellenic Centre for Marine Research (HCMR) in Greece for 12 weeks. In both tests, the growth, feed utilization, and health of the fish were evaluated.

Salmon trial The trial with Atlantic salmon pre-smolt in Chile tested three different diets: a positive control with a typical high-energy diet with more than 24 MJ/kg as gross energy with fish oil and rapeseed oil as fat sources; a negative control, the same diet as the positive control with 4.3% less of rapeseed oil and 1% less of blood meal, with 23MJ/kg gross energy; a third diet, the same as the negative control, with 4.3% less of rapeseed oil and 1% less of blood meal comparing to the positive control, but with 0.1% of Aquatria™ DR, with 23 MJ/kg gross energy. Under the reduction of total energy and lipid in feed formulation, Aquatria™ DR helped to maintain the fish performance at a similar level to positive control. Results showed that the highest average specific growth rate was seen for positive control (0.947) and Aquatria™ (0.945). The lowest value was found for negative control (0.903). The lowest average FCR was for Aquatria™ (1.51) and positive control (1.55) and the highest value was seen for negative control (1.63). The condition factor of the positive control was significantly lower than that of the negative control and Aquatria™ group. The poor condition of the fish in the

Aquafeed: Advances in Processing & Formulation Vol 16 Issue 1 2024

FEED ADDITIVES

positive control was mainly due to their bigger length for similar weight. The application of Aquatria™ along with the reduction of total energy and total lipid, reduced the accumulation of visceral fat without causing a negative impact on liver condition. In protein and energy retentions, results showed the largest retention in the Aquatria group. The reduction of rapeseed oil improved the fatty acid profile in the muscle and improved fish health condition. Aquatria™ also reduced liver fat deposition and improved liver health.

Seabass trial In the trial run on juvenile seabass at HCMR for 12 weeks, four diets were evaluated through in vivo tests in triplicate. Specifically, the commercial-type diets

were formulated as follows: Control 1: Positive control with a high-fat commercial formula (18.9%) with 14% of salmon oil as fat source; Control 2: A low fat commercial formula (16%) with 3% less of salmon oil, with 11% salmon oil as fat source; Diet 3: Control 2 diet with the addition of 0.025% Aquatria™ LQ; Diet 4: Control 2 diet with the addition of 0.05% Aquatria™ LQ. The experimental diets were formulated to be isonitrogenous (~44% crude protein), with different total lipids and energy content. Micronutrients (essential amino acids, phosphorus, essential vitamins, and minerals) were balanced among the experimental diets. The experimental diets were produced by extrusion at HCMR. Results for growth parameters are shown in Table 1 showing that feed consumption increased for the

Table 1. Growth parameters

Control 1

Control 2

Control 3

Control 4

Final weight (g)

49.6 ± 1.1

48.0 ± 0.6

51.8 ± 1.7

51.2 ± 1.1a

SGR (%/day)

1.1 ± 0.0ab

1.1 ± 0.0a

1.2 ± 0.0b

Daily feed intake

2.3 ± 0.4

2.7 ± 0.1

2.6 ± 0.0

2.6 ± 0.1

FCR

1.4 ± 0.1

1.6 ± 0.1

1.4 ± 0.0

1.4 ± 0.1

Aquafeed: Advances in Processing & Formulation Vol 16 Issue 1 2024

FEED ADDITIVES negative control (Control 2) and Diet 3 and 4 due to the lower energy content of the latter compared to the higher fat Control 1. FCR was lower for Diet 3 and 4 (lowfat diets) and Control 1, showing the positive effect of Aquatria™ on feed utilization. Diet 3 and 4 also showed the highest specific growth (% growth/day) rate value. Overall, the growth and feed utilization results showed that Aquatria™ supplemented at a level of 0.025% can support a 3% reduction of dietary fat in seabass juvenile diets with no effect on growth performance and feed utilization. Regarding the liver histology, at the end of the growth period, the liver morphology of the different groups showed significant differences. The effects of including Aquatria™ (Diet 3 and 4) included attenuation of liver lipid deposition and nucleus displacement. Pending analyses will reveal further the role of Aquatria™ on health and metabolism of the experimental population. In the words of Dr. Ioannis Nengas, research director at the Institute of Marine Biology, Biotechnology and Aquaculture of the Hellenic Center for Marine Research, “the results showed that aquafeed mills could reformulate and decrease the total dietary fat level by at least 3% with the addition of Aquatria™ LQ at 0.025%, with no effect on growth performance, feed utilization and health of juvenile seabass for the given experimental conditions and feed formulation.”

Conclusions Digestibility enhancers that optimize the digestive potential give the formulator flexibility in the use of plant ingredients and formulation of least cost and highquality feeds. Therefore, it is important to not only aim for optimizing nutritional inputs at a lower cost but also use alternatives that maximize nutrient absorption and utilization. For instance, Aquatria™ acts as a biological emulsifier and fat utilization enhancer, improving digestion and absorption of fat while promoting fat metabolism in fish bodies. Aquatria™ is a more powerful nutrient utilization enhancer than traditional lysolecithin for aquaculture.

More information: Emmanuel Pruvost Senior Global Sales Manager EMENA Kemin AquaScience™ E: emmanuel.pruvost@kemin.com

Elvira Alcalde Sales Manager Southern Europe Kemin AquaScience™ E: elvira.alcalde@kemin.com

Aquafeed: Advances in Processing & Formulation Vol 16 Issue 1 2024

FEED ADDITIVES

Dietary potassium diformate in bacterially challenged whiteleg shrimp in Latin America Christian Lückstädt, Nicolas Greiffenstein, ADDCON

Intensive production of the whiteleg shrimp, Litopenaeus vannamei (Boone 1931), in Central America and SE Asia is estimated to have reached 5.8 million tonnes in 2022. The global shrimp market size in 2022 was valued at USD 47 billion and is suggested to reach USD 69 billion in 2028, thus having an estimated compound annual growth rate of 6.7%. In Ecuador, the increase in shrimp production between 2021 and 2022 was 30%. In such intensive aquaculture production, bacterial diseases have been identified as a major cause of economic loss to producers. Feeding antibioticmedicated feeds used to be a common practice to treat bacterial infections and the prophylactic use of antibiotics as growth promoters in aquaculture production has also been widely applied. However, growing awareness from both consumers and 30

producers of all species grown in aquaculture has resulted in the current demand for responsible and sustainable aquaculture. Regulatory authorities in most exporting countries now focus on preventing the misuse of antibiotic growth promoters (AGP) in aquaculture, while public attention has shifted towards sustainable production methods. Although state-of-the-art shrimp farm management is often practiced, severe losses have occurred, diagnosed as white feces disease (2010) and early mortality syndrome (EMS, aka acute hepatopancreatic necrosis syndrome AHPNS). EMS is a fatal disease occasionally found in farmed shrimp throughout the world, with an estimated global cost to the aquaculture industry of USD 1 billion. Both diseases are caused by Vibrio spp., a bacterium residing in the shrimp gut and hepatopancreas. In both cases, affected

Aquafeed: Advances in Processing & Formulation Vol 16 Issue 1 2024

FEED ADDITIVES Table1. Effects of potassium diformate (AQUAFORM) on performance and survival parameter on shrimp grow-out against a positive control

Positive control 0.2%

KDF 0.3%

Difference (%)

122 ± 8

128 ± 8

+6 days

39.7 ± 18.8

59.6 ± 8.9

+37.5

Shrimp harvest (n/m )

4.4 ± 2.1

6.0 ± 1.0

+36.4

Shrimp harvest (kg/ha)

823 ± 323

Trial period (d) Survival (%) 2

900 ± 63

+9.4

FCR

2.23 ± 0.11

2.00 ± 0.17

-10.3

Productivity index

35.1 ± 13.7

45.1 ± 5.9

+29.6

organs are ruptured, reducing feed intake and shrimp condition with massive mortalities within a few weeks after stocking. Researchers have meanwhile identified at least four strains of EMS in shrimp farms in Latin America within the last few years. These were isolated either from the stomachs of diseased shrimp or from the sediment of AHPNS-affected farms. Since dietary acidifiers are known particularly to inhibit pathogenic Gramnegative bacteria directly, they may therefore have a supporting action in suppressing the onset of the disease. Dietary organic acids, and especially potassium diformate, which is the most widely tested organic acid salt in aquaculture, are therefore among the various alternatives at the forefront of environmentally friendly and nutritive-sustainable aquaculture approaches, without resorting to the use of AGPs. This formed the impetus for a commercial trial in an affected area in Central America, which tested the use of two different dietary acidifiers.

Methods Fifteen ponds on a commercial farm of approximately 6 ha each were stocked with 110,000 PL of white leg shrimp per ha. The farm had previous issues with Vibrio spp., including Vibrio parahaemolyticus occurrence. While the control diet (containing 0.2% of an additive based mainly on citric acid, fumaric acid and phosphoric acid) was fed to shrimp in 11 ponds, the test diet was used in 4 ponds containing potassium diformate (KDF, 0.3%; Aquaform®, ADDCON). Shrimp were fed to satiation over the day. The trial lasted for 123 ± 8 days. Results on performance and productivity index are expressed as mean ± standard deviation. Data were subjected to statistical analysis and a significance level of 0.05 was used in all tests.

