Aquafeed Magazine Vol 17 Edition 1 2025

Vol 17 Issue 1 January 2025

AQUAFEED Advances in processing & formulation An Aquafeed.com publication

LIMITING EHP EFFECTS IN SHRIMP Disease prevention and management Fish protein hydrolysates The Global Feed LCA Institute

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 17 ISSUE 1 2025

Contents

SOYBEAN PEPTIDES FOR FISH-FREE AQUAFEEDS 42 Recent trials using a soybean peptide showed that fishmeal replacement is feasible.

ALLEVIATING ECTOPARASITE INFECTIONS IN POMPANO 16

EHP PREVENTION IN SHRIMP FARMING 24

Phytobiotic-based additives reduce the severity of ectoparasite infections.

Feed supplements could contribute to limiting EHP impacts.

Aquafeed: Advances in Processing & Formulation Vol 17 Issue 1 2025

FISH PROTEIN HYDROLYSATES 38 Improved quality and processing methods of the unrivaled marine ingredients.

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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

VOL 17 ISSUE 1 2025

Contents

Advertising enquiries: sales@aquafeed.com

6 Interview with Amiya D. Nath 9 News Review 12 New on the market 16 Phytobiotic-based additives to alleviate

Accounts & all other enquiries: info@aquafeed.com

19 Combating a significant threat to whiteleg shrimp growth:

SUBSCRIBE Digital editions are free to industry subscribers.

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ectoparasite infections in pompano farming EHP infection *Cover story

24 EHP prevention in shrimp farming: Spore generation, a key phase in limiting EHP outbreaks and effects

28 The power of krill meal to improve skin health and reduce sea lice

32 Recent trials show that mycoprotein is beneficial to salmon and tilapia immune systems and the reduction of GHG emissions

FOLLOW US

35 Diformates in seabass aquaculture: A proven concept 38 Fish protein hydrolysate from pelagics: A sustainable and efficient solution for aquafeeds

42 Soybean peptides for a fish-free aquaculture 44 Breaking free from the ocean's grip: We are grateful to the following companies for sponsoring this issue of the magazine.

Their support allows us to make our publications available without charge.

Adisseo......................................................................................... 2 Victam......................................................................................... 5 Scoular......................................................................................... 8 Aquaproducts..........................................................................13 Extru-Tech..................................................................................15 Catalysis.....................................................................................23 Nouvelles Vagues....................................................................34 Berg + Schmidt.........................................................................41 China Feed Industry Expo.......................................................51 AquaFarm.................................................................................67

A new era for aquatic feed

48 Driving sustainability in animal nutrition:

The role of the Global Feed LCA Institute

52 Column: Shrimp nutrition & health:

A holistic approach by Albert G.J. Tacon

58 Hacking the line: A systematic review of how

probiotics disrupt bacterial communication through quorum-quenching

63 Innovative sustainable recirculating aquaculture feeds: The InSuRAFeed project

66 Calendar of events

WAS ..........................................................................................68

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Aquafeed: Advances in Processing & Formulation Vol 17 Issue 1 2025

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INTERVIEW with Mr. Amiya D. Nath Mr. Amiya D. Nath, V.P., JAPFA India AQ: When did JAPFA enter the Indian market? When and why did the company start serving the aquaculture industry?

AN: JAPFA India was established back in 1995, proudly pioneering pellet feed technology nearly three decades ago. Initially, the focus was on innovative feed solutions for poultry and broilers, positioning JAPFA India at the forefront of transforming the country’s feed sector.

Growing demand for JAPFA feeds nationwide led to the establishment of world-class poultry feed mills to serve major regions across India.

In recent years, JAPFA India, with a strong distribution and supply chain network, has experienced strong demand for premium-quality aquafeeds. Our aquafeed brand, Suri Tani Premuka (STP), is well-known for

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supporting and empowering farmers in Indonesia, Vietnam, and other countries. To meet this demand, JAPFA India made a significant investment of over $10 million and inaugurated its first aquafeed plant on November 7, 2024, in West Bengal. The new feed mill produces shrimp feeds and extruded feeds.

AQ: Can you describe the new shrimp feed facility?

AN: Our new shrimp feed facility is equipped with worldclass German technology from Bühler, enabling the production of up to 5 tons of shrimp feed per hour. The plant is modeled after our operations in Indonesia, where STP is the top-selling shrimp feed brand produced at five facilities. This replication ensures we deliver the highestquality shrimp feed in India.

Aquafeed: Advances in Processing & Formulation Vol 17 Issue 1 2025

INTERVIEW To meet the local demand, JAPFA India made a significant investment of over $10 million and inaugurated its first aquafeed plant on November 7, 2024, AQ: Can you share more about your facilities and the in West Bengal. The new STP aqua team in India?

AN: We have the capacity to produce over 60,000 MT feed mill produces shrimp of floating fish feed, 25,000 MT of sinking feed, and 35,000 MT of shrimp and polyculture feed annually feeds and extruded feeds. in the first phase, using a state-of-the-art feed mill.

AQ: The new feed mill can also produce extruded feed.

Is there a demand for it in the Indian market?

AN: Over the past three decades, India’s aquaculture sector has seen exponential growth, accompanied by a notable shift from sinking pellet feed to extruded floating feed. JAPFA India has been producing and selling sinking feeds for several years, but farmers have increasingly requested floating feeds for major carps, pangasius, catfish, and other aquaculture species. Our new aquafeed plant is designed to consistently supply highly digestible and cost-effective extruded feeds to meet this demand.

Our operations are supported by top-tier quality control systems and fully equipped laboratories. With warehousing and silos that store raw materials for over three months, we ensure smooth, uninterrupted operations.

Our STP Aqua Team comprises highly qualified professionals, including MFSc graduates and PhD scholars.

Supported by our international teams, we leverage global expertise to maintain our leadership in the market.

Over time, our market research in Indian aquaculture has given us a deep understanding of how to produce the highest-quality feed. JAPFA STP benefits from its corporate headquarters in Singapore and expertise from Indonesia, giving us a competitive edge in delivering superior products. We strictly adhere to all regulations and hold certifications like CAA, reflecting our commitment to sustainability and farmer welfare.

AQ: How are you addressing the recent volatility in ingredient prices?

AN: Ingredient price volatility has been a persistent challenge for STP in India and other markets. While fishmeal and fish oil costs are no longer the main concern, the prices of alternative raw materials have risen sharply. Thankfully, our experience in navigating market fluctuations has allowed us to efficiently manage supply and demand. By proactively stocking raw materials in our warehouses and silos, we ensure that farmers remain unaffected by price instability and continue to receive quality feed at stable prices.

AQ: Tell us a bit more about the fish segment. What challenges and opportunities do you see in India?

AN: India has primarily been a carp-producing nation, with double-digit annual growth over the past three decades. However, fish prices haven’t increased proportionately, leaving farmers seeking affordable, highquality floating feeds.

Species like pangasius and tilapia are gaining popularity, and farmers have recently succeeded in farming highvalue fishlike milkfish and seabass, among others. JAPFA is committed to serving fish farmers with superior feed at competitive prices. Unlike some competitors whose product specifications vary between brochures and packaging, we ensure that our products meet the specifications we advertise.

AQ: Why did you decide to enter the shrimp market?

AN: Our decision to enter the shrimp feed market was driven by the needs of shrimp farmers and final consumers. While shrimp aquaculture in India has grown consistently over the years, the net profits and livelihoods of shrimp farmers have not kept pace. Andhra Pradesh, India’s top shrimp-producing state, has the majority of shrimp feed plants concentrated in its vicinity. As a result, neighboring states face higher feed costs due to transportation and other expenses.

Aquafeed: Advances in Processing & Formulation Vol 17 Issue 1 2025

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INTERVIEW To address this, JAPFA India conducted extensive surveys to understand market dynamics and farmer needs.

We strategically chose West Bengal, India’s second-largest aquaculture-producing state, as the location for our first shrimp feed plant. This decision allows us to serve local farmers and nearby states more effectively.

AQ: The shrimp industry has faced challenges recently.

How can JAPFA help shrimp farmers?

AN: Shrimp farmers face issues like variable growth rates, early harvests, and the high cost of pond and feed supplements. Our STP shrimp feed has gained significant traction in other countries by reducing shrimp production costs and increasing farmer profits. JAPFA India is committed to achieving the same in India through premium feed quality and robust technical support to improve farmer livelihoods.

We also see Penaeus monodon (tiger prawn) making a strong comeback, with growing interest among farmers to cultivate it for domestic consumption. STP has extensive experience formulating feeds for various shrimp species, including vannamei and monodon, and we offer

specialized formulations for both. Additionally, with limited high-quality feed suppliers dominating the market, we see opportunities to expand our presence.

AQ: Tell us about the unique “polyculture fish feed” segment in India. What is it?

AN: Freshwater prawn, also known as scampi, farming was once very popular in India, but its longer crop cycles and higher production costs led many farmers to shift toward vannamei and other species. Polyculture allows farmers to raise carp and other fish alongside scampi in the same pond. This system caters to a variety of species, requiring feed tailored to their diverse dietary needs.

JAPFA has developed a polyculture feed specifically designed to meet these requirements.

AQ: What are your growth projections?

AN: Based on farmer feedback, the demand for our feeds is overwhelming. We anticipate fully utilizing both our fish and shrimp feed mills within three years. We also plan to expand into new regions during this time and aim to double our business annually.

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Aquafeed: Advances in Processing & Formulation Vol 17 Issue 1 2025

NEWS

NEWS REVIEW Highlights of recent news from Aquafeed.com

De Heus opens its fifth production site in Indonesia

Nicovita opens new feed facility in Ecuador

The group strengthened its presence in Indonesia with the acquisition and launch of its fifth production facility in Purwodadi, Central Java.

The new Purwodadi factory will serve its customers and meet the growing demand for high-quality animal feed in Central Java and surrounding areas.

The factory has expanded its production capacity to 15,000 tons per month.

Nicovita opened a state-of-theart facility in Ecuador aiming for sustainable and efficient growth in the country and supporting the future of the industry. The new facility, equipped with cutting-edge technology and high levels of digitalization, ranks among the most advanced shrimp feed manufacturing plants. The new plant joins Vitapro's network of facilities in Ecuador, Peru, Honduras, and Chile.

BioMar develops solution to boost fish health and resilience Cargill revamps its shrimp portfolio with highperformance philosophy

SmartCare Endurance is an innovative feed solution developed to help farmed fish endure challenging conditions, such as SRS outbreaks. It features a unique blend of antioxidants, vitamins, and minerals that work to mitigate oxidative stress, reduce inflammation, and enhance the fish's natural immune response. This results in a more resilient fish, better equipped to withstand threats from pathogens such as Piscirickettsia salmonis and viral diseases like IPN and PD.

Cargill Ecuador launched its revamped Aquaxcel portfolio, now under a high-performance philosophy.

As one of the most significant innovations in the nutritional portfolio, the new Aquaxcel lineup offers improvements in key performance indicators, targeting the production and economic goals of shrimp farmers.

Under this new philosophy, average FCRs have been reduced by 15%, while weekly growth rates have increased by 49%.

Aquafeed: Advances in Processing & Formulation Vol 17 Issue 1 2025

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NEWS Cargill rebrands its micronutrition solutions and services

Cargill Animal Nutrition unveiled a comprehensive range of best-in-class micronutrition solutions and services under its new Micronutrition and Health Solutions brand portfolio. The company evolved its current portfolio moving from a technology-first to species-first offering, transitioning brands like Delacon and Provimi into eight focused product brands. Syrena is the new aqua brand that provides innovative sustainable solutions that support the health and performance of cold and warm water species, including fish and shrimp.

Cooke expands acquires Peruvian fishmeal giant

CPM acquires Jacobs Global CPM acquired Jacobs Global, a premier supplier of aftermarket parts for hammermills and pellet mills. This strategic transaction will enhance CPM’s product offerings, expand its market reach and add capacity to better serve CPM’s growing global customer base.

BioMar Chile to consolidate aquafeed production at Pargua plants by 2025 Starting in the third quarter of 2025, BioMar will concentrate its production strategy on its two Pargua plants, ceasing operations in Castro, Chiloé, in Chile.

The decision reflects BioMar's future-oriented focus on business sustainability across all dimensions and a commitment to efficiency throughout the salmon value chain.

Bühler opens Grain Innovation Center

Cooke, through its subsidiary PF Cayman New Holdco Limited, acquired Corporación Pesquera Inca S.A.C. (Copeinca) of Peru. Copeinca holds the largest anchoveta quota in Peru at 15.9% and processes approximately 21% of the country’s total catch for annual production of approximately 200,000 MT of fishmeal and 23,000 MT of fish oil.

The Swiss technology group inaugurated its new Grain Innovation Center in Uzwil. The center was carefully designed to empower customers from the food and animal nutrition industries to innovate and improve their processes. Customers can conduct tests on food and animal feed and can develop new processes and solutions in various areas, including cleaning, optical sorting, grinding, sifting, mixing, and protein shifting.

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Aquafeed: Advances in Processing & Formulation Vol 17 Issue 1 2025

NEWS dsm-firmenich, Format Solutions forge alliance to unlock the value of sustainable feed dsm-firmenich’s Animal Nutrition & Health division, alongside Format Solutions, a provider of integrated formulation and feed ERP software, signed a strategic alliance aimed at quantifying and managing the environmental impact of animal feed. The collaboration melds Format Solutions’ formulation software with the comprehensive life cycle assessment (LCA) platform, Sustell™. This integration empowers customers to calculate and share their feed footprints effortlessly.

Aker QRILL Company launched Aker QRILL Company, a global standalone entity spun out of Aker BioMarine, was launched and embarked on a new chapter, now supported by new ownership from American Industrial Partners (AIP) and Aker Capital. This transition marks more than just a name change; it’s the start of a new era in the company and the development of its brands, QRILL Aqua and QRILL Pet.

Feed and Feeding for Fish and Shellfish: Nutritional Management A new book relevant to the aquafeed industry has bee released.

Edited by Vikas Kumar, written by an international panel of experts and with an applied focus, the book serves as a reference book for aquaculture industry professionals seeking information on the nutritional requirements and nutritional pathology of key species, metabolism, physiology and concerns associated with feed manufacturing within the aquafeed industry.

It presents foundational knowledge and the most recent advances in aquaculture finfish and crustacean metabolism and nutrient requirements, feed ingredients, nutritional deficiency disorders, and integrated sciences.

This book contains fresh approaches to understanding feed, nutrition, metabolism, and overall health in the aquaculture industry.

Benchmark to sell genetics business to Novo Holdings Benchmark Holdings plc has entered into a binding agreement to sell its genetics business, Benchmark Genetics, to Starfish Bidco, a wholly owned subsidiary of Novo Holdings, valuing the enterprise at up to £260 million. The sale will allow Benchmark Holdings to focus on its Advanced Nutrition and Health business areas.

Aquafeed: Advances in Processing & Formulation Vol 17 Issue 1 2025

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NEW ON THE MARKET

Aquaproducts unveils PEPTITOM: A game-changing shrimp hydrolysate powder for functional animal nutrition 21/01/2025, 10:12

Aquaproducts, an expert company in marine bioextracts, has cemented its position as a key innovator in sustainable seafood byproduct processing through strategic partnerships with global industry leaders. Collaborating with Norway's Scanbio SAS and Indonesia's Maqpro Biotech, Aquaproducts has leveraged decades of expertise to create a robust, ecofriendly supply chain for functional marine proteins.

In 2022, the company expanded its reach by establishing a new facility, Marine Biotech Vietnam (MBV), through a collaboration with the Mirova Sustainable Ocean Fund. It was inaugurated on June 11, 2024, in Vinh Loc 2 Industrial Park, Long An. This cutting-edge hydrolysate factory focuses on producing dry hydrolysate protein powders for the Asian feed and pet food markets, solidifying Aquaproducts’ role as a regional hub for sustainable, high-quality marine ingredients.

Continuing its pursuit of innovation, Aquaproducts recently introduced PEPTITOM, an advanced shrimp hydrolysate powder designed for both aquaculture and livestock markets. PEPTITOM redefines the use of marine resources in feed formulations, aligning with the company's core values of quality, sustainability, and efficiency. Derived from underutilized shrimp byproducts, PEPTITOM offers a highly digestible, nutrient-rich feed solution for aqua and livestock, helping to reduce waste while boosting animal nutrition.

Functional benefits in aquaculture PEPTITOM is designed for marine fish species and supports the nursery and early developmental stages of all fish species, offering numerous functional benefits beyond basic protein supplementation. It contains bioactive peptides and free amino acids that improve feed intake, digestion, and nutrient absorption, leading to efficient growth during critical early stages.

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The bioactive peptides in PEPTITOM exhibit strong antioxidant and antimicrobial properties, reducing oxidative stress and protecting fish from bacterial infections. This dual role enhances health and defends against pathogens, making PEPTITOM a valuable component in fish diets.

PEPTITOM’s high digestibility ensures optimal nutrient utilization, supporting faster growth and better overall health. It helps aquatic animals cope with stress from environmental changes or feed transitions by maintaining consistent feed intake and enhancing resistance to pathogens.

Additionally, PEPTITOM is an environmentally responsible choice, serving as a sustainable alternative to animal protein sources like krill meal, thus alleviating pressure on marine ecosystems caused by overfishing.

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Quality at its core PEPTITOM reflects Aquaproducts’ larger mission to up-cycle seafood byproducts using cutting-edge

Aquafeed: Advances in Processing & Formulation Vol 17 Issue 1 2025

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NEW ON THE MARKET technologies. The hydrolysate powder is produced through a controlled batch hydrolysis process using selected enzymes, which allows for precise peptide control and quality standardization. This stands in contrast to continuous hydrolysis methods that may yield less consistent results.

Strict control over raw material freshness and supply chain allows for keeping histamine and TVBN levels low (below 200 ppm), extending product shelf life. Further ensuring the product's quality, PEPTITOM undergoes a pasteurization step in compliance with HACCP standards to guarantee food safety and maintain a high standard of freshness.

Economic and functional advantages Aligned with Aquaproducts’ commitment to regional sourcing, PEPTITOM offers a reliable supply for both local and regional markets. This consistent availability allows farmers and feed producers to depend on steady volumes and competitive pricing, facilitating better planning and cost management.

Additionally, Aquaproducts has optimized the drying process of PEPTITOM to enhance cost-efficiency while maintaining high quality. By co-drying, the hydrolysate powder with a carrier, production costs are significantly reduced and hygroscopicity is minimized, preventing caking. This presents a notable advantage over pricier pure spray-dried hydrolysates, ensuring that PEPTITOM remains an economically attractive option without sacrificing performance.

The future of functional feed ingredients Aquaproducts continue to play a pivotal role in reshaping the feed industry with products like PEPTITOM. By leveraging sustainable seafood byproducts and transforming them into high-value, functional ingredients, the company is not only reducing waste but also contributing to the health, productivity, and sustainability of global food systems.

PEPTITOM exemplifies how Aquaproducts continues to lead the charge in marine bio-extract innovation, advancing the industry toward more sustainable and responsible resource use. The company’s ability to innovate at scale, combined with its focus on local partnerships and supply chain stability, ensures that it will remain a key player in the feed and pet food markets for years to come. Inquiries can be addressed at contact@aquaproducts.org.

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NEW ON THE MARKET New Tietjen’s hammer mill development for the fine grinding of fish feed

Biochem expands portfolio with advanced organic trace minerals

The requirements for throughput rates and fineness in the grinding of fish feed and pet food for extrusion have increased significantly in recent years, while the recipes are becoming ever richer in fat and protein. With the FD 32 Pro hammer mill, Tietjen is presenting a completely new development that fits the throughput rates and fineness in the grinding for the extrusion of fish feed and pet food.

BetaTrace® is a unique, patent-pending source of organic zinc, copper, manganese and iron designed for every animal species. It brings together the added value of betaine with an improved trace mineral bioavailability. BetaTrace® is a powerful supporter of animal performance and well-being to meet the challenges of modern livestock farming.

Olmix develops mycotoxin risk management platform Olmix launched MycoKingdom.net, a website platform dedicated to the management and understanding of mycotoxin risk. Myco’Kingdom gathers seven functionalities for identifying, assessing, preventing and managing mycotoxin risk, in order to minimize economic losses linked to contamination.

BinSentry develops platform for commercial grain inventory management

ProSense HD is a new, artificial intelligence-powered inventory management platform aimed at commercial grain handlers and feed mills. It delivers precise, real-time inventory monitoring, so operators always know what is in their commercial bins and silos at any moment in time. This inventory monitoring is carried out using high-definition 3D sensors that are designed to see past dust and debris.

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Aquafeed: Advances in Processing & Formulation Vol 17 Issue 1 2025

PEOPLE IN THE NEWS

PEOPLE IN THE NEWS Paulo Alarcón

Hai Dang Do Vitapro Chile, through its brand Salmofood, has announced a strategic restructuring of its Feed Technology division. Paulo Alarcón Bruce has been appointed as feed technology manager, succeeding Sergio Castillo Alvarado, who will take on new responsibilities within the corporation in Ecuador’s shrimp industry.

Aedrian Ortiz Johnson

INVE Aquaculture appointed Mr. Hai Dang Do as area manager Vietnam. Hai will oversee operations in Vietnam, leading the team and reporting directly to Amir Khalil, regional sales director APAC & China.

Evy Vikene

Zeigler Bros. appointed Aedrian Ortiz Johnson as director of aquaculture sales. Aedrian will spearhead the company’s efforts to grow and maintain its market share, driving sales and delivering nutritional innovation and value creation to customers worldwide.

Ramakanta Nayak

Nuseed® Nutritional appointed Evy Vikene aquaculture lead for Europe/Norway. This is a new role that will support the adoption of Aquaterra omega-3 canola in the Norwegian, UK and EU aquaculture markets.

LaVon Wenger

Ramakanta Nayak was appointed as Orffa's managing director for the Asia-Pacific region.

Ramakanta will spearhead the company’s initiatives to provide science-driven feed solutions that address the specific needs of Asian livestock producers.

Wenger Manufacturing announced the passing of LaVon Wenger on December 29, 2024. He was 89. LaVon was the son of Lou Wenger, one of the company’s original founders. Through the years, LaVon used his intuition and foresight to develop numerous innovations for extrusion cooking technology. LaVon is the inventor on 44 US patents.

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Aquafeed: Advances in Processing & Formulation Vol 17 Issue 1 2025

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DISEASE PREVENTION AND MANAGEMENT

Phytobiotic-based additives to alleviate ectoparasite infections in pompano farming I-Tung Chen, Adisseo, Tran Vi Hich, Nha Trang University, Khuong Duy Nguyen, Maria Mercè Isern-Subich, Waldo G. Nuez-Ortín, Adisseo

The warm water tropical finfish Pompano (Trachinotus blochii and Trachinotus carolinus), valued for their rapid growth rates and excellent market potential, are farmed primarily in Southeast Asia, the United States, and Latin America. However, their production faces significant hurdles due to the presence of ectoparasites, including trematode monogeneans (Benedenia spp., Neobenedenia spp.) and protozoans (Trichodina spp., Cryptocaryon spp.). These parasites inflict severe health issues, leading to high mortality rates and reduced profitability (Weirich et al., 2021).

In Vietnam, where pompano farming is a significant part of the aquaculture industry, mortality rates due to parasites are a recognized challenge during the first four months after fish are transferred to rearing

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cages (Nguyen et al., 2021). Similar cases reported in Taiwan and the Philippines highlight mortality rates reaching 30-40% during this period (Yen et al., 2024; Palma et al., 2022). Parasites weaken fish health and facilitate opportunistic bacterial infections from Vibrio alginolyticus and Vibrio harveyi, which cause ulcers and other health issues, further compromising production.

Traditionally, parasiticides such as praziquantel and fenbendazole have been used to control these infestations (Reed et al., 2009). However, these chemicals pose environmental concerns and the risk of parasite resistance, prompting the exploration of alternative solutions. Effective health management practices, including the supplementation of health additives specifically formulated to reduce ectoparasite

Aquafeed: Advances in Processing & Formulation Vol 17 Issue 1 2025

DISEASE PREVENTION AND MANAGEMENT

Figure 1. The experimental design schematic illustrating the health additive strategy to mitigate ectoparasite infections in pompano, Trachinotus blochii.

load, can mitigate these issues and ensure the health and productivity of pompano culture.

Feeding trial and parasite infection This article evaluates the efficacy of two phytobioticbased additives in mitigating ectoparasite infections to sustain growth in pompano, Trachinotus blochii.

In collaboration with Nha Trang University, Vietnam, the study explored the supplementation strategy based on Sanacore® GM (SNGM) and APEX® (APEX) to reduce the severity of parasite infection under experimental conditions. The study focused on a high-risk period when fish were transferred from nursery to open cages, during the summer months (June to August), when the water temperature rises. SNGM promotes fish gut health and overall immunocompetence, while APEX enhances skin integrity and mucus defensiveness.

The trial was conducted for 70 days, with 60 pompano (initial body weight, IBW 10g) divided into two groups, each with three replicates. For each replicate, 10 fish were housed in a 250L tank, receiving untreated water, and cohabitated with naturally infested carrier fish

to simulate farming conditions where parasites are transmitted via water and from fish to fish. During the transition to the grow-out stage, from day 1 to day 30, the treatment group received feed top-coated with 0.3% SNGM and 0.3% APEX. From day 31 to day 70, as parasite pressure was expected to increase due to fish size growth and environmental stressors such as poorer water quality and crowding, the application dosage was adjusted to 0.1% SNGM and 0.4% APEX. This increase in APEX during the risk period aimed to further strengthen skin immunocompetence and integrity for enhanced capacity to reduce the severity of infection. Fish were monitored regularly, and weight, length, and parasite prevalence were recorded to evaluate the efficacy of the health additive strategy (Fig. 1).

Results Parasite infestations started to be seen after the first week of culture, with protozoans such as Trichodina spp. and Cryptocaryon spp. appearing early. Trematode monogeneans became prominent by day 28, peaking on day 42, and continued to increase until the end of the trial at day 70. By the end of the trial, the combination

Figure 2. The effect of SNGM+APEX supplementation on reducing the prevalence of ectoparasites: monogeneans (A), Trichodina spp.

(B), and Cryptocaryon spp. (C) over 70 days of cultivation. Results were analyzed using one-way ANOVA, and asterisks indicate significant differences (P < 0.05).

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DISEASE PREVENTION AND MANAGEMENT

Figure 3. The effect of SNGM+APEX supplementation on reducing mortality (A) and promoting growth in body length (B) and body weight (C) of pompano over 70 days of cultivation. Results were analyzed using one-way ANOVA, and asterisks indicate significant differences (P < 0.05).

of SNGM and APEX significantly reduced the prevalence of trematode monogenean by 50%, decreasing from 80% in the control group to 40% in the treatment group. Reduction of the prevalence of protozoan infections were more modest, with Trichodina spp. decreasing by 7%, from 58% in the control group to 54% in the treatment group. Similarly, Cryptocaryon spp. prevalence dropped by 2% from 55% in the control group to 52% in the treatment group (Fig. 2).