Results Results (Table 1) showed a significantly higher (P=0.03) number of harvested shrimp in the ponds fed with KDF (368,000 vs. 246,000). Therefore, the overall survival rate tended (P=0.09) to be increased in shrimp on diets containing dietary potassium diformate (54.6% vs. 39.7%). Shrimp in the KDF-fed ponds had a significantly (P=0.02) lower final weight but achieved a highly significantly (P < 0.01) improved feed conversion (2.00 vs. 2.23). The harvested biomass per ha was numerically increased by almost 80 kg/ha when fed with KDF (900 kg vs. 822 kg). Overall, this led to an increased productivity index, based on weight gain, survival rate and FCR (P<0.1) by almost 30% (45.5 vs. 35.1) in shrimp fed with KDF. Conclusion Using dietary diformates poses a promising nutritional alternative to reduce bacterial-related losses in modern shrimp farming, even when tested against a positive control and contributes to an economically and ecologically sustainable grow-out operation. *Partly published at World Aquaculture 2022 in Singapore and AquaExpo 2023 in Guayaquil

More information: Christian Lückstädt Technical Director ADDCON E: christian.lueckstaedt@addcon.com

Aquafeed: Advances in Processing & Formulation Vol 16 Issue 1 2024

FEED PROCESSING

Dietary browning products and mineral availability in rainbow trout Lorenzo Márquez, Xavier Serrano, Adrián J. Hernández, Majorie Larson, Patricio Dantagnan, CIIC/NIPA/Facultad de Recursos Naturales, Universidad Católica de Temuco, Chile

Aquafeed production involves heating processes that alter the composition of raw ingredients and formulated diets. Notably, specific ingredients such as distilled dried grains with solubles (DDGS), exhibit an apparent color shift towards brown due to these heating processes. This brown coloration is also observed in aquafeed-like formulations subjected to heat. Furthermore, plant ingredients presently included in commercial fish diets can increase the content of sugars in the dietary matrix, potentially initiating Maillard reactions and producing brown-colored products. Within this context, melanoidins emerge as a significant component of the browning process, characterized by high molecular weight, chemical groups with negative charges, and antioxidant properties. These molecules can interact with mineral cations, influencing mineral availability. While this kind of interaction has been proven in terrestrial animals, to our knowledge, there is no literature on fish. This study aims to address this knowledge gap and explore the potential impact of melanoidins on mineral availability in fish.

Exploring the effects of melanoidins on mineral availabilities in fish An experiment was conducted at Universidad Católica de Temuco (UCT, Chile) to disclose the effects of a model melanoidin on the apparent availability coefficients (AAC) of dietary minerals in the rainbow trout, Oncorhynchus mykiss. This study was supported by ANID - Chile, through the project Fondecyt Regular 1150147 (approved by Comité de Ética de la Investigación of UCT).

Two diets were produced with non-browned ingredients: freeze-dried Hoki muscle, fish oil, sunflower oil, gelatin, casein, starch, cellulose, and vitamin and mineral supplements for trout diets. The control diet contained no melanoidins, whereas the experimental diet contained 1.2% (dry basis) of glycine-glucose melanoidins (Gly/Glu melanoidins hereafter). Chromium oxide was included at 0.75% as an inert marker. Gly/Glu melanoidins were produced by heating (103°C, 24 h) a solution of glycine (1 M), glucose (1 M) and NaHCO3 (0.1 M). Browning products (melanoidins) were precipitated by acidification with HCl (pH 2.5) and centrifuged at 4,800 g for 25 min. Diets were pelleted without heating, and the pellets were freeze-dried and stored at -80°C. Diets were isoproteic (50.3-50.6%) and isolipidic (19.3-19.4%). Mineral contents in the control diet were: 1.70, 7.15, 1.25, 7.20, 0.005, 0.037, 0.202, and 0.144 g kg-1 for Na, K, Mg, Ca, Cu, Mn, Fe and Zn, respectively. In the case of the browned diet, mineral contents were: 2.00, 7.15, 1.20, 7.15, 0.004, 0.036, 0.270, and 0.143 g kg-1 for Na, K, Mg, Ca, Cu, Mn, Fe and Zn, respectively. Each diet was supplied to three tanks, 100 L in volume, for 17 days at the experimental hatchery of UCT (Chile). Each tank contained 33 juvenile rainbow trout initially weighing 27.8 ± 1.9 g. Fish handling was in accordance with Chilean law for the welfare of experimental animals (Ley 20.380, Ministerio de Salud de Chile). Temperature was close to 14°C throughout the experimental time. Animals were fed to apparent satiation twice daily (9:00 and 16:00). Feces from each tank were collected every day, using Guelph-type settling columns. At day 17, fecal materials

Aquafeed: Advances in Processing & Formulation Vol 16 Issue 1 2024

FEED PROCESSING

Figure 1. Browning sequence of a casein-gelatin-dextrin-starch experimental diet subjected to heating.

collected during the last week were freeze-dried and milled to a fine powder. Apparent availability coefficients (AAC) of minerals were calculated for each tank according to the indirect method based on chromium oxide as an inert marker. Mineral compositions of diets and feces were analyzed at Instituto de Agroindustria (Universidad de la Frontera, Chile). The contents of minerals were determined through atomic absorption spectrophotometry (Spectrophotometer UNICAM 969) according to Sadzawka et al. (2007). Chromium oxide was determined at Universidad Católica de Temuco according to the Furukawa and Tsukahara method.

Dietary melanoidins can affect mineral availability in fish No mortality or signs of disease were observed during the experiment, and fish grew and displayed a very active eating behavior. Mineral concentrations in feces (Table 1) were in agreement with dietary mineral contents, and Zn was significantly higher in feces from the control treatment. Mineral AAC’s were below 100% and positive, but iron AAC showed a negative value in the control diet (Table 2). No significant effect was found for the majority of the minerals studied, but iron and zinc AAC’s were higher in animals fed the browned diet (Table 2).

Table 1. Mineral composition of diets and feces (mean ± sem) from rainbow trout fed diets with or without Gly/Glu melanoidins. Different letters in superscripts indicate significant differences.

Mineral

Control diet

Browned diet

-1

Na (g kg )

0.6 ± 0.15

0.6 ± 0.06

K (g kg )

0.3 ± 0.00

Mg (g kg-1)

1.7 ± 0.06

1.6 ± 0.12

Ca (g kg )

2.4 ± 0.12

2.4 ± 0.20

-1

Cu (mg kg-1)

15 ± 0.6

15 ± 1.5

Mn (mg kg-1)

140 ± 0.9

140 ± 0.3

Fe (mg kg-1)

1,065 ± 30.3

1,001 ± 32.4

Zn (mg kg )

469 ± 3.2

444 ± 4.7b

-1

Aquafeed: Advances in Processing & Formulation Vol 16 Issue 1 2024

FEED PROCESSING Table 2. Apparent availability coefficients (%) (mean ± sem) of minerals in rainbow trout fed diets with or without Gly/Glu melanoidins. Different letters in superscripts indicate significant differences.

Mineral

AAC for control diet

AAc for browned diet

91.7 ± 3.8

93.3 ± 1.1

99.0 ± 0.0

99.1 ± 0.0

68.4 ± 2.9

69.8 ± 3.8

22.9 ± 3.0

24.5 ± 0.8

27.9 ± 2.6

32.6 ± 12

12.1 ± 2.6

13.8 ± 0.2

-21.9 ± 5.1a

17.7 ± 4.8b

24.5 ± 1.6a

31.0 ± 1.5b

Figure 2. Color of freeze-dried, milled fish muscle, a non-heated pelletized diet with chromium oxide, and the same diet but including 1.2% of glycine-glucose melanoidins.

The increased iron availability can be explained by the properties of Gly/Glu melanoidins. Firstly, the rainbow trout intestine preferentially transports the reduced ion, Fe2+ (Kwong & Nigoyi, 2008), and antioxidant compounds promote the accumulation of iron into the intestinal epithelium of the fish Opsanus beta (Cooper et al., 2006). Accordingly, the antioxidant capacity of Gly/ Glu melanoidins in the diet (Serrano et al., 2018) could have promoted iron absorption. Secondly, Cooper and Bury (2007) reported that iron chelated with humic acids (a soil substance chemically similar to Maillardreaction brown products) can be absorbed by the rainbow trout through the gills. In mammals, dietary browning products have been reported to increase iron retention in the rat (Delgado-Andrade et al., 2016), but also to decrease iron retention in male adolescents (Mesías et al., 2012). The effect of melanoidins on zinc was smaller than what we found on iron. In rats, dietary Maillard reaction products from bread crust do not seem to have any effect on Zn retention (DelgadoAndrade et al. 2016), however, the physiological similarities and differences between the digestive tracts of the rat and the rainbow trout are not well known.

Conclusions While dietary melanoidins exhibit no significant impact on the availability of the majority of studied minerals, there are exceptions. The availabilities of iron (Fe) and zinc (Zn) show a tendency to increase. However, the consequences of this phenomenon remain to be thoroughly investigated. The present study underscores the convenience of further research into the effects of dietary Maillard reaction products on fish nutrition. References available on request.

More information: Lorenzo Márquez Researcher at Centro de Investigación, Innovación y Creación Universidad Católica de Temuco, Chile E: marquez.lorenzo728@gmail.com

Aquafeed: Advances in Processing & Formulation Vol 16 Issue 1 2024

FEED PROCESSING

Extrusion of premium shrimp feeds on single screw extruders: A review Dana Nelson, Extru-Tech Shrimp farming has been and continues to grow around the globe. Recent trends and improvements have led to continued optimism. Significant growth and investment continue, but several challenges threaten future stability and sustainability. This discussion will only attempt to address one of the issues: improved feed quality.

Introduction Shrimp farming – the practice of raising and harvesting shrimp in a controlled environment – has been going on for a long time. The earliest examples likely date back to the 13th century in China. Others point to shrimp farming in Japan in the late 1930s as the beginning (Chamberlain, 2011). The modern practice of shrimp farming was boosted during the 1970s when researchers and academics began research in which they raised shrimp in ponds and tanks to refine captive shrimp “aquaculture." During the 1980s, shrimp farming became increasingly popular in Asia. This growth was easily identifiable in Thailand, Indonesia, and Vietnam. These countries quickly became significant producers of farmed shrimp; by the 1990s, shrimp farming had become a substantial industry in many parts of the world. Unfortunately, the rapid expansion led to several environmental and social issues. The use of sensitive coastal areas, valuable mangrove forests, and valuable habitats, combined with chemicals, antibiotics, and pollution from waste, had significant negative impacts (Suzuki & Nam, 2023). In response to these issues, many governments and organizations began promoting more sustainable and responsible practices in shrimp farming. These new practices included better management techniques, improved water quality management, disease prevention measures, and more environmentally

0.8 mm ADT shrimp die

friendly farming methods, such as integrated multitrophic aquaculture. Today, shrimp farming is a global industry, with major producers in many countries, including key locations in Ecuador, India, and China. The list of countries growing shrimp commercially is quite impressive. While challenges remain, the industry has made and continues to make significant strides in promoting more sustainable and responsible practices over the past few decades.

Aquafeed: Advances in Processing & Formulation Vol 16 Issue 1 2024

FEED PROCESSING The importance of feed quality Improving feed quality is crucial in ensuring shrimp aquaculture's continued growth and sustainability. These improvements include using high-quality ingredients and sustainable and environmentally friendly feed ingredients. Promoting alternative protein sources, such as protein-concentrated products, single-cell proteins, insect meals, and other raw materials, will help reduce the reliance on fishmeal and other unsustainable feed sources. Overall, improving feed quality has been and should continue to be a primary focus to ensure the longterm success and sustainability of shrimp aquaculture. One sometimes-overlooked issue directly relates to the manufacturing process used to prepare these valuable nutrients into a form ideal for shrimp to consume in the habitat they are being reared in from their larval stages to harvest. Pelleted feeds from the past have improved significantly. The process steps have changed or been modified, incorporating innovations focused on pellet durability, water stability, and predictable density. Although quality varies among these pelleted feeds, they differ from those produced a decade ago. One improvement involves significantly reducing the use of artificial binders to hold ingredients together. While these binders improved water stability, they also negatively impacted shrimp health and the environment. Perhaps most importantly, growers are becoming much more sophisticated with their practices. More and more farmers are becoming aware of and have gained more control over the variables that affect their facilities. The relationship among water quality, feeding practices, and a lengthy list of variables far too long to include in this article are becoming much better understood. Without offense to growers, it is reasonable to suggest that many more facilities can isolate and measure feed performance today in ways that were not the case just a few years ago. Although these gains are significant, much more improvement is needed. Continued feed production technology, feeding practices, and discovering the ideal physical feed characteristics, including ideal feed sizes for individual rearing facilities, are all necessary. Extrusion technology Cooking extrusion technology has broadly been accepted as the preferred method for forming most

aquaculture formulations into pellets. This acceptance began over 50 years ago. The earliest extruders for aquafeed production were borrowed from the pet food industry. These extruders had a natural ability to form very durable expanded pellets that proved to be incredibly valuable in many ways. The most obvious was the feed's improved ability to maintain its size and integrity during manufacturing, packaging, transport, storage, and feeding. The second was the new ability to use feeds that float to improve feeding practices. The simple ability to monitor feed intake and acceptance by the target species proved valuable. This floating feed characteristic alone justified using extrusion for many end users. When combined with the improved water quality via the reduction of unutilized and consumed nutrients, the acceptance became quick as capital resources became available to gain this advantage in most parts of the world. Salmon feed producers were among the first to adopt extrusion technology. Salmon requires a high-protein diet, and like those of other species, their feed must also contain a specific balance of nutrients, including essential fatty acids, vitamins, and minerals. These high-protein and high-fat formulations proved to be problematic for steam pelleting systems. Pet food extrusion technology, combined with modifications and specialized ancillary equipment, better meet these requirements by producing high-quality, nutritionally complete feeds that satisfy the specific needs of salmon in a much more durable form. In addition, more precise control of the extrusion process led to better control over the feed's cell structure and bulk density. These products were then introduced to vacuum coating mixers and other proprietary innovations; unprecedented protein and lipid concentrations were achieved in a durable dry feed. These feeds were also engineered to be "slow sinking." Feed that stayed in the water column longer offered tremendous value in net pen systems. As the salmon industry grew, so did extrusion equipment suppliers' ability to meet the need for large-scale, efficient production. High-capacity continuous production facilities soon became the norm. Finally, salmon feed producers were also among the first to adopt extrusion technology because of the need for product consistency. Salmon farming evolved very quickly. Shortcomings in the physical

Aquafeed: Advances in Processing & Formulation Vol 16 Issue 1 2024

FEED PROCESSING feed characteristics were quickly identified – feed needed to be as consistent in physical quality as possible to avoid waste. Regarding shrimp, single-screw extruders have been studied and determined to produce shrimp feed with the desired physical and nutritional attributes effectively. The advantages of extruded feeds over steam-pelleted products have been demonstrated in controlled studies (Tacon, 2003). Many of these studies have included production costs in their conclusions. For some of the reasons discussed and others, commercial growers have experienced difficulty in identifying and measuring improved performance.

Specific challenges of shrimp feeds Twin- and single-screw extruders work by forcing a mixture of raw materials, including proteins, carbohydrates, and fats, through a heated barrel using a single or pair of rotating screws. The heat and pressure generated in the barrel cause the mixture to undergo a series of chemical and physical transformations that eventually produce a uniform and durable product (Camire, 2000). Restated in another way, when the raw material mix is subjected to enough moisture and heat, some transition from a solid to a liquid; some ingredients melt. This mixture of solid and melted components transitions rapidly into uniform solid pellets when the conveying action of the extrusion system forces the product through a die that separates the pressurized process and ambient conditions outside the barrel. The primary ingredients that undergo this transition from solid to liquid and then back to a solid that bonds all the ingredients together are the starch faction of the carbohydrates. So, in review, the primary tasks performed during extrusion are cooking or gelatinizing the starch and bonding all the ingredients into specific-sized pellets as they exit the process. Producing shrimp feed on extruders has been more challenging than other aquatic feed in one fundamental way. Most of the shrimp feeds used globally are steam-pelleted. Extruders' use of heat and moisture correlate nearly linearly with pellet durability and water stability, as do ingredient pasteurization, improved bioavailability, digestibility, and reduced anti-nutritional components in the ingredients. This use of heat and moisture is fundamental to their superior performance.

Unfortunately, the typical operating parameters influencing these advantages promote pellet expansion. Shrimp farmers have always perceived the bulk density of feed as a good indicator of its quality. Heavy pellets sank quickly, helped deter feeds from migrating to unintended locations, did not accumulate on shorelines, and did not attract competing birds, animals, or other pests. They also absorb water more slowly (water absorption), contributing to their ability to hold their shape longer over time (physical water stability) and deter water-soluble nutrients from leaching out of the feed and into the water (nutrient water stability). The stocking densities and feeding habits of farmed shrimp dictated that the feeds had to hold their shape long enough to be discovered and ingested. Some growers focused on water stability characteristics lasting longer than four hours. Initial use of extruded shrimp feed was plagued by an increased risk of a percentage of floating pellets and perceived lower quality due to a much lower bulk density than steam pellets. To increase density and reduce the risk of floating pellets, manufacturers reduced heat and moisture during processing – the root cause of the undesirable expansion of the pellets. Unfortunately, this practice undermined the strengths of the extrusion process. One problem was exchanged for a different one. Many of these first extrusion systems sadly evolved into expensive and inefficient pellet mills during this time. Gelatinization levels were low, pellet durability was inconsistent, and the constant risk of outlier floating pellets never seemed resolved. Although there were other challenges, simple retrospection reveals shrimp required a product that cooking extruders had difficulty producing easily. Moreover, when quality feeds were successfully produced, most growers could not see or document a significant improvement in feed performance. When these challenges were combined with higher initial capital requirements, increased complexity for operators, increased energy requirements, and a costly drying step, it is not much of a surprise that their popularity was never embraced as it had been for other species. When twin screw extruders were offered to be the solution due to their ability to better control expansion without compromising gelatinization at a higher initial and operational cost, the quick and easy answer was, "No, thank you."

Aquafeed: Advances in Processing & Formulation Vol 16 Issue 1 2024

FEED PROCESSING

Many have asked me to clarify this historical review. This review may need to be clarified for some who have watched aquaculture over a long period. Why? Salmon feeds were sinking feed and among the highest-quality feeds found anywhere. They were being successfully produced on single- and twin-screw extruders for decades in many parts of the world while shrimp feeds continued to be steam-pelleted. The answer is quite simple. Salmon feeds do not sink due to extrusion. Pellets float after the extrusion and drying steps. However, salmon feed is processed at high moisture and heat levels to promote an extremely fine and expanded cell structure, enabling high levels of lipids to be applied outside after the extrusion and drying steps are complete. This vacuum coating step results in pellets that sink slowly but only after the tiny voids in

the pellet are filled with oil. High-quality salmon feed requires expansion. Shrimp feeds do not.