Under lower severity of infection, the treated fish had a notable reduction in mortality rates. By the end of the 70-day culture period, the health additive strategy reduced mortality by 50%, from 60% in the control group to 30% in the treatment group. Significant growth differences were observed from day 42 of the trial, when parasite loads peaked, and differences between groups became apparent. The additive strategy exhibited better growth performance by 19%, with an average body weight of 76 grams compared to 64 grams in the control group. Similarly, supplementation increased the average total length of fish by 13%, from 15 cm in the control group to 17cm in the treated group. These outcomes suggest that reducing parasite burdens allowed pompano to allocate more energy toward growth and development rather than combating infections (Fig. 3).

Conclusion The present study demonstrated a health additive strategy to effectively support fish growth and survival under parasite infection. The supplementation of phy tobiotic-based Sanacore® GM and APEX® provides overall immunocompetence and skin health to reduce the severity of ectoparasite infections during the early and most susceptible

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stages of the grow-out period. This strategy not only promotes health status and growth performance but also offers a sustainable option alternative to chemical antiparasitic treatments.

References available on request.

More information: I-Tung Chen Global Technical & Scientific Support Manager, Aquaculture Adisseo SAS

Khuong Duy Nguyen Business Development Lead for Health & Farm Care Adisseo SAS

Maria Mercè Isern-Subich Global Product Manager Aquaculture Health Adisseo SAS

Waldo G. Nuez-Ortín Head of Aqua Technical and Scientific Support Adisseo SAS

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Combating a significant threat to whiteleg shrimp growth: EHP infection Bulent Kukurtcu, Catalysis, Loc Tran, Phuc Hoang, Giang Vo, Khanh Nguyen, Ei Mon Ko, Saw Carris Htun, ShrimpVet Laboratory

Credits: ShrimpVet Laboratory

The global shrimp industry, particularly the production of whiteleg shrimp (Penaeus vannamei), has seen remarkable growth over the past decade, especially in Southeast Asia, China, India, and Latin America.

This growth has been accompanied by advancements in healthcare and productivity, driven by the adoption of new technologies and sustainable farming practices for cultured shrimp.

Despite these advancements, the industry continues to face significant challenges, particularly disease outbreaks and the environmental impacts of production systems. To tackle these issues, innovative strategies – such as the development of novel supplements – are being pursued to improve shrimp health, enhance growth performance, and strengthen disease resistance.

EHP, a growing threat Enterocytozoon hepatopenaei (EHP) currently poses a significant threat to the shrimp industry. This intracellular parasite can cause severe damage to shrimp, leading to reduced growth rates, poor feed conversion, and increased mortality rates during the production cycles. EHP can be transmitted horizontally from water sources, cross-contamination from infected farms or shared equipment, and/or vertically via the broodstock and nursery batches. Besides, the EHP-infected population is easily prone to a secondary infection, such as vibriosis and white feces syndrome, as reported in numerous studies. These co-infections could further compromise shrimp physiology, metabolism, growth rate, and even survivability. The severity of these impacts presents a formidable challenge for aquaculture researchers, who are working to identify effective and sustainable solutions to inhibit pathogens and mitigate their negative effects on shrimp health and productivity.

Application Protocol for VIUSID DETOX Vet & ASBRIP Vet in Aquaculture Aquaculture faces significant challenges from a range of pathogens, including Enterocytozoon hepatopenaei (EHP). VIUSID DETOX Vet & ASBRIP Vet provides a natural, innovative solution to mitigate these threats, enhance immunity, and improve growth and productivity.

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DISEASE PREVENTION AND MANAGEMENT VIUSID DETOX Vet combines probiotics and bioactive molecules to combat pathogens, improve gut health, and enhance resilience against environmental and infectious challenges. It promotes better growth, feed conversion, and survival rates while improving water quality. ASBRIP Vet utilizes essential oils like thyme and eucalyptus to improve gut health, digestion, and feed efficiency. It strengthens the shrimp's immune response and protects against pathogens like EHP and Vibrio. When used together, these products provide a comprehensive approach to combating EHP, Vibrio, and viruses, ensuring sustainable and productive aquaculture systems.

7 days-challenge, the contaminated feed was prepared and renewed daily. Negative control tanks were fed by TSB+ top-coated feed.

After a feeding period with treatment diets for 14 days, shrimp entered into a seven-day EHP challenge period using per os approach. During the 21-day post-challenge, shrimp survival, growth and feed consumption rates were measured, as well as an immuno-analysis performed across different sampling points: day 7 and 21 of the post-challenge period. Total Hemocyte Count (THC), Differential Hemocyte Count (DHC) and Phenoloxidase Activity (PO) of shrimp were key interests in this study.

Immune response of shrimp to EHP Infection A fresh in vivo study conducted by Catalysis, in partnership with ShrimpVet Laboratory in Vietnam, aimed to evaluate the efficacy of VIUSID DETOX Vet & ASBRIP Vet in the improved immune function of P. vannamei juveniles under induced EHP infection.

The study was set up in 350-liter aquariums, utilizing a population of 40 shrimp per aquarium. Shrimp were approximately 1.78 grams and were fed with diets topdressed with VIUSID DETOX Vet & ASBRIP Vet at 5 mL, 10 mL and 15 mL per kilogram of feed. Control groups (challenge and unchallenged conditions) were set up as a comparison to the testing groups as the groups are shown in Figure 1. Only the negative control groups were absent of challenge.

Feces from EHP-infected shrimp were mixed with 5% seawater and 0.3% oxytetracycline to form the original EHP suspension stock. This stock was diluted 100 times and applied to feed pellets as a topcoat. During the

Data analysis Data for survivability, water quality, and analysis results were analyzed using SPSS version 25.

For immunology analysis, SPSS version 27 was used.

Statistical significance was determined by P-values < 0.05. ANOVA and Duncan's test were used for multiple comparisons. Independent samples and T-test were used for differences among sampling time points.

Percentage data was arcsine-transformed before analyzing statistics.

Results: Survival and growth At the end of the trial, the survival rates in the application of VIUSID DETOX Vet & ASBRIP Vet at higher doses (10 ml and 15 ml per kg) were significantly higher than the positive control and the lowest applied dose (P < 0.05). As reported in Figure 1, the shrimp survival in those two groups was improved by 18% and 11%, respectively.

Figure 1. Survival rate on day 21 of EHP post-challenge. Values are presented as mean ± standard deviation. Statistically significant differences are expressed in different small alphabets (a, b, c). (N = 4, P < 0.05).

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G1 Negative control

G2 Positve control

G3 5 mL/kg

G4 10 mL/kg

G5 15 mL/kg

Initial mean weight (g)

1.78 ± 0.00a

1.78 ± 0.00a

1.78 ± 0.00a

1.78 ± 0.00a

1.78 ± 0.00a

FMW (g)

10.44 ± 0.12a

6.91 ± 0.09d

7.09 ± 0.08c

7.36 ± 0.13b

7.39 ± 0.14b

MWG (g)

8.66 ± 0.12

5.13 ± 0.09

5.31 ± 0.08

b

5.58 ± 0.13

5.61 ± 0.14b

ADG (g/day)

0.206 ± 0.00a

0.122 ± 0.00c

0.126 ± 0.00bc

0.133 ± 0.00b

0.134 ± 0.00b

SGR (%/day)

4.21 ± 0.03

3.23 ± 0.03

3.29 ± 0.03

3.38 ± 0.04

3.39 ± 0.05b

Group

a

d

a

c

d

c

Final biomass (g/tank)

a

372.53 ± 4.48

235.30 ± 6.36

Total feed consumption (g)

368.60 ± 5.57a

FCR

a

1.22 ± 0.02

241.1 ± 14.06

b

257.50 ± 4.16

247.20 ± 11.02bc

295.44 ± 6.62d

298.80 ± 4.56cd

307.79 ± 5.11bc

315.43 ± 9.02b

c

1.80 ± 0.03

c

1.77 ± 0.13b

b

1.65 ± 0.04

1.80 ± 0.09c

24.02 ± 2.64

b

CV of shrimp size (%)

14.48 ± 0.75

25.59 ± 3.94

23.18 ± 2.31

23.30 ± 1.26b

c

a

b

c

b

b

Values are presented as mean ± standard deviation. Statistically significant differences are expressed in different small alphabets (a, b, c, d). (N = 4, P < 0.05).

In terms of final mean weight (FMW), mean weight gain (MWG) and specific growth rate (SGR), all treatment groups significantly showed improved results than the positive control (P < 0.05; Table 1). Regarding average daily growth (ADG), G4 and G5 had significantly higher rates than G2, however not from G3. Regarding the final biomass, G4 was followed by G5 and significantly different from other challenge groups of G2, G3 as it gained the highest survival of shrimp after EHP infection. Similarly, the FCR (feed conversion ratio) in G4 showed the lowest at 1.65, being significantly different from the positive control (P < 0.05), showing a notable improvement in FCR.

Results: Shrimp immunity THC and DHC results indicated that the treatment groups maintained stable hemocyte counts and effectively managed EHP infection, particularly in the 10 mL/kg Table 2. Immunological assays results

Parameter

Time point

Total hemocyte count (THC) (x106 cells/mL)

G1

G2

G3

G4

G5

1wpc 3wpc

aA

20.39 ± 11.16 35.54 ± 13.11bA

aA

9.85 ± 7.19 16.17 ± 7.13aA

aA

24.34 ± 13.8 17.30 ± 6.95aA

aA

30.35 ± 10.97 22.15 ± 3.52abA

21.39 ± 8.55aA 18.68 ± 6.3aA

Differential hemocyte countSemi Granular cell count DHC-SGC (%)

1wpc 3wpc

18.08 ± 3.84bA 15.33 ± 2.16dA

13.33 ± 2.46aB 8.33 ± 1.39aA

17.67 ± 1.78bB 13.75 ± 1.6cdA

13.33 ± 1.05aA 12.33 ± 0.72bcA

12.42 ± 0.79aA 10.92 ± 1.1bA

Differential hemocyte countGranular cell count DHC-GC (%)

1wpc 3wpc

5.50 ± 2.78bA 5.75 ± 1.17cA

1.75 ± 0.57aB 0.75 ± 0.42aA

3.50 ± 1.6abA 5.33 ± 1.91cA

2.17 ± 0.64aA 2.17 ± 0.58bA

2.00 ± 0.86aA 2.17 ± 1.29bA

Differential hemocyte countHyaline cell count DHC-HC (%)

1wpc 3wpc

76.42 ± 6.24aA 78.92 ± 2.59aA

84.92 ± 3.00bA 90.92 ± 1.00cB

78.83 ± 3.10aA 80.92 ± 1.52aA

84.50 ± 1.17bA 85.50 ± 0.84bA

85.58 ± 1.48bA 86.92 ± 2.33bA

Phenoloxidase activity (PO) (U/min/mL)

1wpc 3wpc

68.55 ± 18.84aA 81.00 ± 17.88aA

48.00 ± 19.23aA 66.00 ± 11.71aA

83.70 ± 48.42aA 60.75 ± 14.58aA

66.75 ± 17.92aA 59.10 ± 53.18aA

69.60 ± 32.36aA 63.30 ± 33.79aA

Results are expressed as mean ± standard deviation. Identical superscripts indicate no significant difference (P > 0.05) between the groups and among the two sampling time points within each group (uppercase). 1wpc: 1-week post-challenge; 3wp: 3-week post-challenge.

Similar superscript letters indicate that there is no statistically significant difference (P>0.05) between the groups and among the two sampling time points within each group (uppercase).

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DISEASE PREVENTION AND MANAGEMENT group (Table 2). The PO activity data showed that all doses positively influenced the regulation and response of the shrimp’s humoral immune system. Phenoloxidase activity was initially elevated in all treatment groups compared to the positive control suggesting that the applied doses of the test product effectively activated the shrimp’s humoral immune system.

The decrease in PO activity within treatment groups over time indicates that the test product can effectively manage severe EHP infection.

These results showed that the application of VIUSID DETOX Vet & ASBRIP Vet had an influential effect on shrimp immunity within an EHP infection.

Conclusion The results of our study suggest that VIUSID DETOX Vet & ASBRIP Vet yielded the best overall outcomes, significantly enhancing shrimp survival, boosting growth performance, and positive effects on shrimp’s immune response. The 10 mL/kg dose was identified as the most effective treatment under the tested conditions.

Catalysis’s VIUSID DETOX Vet & ASBRIP Vet can be considered for practical application in shrimp farming as a means to improve survival and overall health when facing EHP challenges. Further research, including field trials and mechanistic studies, is warranted to fully evaluate its potential and optimize its application in commercial shrimp farming.

Acknowledgements We would like to express our appreciation to ShrimpVet, Vietnam, for conducting the trial.

References available on request.

More information:

Figure 2. (A) Total hemocyte count (THC), Differential hemocyte count (DHC) as (B) semi-granular cell, (C) granular cell and (D) hyaline cell and (E) Phenol Oxidase activity. Similar superscript letters indicate that there is no statistically significant difference (P value > 0.05) between the groups and among the two sampling time points within each group (uppercase).