Single screw extruders today Although steam-pelleted shrimp feeds are still much more common than extruded shrimp feeds, a shift back to extrusion has been renewed. There are several contributing factors. First and foremost, experienced producers in the salmon sector have made a significant effort to diversify their activities into shrimp feed. These companies are well-capitalized and well aware-of the strengths and weaknesses of extrusion. Second, single-screw extruders have evolved dramatically in the last decade. Single-screw extruder manufacturers familiar with the challenge of producing dense pellets have developed hardware innovations

Aquafeed: Advances in Processing & Formulation Vol 16 Issue 1 2024

FEED PROCESSING

to achieve higher bulk density without compromising cook and gelatinization. These innovations have evolved to the point that they are no longer theoretical. The highest quality feeds found in Ecuador, Peru, and Mexico are currently produced on single-screw extruders. Directly extruded pellets down to 0.8 mm are successfully produced at unprecedented capacity. Several manufacturers produce these small feeds at capacities higher than five metric tons per hour. Production size feeds between 1.2 and 2.0 mm are successfully produced at two to three times that of these micro pelleted feeds (0.8 mm). All these feeds are highly gelatinized and have water stability characteristics considered impossible without nonnutritive binders just a few years ago. Some key manufacturers have now embraced extrusion completely. Their steam-pelleted products are no longer available. Steam pelleting lines are being removed, and additional modern extrusion equipment is being commissioned. It should be noted that these premium feeds have slightly different physical

characteristics. Although they soften more rapidly than a steam pellet, they maintain their shape for long periods and become more elastic. These pellets take on a "meat-like" feel and texture. In addition, the results have been impressive when these feeds are delivered to shrimp using automated feeding systems that monitor feeding activity via sound (Poveda, 2021). Although this transition to extruded feeds and more complex feeding systems comes at a cost, it appears that growers and feed producers are not interested in returning to past practices. Extru-Tech has been involved in the challenges associated with shrimp feeds from the very beginning. Single-screw extruders have always had lower initial capital and operational costs than twins. Few would dispute this. Some end users have embraced the more expensive and complex twins deemed a better value, with all things considered. ExtruTech consciously decided to improve the performance of a single-screw system rather than offer both extrusion types. This decision demanded real innovative improvements.

Magnified image of 0.8 mm high quality shrimp feed (up). 0.8 mm shrimp feed water submerged for 4 hours (down)

Aquafeed: Advances in Processing & Formulation Vol 16 Issue 1 2024

FEED PROCESSING A brief review of single screw limitations should provide a backdrop to discuss the improvements that have been developed. Single-screw extruders have been evolving for a long time, not just in the past few years. Using an intensive preconditioning step before the extruder is undoubtedly not new. Although most manufacturers have made recent improvements, the primary goals remain the same. Pre-wetting, precooking, and thoroughly mixing the ration before the extruder improves the extrusion step in terms of capacity and product quality (Strahm, 2000). The simple truth is that preconditioners can begin and even complete these tasks better than an extruder alone. In past and recent years, there have been other innovations in single-screw extruders that have improved their capabilities and made them function similarly to twin screws. One such innovation is the use of a multi-stage (segmented) screw configuration that permits changes to the type and location of conveying, cooking, mixing, and kneading elements (Riaz, 2000). This configuration flexibility facilitates much better control of the process. This change also paved the way for the development of new screw geometries and flow restriction devices at not only the die but mid-barrel locations as well. Externally controlled flow restriction devices like Extru-Tech's Mid-Barrel-Valve have pushed the cooking and shear zone further away from the die than has been possible in the past. Novel die designs have also improved the quality and capacity of shrimp feeds. These changes, when combined, have proven to be significant. Products once considered squarely in twin screw territory have now been standardized on single screw architecture. Pet food manufacturers challenged with developing new products like limited

ingredient diets, grain-free diets, and high-inclusion fresh meat diets have all been achieved on these modern single-screw systems. Lessons learned from these developments, and others, have enabled these more advanced single screw systems to excel at producing shrimp feed with lower energy, maintenance, and production costs.

Conclusion Single-screw extruders have evolved considerably over time. The list of improvements is lengthy. The use of an independently controlled flow device (mid-barrel valve) early in the extrusion process, combined with more comprehensive preconditioning, novel final screw geometry, and novel die designs, have successfully moved the mixing and cooking steps far enough away in time from the forming step at the die that expansion can now be satisfactorily controlled. The days of compromising capacity or gelatinization on single screw extruders to control expansion can be left behind. All the advantages realized for other species utilizing feeds produced on single screw extruders are now available to shrimp.

More information: Dana Nelson Aquaculture Specialist Extru-Tech E: dnelson@extru-techinc.com

SINGLE SCREW MULTIPLE MARKETS SINGLE SCREW EXTRUSION SYSTEM OPTIMIZES

AQUA FEED

Optimize the return on your capital investment FLEXIBILITY AND MARKET OPPORTUNITIES with a single screw extrusion system. Buy one system and cost-effectively deliver product to multiple market opportunities. Contact Extru-Tech today

PREMIUM PETFOOD

• One system capable of economy up to super premium fresh meat petfood • Aquatic feeds that range from floating to sinking shrimp feed • Capitalize on high margin petfood treat opportunities • Significantly lower operating cost per ton versus competitive systems

PETFOOD SNACKS

An Extru-Tech Single Screw Extrusion System provides all the flexibility and production efficiencies at around half the cost of competitive extrusion systems with high operating costs.

ET-337C.indd 1

Aquafeed: Advances in Processing & Formulation Vol 16 Issue 1 2024

at 785-284-2153 or visit us online at extru-techinc.com P.O. Box 8 100 Airport Road Sabetha, KS 66534, USA Phone: 785-284-2153 Fax: 785-284-3143 extru-techinc@extru-techinc.com www.extru-techinc.com

1/28/21 8:32 AM

FEED PROCESSING

Is water killing your fish? Benefits of moisture measurement and control in aquafeed production Neal Cass, Hydronix The key to optimizing aquafeed production processes is to ensure stability and repeatability. Other than the quality of ingredients and process equipment, the largest unknown variable in the material is water. The moisture content within each ingredient fluctuates over time. These fluctuations create various challenges for the producer. It is very advantageous to continually measure and correct for the changing moisture contents. Aquafeed producers mix many ingredients including plant and fishmeal, oils, and other proteins. Feed formulations are defined according to the species, age of the animal being fed and other factors such as seasonal availability, and quality of each ingredient. Frequently shortages of fishmeal or fish oil cause fluctuations in prices, so adjustments are made to the formulation to reduce cost while maintaining quality and nutritional value. Ingredients are substituted and expensive additives may be required to maintain nutritional value. Raw materials such as meat and bonemeal may have an average moisture of 5%, while soybean has 12.7% (Rezaya Rabbi Bhuiyan et al., 2018). As formulations are designed to give specific ratios of each material, when the moisture continuously varies at different rates between the materials, this leads to changes in the proportions of the dry materials. A system producing a recipe specifying 1,000kg of dry soybeans may have an input material at 12.7% moisture (wet basis), so this should be controlled as accurately as possible to dose exactly 1,127kg. This would provide the required 1,000kg of dry soybean meal specified along with 127kg of water. If the soybean meal moisture is reduced to 10%, the same dosed weight (1,127kg)

would actually result in 1,024kg of dry material in the measurement and only 102.4kg of water. If the proportions of the dry materials are not correctly adjusted for the constant change, then this will cause variations in protein and fat, and the nutritional value of the final feed pellet. This can lead to noncompliant products resulting in growth problems or animal health issues, and affect the pelleted product’s market value. With the improved technologies, now available at lower costs, which are simpler to use, easier to install and have very low maintenance requirements, measurement and control of the feed processing operations is now a logical next step in plant

Aquafeed: Advances in Processing & Formulation Vol 16 Issue 1 2024

FEED PROCESSING development. It is now cost-effective and efficient to implement end-to-end, in-line moisture measurement throughout the entire plant to optimize the entire process, from ingredient reception and storage to mixing, drying, and final storage.

Types of moisture measurement Each type of measurement has its own advantages and disadvantages. The advantages of measuring in-line are considerable when controlling a running process. The use of multiple sensors enables real-time adjustments to be made to the process with no delay or lag. In-line sensors also allow for both feedforward and feedback process control. While there are significant advantages in installing a sensor in a single location, there are even greater advantages to achieving end-to-end control. This requires more sensors, meaning that it is even more important that the sensors used are low-cost, and simple to install and maintain. Selecting a sensor that is repeatable and reliable will enable a producer to truly benefit from the anticipated gains. Introducing a sensor that is fragile or requires complex calibration or that may drift over time simply introduces another unknown variable into the process, the exact opposite of what should be happening. The objective is to consistently measure moisture and make meaningful adjustments based on reliable and repeatable input data. This should remove a single,

and very important variable, moisture, from unduly influencing the processes and product quality. When choosing a sensor, it is also important to consider the total cost of ownership, the capital cost and ongoing maintenance costs including anticipated reliability and the time required from engineers to maintain the system. Selecting an incorrect sensor for a given situation results in more anomalies in the process, frustration from engineering staff and often leads to the equipment eventually being abandoned. Hydronix produces a range of microwave sensors that meet all these requirements while also being robust enough to withstand the toughest industry conditions and can offer worldwide support, training, and service.

Where to measure moisture? End-to-end moisture control can now be achieved, efficiently and cost effectively, by installing sensors throughout the process, in locations such as shown in Figure 1. 1. Raw material intake: Materials delivered from the supplier Measurement between the raw material intake and storage silos enables quality control of incoming materials. This also allows for material storage to be managed, or material to be rejected, to reduce the risk of spoilage by mycotoxins. This also allows supplier performance to be automatically monitored, for example, when purchasing by dry weight.

Figure 1. Suggested moisture measurement locations in a typical aquafeed production plant

Aquafeed: Advances in Processing & Formulation Vol 16 Issue 1 2024

FEED PROCESSING 2. Weighing: Materials weighed to achieve the correct formulation Measuring between the storage silos and weigh hoppers ensures raw materials are correctly proportioned, compensating for the varying moisture levels in individual materials. 3. Mixing: Water addition to the mixer Measurement in the mixer allows the correct addition of water and other moisture-dependent additives to ensure material sent to the hammer mill has the correct moisture. 4. Milling: Pre and post milling Measuring before milling ensures that optimum conditioning has been achieved to maximize milling performance, as the moisture content affects the final size and shape of the milled material. As milling is an energy-intensive process that affects the moisture in the material, measuring after the mill also allows the moisture loss to be monitored. Measuring both before and after allows for complete monitoring of the realtime efficiency of the process. 5. Conditioning: Correcting the moisture of raw materials Incorrect conditioning will cause issues with the pelletizing process with variations in moisture affecting equipment wear and tear and energy consumption of the machines. High moisture can result in soft or poorly formed pellets making them more likely to block the pellet machine or create sticky pellets, often with drastic consequences, such as damange to the die. Low moisture gives reduced binding properties and pellet breakage. 6. Drying: Pellets dried for durability Measuring the input to the dryer enables feed-forward calculation of dryer parameters to ensure optimized performance and minimal energy consumption. Pellets must be under 12% internal moisture for maximum pellet durability and storage life. 7. Coating: Fat coating added Measuring before fat coating, at the output of the dryer, enables PID feedback control using the error from the target moisture value to adjust the process

appropriately. 8. Cooling: Cool to storage temperature During cooling some moisture is lost, so the pellets finish at ~10% but this is often not monitored. Measuring the output of the cooler allows control of cooling parameters and also enables a final quality control check prior to packaging or storage.

Moisture control in action Figure 2 shows an example of a moisture control installation on a holding hopper before a fat coater. This holds material at around 60°C and is fed from the dryer. This allows a buffer of material to build up to ensure the coater has a constant feed of material while in operation. In this installation, a Hydronix sensor is used to measure the moisture of the dosed material. The sensor has built-in averaging (and filtering) functions making this straightforward to achieve without needing complex changes to the control system. The average function may be triggered in various ways, in this case, starting when the high-level switch is covered during filling and stopping when the low-level switch is uncovered. Additionally, a successful application using a Hydronix sensor after the dryer, in aquafeed, or other applications, is to monitor the dryer output moisture level. This may then be used to control the dryer temperature. This is proven to improve the consistency and repeatability of the final product moisture, and hence improve the final product quality. Problems associated with numerous formulations and sensor calibration The major issue associated with multiple formulations, as required by aquafeed manufacturers, is to ensure the correct calibration of sensors for each formulation, or family of formulations. It is impossible to manage these calibrations if using a sensor that varies its output due to material or ambient temperature variations, wear and tear, or other installation issues such as an optical sensor becoming contaminated by dust or being affected by vibration. As the number of sensors used across the process increases, it becomes even more important that each one works accurately and that all sensors used have a repeatable and reliable output and have identical measurement characteristics.

Aquafeed: Advances in Processing & Formulation Vol 16 Issue 1 2024

FEED PROCESSING

Figure 2. Moisture control between the dryer and coater

A significant advantage of the Hydronix sensors is that each sensor will output an identical default factory calibrated value. This may be used as a direct linear input into a standard PLC feedback control loop and does not require any material calibration. The requirement for an absolute moisture value, requiring calibration, is redundant. Advanced independent system integrators are now able to use sophisticated AI-based algorithms linked to Hydronix sensors to continuously monitor and fine-tune the calibrations in real-time. Using stable sensors that all have identical measurement characteristics allows users to reliably record and re-use calibrations. One further indirect advantage of this is that any sensor may be removed and replaced without any need for re-calibration. The Hydronix sensor’s default linear measurement can output a value between 0 and 100 units in increments of 0.1. This output is known as the Unscaled Value and is identical between sensors. A user can simply set a target Unscaled Value for each formulation. If production switches between formulations, it is simple to change to the appropriate Unscaled Value target. Using sensors with identical output characteristics makes it possible for precise control loop control, enabling more and more users to adopt AI generated control loops for the best possible performance.

Conclusion With the Hydronix measurement, there is confidence that the measurements are precise, accurate, and repeatable, allowing their use to improve the AI model and constantly improve efficiencies. This allows the feed producers to achieve consistency in their process, while working with many formulations, reducing variations in the individual process stages, and allowing them to target a higher final product moisture, while concurrently improving quality. Quality testing by one of Hydronix’s automation partners demonstrated a 40% reduction in Standard Deviation, achieving +/-0.4% moisture. This allowed their customer to increase their final moisture target to 10.6% to ensure the product is always below 11%. Overall, Hydronix moisture sensors offer accurate, precise, and repeatable measurement, giving an improved and consistent product, a reduction in spoiled and wasted material, better plant efficiencies and reduced process downtime. More information: Neal Cass Sales Manager Hydronix E: enquiries@hydronix.com

Aquafeed: Advances in Processing & Formulation Vol 16 Issue 1 2024

PROTEINS

Slow-release amino acids: How they work in aquafeeds Dr. Antonios Chalaris, Devenish

The challenge Increasing levels of plant protein in aquafeeds often causes essential amino acid imbalance, leading to compromised growth, reduced feeding efficiency and increased feeding cost. Most commonly, lysine and methionine are the first limiting amino acids in plant formulations where fishmeal has been replaced by plant derivatives. Synthetic versions of essential amino acids are gaining popularity especially in reducing dietary protein levels and balancing the nutritional requirements of fish and shrimp. The technology of slow-release amino acids Protein is the most expensive component in animal feeds. As there is no mechanism for storing limiting amino acids in animals, the use of synthetic amino acids can lead to high levels of wastage. A slow release of the synthetic amino acids from the diet enables the supply and demand for protein to be better matched.

Devenish has invested significantly in this area for the past 20 years, responding to demand from the livestock production industry to reduce nitrogen output and fishmeal usage without compromising on animal performance. The result: DevAmine™. DevAmine™ technology is an alternative, nutritionally enhanced vegetable protein that allows more controlled release, efficient absorption and utilization of amino acids in the digestive tract of animals. The technology behind DevAmine™ is based on amino acids being glycated to sugars, with glycation being the covalent attachment of sugar to a protein or lipid. In other words, there is a sugar carrier used to achieve this consistent release of amino acids over time (Fig. 1). The efficacy of DevAmine™ technology has also been validated in fish using Devenish’s commercially available product NatuPro®. Trials with tilapia and European seabass confirmed that NatuPro® can successfully reduce crude protein without compromising fish

Figure 1. Typical release pattern of conventional dietary crude protein, synthetic and glycated (slow release) amino acids.

Aquafeed: Advances in Processing & Formulation Vol 16 Issue 1 2024

PROTEINS performance. Additionally, this technology offers an opportunity to reduce stress during periods of high-water temperature, as the oxygen demand to metabolize protein for energy use is higher compared to starch or fat.

The evidence The continuously increasing pressure to use alternative protein sources, better nutrient utilization and sustainability concerns are priorities in aquafeeds. Commercially farmed fish species are facing challenging times with limited options in their raw material baskets and formulation costs. Trials to assess the validity of DevAmine™ technology were conducted with tilapia and European seabass. Tilapia trial In collaboration with Pontus Research Ltd. in the UK a feeding trial was designed to assess whether a 2% inclusion of NatuPro® can allow improvements in performance over standard formulations in Nile tilapia (Oreochromis niloticus). Four experimental diets were tested in triplicate for 7.5 weeks: (a) positive control (standard feed, CTRL), (b) negative control (2% protein reduction) (NG CTR) and (c)(d) test diets; 2% protein reduction + product inclusion at 2% (NP2) and 4% (NP4). Data was analyzed using Analysis of Variance command, with a Shapiro-Wilk test for normality and a Levene’s test for homogeneity of variance. Dietary inclusion of NatuPro® had a positive effect on feed conversion ratios (-6.6%) and specific growth rates (+3.9%) when compared to positive controls, although not statistically significant (Fig. 2). The formulation

saving during this trial was calculated at €16 per tonne of feed.

European seabass trial A feeding trial was conducted in collaboration with the Agricultural University of Athens in Greece to investigate the use of NatuPro® in juvenile European seabass (Dicentrarchus labrax), using three experimental diets: (a) a diet formulated according to current commercial diets for European seabass (Control diet, crude protein 46.9%, fishmeal 15%), (b) a negative control diet, i.e. a diet with 1 % lower crude protein level by 1.5% lower fishmeal level (C1, crude protein 45.7%, fishmeal 13.5%), and (c) a C1 diet with NatuPro® included at 2% (C1-N2, crude protein 46.0%, fishmeal 13.5%). Experimental diets were fed, for 140 days, to quadruplicate fish groups (mean initial body weight ± s.e.: 38.3 ± 0.08 g; fourteen fish per group; recirculating seawater system). After the completion of the main rearing period and growth performance assessment, fish were fed the same diets including chromic oxide (1.0%) as a marker, once daily to satiation. Feces were collected by stripping every two days. At the end of the digestibility trial all fish were sacrificed; liver and carcass were weighted to calculate hepato-somatic index (% body weight) and carcass yield (% body weight). Diets and lyophilized feces were analyzed for chromium, protein and amino acid content. Data were analyzed by one-way ANOVA and tank was the experimental unit (n=4). No significant differences were detected among experimental diets for growth performance (i.e. final body weight, condition factor, specific growth rate,

Figure 2. Performance data for each experimental group in red tilapia. No significant differences were observed (P<0.05).