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Bulent Kukurtcu Scientific Manager Catalysis E: bulent@catalysis.es

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EHP prevention in shrimp farming: Spore germination, a key phase in limiting EHP outbreaks and effects Sophie Reys, Pierre Fortin, Nicolas Tanrattana, Techna France Nutrition

Parasites impact on aquaculture Parasites represent a significant threat to the aquaculture industry today. Among the most concerning are sea lice (Lepeophtheirus salmonis), which infest the skin of salmon; Sparicotyle chrysophrii, which affects the gills of European seabream; or gregarines, which compromise the gut health of shrimp (Buchmann, 2022). These parasites individually cause economic losses estimated at hundreds of millions of dollars annually, with sea lice alone accounting for losses of over half a billion USD. The economic impacts arise not only from direct factors, such as animal mortality and treatment costs, but also from indirect consequences.

These include reduced market value due to smaller size or poor appearance, slower growth rates, increased feed conversion ratios (FCR), and elevated fixed costs.

Overall, the aquaculture industry is estimated to lose several billion Euros or USD annually due to parasitic infections (Costello, 2009; Rigos et al., 2023).

Among these parasites, the microsporidian Enterocytozoon hepatopenaei (EHP) has emerged as a particularly severe threat to shrimp aquaculture. This parasite predominantly impacts Asia’s major shrimpfarming regions. EHP infection does not cause any specific clinical signs, but infected shrimp typically exhibit reduced growth rates, increased FCR, and increased mortality. All of which directly impair the productivity and profitability of shrimp farms. The growth retardation associated with EHP is due to the damage it causes to hepatopancreatic cells, which impairs nutrient absorption and metabolic efficiency.

Consequently, infected shrimp require significantly more time to reach market size, leading to higher operational costs. Furthermore, EHP adversely affects FCR, a critical parameter for evaluating farming

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efficiency, thereby driving up production expenses (Aldama-Cano et al. 2024).

EHP life cycle and effects EHP specifically targets the hepatopancreas, a vital organ responsible for digestion and metabolism in shrimp. Its life cycle, typical of microsporidians, involves a highly resistant spore stage that facilitates environmental persistence and transmission. EHP spores are released into pond environment via the feces of infected shrimp, enabling waterborne transmission. Other transmission routes include cannibalism and contamination from external sources.

Once ingested by a shrimp, the spores germinate in the digestive tract and inject their contents into the epithelial cells of the hepatopancreas. Within these cells, the parasite proliferates, producing new spores that are eventually released back into the gut and environment, perpetuating the infection cycle (Fig. 1).

(Chaijarasphong et al., 2021).

Outbreaks of EHP are often associated with environmental and management factors in shrimp farming. Poor water quality, high stocking densities,

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Figure 1. From Thawatchai Chaijarasphong et al., Journal of Invertebrate Pathology, https://doi.org/10.1016/j.jip.2020.107458z

and inadequate biosecurity measures create ideal conditions for the spread of the parasite. Additionally, EHP frequently co-occurs with opportunistic pathogens such as Vibrio bacteria, which exacerbate the severity of the disease and complicate treatment efforts. Asymptomatic shrimp can also act as reservoirs, silently spreading the parasite to uninfected stocks. Moreover, EHP spores are highly resistant to environmental stresses, allowing them to persist in pond sediments for extended periods even after disinfection or drying measures. This resilience makes eradication extremely challenging (Chaijarasphong et al., 2021).

Active ingredient selection, knowledge and in vitro testing The persistent prevalence of EHP poses a significant challenge for shrimp farmers globally. Addressing this issue and developing innovative, practical solutions to safeguard shrimp health and ensure the economic sustainability of aquaculture operations remains a critical priority for the industry.

Leveraging its expertise in active ingredients and molecules, particularly those derived from plants, TECHNA has identified antiparasitic compounds.

Monoglycerides of fatty acids, including short- and medium-chain variants such as capric and caprylic acids, have demonstrated antiparasitic effects by disrupting

the lipid membranes of parasites and interfering with their metabolism. Additionally, essential oils are recognized for their repellent properties, targeting the hormonal and nervous systems of parasites to reduce their activity, such as feeding and reproduction. Other plant-based molecules, including phenolic compounds and terpenes, inhibit crucial enzymatic processes vital for parasite survival.

In vitro testing conducted at ShrimpVet Laboratory allowed for ranking three plant-based ingredients based on their efficacy in inhibiting EHP spore germination.

EHP spores were isolated from the hepatopancreas of infected shrimp and cultivated in Hank’s Balanced Salt Solution. Under these conditions, at room temperature (25-28°C), spores germinated. Visual examination under a microscope (x1000) determined germination by observing flagella development. The proportion of flagellated versus non-flagellated EHP spores was calculated for germination rates. The trial set-up included a control group (only Hank’s solution) and treatments with three different products were included at a concentration of 0.0625% in Hank’s solution + EHP Spores.

Results indicated that phenolic compounds were the most effective in controlling EHP spore germination, followed by Essential Oil Blend #2, while Essential Oil Blend #1 exhibited minimal impact on germination rates (Fig. 2).

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Figure 2. In vitro EHP spores germination with the inclusion of 0.0625% plant extract blends (ShrimpVet Laboratory, 2023)

In vivo challenge with EHP The best ingredients were incorporated together with others in a commercial product that aims at reducing the outbreaks of EHP and limiting its impacts.

This product, called EHPROTECT, has undergone testing to validate its efficacy.

An in vivo trial conducted at the ShrimpVet Laboratory demonstrated a positive dose-response effect of EHPROTECT (2 to 4 kg/MT) on EHP-infected

shrimp. Shrimp with an initial weight of 1.6 g were raised in 250 L tanks for 50 days, which included an EHP challenge from day 16 to day 23, followed by an observation phase. Each treatment was replicated across six tanks, each containing 50 shrimp. EHPROTECT was added to the diet prior to production.

Shrimp infected with EHP (positive control) exhibited reduced growth, increased mortality, and higher feed conversion ratios (FCR) compared to unchallenged

Figure 3. Shrimp mortality at the end of the EHP challenge trial

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Figure 4. Shrimp final weight and FCR at the end of the EHP challenge trial

shrimp (negative control). The inclusion of EHPROTECT at 2 kg/MT and 4 kg/MT significantly improved these parameters. Mortality was reduced by 27.1% and 52.9%, FCR improved by 41.8% and 56.0%, and final weights were 16.1% and 45.7% higher for shrimp treated with EHPROTECT at 2 kg/MT and 4 kg/MT, respectively (Fig. 3, 4).

Conclusion Enterocytozoon hepatopenaei remains a significant threat to shrimp farmers, particularly in Asia. Feed supplements, such as EHPROTECT, could contribute to limiting EHP impacts through various mechanisms.

The above results show that EHPROTECT could help in limiting EHP outbreaks and probably prevent them.

Robust biosecurity and prophylactic measures are other milestones to limit EHP impacts.

References Buchmann K. Control of parasitic diseases in aquaculture. Parasitology. 2022, Dec; 149(14):19851997. doi: 10.1017/S0031182022001093.

Costello, MJ. The global economic cost of sea lice to the salmonid farming industry. Journal of Fish Diseases, 2008, 32(1), 115-118. https://doi.org/10.1111/j.13652761.2008.01011.x Aldama-Cano DJ et al. The Microsporidian Enterocytozoon Hepatopenaei in Shrimp Culture, The shrimp book II, Chapter 22. 2024.

Rigos G et al. Antiparasitic approaches and strategies in European aquaculture, with emphasis

on Mediterranean marine finfish farming: Present scenarios and future visions. Reviews in Aquaculture, 2023, https://doi.org/10.1111/raq.12857 Chaijarasphong T, Munkongwongsiri N, Stentiford GD, Aldama-Cano DJ, Thansa K, Flegel TW, Sritunyalucksana K, Itsathitphaisarn O. The shrimp microsporidian Enterocytozoon hepatopenaei (EHP): Biology, pathology, diagnostics and control. Journal of Invertebrate Pathology, 2021 Volume 186, 107458, ISSN 0022-2011, https://doi.org/10.1016/j.jip.2020.107458 More information: Pierre Fortin Aquaculture Manager Techna E: pierre.fortin@groupe-techna.com

Nicolas Tanrattana Aquaculture Nutritionist Techna

Sophie Reys R&D Project Manager Techna

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The power of krill meal to improve skin health and reduce sea lice Kiranpreet Kaur, Aker QRILL Company

For Atlantic salmon farmers, the health and welfare of the fish remain a constant challenge. Key among those challenges is skin health, which directly impacts fish resilience against parasitic sea lice infestation.

Though traditional delousing methods can be effective and have been heavily relied upon in the past, these methods often lead to unintended consequences, such as excess stress or injury to the fish. Recent studies point to fish diet and nutrition as a possible solution, specifically through the inclusion of functional ingredients such as krill meal in the aquafeed – a more sensitive tool that requires no extra handling of fish.

It’s an approach that can serve to strengthen the skin’s mucosal barriers and reduce sea lice while boosting the overall welfare of the fish.

The cost and consequences of sea lice in salmon farming Skin is the first line of defense against environmental issues for most creatures, including farmed salmon.

When skin health is less than optimal, it exposes the fish to pathogens that can result in infections and diseases. The salmon become more susceptible to sea lice when skin conditions are poor, and their overall welfare deteriorates.

Sea lice infestations are considered one of the costliest challenges for salmon farmers. The parasites latch onto the salmon to feed on their skin and mucus, which compromises fish health. Chemical solutions take a significant environmental toll and can lead to resistance development in sea lice and can cause potential harm to non-target species. There are non-medicinal solutions that are less invasive than chemical solutions, such as thermal delousing, but these can still cause stress and negatively affect the welfare of salmon. In Norway alone, delousing costs exceed USD 576 million (NOK 6 billion) per year.

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This challenge also goes beyond cost, as it is also in conflict with industry-wide aquaculture ambitions to improve fish welfare by practicing more humane farming. Typically, farmers experience poor skin health that leads to less robust fish, reduced quality, and higher mortality rates.

Krill meal can be a part of a nutritional solution to skin health and sea lice challenge Functional diets have emerged as a promising tool in the fight against sea lice – using nutrition to achieve improved health. Traditional aquafeeds often lack the comprehensive nutritional profile needed to address complex challenges like skin health and parasite resistance. Krill meal, derived from Antarctic krill, is a standout ingredient due to its unique nutritional profile.

Within krill meal, there’s a bounty of phospholipids, omega-3 fatty acids, amino acids, trimethylamine N-oxide (TMAO), vitamins, minerals, and chitin – all of which support the integrity of salmon skin and mucus layers and help form a stronger barrier against parasites. Recently, TMAO has gained attention for its role in protecting fish from dehydration, strengthening their barriers, improving skin health, reducing ulcers, and ultimately enhancing their overall resilience. Unlike plant-based ingredients, which can require supplementation to balance amino acids, krill meal offers a natural, good balance of essential and non-essential amino acids to meet the needs of salmon. Its rich phospholipid composition enhances the bioavailability of omega-3s, especially EPA and DHA, ensuring that the fish get the maximum benefit from these important fatty acids. This is particularly important for strengthening the structural integrity of the skin and mucosal barriers.

In addition, chitin, a component of krill exoskeletons, positively influences gut microbiota, which helps

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Figure 1. Mean and standard deviation number of preadult sea lice per fish in the three diet groups. The 8% KM provided a significant reduction in mean number of sea lice numbers per fish.

improve immune responses and overall health. On top of that, krill meal’s astaxanthin content delivers antioxidant properties that reduce oxidative stress, which can be common in densely populated aquaculture environments. When served in combination with krill meal, the result is an effective and multi-faceted ingredient that improves skin resilience and supports overall health.

Putting krill meal to the test New research from Aker BioMarine compared two krill meal-enriched diets with a typical commercial salmon feed to measure the effects on skin health.

The study showed that 8% krill meal in the salmon diet can significantly enhance skin epithelial thickness and mucosal health – with a significantly lower sea lice number recorded. It’s an improvement that resulted in a 25% reduction in sea lice infestations when compared to a standard commercial diet.

However, not just any amount of krill meal will do the trick. Getting the right inclusion level in the feed is crucial for ensuring maximum health benefits and optimal growth performance. Studies suggest that an 8-10% inclusion rate is optimal for young salmon, especially during the seawater transfer phase. During the grower and pre-slaughter phases, when the fish are

Figure 2. The average dorsolateral skin epithelial thickness. The epithelial skin thickness (in microns) in stockfish before feeding with the test diets and in test groups 8 weeks after feeding with a control diet, a diet with 8% KM, or a diet with 12% KM.

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Figure 3. The changes in dorsolateral skin mucosa after 8 weeks of feeding on the test diets. mean mucus cell area in square microns (a), the mucous cell volumetric density in the epithelium (x 100 = %) (b), and the defence activity of the mucosa (c). The value in the boxplot is the mean + standard deviation values per group.

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larger, studies suggest an inclusion rate of around 12% to get the best health benefits.

A sustainable future for salmon farming The inclusion of krill meal as a functional feed ingredient for salmon not only addresses critical health challenges in aquaculture but also aligns with the industry’s sustainability goals by promoting more environmentally friendly feed practices. Farmers today are seeking ways to reduce their dependence on chemical treatments to combat sea lice, as well as to minimize handling stress during delousing, which can lead to reduced fish welfare, greater mortality, and hinder efficiency.

Krill meal offers a natural, sustainable, and effective alternative to these methods.

Aker Qrill Company is committed to sustainable krill harvesting practices, as evidenced through their operations which are certified by the Marine Stewardship Council (MSC). This certification is a testament to the krill fishery’s commitment to responsible harvesting and minimal environmental impact. Krill meal comprises one of the largest biomasses on Earth to this day, and a decade of research from the Commission for the Conversation of Antarctic Marine Living Resources (CCAMLR), in cooperation with Aker Qrill Company, shows that its population remains

stable, even exhibiting growth in some years. This has earned krill the status of a highly renewable resource with a relatively low carbon footprint compared to other feed ingredients, making it a more sustainable choice for aquafeed formulations.

Krill meal is a powerful nutritional strategy for the future of aquaculture As the aquaculture industry continues its hunt for solutions to its most urgent challenges, krill meal offers a compelling answer. Its ability to enhance skin health and reduce sea lice infestations shows its value as a part of a nutritional strategy to not only support fish welfare but also to improve overall productivity and ensure the long-term sustainability of salmon farming.

The power of krill is far more than a scientific discovery – it’s a step towards a brighter and more resilient future for aquaculture.

More information: Kiranpreet Kaur, PhD Director R&d Aker QRILL Company E: Tormod.sandsto@qrill.com

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Recent trials show that mycoprotein is beneficial to salmon and tilapia immune systems and the reduction of GHG emissions Heikki Keskitalo, Enifer

Dealing with salmon disease and welfare Infectious diseases are the cause of the second-largest loss in salmon farming. The single largest infectious disease loss is caused by Moritella viscosa bacteria, which causes a disease commonly called winter ulcer.

For almost 40 years, the industry has been aware of the problem of winter ulcers. However, due to their growing frequency, frequently attributed to the increase in mechanical sea lice treatments, they are a hot topic of debate in the salmon business. Winter

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ulcer issues are estimated to cause USD 750 million in damages annually.

It is possible to vaccinate against these winter ulcer bacteria, but the problem is that the bacteria have many different strains. Therefore, ulcer development can occur even in fish that have been vaccinated. Vaccines aren’t perfect and new solutions are needed.

Pekilo Aqua® mycoprotein contains immune systemand vaccine-boosting betaglucan, nucleotides, and spermidine. Therefore, it could act as a vaccination

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booster, besides being a source of highly digestible and sustainable protein. Application is easy since Pekilo protein can be easily mixed into a commercial diet.

There is already scientific evidence on this.

According to a recent peer-reviewed scientific study (Mensah et al., 2024), Pekilo mycoprotein helped to prime and regulate the immune function in cells from Atlantic salmon. Interestingly, the co-stimulation of salmon spleen leukocyte cells with Pekilo mycoprotein and inactivated M. viscosa cells induced both immune homeostasis and antimicrobial response.

High concentration of polyamine spermidine (~1,500 mg/kg) is probably one of the main reasons besides nucleotides why Pekilo Aqua® improves immune homeostasis and the effect of vaccines (Jeong et al., 2018). In the immune cells of elderly humans, spermidine has also been shown to boost antibody production.

Recently, Norway's Ministry of Agriculture and Food published a 20-year update to their animal welfare report and presented it to the country's parliament.

Norway aims to have "the top animal welfare in the world" with a 5% target for farmed salmon mortality.

The industry must figure out a way to drastically cut its numbers from the 16% recorded in 2023 to reach

that goal. One major part of the solution might be the regular use of immune- and vaccination-boosting compounds like Pekilo Aqua®.

An effective tool for tilapia Streptococcus agalactiae (Group B Streptococcus, GBS) has emerged as a significant pathogen affecting mainly warm water aquatic species like tilapia. Outbreaks of this causative agent of this zoonosis are causing alarm worldwide. Infection rates have already reached 50%, and mortality rates have surpassed in some cases 95% (Ye et al., 2011). S. agalactiae has caused losses for Chinese growers ranging from 30-80%. In China alone, that amounts to almost USD 1 billion in lost revenue, assuming an average loss of 40% annually.

A trial performed in Brazil showed that the oral administration of Pekilo Aqua® at a concentration of 2.5% in tilapia diet resulted in specific IgM anti-S. agalactiae titration values that were higher than those of the control group (unpublished data). Higher levels of specific antibodies might lead to increased survival rates of the animals, demonstrating the product's efficacy against potential outbreak occurrences. Therefore, the addition of Pekilo Aqua® to the diet of tilapia is justified, even though the price of Pekilo Aqua® is higher than the price of bulk proteins.

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DISEASE PREVENTION AND MANAGEMENT Reducing GHG emissions in fish farming Margareth Øverland's research group showed that the inclusion of Pekilo mycoprotein in salmon feed exhibited favorable outcomes concerning environmental impacts, food-feed competition, and solid phosphorus and dissolved nitrogen waste outputs (Hooft et al., 2025).

Pekilo protein exhibited efficient environmental performance associated with GHG emissions reduction compared to other feed ingredients, specifically soy protein concentrate.

Conclusion Increasing the usage of innovative ingredients like Pekilo protein is imperative to meet increasing seafood demands while addressing critical issues linked to global warming, limited natural resources, food safety, and self-sufficiency.

References Mensah DD, Morales-Lange B, Øverland M, Baruah K, Mydland LT. Differential expression of immune-related biomarkers in primary cultures from Atlantic salmon (Salmo salar) exposed to processed Paecilomyces variotii with or without inactivated Moritella viscosa.

Fish & Shellfish Immunology, Volume 148, 2024, 109506, ISSN 1050-4648, https://doi.org/10.1016/j. fsi.2024.109506 Jeong JW, Cha HJ, Han MH, Hwang SJ, Lee DS, Yoo JS, Choi IW, Kim S, Kim HS, Kim GY, Hong SH, Park C, Lee HJ, Choi YH. Spermidine Protects against Oxidative

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Stress in Inflammation Models Using Macrophages and Zebrafish. Biomol Ther (Seoul). 2018 Mar 1;26(2):146156. doi: 10.4062/biomolther.2016.272. PMID: 28365977; PMCID: PMC5839493.

Ye X, Li J, Lu M et al. Identification and molecular typing of Streptococcus agalactiae isolated from pond-cultured tilapia in China. Fish Sci 77, 623–632 (2011). https://doi. org/10.1007/s12562-011-0365-4 Digestibility of Paecilomyces variotii for juvenile Nile Tilapia and zootechnical performance of juvenile tilapia fed diets containing Fungal Single-Cell Protein Paecilomyces variotii. Unpublished study performed at the Bioassay Laboratory of the Fishery Institute in São José do Rio Preto, SP, Brazil.

Hooft JM, Tran HQ, Montero R, Morales-Lange B, Stejskal V, Mydland LT, Øverland M. Environmental impacts of the filamentous fungi Paecilomyces variotii (PEKILO®) as a novel protein source in feeds for Atlantic salmon (Salmo salar). Aquaculture, Volume 596, Part 1, 2025, 741779, ISSN 0044-8486, https://doi. org/10.1016/j.aquaculture.2024.741779

More information: Heikki Keskitalo Business development manager Enifer E: Heikki.Keskitalo@enifer.com

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Diformates in seabass aquaculture: A proven concept Christian Lückstädt, ADDCON

According to FAO, aquaculture diversification is one of the most important strategies to strengthen and consolidate expansion in this fast-growing, global industry. Besides the regular candidates for top-aquaculture species – Chinese carps, shrimp, tilapia or salmon for example, certain seabass species have emerged as interesting contenders in this quest.

Production of both the European (Dicentrachus labrax) and Asian seabass (Lates calcarifer) has increased in recent years worldwide, reaching almost 500,000 tonnes of total annual production in 2023.

However, with such rapid growth, especially for the Asian seabass, special attention is now being given to the application of natural, sustainable and safe antimicrobial feed additives to reduce the impact of stress and/or disease challenges. This is especially important since bacteriological vibriosis is the most frequent disease affecting the Asian seabass, with losses accounting for more than 7% of the total production cost. Therefore, there is an understandable willingness from the industry to test new concepts.

Growth and health status in farmed fish may be improved through the application of feed additives to high-quality feeds. This management strategy is one of the main factors ensuring the future success of aquaculture. Among such additives, acidifiers have been increasingly used in many fish species over the last decade – they have for instance been successfully tested in salmon, rainbow trout, African catfish, European seabass, Asian seabass, milkfish, tilapia and pangasius as well as shrimp.

In animal nutrition, acidifiers exert their effects on performance via three different ways: (i) in the feed; (ii) in the gastrointestinal tract of the animal; and (iii) in the animal’s metabolism. A wide range of different organic acids and salts have been tested so far. Diformates in particular are used widely in both tropical and cold-water aquaculture, because

of their high load of active ingredients as well as their stability and handling properties in extruded feeds. Dietary potassium diformate (KDF, traded as Aquaform®), as well as sodium diformate (NDF, traded as Formi® NDF), have been tested in various aquaculture species from tropical and ambient regions since 2005 and several meta-analyses, as well as an independently published review article on their use in aquaculture, are available.

This article examines the impacts of both diformates in European and Asian seabass.

Trials in Asian seabass Two trials with the Asian seabass were carried out under university conditions using commercial feeds in Thailand.

The first trial (Lückstädt & Kühlmann, 2014) used 120 juvenile Asian seabass with a mean weight of 16.50 ± 3.97 g and stocked into twelve 240 L freshwater aquaria in a completely randomized design. Fish were divided into two groups (negative control and KDF) and reared for 75 days at 26-30°C water temperature.

The crude protein content of the Australian-formulated diet was 54%. The fish in both the control and KDF treatments were given the appropriate feed with a daily ration equivalent to 3-5% of their body weight. Feed was dispensed three times a day. Water parameters and growth performance of fish were monitored regularly.

Data was analyzed using the t-test and a confidence level of 95% was defined for these analyses.

The diet supplemented with 0.5% KDF showed improved growth data (P<0.05; Fig. 1). Seabass in the control group reached a mean final body weight of 54.5 ± 7.5 g, while fish fed with potassium diformate reached an average weight of 62.7 ± 2.8 g (Table 1). Additionally, feed conversion ratio and protein efficiency ratio tended (P=0.06) to improve. Finally, the survival rate as well as the fish productivity index, which is calculated based on weight gain, survival rate

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DISEASE PREVENTION AND MANAGEMENT and feed conversion - thus combining the three most important parameters in any fish production, improved significantly (P<0.05).

A subsequent trial (Arreza et al., 2014), using the same methodology, but with a lower diformate dosage (0.3% KDF) showed similar results (Table 2).

With an increase of the fish productivity index by 60% and 100% for 3 kg/t and 5 kg/t potassium diformate inclusion, respectively, both trials proved that the dietary acidifier can enhance growth performance and thus achieve economic and sustainable fish production.

A more in-depth view of the production of juvenile Asian seabass was provided

by Reyshari et al. (2019). These researchers investigated growth performance, gut microflora, digestive enzymes and innate immunological parameters of seabass juveniles.This time, dietary sodium diformate (NDF) was used as a gut health-promoting feed additive (Table 3).

Figure 1. Asian seabass juveniles after the trial

Table 1. Production parameters of Asian seabass (Lates calcarifer) after 75 days of rearing with or without dietary potassium diformate (KDF, Aquaform®) – modified after Lückstädt & Kühlmann, 2014

Parameter

CON

0.5% KDF

P-value

Initial weight (g)

15.8 ± 3.2

16.2 ± 4.2

0.430

Final weight (g)

a

54.5 ± 7.5

b

62.7 ± 2.8

0.026

Weight gain (g)

38.8 ± 6.6a

46.5 ± 3.8b

0.028

Survival (%)

82.0 ± 13.3a

98.3 ± 3.7b

0.014

FCR

2.72 ± 0.94

1.95 ± 0.30

0.061

FPI* PER**

0.75 ± 0.22

b

234 ± 56

0.026

0.96 ± 0.14

0.055

a,b: Means with a different superscript differ significantly at P<0.05 *FPI = Fish Productivity Index: Weight gain (g) × Survival (%) / (FCR × 10) **PER = Protein Efficiency Ratio: Weight gain (g) / Protein intake (g) Table 2. Production parameters of Asian seabass fed with or without dietary KDF – modified from Arreza et al., 2014

Parameter

CON

0.3% KDF

% Difference

Final weight (g)

54.5 ± 7.5

61.8 ± 6.2

+13.4

Weight gain (g)

38.8 ± 6.6

44.9 ± 6.1

+15.7

Survival (%)

82.0 ± 13.3

88.3 ± 6.9

+7.7

FCR

2.72 ± 0.94

2.12 ± 0.35

-22.l

FPI

36

+59.8

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DISEASE PREVENTION AND MANAGEMENT Table 3. Production parameters of Asian seabass fed over a period of 6 weeks with or without dietary NDF – modified from Reyshari et al., 2019

Parameter

CON

0.5% NDF

% Difference

Initial weight (g)

12.9 ± 0.2

13.5 ± 0.6

+4.7

Final weight (g)

45.9 ± 0.8

67.3 ± 2.7

+46.6

Weight gain (%)

255.8 ± 8.7a

398.5 ± 33.6b

+55.8

FCR

1.2 ± 0.1

1.0 ± 0.1

-16.7

FPI

275

538

+95.6

a

b

Table 4. Growth and feed utilization indices of European seabass fed with sodium diformate (Formi® NDF)-supplemented diets for 13 weeks – modified from Wassef et al., 2017

Parameter

CON

0.3% NDF

0.4% NDF

Final weight (g)

a

22.8 ± 0.2

b

25.2 ± 0.3

24.2 ± 1.5b

Weight gain (g)

17.9 ± 0.3a

20.5 ± 0.3b

19.4 ± 1.5b

FCR

1.59 ± 0.00

1.53 ± 0.11

1.43 ± 0.08

95.0

98.3

96.7

Survival (%) PER

a

1.00 ± 0.0

2.00 ± 0.0

b

2.00 ± 0.0b

PPV*

29.9 ± 0.6a

33.2 ± 2.0b

34.9 ± 1.7b

FPI

107

132

131

*PPV = Protein Productive Value: 100 × (Protein gain (g) / Protein fed (g))

Results proved, as with KDF, that dietary NDF is able to improve the performance of Asian seabass significantly. Furthermore, the gut total viable bacterial count decreased, while the lactobacilli count increased – which is seen as a move towards eubiosis.