Aquafeed: Advances in Processing & Formulation Vol 16 Issue 1 2024

PROTEINS Table 1. Amino acids apparent digestibility coefficients (ADC, %) of European seabass

Diets

Control 1

C1-N2

Arginine

84.1 ± 0.73

84.2 ± 0.56

82.8 ± 0.38

Histidine

82.6 ± 0.91

78.6 ± 1.91

78.4 ± 0.36

Isoleucine

79.9 ± 0.27

79.9 ± 0.41

82.1 ± 0.63

Leucine

76.1 ± 0.65

77.6 ± 1.57

75.0 ± 0.11

Lysine

80.2 ± 0.16

79.8 ± 0.39

82.3 ± 0.19

Methionine

89.9 ± 0.87

83.9 ± 0.47

95.8 ± 0.63

Phenylananine

79.0 ± 0.12

77.8 ± 0.87

79.0 ± 0.27

Threonine

82.2 ± 0.97

77.1 ± 1.59

81.7 ± 1.13

Valine

80.8 ± 0.58

77.3 ± 1.43

81.3 ± 0.42

Essential amino acids

Non-essential amino acids Alanine

84.0 ± 0.31

84.3 ± 0.42

84.5 ± 1.34

Aspartic acid

74.7 ± 0.58

72.6 ± 1.35

68.9 ± 1.53

Glutamic acid

88.9 ± 0.29

87.1 ± 0.03

90.2 ± 0.11

Glycine

85.1 ± 0.06b

78.2 ± 1.74a

86.9 ± 0.95b

Proline

83.8 ± 0.32

84.2 ± 0.64

85.4 ± 0.36

Serine

83.8 ± 0.01

82.5 ± 0.62

84.9 ± 0.19

Tyrosine

82.9 ± 1.33

81.1 ± 0.92

81.8 ± 1.31

thermal growth coefficient, weight gain) and feed efficiency (i.e. feed conversion ratio, protein efficiency ratio, economic conversion ratio) (Table 1). Also, experimental diets did not affect hepato-somatic index and carcass yield. Lysine (Lys) digestibility was significantly higher in C1-N2 diet compared to C1 and control diets. Methionine (Met), glutamic acid (Glu) and glycine (Gly) digestibility was significantly reduced in C1 diet compared to Control diet; the inclusion of NatuPro® (C1-N2 diet) highly improved Met and Glu digestibility in higher levels than those observed in Control diet, while Gly digestibility was back at Control diet levels. Obtained results indicate that the inclusion of NatuPro® in European seabass diets does not compromise fish performance and feed efficiency. The improvement of some essential and non-essential amino acids digestibility suggests that product’s potential use in fish diets.

Application in aquafeeds Trials have highlighted that the inclusion of slow-release amino acids in aquafeeds can provide a solution to

increase feed efficiency and reduce formulation costs, without compromising fish performance. NatuPro® is a commercially available product that utilizes the slow-release amino acids technology, supplied by Devenish. NatuPro® should be considered as part of a strategy to reduce formulation costs in aquafeeds, with estimated savings of approximately €10-20 per tonne. It provides the ability to reduce formulated protein levels without compromising on fish performance and should also be considered as part of a strategy to reduce stress during periods of high water temperature.

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

Aquafeed: Advances in Processing & Formulation Vol 16 Issue 1 2024

NOVEL INGREDIENTS

How AI and big data discovered new optimal EPA + DHA levels for salmon Tony Chen, Manolin

The Amazon paradigm in aquaculture In the early days of the internet, Jeff Bezos saw a unique opportunity to transform the way books could be cataloged for retail buyers. His solution? Leveraging technology to simplify and streamline the categorization of millions of books into a single online marketplace. Bezos recognized the future viability technology had in vastly improving our ability to discover books more easily and systematically. In a 1997 interview, Bezos stated, “When you have that many items [books], you can literally build a store online that couldn't exist any other way. And that's important right now because the web is still an infant technology.” Amazon’s approach, prioritizing innovation over traditional methods in the face of complex challenges, is not just confined to retail. There’s a parallel in modern aquaculture, where technology and data are becoming pivotal in answering many lingering questions industry leaders have – questions, that are critical to understanding how aquaculture will exponentially advance in the next decade. Economic and market analysis: The shift to sustainable sources Like the internet's rapid expansion in Amazon's early years, aquaculture has seen significant growth over the past three decades, averaging a 6.7% annual growth rate from 1990 to 2020. As one of the fastestgrowing sectors in the global food market, it now faces challenges from macroeconomic shifts, especially rising commodity prices that have increased global feed costs and operational expenses for farms. This demands a better understanding of fish growth and the strategic use of sustainable alternative feed ingredients.

Central to this ingredient challenge is the role of lipids, especially omega-3 fatty acids, in fish feed. The cost of fish oil, the primary source of these lipids, has doubled in the last two years, escalating from $2,000 to over $4,000 per metric ton. This stark increase highlights the critical need for sustainable and cost-effective alternatives. In response, the global algae oil market in aquaculture is projected to reach USD 1.23 billion by 2030, growing at a CAGR of 25.4%. This growth trajectory underscores the increasing demand for sustainable options and the urgency for the aquaculture industry to embrace innovative technologies to sustain its growth and meet evolving consumer demands.

Implementing AI and big data to advance research The volatile commodity landscape in the industry necessitates the adoption of advanced technologies like Artificial Intelligence (AI) and big data. These are crucial for addressing complex research questions in the sector. The use of big data, in particular, has significantly grown in food science, increasing by nearly 300% every five years since 2010. This highlights its essential role in linking laboratory research with commercial-scale applications in aquaculture. Echoing Bezos's realization that a vast bookstore is only feasible online, the most helpful solutions for farmers today lie in technology. This is particularly true in the realm of feed research for alternative ingredients, which presents key challenges. • Robust quality data doesn’t exist: For the most part, alternative ingredients haven’t been tested at scale in many commercial settings, meaning that the confidence behind products doesn’t exist yet, and

Aquafeed: Advances in Processing & Formulation Vol 16 Issue 1 2024

NOVEL INGREDIENTS

there is a huge risk for farmers testing new products at scale. • Farms are changing too quickly: For each generation of fish, the farming method, cages, genetics, and water conditions are fluid and quite different. That means that knowledge and tests that were done recently may not even be applicable to the farm this generation. Big data methods offer solutions to these challenges by enhancing field trials and lab results. They improve confidence in applying research findings in real-world scenarios, uncover hidden insights, and provide near real-time answers. With today's data capabilities, it's possible to address the myriad of conditions farmers encounter in actual environments.

EPA + DHA study: A leap in aquaculture research In aquaculture research, turning lab research into practical farm applications is challenging due to factors like limited sample size, cost, and diverse farm variables. When Veramaris partnered with Manolin to study the effects of different dietary levels of Eicosapentaenoic Acid (EPA) and Docosahexaenoic Acid (DHA) on fish, Manolin's technological expertise and

thorough research approach were key in addressing these challenges. Manolin's technology and thorough research approach enabled a smoother transition of lab results to actual aquaculture practices and bridged a vital industry gap. Together, Veramaris and Manolin investigated the impact of varying EPA and DHA levels in fish diets, using big data analytics to explore this intricate topic. EPA and DHA, vital for salmon feed, are essential for fish health, supporting growth, organ development, and immune response. The study aimed to determine the optimal dietary levels of these omega-3 fatty acids and their effect on fish performance and quality. This joint effort enhanced our knowledge of fish nutrition and established new standards in aquaculture research methods. Methodology The study was meticulously structured into three distinct phases, wherein the feed products utilized during the designated study period were systematically tagged with their specific EPA+ DHA levels. To ascertain the comprehensive impact of these nutritional components, individual EPA and DHA levels were

Aquafeed: Advances in Processing & Formulation Vol 16 Issue 1 2024

NOVEL INGREDIENTS Table 1. Feed batches tested with specific median EPA+DHA levels

EPA+DHA category

EPA+DHA level

Total generations

Above average

> 8.0%

Average

7.0-8.0%

110

Below average

< 7.0%

calculated for each generation of fish, thereby determining the overall average level employed throughout the generational cycle. To rigorously evaluate the efficacy of varying EPA + DHA levels, a detailed statistical significance analysis was conducted. This analysis encompassed a broad spectrum of fish welfare variables, including growth rates, mortality rates, prevalence of disease, feed efficiency, and overall harvest quality. Such a comprehensive approach ensured a thorough measurement of the results, providing a holistic understanding of the influence of EPA and DHA levels on aquaculture productivity and fish health.

Data pool For this collaborative study, the impact of three distinct EPA+DHA levels in feed was evaluated using real production data from 232.6 million smolts across 99 active Atlantic salmon farms along the Norwegian coast from 2013 to 2022. Each feed batch was tagged with specific median EPA+DHA levels, averaged per generation, to categorize into three groups (Table 1). Key performance indicators, including Atlantic salmon mortality (%), economic Feed Conversion Ratio (eFCR), and superior harvest percentage (%), were calculated for each group, providing insights into the effects of varying EPA+DHA levels in salmon feeds.