The specific activity of chymotrypsin improved in fish-fed NDF, leading to a significantly higher protein efficiency ratio. Finally, seabass fed with sodium diformate showed a higher serum lysozyme level, indicating an improved immune status. Based on a break-point regression analysis, the optimal dosage of NDF for Asian seabass was estimated to be between 4.6 and 5.1 kg/t; this is in the same range as with KDF.

Trials in European seabass To see whether the diformate-additive can also be applied to European seabass, a trial was carried out in conjunction with an Egyptian university (Wassef et al., 2017). Juvenile (IW: 4.8 ± 0.0 g) Dicentrachus labrax were fed with either a control or NDF-containing diet over the period of 13 weeks. Results showed a prominent growth-promoting effect of 0.3% NDF in the seabass diet, which resulted in the highest weight gain, specific growth rate and feed utilization (Table 4). Fish from the same treatment showed also a significant elevation in health and immune indices.

Conclusions It can be summarized, also for the European seabass, that the inclusion of diformate, leads to improved performance – with significantly improved protein usage. This is particularly noteworthy, as carnivorous fish species are often viewed critically for this parameter and public opinion has shifted towards demanding information about the “fish in – fish out” factor. The overall performance based on the fish productivity index, for both dosages, improved by around 23%.

The addition of either potassium diformate or sodium diformate in the diets of both seabass species improved growth and other health and digestibility parameters significantly. These findings are in line with several other studies testing the effect of diformates in various tropical and temperate fish/shrimp species and are, therefore, an interesting option for sustainable fish production.

As such it can finally be concluded – diformates in seabass production are indeed a proven concept.

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

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Fish protein hydrolysate from pelagics: A sustainable and efficient solution for aquafeeds Dominique Corlay, Aquaculture Natural Solutions

There is no doubt the aquafeed industry needs to do “more and better” in terms of sustainable use of marine ingredients. Besides price volatility, the sector is facing several major challenges when sourcing marine ingredients: extreme variability in the quality of fishmeal according to origin, species, season, process, etc.; safety concerns with high levels of contaminants, such as heavy metals and microplastics; and sustainability issues with some poorly managed fisheries.

In this context, the limited supply of wild fish stocks should drive the optimization in their process preserving the unrivaled nutritional values of marine ingredients compared to other alternatives. Thus, Fish Protein Hydrolysates (FPH) from pelagic species feature many key advantages that we will review through the case study of the BLUE FISH FACTORY project, the first and new largest FPH plant in Morocco.

Sustainability and traceability The fishing industry is considered a key activity for the Moroccan economy (2-3% of the GDP). Pelagic species

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represent around 85% of the 1.35 million tonnes total landings (ONP, 2023), with around 65% being sardines (Sardina pilchardus). The pelagic stocks are considered in good condition and as “not fully exploited” (FAO/CECAF/ MarinTrust) while Moroccan authorities implement regular controls and fish stock assessments. Since 2023, biological rests have been put in place in the major South Atlantic ports to preserve the resource. At the BLUE FISH FACTORY plant, only byproducts of pelagics from their own cannery are used, such as discarded batches for technical reasons (visual appearance, stacked fish, freshness grade, etc.) or trimmings, including heads and viscera. Finally, all raw materials are fully traced up to the fishing boats and controlled by a third party, with under-progress certifications including GMP+, MarinTrust, and ISO 22000.

Safety and conformity to regulations In the European Union, there are strict regulations for the maximum levels (MLs) of contaminants such as heavy metals, authorized in fishmeal and fish oil.

Regulators view these levels in terms of safety for the farmed animal, and ultimately, food for the consumer. However, the relevance of the MLs and the declining inclusion of fishmeal in modern aquafeeds need urgent review. Parameters such as arsenic or mercury are unnecessarily restrictive, affecting the opportunity to use valuable marine byproducts in total contradiction with the EU’s Circular Economy and Blue Growth initiatives.

Fortunately, pelagics like sardines are placed in the mid-low Trophic Level (TL around 3) consuming prey such as copepods (Mounir, 2022). Hence, sardines have a naturally low level of contaminants as described in Table 1.

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PROTEINS Table 1. Heavy metal values in Blue Fish Factory Fish Protein Hydrolysate

Heavy metals

FPH from sardine, Blue Fish Factory (as is - 5% moisture)

Maximum levels based on 12% moisture (EU regulation on fishmeal)

Cadmium

<0.9 ppm

2 ppm

Mercury

<0.02 ppm

0.5 ppm

Lead

<0.04 ppm

10 ppm

Arsenic

<7 ppm

25 ppm

High nutritional values of fish protein hydrolysate The enzymatic hydrolysis method ensures and preserves the full value of marine proteins, being more gentle in terms of temperature compared to the standard fishmeal process, keeping a more stable amino acid profile. Most FPH feature high crude protein levels from 75-85% (as is) in comparison to fishmeal (55 to 70%). Figure 1 describes the content of essential amino acids of FPH from sardines compared to vegetable proteins such as soy. It is also

worthwhile to mention the high content of non-essential amino acids such as alanine, glycine, and proline, which will participate in bioactive peptides, or the presence of taurine, reconsidered as essential for some species with key metabolic functions (Salze, 2015).

The freshness of the byproducts should be effective and a reality for pelagics dedicated to human consumption: daily landings to the canneries with no long storage on board including cold storage conditions to prevent

Figure 1. Comparison of essential amino acid profiles of fishmeal (anchovy), fish protein hydrolysates (sardine), soy protein concentrate and soybean meal (dehulled, standard, 48%)

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PROTEINS Table 2. Biogenic amines and total volatile nitrogen values in fish protein hydrolysate

Parameter

Indicative value

TVN (Total Volatile Nitrogen)

<400 mg N/100 g

Histamine (from histidine)

<100 mg/kg

Cadaverine (from lysine)

<300 mg/kg

Putrescine (from arginine)

<200 mg/kg

spoilage. This freshness is expressed in Table 2.

Biogenic amines are heat stable while Total Volatile Nitrogen (TVN) by definition considered volatile compounds. TVN is considered mostly under two forms: TMA (Trimethylamine) and ammonia. TMA comes from the microbiota and bacterial growth while ammonia comes from the amino acid decomposition.

Therefore, TVN could be lower in fishmeal dried at high temperatures. However, in hydrolysates, TVN is mostly under the trimethylamine form and might be higher than in fishmeal. Recent studies highlight the key role of metabolites such as TMAO in osmoregulation and stress (Mengge, 2023).

High protein solubility and molecular weight distribution There is increasing evidence that some components present in the water-soluble fraction of marine

protein sources are important for fish performance, especially in high plant protein/low fishmeal diets (Kousoulaki, 2009). The FPH method allows us to keep all the soluble proteins within the product. Table 3 details the water protein solubility of various marine ingredients including live food. During the conventional fishmeal process, the stick water rich in soluble proteins might be insufficiently collected, concentrated and injected back into the fish cake. Furthermore, harsh drying conditions induce protein denaturation and Maillard reaction. The high content of watersoluble protein fractions will clearly enhance the feed palatability in all situations, including the better acceptance of medicated feeds. The protein digestibility coefficient of FPH is considered to be above 95% for most species.

By definition, enzymatic protein hydrolysis consists of water addition and a selected protease to cleavage the peptide bonds of large proteins into low molecular weight peptides and free amino acids. Figure 2 compares the average Molecular Weight (MW) distribution of fishmeal to the BLUE FISH FACTORY FPH. While FPH is 90% below 5,000 Da (average 3,300 Da) with 25% as free amino acids and di or tri-peptides, the fishmeal MW distribution is 90% below 60,000 Da (average 30,000 Da) with no detected free amino acids.

Table 3. Values of protein solubility (%) in some marine ingredients

Ingredient

References

Artemia (nauplii)

54 to 67

Carvalho, 2003; Tonheim, 2007

Rotifers

54 to 61

Carvalho, 2003; Nordgreen, 2009

54

Tonheim, 2007; Nordgreen, 2009

Calanus copepod Fishmeal Fish protein hydrolysate (sardine)

40

Protein solubility

(% of total crude protein, average values)

11 to 28 > 90

Tonheim, 2007; Folador, 2006; Opstvedt, 2003 Blue Fish Factory, 2024

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Figure 2. Average molecular weight distribution in fishmeal and fish protein hydrolysate. Adapted from Chen, 2022; Blue Fish Factory, 2024.

Molecular weight distribution is related to the protein water solubility. It correlates also with higher protein digestibility, meaning faster and easier intestinal assimilation, well assessed for larvae and juveniles.

Functional properties of fish protein hydrolysates Bioactive peptides are defined “as the fragments of amino acid sequences in a protein that confer biological functions beyond their nutritional value”. These bioactive peptides are usually 2-20 amino acid residues in length with the frequent presence of arginine, lysine or proline. They have well-known properties in fish and shrimp, such as antimicrobial, antioxidant and immunomodulatory activities. FPH from sardines contains specific bioactive peptides that help sustain animal health against pathogens or reduce the antinutritional side effects of high-vegetable diets (Costa et al., 2020).

Conclusion In the challenging context of sustainability, availability and price, the aquaculture industry needs to improve the quality and processing methods of the unrivaled marine ingredients. Fish protein hydrolysates from pelagic species such as sardines feature some key technical advantages and offer the opportunity to do “better with less”.

References available on request.

More information: Dominique Corlay Aquaculture Natural Solutions E: ansaqua.dc@gmail.com Web: bluefishprotein.com

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Soybean peptides for a fish-free aquaculture Liang Chao PhD., Jiangsu Fuhai Biotech Co., Ltd

In the global aquaculture industry, stakeholders are actively seeking methods to reduce reliance on oceanderived materials. A groundbreaking trial in China has captured the interest of domestic and international aquaculture professionals alike. On January 3, 2025, Jiangsu Fuhai Biotech Co., Ltd (Fuhai Biotech) concluded an 80-day "fish-free" soybean peptide feed trial for largemouth bass in Zhongshan, Guangdong Province.

The trial culminated in a fish harvesting event attended by over 40 guests, including 10 renowned researchers from universities and institutes, notably Academician Professor Mai Kangsen of the Chinese Academy of Engineering. More than 30 technical managers from leading Chinese aquafeed companies also participated.

The event was live-streamed, attracting tens of thousands of viewers online.

Trial design The trial ran from June 25, 2024, to January 3, 2025.

Two ponds were used: the control pond (0.57 acres)

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and the trial pond (0.38 acres). Initial fish weights were approximately 4.35 grams, with densities of 31,930 fish per acre in the control group and 41,763 fish per acre in the trial group.

The control group was fed conventional feed containing 40% premium Peruvian fishmeal. For the trial group, conventional extruded feed was used for 111 days, followed by an 80-day fish-free feed trial incorporating 50% Fuhai products. Both ponds were managed under identical conditions.

On January 3, 2025, 60 fish were randomly selected from each pond for measurement.

Results Trial results are summarized in Table 1. Dr. Liang Chao provided a detailed analysis of the trial results, addressing the observed differences in liver condition, higher Hepatosomatic Index (HSI), and Feed Conversion Ratio (FCR) in the Fuhai group compared to the control group. FCR differences may be due to the different

Aquafeed: Advances in Processing & Formulation Vol 17 Issue 1 2025

PROTEINS Table 1. Final results

Final average weight (g)

Condition factor

Hepatosomatic index (HIS,%)

Viscerosomatic index (VSI,%)

FCR

Control group

454.5

3.03

4.02

10.31

1.06

Fuhai Group

456.3

2.96

4.49

10.62

1.24

Group

Sampling records

protein level, over 50% in the control feed and 45% in the trial feed. He identified areas for improvement in the trial group’s feed formulation, specifically regarding protein and fat content and composition, as well as micronutrient supplementation. The trial’s conditions – such as high summer temperatures, fish being fed to satiation, and constant water quality adjustments – placed significant metabolic pressure on the fish.

Additionally, the trial group’s much higher stocking densities contributed to these differences.

Global significance of the fish-free feed trial This trial carries global significance, showcasing a revolutionary feed formulation that replaces fishmeal with 50% soybean peptides. As Dr. Liang Chao remarked, a conventional fishmeal replacement of just a few percent would not have generated such widespread interest.

At the trial’s conclusion, researchers and industry representatives sampled steamed fish prepared without spices or sauces. They unanimously praised the fish for its delicious flavor, firm texture, and lack of any unpleasant aftertaste. Academician Mai and other experts shared valuable insights, encouraging Fuhai Biotech to expand its solutions to meet the needs of the global aquaculture industry.

Attendees highlighted two key takeaways: • Fishmeal replacement is feasible: Fuhai Biotech’s fish-free feed trials, including a large-scale trial on Seagull Island in Guangzhou in 2023, and the recent verification in Zhongshan, consistently demonstrated positive results. Fish fed with the alternative diet exhibited excellent feeding rates, robust health, and no signs of enteritis, signaling to the aquaculture community that fish-free feed is a viable option.

• High-proportion soybean peptides are safe and effective: The use of 50% soybean peptides in the trial diet, an approach akin to a high-stress test, resulted in healthy fish with normal growth and digestion. This reassures the industry that lower inclusion levels of soybean peptides are even more secure and practical for broader adoption.

Uniqueness of Fuhai products Fuhai products feature a unique peptide-coated lipid capsule structure that is highly digestible and protects lipids from oxidation. With lipid content reaching up to 20%, the need for external oil spraying can be reduced or eliminated, minimizing the risk of oil oxidation during storage, especially at farming sites when applied correctly. Furthermore, Fuhai soybean peptides offer excellent extrusion properties, allowing for reduced starch content in the feed. This, in turn, helps lower blood sugar levels in fish, promoting better overall health and reducing related issues.

With advanced technologies and high-quality products, Fuhai is dedicated to providing innovative solutions to promote the global aquaculture industry toward a sustainable and prosperous future.

More information: Liangchao Phd Director of Aquafeed Jiangsu Fuhai Biotech Co., Ltd. E: info@fatide.com

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Breaking free from the ocean's grip: A new era for aquatic feed Federico Melenchón Ramírez, Nana Wentzel Thorringer, Olivier Hartz, Unibio The aquaculture industry faces growing challenges due to its reliance on traditional feed ingredients like fishmeal, which are subject to supply volatility and environmental concerns.

Novel solutions, such as Single Cell Protein (SCP) produced through gas fermentation, are revolutionizing the sector. Uniprotein®, an SCP developed by Unibio offers a sustainable, reliable, and functional alternative, providing a consistent protein source with minimal environmental impact. Beyond basic nutrition, SCP enhances fish health and growth through its functional properties, representing a breakthrough for a resilient and eco-friendly aquaculture industry.

This innovation is paving the way for a sustainable future, meeting the rising global demand for protein.

Uniprotein® is provided both as pellets and powder

Imagine an aquaculture industry unshackled from the whims of the ocean. A future where feed production isn't held to dwindling fish stocks and unpredictable price swings. A world where sustainable, reliable, and even functional ingredients power the growth

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of healthy, thriving aquatic species. Could such a reality exist?

For decades, the aquaculture industry has been tethered to a single, volatile resource: fishmeal.

This reliance has created a precarious situation,

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PROTEINS leaving the industry vulnerable to supply shocks, price fluctuations, and the ever-present threat of overfishing. While alternative protein sources like vegetable proteins have offered some respite, their own set of challenges, including reliance on agriculture and susceptibility to market disruptions, have limited their long-term viability.

The growing pressures of climate change, resource scarcity, and fluctuating markets demand innovative solutions to secure a stable, efficient, and environmentally responsible ingredient supply for aquaculture.

Benefits of novel proteins Novel proteins such as insect meal, algae, fermented soybean meal, corn fermented protein, and even bacteria are beginning to take center stage.

Novel protein alternatives demonstrate promising nutritional profiles, enabling growth performance comparable to fishmeal. Moreover, these alternatives often exhibit the potential to enhance specific aspects of fish physiology, acting as functional ingredients.

This functional capacity is evident in their ability to influence factors such as antioxidant status, immune response, and overall health and welfare.

A significant advantage of many novel protein sources is their potential to function beyond merely providing essential nutrients. These “functional ingredients” can exert beneficial effects beyond basic nutrition, enhancing various aspects of fish physiology.

For instance, research has demonstrated that insect meals, microalgae, and methanotrophic bacteria can enhance antioxidant capacity and modulate the immune response in fish and shrimp.

Federico Melenchón Ramírez, Product Manager Functional Health Ingredients, explains, "Novel protein alternatives offer a chance to move beyond basic nutrition. They can compete with fishmeal in terms of protein content and growth results, and even offer additional benefits. Some novel ingredients can act as functional ingredients, influencing fish physiology beyond just providing basic nutrients. A great analysis

by Hatch Blue gives an overview of some of these alternatives. Using a scoring system based on volume potential, price/T, amount of crude protein and digestibility, methanotroph bacteria and insect meals were considered as the highest scoring and most promising among these alternatives.”

What if a revolutionary technology could provide something more?

These developments give place to allow the aquaculture industry to break free from the ocean and agriculturalcentric dependency. What if a revolutionary technology could provide something more, consistent, sustainable, and even functionally beneficial source of protein for aquaculture? SCP produced through gas fermentation.

This groundbreaking technology, pioneered by Unibio, offers a compelling alternative to traditional feed ingredients, promising a future where aquaculture can thrive sustainably and profitably. This gas-to-protein technology represents a paradigm shift in protein production. Instead of relying on finite agricultural resources or depleting marine ecosystems, Unibio harnesses the power of microorganisms to convert methane into high-quality protein.

This innovative approach offers several key advantages: Guaranteed supply: This technology enables on-demand production, ensuring a consistent and reliable supply of high-quality protein, independent of weather patterns, crop yields, or geopolitical instability.

Imagine a world where feed manufacturers can confidently plan their investment in new growth and their production schedules, knowing that a stable and reliable protein source is always available.

Sustainable production: The process minimizes environmental impact. Producing 1kg of Unibio’s Uniprotein® requires significantly less water and land compared to traditional soy protein sources, reducing the industry's ecological footprint. This aligns perfectly with the growing demand for sustainable and environmentally responsible aquaculture practices.

Consistent quality: Uniprotein® offers a standardized and consistent protein source, eliminating the quality variations often associated with plantbased alternatives. This consistency is crucial for optimizing feed formulations and achieving predictable and consistent growth performance in farmed aquatic species.

Beyond basic nutrition: Functional ingredients But Unibio's innovation extends beyond simply providing a sustainable protein source. Uniprotein® has the potential to go beyond basic nutrition, acting as a functional ingredient that enhances the health and wellbeing of farmed animals.

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PROTEINS

Preliminary research conducted by Unibio A/S and Matis has revealed an exciting potential for Uniprotein® as a functional ingredient. Studies have shown that incorporating Uniprotein® into the diets of Atlantic salmon may shorten the time required for the fish to develop an effective immune response after vaccination against Vibrio salmonicida.

This suggests that gas-fermented protein could act as a functional ingredient, "teasing" the fish's immune system in a controlled way, similar to a pathogen but without causing harm. This can translate to faster and more effective vaccination, leading to a stronger immune system, reduced susceptibility to disease, and lower antibiotic use.

Imagine a future where aquaculture practices not only minimize environmental impact but also enhance the health and well-being of farmed animals. A future where robust immune systems and reduced disease outbreaks contribute to a more sustainable and profitable industry. Gas fermentation has the potential to make this vision a reality.

A new era of collaboration and innovation Unibio is at the forefront of this transformative shift, collaborating with numerous companies and research institutions worldwide to unlock the full potential of

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Uniprotein®. Ongoing trials and positive client feedback paint a bright future for the world of alternative protein sources in aquaculture.

"At Unibio, we are committed to offering the best possible product and establishing ourselves as the leading provider of methanotroph-based protein solutions," adds Melenchón Ramírez. "We collaborate with numerous companies and research institutions worldwide to continuously improve Uniprotein® and explore its full potential. Ongoing trials and positive client feedback paint a bright future for the world of alternative protein sources in aquaculture."

The future of aquaculture: Sustainable, efficient and resilient The future of aquaculture lies in embracing innovation and embracing sustainable solutions. By harnessing the power of gas-to-protein technology, the industry can break free from the constraints of traditional feed ingredients, reduce its reliance on finite resources, and build a more resilient and sustainable future.

Moreover, the nutritional profile of gas to protein is remarkable and brings the protein content similar to fishmeal while having 5% more total amino acids and, additionally, 9% more essential amino acids compared to fishmeal. The nutritional profile of SCP

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PROTEINS cannot stand alone, however, the protein digestibility of Uniprotein® in aquatic species stands out with 81-88% dependent on species.

“In today's market, characterized by rapid technological advancements and the swift exchange of information, staying abreast of emerging trends has become increasingly challenging, often disrupting business predictability. For feed producers, raw material price volatility remains a critical concern, as demonstrated by fluctuations in the costs of fishmeal and soybean meal, among others,” says Nana Wentzel Thorringer, Technical Development Manager, Animal Nutrition at Unibio.

She continues: “In this context, the reliable availability of methane presents a significant advantage for Unibio A/S as producers of methanotrophic SCP. This resource enables Unibio to decouple production processes from the typical annual cycles of raw material pricing, thereby offering greater price stability and predictability.

Uniprotein® expands the basket of high protein ingredients for feed producers without compromising animal health, digestibility or growth delivering tomorrow's protein demand in a VUCA (Volatility, Uncertainty, Complexity, Ambiguity) world.”

Unibio’s Chief Commercial Officer Olivier Hartz summarizes that “Unibio is a leading player in gasto-protein fermentation and is poised to usher in a new era of sustainable protein production.

Our innovative technology produces high-quality SCP with several key advantages: • Predictable and consistent supply through long-term contracts.

• Year-round availability, unaffected by climate variability.

• Minimal environmental impact, requiring 2x less water and 25x less land per kg of protein compared to soybean meal.

• Scalable production to meet evolving market demands.

With Unibio’s Uniprotein®, we offer a sustainable solution to the growing need for high-quality protein sources worldwide.”

This is not just about replacing fishmeal; it's about unlocking the full potential of aquaculture – a sector that can provide a vital source of protein for a growing global population while minimizing its environmental impact. The time for change is now. The future of aquaculture is bright, and Unibio is leading the way.

Uniprotein®

References FAO (2022). The State of World Fisheries and Aquaculture 2022.

Hatch Blue (2024). Emerging Protein-Rich Ingredients for Aquaculture.

Global Metrics for Sustainable Feed. GFLI database.

More information: Federico Melenchón Ramírez Product Manager Functional Health Ingredients Unibio

Nana Wentzel Thorringer Technical Development Manager Unibio

Olivier Hartz Chief Commercial Officer Unibio E: commercial@unibiogroup.com

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INTERVIEW

Driving sustainability in animal nutrition: The role of the Global Feed LCA Institute Lucía Barreiro The Global Feed LCA Institute (GFLI) is transforming sustainability in animal nutrition by developing a comprehensive, globally accessible database for Life Cycle Assessment (LCA), enabling the feed and food industries to reduce environmental impacts and enhance transparency.

For food producers, embracing sustainability is not merely a trendy marketing strategy; it’s about making responsible decisions that significantly impact the environment. Feeds are one of the primary contributors to the carbon footprint in aquaculture production, making it essential to address sustainability in recent years.

A key resource in the quest for sustainability is the Life Cycle Assessment (LCA). However, as a relatively new tool, LCA methodologies have varied widely, leading to inconsistent and non-standardized data across the industry.

In this context, the Global Feed LCA Institute (GFLI) was established as an independent animal nutrition and food industry institute. Its mission is to develop a globally accessible, evolving animal nutrition Life Cycle Analysis (LCA) database that supports meaningful environmental assessments of animal nutrition products while driving continuous improvement in environmental performance across the animal nutrition and feed industry.

The organization The GFLI is a non-profit, member-based organization founded by multiple associations within the feed industry. It was formally established in 2019 as an independent entity.

“Before that, there was a ‘coalition of the willing’, consisting of companies and associations that envisioned collecting data to help the feed industry

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become more sustainable,” explained Arjen Voortman from the GFLI Secretariat in an interview with Aquafeed. com. European companies began gathering carbon footprint data around 2010, and the effort evolved organically until 2019 when GFLI was officially founded.

With its headquarters in the United States and a secretariat in Europe, GFLI operates globally.

Initially managed by a five-member board, the institute has since expanded to include a Technical Management Committee made up of LCA experts elected by GFLI members, and a Scientific Advisory Council, which ensures the accuracy of methodologies through external researchers.

Membership spans national and international associations like IFFO, IFIF, and the U.S. Soybean Export Council, as well as corporations like Nutreco, De Heus, ADM, Adisseo, and Zinpro, among others.

Currently, all members are connected to the feed industry, but GFLI envisions broader value chain participation in the future.

“We all agree that feed is a critical part of the supply chain in becoming more sustainable. We see a clear role for engaging downstream partners, such as retailers, as essential for the future,” Voortman noted.

While GFLI aims for comprehensive coverage of the entire animal industry, progress varies across regions.

“The main drivers are Europe and the U.S., with Asia a key target to expand our global representation.

However, corporate membership in Asia is currently

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INTERVIEW low, so we invite Asian stakeholders to join GFLI and engage with us and our members,” said Voortman.

Membership benefits include database access, discounts on data usage, the ability to nominate members to the Technical Management Committee, board participation, and the opportunity to act as ambassadors. “For companies aiming to enter European markets, where specific reporting guidelines are in place, data provision can be particularly advantageous,” added Voortman.

agencies, and companies aiming to reduce their environmental footprint.

Initially, GFLI envisioned its data being used for the company’s internal calculations, customer support, and academic research. As environmental calculations evolve, so do GFLI’s use cases. “For instance, we now work with associations that aim to distribute data among members, which represents a totally different use case,” Voortman explained.

Figure 1. GFLI member growth in total

Figure 2. GFLI datasets

The database The GFLI database provides comprehensive environmental impact data, including greenhouse gas emissions, using LCA metrics from a cradle-togate of the feed mill. It is comprised of LCA data from various sources, which are contributed by so-called “data providers”, ranging from research institutes, universities, industry consortia, and other entities interested in modeling or contributing data.

The GFLI database contains default datasets based on statistics, and sectoral datasets based on primary company data, and is the acknowledged secondary database for feed ingredients by and for the feed industry. As of September 2024, the database contained 1,830 datasets, with the majority from Europe (57.3%), followed by North America (15.1%) and Asia (14.1%).

The data is available in three formats, catering to different users such as nutritionists via feed formulation software, LCA practitioners, researchers, government

GFLI follows FAO-LEAP guidelines, and its methodology is compliant with the EU Product Environmental Footprint Category Rules (PEFCR) for feed ingredients. “Every update we implement ensures compliance with LEAP and PEFCR standards,” Voortman emphasized.