Aquafeed: Advances in Processing & Formulation Vol 16 Issue 1 2024

NOVEL INGREDIENTS

Key analysis and findings The research conducted revealed valuable insights for feed managers to better understand the benefits of increased levels of EPA and DHA in the diets of farmed Atlantic salmon. One important key finding was that generations of fish supplied with EPA+DHA levels >8% showed 21% less variability in the total mortality and an 11% improvement in the eFCR. For farmers, this translated into a 27% higher chance of obtaining a superior harvest higher than 90%. These findings are particularly useful for Atlantic salmon farmers as they provide a way to quantify the advantages of including higher levels of EPA and DHA in feed formulations. This approach leads to an improved return on investment (ROI) and reduced risk, as the outcomes are based on empirical data from millions of fish. Results To the best of our current knowledge, this study presents the first approach of utilizing big data to provide compelling evidence that optimal nutrition, specifically EPA+DHA levels exceeding 8%, can lead to significant improvements in eFCR, mortality rate, and superior harvest quality. By achieving more fish of superior quality and improved feed conversion, the potential for a higher return on investment becomes evident. Moreover, while this study focuses

on EPA+DHA levels, the BD method holds promise for evaluating other feed ingredients, functional additives, as well as various zootechnical and well-being factors.

Charting a sustainable future in aquaculture with big data Veramaris' study is more than just an academic exercise; it's a new way of guiding the industry toward a sustainable future. Big data and innovative technologies give aquaculture a new strategy for companies to address their growing challenges – from ensuring fish health and quality to minimizing environmental impact. This approach, reminiscent of Jeff Bezos' vision for Amazon, underscores the transformative power of technology in tackling complex tasks. As the industry continues to evolve, it's clear that the path to sustainable growth in aquaculture lies in the intelligent integration of technology, data, and a commitment to environmental stewardship. References available on request.

More information: Tony Chen CEO Manolin E: tony@manolinaqua.com

Aquafeed: Advances in Processing & Formulation Vol 16 Issue 1 2024

NOVEL INGREDIENTS

Low-footprint insect-based feed shows positive effects on salmon health Ikram Belghit, Gopika Radhakrishnan, Institute of Marine Research, Michel van Spankeren, Protix A new study (Radhakrishnan et al., 2023) published by the Institute of Marine Research (IMR) in Norway has examined the effects of insect-based feed on the health of salmon grown at sea, with highly promising results. The top finding of this research indicates that a diet incorporating Protix’s ProteinX© insect-derived meal at an inclusion rate of up to 10% has positive effects on dealing with stress, the mucosal barrier, wound healing and liver health. At the same time, growth performance and mortality remain comparable with a commercial reference feed. These results spell good news in the quest to serve up more sustainable salmon on consumers’ plates.

Credits: Helge Skodvin, Institute of Marine Research

Challenges Oily fish contain essential nutrients that support the nervous system, heart and brain health and are very popular. Atlantic salmon is one of the most commonly reared species. Large salmon farms seek to create a

healthy environment and maintain fish well-being as far as possible, for example through fish feed. Atlantic salmon were traditionally fed a diet of predominantly fishmeal and fish oil, placing a burden on marine ecosystems. Today, fish feeds have high inclusion

Aquafeed: Advances in Processing & Formulation Vol 16 Issue 1 2024

NOVEL INGREDIENTS levels of plant-based ingredients such as soy protein concentrate. However, soy protein concentrate has also been linked to environmental issues. Increasingly, salmon farmers are looking for locally sourced sustainable ingredients that align with the principles of the circular economy.

A sustainable vision Switching to insect-based feed is perceived as a promising alternative that can promote piscine health and well-being, and is more sustainable than conventional alternatives through a circular production process that utilizes organic waste materials as feed for the insects and minimizes environmental impact. IMR has been working with Protix, the leading insect ingredients company worldwide, for over a decade to further study insect-based feed as a potentially positive solution. IMR is a leading advisory and monitoring research institute that supplies knowledge related to the sustainable management of resources in our marine ecosystems and the whole food chain from the sea to the table. Established in 2009, Protix is at the forefront of creating a sustainable food system by developing ingredients from insects, namely the larvae of the black soldier fly (BSF). The two organizations have a shared mission to make the feed and food chain more sustainable. Their ethos is based on a deep scientific foundation, with ongoing studies since 2012 to build on existing data. The study The most recent study was part of the SUStainable INsect CHAIN (SUSINCHAIN) project which aims to contribute to novel protein provision for feed and food in Europe. Atlantic salmon were grown and monitored for a full year in challenging “commercial” sea conditions, with one group fed a diet including 5% ProteinX, another group fed a diet including 10% ProteinX, and the third group receiving a commercial control diet. The salmon in this study faced stress factors typical of marine life, such as water temperature fluctuation, parasites and pathogens as well as handling. ProteinX is a high-quality insect-based protein meal by Protix. It is a careful balance of essential amino and fatty acids, and macro and micro minerals designed to support fish growth and good health. It uses less water

and land than most alternatives and also reduces the carbon footprint.

The results Results showed that a diet incorporating ProteinX can improve the health of salmon reared at sea, while keeping performance on par with conventionally fed fish. As well as a positive impact on well-being, the circular production of the BSF larvae means the feed is more sustainable. The key findings of the trial are detailed below. Better at dealing with stress Fish under stress produce cortisol – a “fight or flight” hormone that renders them brighter and more alert. However, raised cortisol over a longer period can have a negative impact on fish well-being. Atlantic salmon fed an insect-based diet showed an almost 50% reduction in cortisol response along with an improved number of red blood cells (RBC) and 34% higher hemoglobin (Hb) when stressed. These results indicate that salmon fed with insectbased diets are less stressed under demanding conditions than regularly fed fish, thus resulting in overall better fish health when fed diets containing ProteinX (Fig. 1). Improvement of the mucosal barrier The mucus of the skin and gills provide an effective barrier against invading pathogens. In this study, the salmon fed with diets containing ProteinX were found to have a higher skin mucus DNA concentration and increased expression of genes related to inflammatory response in the gills. These results show that dietary insects can change the mucus secretion and improve the inflammatory response in salmon under stressful conditions (Fig. 2). Effective wound healing Atlantic salmon in the study fed with ProteinX demonstrated an increase in the genes related to wound healing and tissue remodeling, allowing them to heal more effectively. This confirms earlier findings using ProteinX in Atlantic salmon (Li et al., 2019). Liver protective effect The liver is an important metabolic organ in salmon.

Aquafeed: Advances in Processing & Formulation Vol 16 Issue 1 2024

NOVEL INGREDIENTS

Figure 1. Improved stress response in Atlantic salmon fed insect-based diets. Plasma cortisol (ng/mL), Red blood cells (RBC, measured number * 1012 cells/L) and Hemoglobin (Hb, g/100mL) of Atlantic salmon fed different diets at two-time points. The data are represented as mean of 6 fish/cage (± SE, triplicate cages per diet). The letters a and b denote the statistical difference among the dietary groups, and letters A and B denote the statistical difference among the two sampling points. Factorial ANOVA was performed with diet, block and time as three factors and cage as random factor, followed by Tukey multiple comparison tests.

Figure 2. Improved mucosal barrier and inflammatory response in Atlantic salmon fed insect-based diets. DNA concentration (ng/mL) and normalized gene expression (NGE) of interleukin 1β (il1β). The data are represented as mean of 6 fish/cage (± SE, triplicate cages per diet). The letters a and b denote the statistical difference among the dietary groups, and letters A and B denote the statistical difference among the two sampling points. Factorial ANOVA was performed with diet, block and time as three factors and cage as random factor, followed by Tukey multiple comparison tests.

The study showed that salmon-fed insect-derived feed can have an improved liver function by reducing the level of alanine aminotransferase (ALT) by half, while the level of aspartate aminotransferase (AST) was reduced by more than 70% in plasma. These results indicate that dietary inclusion of insect meal can reduce inflammatory tissue damage in the liver. The decrease in AST after feeding a diet containing ProteinX was also

confirmed in previous work on Atlantic salmon grown at sea (Belghit et al., 2019), where AST levels were reduced by a quarter. These positive results are supported by findings from earlier studies on antioxidant effects (MouithysMickalad et al., 2020; Ushakova et al., 2021) and liver health (Belghit et al., 2019). Based on these studies ProteinX shows a liver protective function.

Aquafeed: Advances in Processing & Formulation Vol 16 Issue 1 2024

NOVEL INGREDIENTS

Credits: Helge Skodvin, Institute of Marine Research

A sustainable future for salmon Looking ahead, the aim is to satisfy the nutritional requirements of the fish while minimizing the direct environmental effects around the farm and the environmental footprint of the feed materials. Sustainable, high-quality Atlantic salmon is very much on the menu. For more information email eva.wilders@protix.eu

References Radhakrishnan, G., Liland, N. S., Koch, M. W., Lock, E. J., Philip, A. J. P., & Belghit, I. (2023). Evaluation of black soldier fly larvae meal as a functional feed ingredient in Atlantic salmon (Salmo salar) under farm-like conditions. Frontiers in Aquaculture, 2, 1239402. Li, Y., Kortner, T. M., Chikwati, E. M., Munang'andu, H. M., Lock, E. J., & Krogdahl, Å. (2019). Gut health and vaccination response in pre-smolt Atlantic salmon (Salmo salar) fed black soldier fly (Hermetia illucens) larvae meal. Fish & shellfish immunology, 86, 1106-1113.

Belghit, I., Liland, N. S., Gjesdal, P., Biancarosa, I., Menchetti, E., Li, Y., ... & Lock, E. J. (2019). Black soldier fly larvae meal can replace fish meal in diets of sea-water phase Atlantic salmon (Salmo salar). Aquaculture, 503, 609-619. Mouithys-Mickalad, A., Schmitt, E., Dalim, M., Franck, T., Tome, N. M., van Spankeren, M., … & Paul, A. (2020). Black soldier fly (Hermetia illucens) larvae protein derivatives: Potential to promote animal health. Animals, 10(6), 941. Ushakova, N. A., Dontsov, A. E., Marsova, M. V., & Bastrakov, A. I. (2021). Antioxidant properties of an extract of Hermetia illucens larvae. Biology Bulletin, 48, 118-121. Belghit, I., Waagbø, R., Lock, E. J., & Liland, N. S. (2019). Insect‐ based diets high in lauric acid reduce liver lipids in freshwater Atlantic salmon. Aquaculture Nutrition, 25(2), 343-357.

Gopika Radhakrishnan PhD candidate Institute of Marine Research

Aquafeed: Advances in Processing & Formulation Vol 16 Issue 1 2024

NOVEL INGREDIENTS

Reshaping aquafeed formulation with black soldier fly larvae meal: Why the time is now Gil De Clercq, Entobel

In previous editions of Aquafeed.com, the growing pressure on existing protein raw materials, considering factors such as availability, sustainability, and price volatility, has already been widely addressed. The search for high-quality, economically viable protein alternatives continues, with the Black Soldier Fly (BSF) protein meal gaining popularity and experiencing year-on-year market growth. While some pioneers in the industry have already incorporated BSF meal into their animal formulations as a valuable protein source, there remains

skepticism about whether this alternative protein source will secure its place in the diets of aquatic species. This skepticism does not revolve around the concept of sustainable BSF protein meal production based on circular economy principles, which aim for a reduced ecological footprint. Rather, part of this skepticism is rooted in apprehensions regarding volume reliability and consistent quality, particularly in terms of protein level and amino acid profile. Addressing these concerns can be achieved through scaling up production facilities

Aquafeed: Advances in Processing & Formulation Vol 16 Issue 1 2024

NOVEL INGREDIENTS and emphasizing a reliable supply for larvae diets, as demonstrated by Entobel in 2023. With a new production facility in Vung Tau (Vietnam), boasting a capacity of 10,000MT and long-term contracts with suppliers, these challenges can be effectively addressed. Economic viability is another major concern, but it is partially addressed by the previously mentioned parameters. Moreover, at Entobel, we strive to mitigate the volatility of BSF meal prices by establishing long-term contracts with our customers as well. This positions the BSF meal produced by Entobel favorably against other protein sources, especially when fluctuations in demand/supply cause price increases, as observed with premium fishmeal in the second half of 2023. In addition to considering price volatility, it is crucial to factor in the financial gains attributed to enhanced growth performances, as illustrated in the trial description below. The lingering question revolves around whether BSF meal can effectively serve as a replacement for premium protein sources at a nutritional level. It’s established that BSF meal boasts high digestibility, palatability, and appeal as a protein meal with hypo-allergenic capacities. These qualities stem from the molecular weight properties of its protein content. The right balance of free amino acids, di-and tripeptides present in the BSF meal contributes to these additional functionalities. Beyond this, other notable features include the presence of anti-bacterial properties. BSF meal contains around 60 identified anti-microbial peptides acting as a natural defense system for the larvae in the form of long-chain proteins. Another distinctive aspect, unlike other complete protein sources, is the high concentration of the Gram-positive antibacterial lauric acid, that is present in the fat content of the BSF meal. An assessment at Nong Lam University, with the aim to explore the impact of substituting premium fishmeal

with BSF meal on the growth performance of juvenile whiteleg shrimp (Litopenaeus vannamei), is detailed below. BSF meal shows a comparable amino acid profile as premium fishmeal and incorporates all the unique functionalities discussed earlier. Due to BSF meal’s lower protein content in comparison to premium fishmeal, there was a reformulation of the plant proteins, soybean meal and potato protein concentrate 75%, along with the inclusion of synthetic amino acid L-threonine. This adjustment aimed to ensure that the total amino acid profile remained consistent across all treatments. The other protein sources like poultry meal (5%) and wheat gluten (1.8%) were kept constant across the treatments.

Materials and methods Whiteleg shrimp (60 shrimp/diet x 4 replicates) were subjected to the 5 diets, with the primary protein sources outlined in Table 1. BSF meal, produced by Entobel and hereinafter referred to as H-Meal, contains 55% crude protein and has a fat content of 13%. In each diet, premium fishmeal was replaced on a 1:1 basis with H-Meal, except for diet 2 where the substitution was a 2:1 ratio. Reformulation of the ingredients shown in Table 1, ensures that each diet has the same protein content and amino acid profile. After 8 weeks, the final weight, specific growth rate and feed conversion ratio were measured for each treatment. Also, the feed cost per kg shrimp produced was calculated to determine the efficacy of the feed compositions. Results In Table 2, we delve into the performance results across the various dietary treatments. The survival rate across treatments did not show significant differences (P>0.05). However, as outlined in Table 2, after 8 weeks, shrimp fed with diet 1 (3% fishmeal replacement with 3% H-Meal) and diet 2

Table 1. Primary protein resources

Ingredient/treatment

Ctrl

Diet 1

Diet 2

Diet 3

Diet 4

Fishmeal 67%

15%

12%

H-Meal 55%

Soybean meal

32%

30%

31%

27%

26%

Corn protein concentrate

L-Threonine

0.10%

0.13%

0.14%

Aquafeed: Advances in Processing & Formulation Vol 16 Issue 1 2024

NOVEL INGREDIENTS

Table 2. Performance results

Treatment

Fishmeal (%)

H-Meal (%) Final weight (g/fish) SGR (%)

FCR

Feed cost (USD/kg shrimp)

CTRL

21.10a

4.62a

1.93b

1.92

Diet 1

22.80

4.78

1.67

1.69

Diet 2

22.90b

4.75b

1.72ab

1.70

Diet 3

24.30

4.87

1.56

1.63

Diet 4

24.30

4.86

1.65

1.73

(6% fishmeal replacement with 3% H-meal), as well as diet 3 (6% fishmeal replacement with 6% H-Meal) and diet 4 (9% fishmeal replacement with 9% H-Meal), demonstrated notably higher final weights and specific growth rates in comparison to the control group containing 15% premium fishmeal without H-Meal. Specifically, the final weight for Diet 2 and 3 was approximately 8.5% higher compared to the Control group, with a specific growth rate increase of approximately 2.8%. For Diet 3 and 4, the final weight was approximately 15% higher, accompanied by a specific growth rate increase of approximately 5%. Moreover, the Feed Conversion Ratio (FCR) exhibited significantly improved results when fishmeal was partially replaced with H-Meal across all diets. Notably, Diet 2 stands out, involving a 2 to 1 ratio substitution, achieving superior and statistically significant performance results. Overall, the results of the growth parameters resulted in a lower feed cost per kilogram of shrimp produced for all the treatments that included H-Meal in the diets. The feed cost per kilogram of shrimp was the highest in the Control group containing only premium fishmeal and the most cost-effective in Diet 3 with the partial replacement of 6% fishmeal by 6% H-Meal.

Conclusion In conclusion, the trial’s favorable results in growth performance suggest that nutritionally, replacing premium fishmeal with BSF insect meal is a viable option resulting in a lower feed cost per kilogram of fish produced. This underscores the potential of BSF insect meal as a promising and sustainable feed ingredient in aquafeed formulations. Entobel is actively contributing to the revolution in animal nutrition with its Black Soldier Fly insect meal. Achieving cost-effectiveness and establishing a dominant position for BSF meals in the nutritional landscape requires collective efforts from suppliers and customers. Together, we can propel this new sustainable alternative to new heights and secure a reliable protein supply, a necessity for the future growth of aquaculture.

More information: Gil De Clercq International Sales Manager Entobel

Aquafeed: Advances in Processing & Formulation Vol 16 Issue 1 2024

Industry Events 2024 JANUARY 30 – Feb 1:

2024 IPPE, USA

www.ippexpo.org

SIMEC Expo 2024, Saudi Arabia 33rd Annual Practical Short Course on Feeds, Pet Food and Aqua Feed Extrusion, USA AquaFarm 2024, Italy Aquaculture America 2024, USA Fish International, Germany

www.simec-expo.com web.cvent.com

Victam Asia 2024, Thailand AFIA Purchasing & Ingredient Suppliers Conference, USA Aqua Sur 2024, Chile

victamasia.com www.afia.org www.aqua-sur.cl

Seafood Expo, Spain

www.seafoodexpo.com

Aquaculture UK XXI International Symposium on Fish Nutrition and Feeding, Mexico Ildex Vietnam 2024

aquacultureuk.com www.isfnf2024.com

Insects to Feed the World, Singapore Shimp Summit, India

www.ifw2024.com responsibleseafood.org

Asian-Pacific Aquaculture 2024, Indonesia

www.was.org

AQUA 2024, Denmark

was.org

FEBRUARY 4 – 6: 5 – 9: 14 – 15: 18 – 21: 25 – 27:

www.aquafarm.show was.org fishinternational.de

MARCH 12 – 14: 12 – 14: 19 – 21:

APRIL 23 – 25:

MAY 14 – 15: 27 – 31: 29 – 31:

ildexvietnam.com

JUNE 19 – 22: 27 – 29:

JULY 2 – 5:

AUGUST 25 – 29:

SEPTEMBER 3 – 5: 9 – 12:

Global Shrimp Forum, The Netherlands larvi 2024, Belgium

shrimp-forum.com aquaculture.ugent.be

AQUA EXPO Guayaquil, Ecuador

aquaexpo.com.ec

OCTOBER 21 – 24:

NOVEMBER 12 – 15:

EuroTier, Germany

www.eurotier.com

Aquafeed: Advances in Processing & Formulation Vol 16 Issue 1 2024