Branded data A unique GFLI initiative is the development of branded data, enabling feed ingredient producers to showcase the environmental impact of their specific products.

This approach provides transparency, credibility for customers, and the ability to compare products against sector averages.

As branded data is currently not widely available in, or accepted by, existing life cycle databases, a pilot phase was initiated by GFLI to test the robustness of the draft methodology about different types of feed ingredients and ultimately determine the feasibility of including this kind of data in GFLI.

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INTERVIEW Branded data, not widely available or accepted in existing databases, requires rigorous quality standards of level of primary data necessary and its quality rating as aligned with the EU PEFCR feed (which ranges from 1 to 5, whereas 1 is seen as best available data, and 5 being either no longer representative in time or technological representativeness). “To participate in the branded data project, data quality ratings must be below 1.6, compared to the 3.0 standard for sectoral studies. This ensures companies cannot misuse data for commercial claims,” Voortman explained.

By aggregating branded data, GFLI aims to continuously update sectoral averages, keeping the industry equipped with the most accurate data.

“The industry is shifting from requesting general information, such as the average environmental impact of soy, to specific, real-time data about the soy the company uses,” Voortman noted.

Figure 3. GFLI member growth per group

The aquafeed industry The aquafeed industry is highly engaged in sustainability efforts and actively supports GFLI’s initiatives. “Aquafeed companies are well-organized and collaborate across the value chain, which is rare in other feed sectors,” said Voortman. One of the founding members of the GFLI is the Norwegian Seafood Federation, which demonstrates the aquafeed industry’s commitment to sustainability.

Another active member is IFFO - The Marine Ingredients Organisation, which not only disseminates sustainability facts and information

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about LCA for their member on their website but also actively engages their global members to collect data and participate in GFLI’s “Data-in projects” to have accurate and representative data of their sector in the GFLI database.

Looking ahead GFLI’s goal is to harmonize global data collection and provide tools for the industry to achieve greater sustainability.

“As a non-profit, all funding is reinvested into expanding the database, improving technical possibilities, and fostering collaboration across the value chain. There is no profit motive – only the goal of making the industry more sustainable,” Voortman explained. “The industry itself sees value in the data but in the end, somebody needs to pay for an investment as well. We are trying to find a balance on how we can monetize that in a way that we know we can provide that additional value to the value chain.”

Building this database has not been an easy task.

Data providers face high costs for data collection, data modeling, and critical external reviews. GFLI seeks to lower this barrier to attract more contributors.

“We also have some limitations in terms of IT infrastructure that have prompted plans for upgrades including APIs for enhanced interactivity. As the number of members increases, so does the funding that will allow us to become more robust,” Voortman said.

Europe has embraced sustainability, especially with directives like the Corporate Sustainability Reporting Directive (CSRD), reinforcing the need for harmonized global standards. GFLI aims to set these standards, offering a global methodology that can be also used for benchmarking and regulators. “We’re proud to be an industry-led initiative, and that value is recognized by legislators. We are also eager to expand membership out of Europe, particularly in Asia and South America,” Voortman said.

Call to action “Our database and services offer immense possibilities, but we have limited resources and funding. We call on stakeholders to join as members, sponsors, or collaborators,” Voortman concluded. “By working together, we can achieve sustainability goals and create a brighter future for the industry.”

For more information, contact info@globalfeedlca.org.

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COLUMN

Shrimp nutrition & health: A holistic approach Albert G. Tacon, Ph.D. Aquahana LLC, agjtacon@aol.com In marked contrast to the salmon farming industry where animals are universally reared at high stocking densities within open-net cages and fed exclusively on nutritionally complete high-fat extruded feeds using automated feeding systems, the shrimp farming industry is characterized by its wide diversity of different farming systems, feeds, and feeding regimes used (Fig. 1).

Figure 1. Shrimp farming problems: Different culture systems, genetics and feeds used

Figure 2. Another major issue: Need to reduce stress and disease during the culture cycle

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COLUMN While this may be a blessing from a shrimp farmer’s perspective, because of the ability of shrimp (unlike salmon) to filter live food organisms from the water column, (depending upon the farming system employed), this creates a major issue for shrimp feed compounder when formulating feeds for specific farming systems and/or stocking densities. Moreover, as shrimp stocking densities have increased with the progressive intensification of farming systems, there is an important need to reduce shrimp stress and disease occurrence through the use of improved farm biosecurity and formulating feeds for optimum health and disease resistance (Fig. 2). Figure 3 shows the generalized relationship between different environmental and nutritional stressors on shrimp nutrition and health.

Figure 3. Nutrition and health interactions (modified after Tacon & Tran, 2022)

In particular, as global feed ingredient prices have risen, there has been increasing pressure on the shrimp feedmanufacturing sector toward the increased use of lower cost plant-based feed ingredient sources, including plant oilseed meals and legumes (25-50% by weight, including soybean meal, soybean protein concentrate, rapeseed meal, canola protein concentrate, canola meal, sunflower seed meal, cottonseed meal, peanut meal, pea protein meals, and lupin kernel meal) and cereals (15-35% by weight, including wheat flour, corn flour, rice polishings, wheat middlings, wheat gluten, wheat gluten meal, corn gluten meal, corn protein concentrates, distillers dried grains with solubles; Fig. 4).

Figure 4. Increasing use of plant proteins, meals and oils in shrimp feeds

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COLUMN Notwithstanding their lower-cost compared to animal-based feed ingredient sources, almost all plant-based feed ingredients contain endogenous anti-nutritional factors (ANFs) and mycotoxin contaminants, which unless removed through processing or inactivated can have a negative effect on feed intake, growth, gut health, nutrient digestion, immune response, and consequent disease susceptibility and health. ANFs and mycotoxins are secondary metabolites produced by plants and filamentous fungi which serve protective functions within plants against insects and animal predators for maximize plant survival (Fig, 5, 6).

Figure 5. Endogenous anti-nutritional factors within common plant feed ingredients

Figure 6. Mycotoxins, the hidden threat

The gastro-intestinal tract of shrimp is the first line of defense from the viewpoint of shrimp ingesting compound feeds, and so it follows that maintaining gut integrity and gut health is of paramount importance. Figure 7 shows the negative effect of different ANFs and mycotoxins on gut health.

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Figure 7. Negative effects of ANFs and mycotoxins in aquaculture species

The commercial aquafeed sector uses the following strategies to minimize or reduce the effect of mycotoxins: • Limiting use of lower cost/quality feed ingredients: cereal byproduct meals, use of expired ingredients, use of floor sweepings; • Use of organic acids and feed additives to prevent/limit mold growth in finished feeds: propionic acid, sorbic acid, herbs and spices, essential oils, phenolic antioxidants; • Use of mycotoxin binders to prevent the absorption of mycotoxins: clay minerals, bentonites, zeolites, activated charcoal, yeast and yeast cell wall derived products, algae extracts, chitin oligosaccharides, and synthetic polymers; • Use of microorganisms/dietary enzymes to bio-transform mycotoxins in GI-tract; • At the farm level: store feeds under cool well-ventilated conditions, use on a first-in first-out basis, not subjected to adulteration by top-dressing prior to use, ensure feed bags are not left outdoors (hours) prior to feeding, following good BMPs Figure 8 shows strategies to manage ANFs. Clearly, feeds need to be formulated for optimum growth performance and feed efficiency, but also for optimum gut health, microbiome development, and shrimp immune capability.

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COLUMN

Figure 8. Commercial methods used for ANFs removal and/or reduction

In conclusion, although many specific nutrients and feed additives have been reported with beneficial effects on shrimp health and well-being (Fig. 9), many of these feed additives have not been proven under practical farming conditions within market-sized animals, and their use efficacy economically clearly demonstrated.

Figure 9. Feed additives with reported health benefits – direct or indirect

For example, Figure 10 shows the important role played by butyric acid and tributyrates on gut health and integrity, and Figure 11 shows the beneficial effect of using tributyrates within independent shrimp feeding trials conducted under field conditions.

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RESEARCH

Figure 10. Reported benefits of butyric acid and tributyrates in animal gut health

Figure 11. Beneficial effects of tributyrates in hepatopancreas and intestine of Pacific white shrimp at 10 weeks (Loc Tran, personal communication)

The real cost of mycotoxins and ANFs to the feed compounder, the farmer, and the consumer are real and should be viewed at three levels: • The real cost to the aquafeed compounder, in terms of ingredient sourcing, quality control, additional processing requirements, the use of specific feed additives to mitigate the effects of ANFs and mycotoxins • The real cost to the farmer, in terms of reduced growth, feed efficiency, gut health, immune capability, disease resistance, survival, and farm profitability • The real cost to the consumer, in terms of the possibility of mycotoxin carry-over into edible tissues and possible danger to human health and wellbeing It is hoped that this paper will stimulate renewed research interest in ANFs and mycotoxins, and the mitigation strategies used for their removal and/or deactivation.

The current paper is based on the presentation made by the author in Natal (Brazil) at Fenacam from November 19-22, 2024, and is summarized here.

References Tacon, A.G.J. & L. Tran. (2022). Nutritional fish and shrimp pathology: A handbook. 5m Books Ltd (Publishers), Lings, Great Easton, Essex, UK: 162p. ISBN-13:9781789181319

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RESEARCH

Hacking the line: A systematic review of how probiotics disrupt bacterial communication through quorum-quenching Edeth Novia Tubal, Brett Laudato, VDS, N.V.

Communication has long been an evolutionary survival tool for plants and animals. These living organisms communicate constantly, either by gestures, sounds, actions, colors, smells, or pheromones. Meanwhile, bacteria can also communicate. A single cell can literally send messages to nearby cells and is also capable of listening back for feedback. These constant two-way communications influence the cell’s individual behavior to create an organizational behavior that is well aligned with a common goal. The bacterial common goals for survival are, to name a few, biofilm formation, virulence factor production, and bioluminescence (Miller & Bassler, 2001).

The bacterial dialogue Bacteria can still interact with other bacteria thanks to a mechanism called Quorum-Sensing (QS). This process enables bacterial cells to exchange information with one another, which can manage their virulence (Rutherford & Bassler, 2012). The key element in the QS system is the production of “autoinducer”, a chemical molecule that diffuses outside of a bacterial cell. This autoinducer is usually a unique molecule produced by the same family (Rutherford & Bassler, 2012). In simple terms, bacterial cell releases (endogenous) and detects (exogeneous) autoinducers to "sense" how many bacterial cells are present nearby for a "quorum" to happen on a common goal (Sarah, 2024).

In the case of a high same-family bacterial population, there will be a consequent high concentration of autoinducers in a certain region of space. This high concentration is detected by each of the bacterial cells

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and causes individual biochemical reaction cascades that result in the regulation of gene expressions (Xiao et al., 2022). As a consequence, the individual gene regulation may trigger the whole bacterial community to activate some of its detrimental phenotypes: toxin secretion, biofilm formation, and other virulence factors (Miller & Bassler, 2001) which causes severe tissue damage to its host.

On the other hand, if a bacterium detects no or low concentration of autoinducer, particularly during an unfavorable condition, some species can undergo sporulation (Sarah, 2024), modifications to their cellular membranes, some produce damage-repairing enzymes, or synthesize molecules to alleviate stress (Storz & Hengge, 2010).

Quorum-sensing: Gram-positive vs. Gram-negative Bacteria are classified as either Gram-positive or Gram-negative. Its cell walls are one of its primary distinguishing characteristics. Gram-negative bacteria (GNB) have thinner peptidoglycan cell walls than Gram-positive bacteria (GPB). The former has an outer membrane composed of lipopolysaccharide, while the latter lacks one (Xiao et al., 2022).

GPB and GNB have distinct quorum-sensing mechanisms: GNB uses N-acyl homoserine lactone (AHL) molecules (autoinducer-1, AI-1), while GPB uses autoinducing peptides (AIPs) (Taga & Bassler, 2003). Due to the small size and lipophilicity of AHL autoinducers, they readily pass the cell membrane using passive diffusion (Sifri, 2008). Therefore, GNBS have a more effective QS-system than GPBs.

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Figure 1. How quorum-sensing works in a high-dense bacterial population.

Figure 2. How quorum-sensing works in a low-dense bacterial population.

Figure 3. The different types of molecules responsible for quorumquenching exhibited by Gram-positive bacteria.

Figure 4. Examples of Quorum-Quenching Enzymes.

Source: Rémy et al., 2018

Figure 5. Examples of quorum-sensing inhibitors.

Source: Rémy et al., 2018

Aquaculture bacteria community Pathogenic bacteria, usually GNBs, are typically responsible for disease outbreaks while GPBs are only responsible to a lesser extent (Bacterial Fish Diseases, 2022). The most common examples of GNBs that continuously pester the aquaculture sector are

Aeromonas salmonicida, Pseudomonas anguilliseptica, Vibrio harveyi and V. anguillarum, Moritella viscosa, and Tenacibaculosis (Irshath et al., 2023).

On the other hand, a number of GPBs are commercially used in aquaculture (known commonly as probiotics) because of their benefits: i.e., improved feed efficiency,

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RESEARCH disease resistance, and the growth of culture animals, to name a few. Some of these species include Bacillus sp., Lactobacillus sp., Enterococcus sp., Carnobacterium sp., and the yeast Saccharomyces cerevisiae among others (Martínez Cruz et al., 2012).

Quorum-quenching: Hacking the communication network Recent studies have shown that GPBs have a QS-suppression ability (Prazdnova et al., 2022).

This mechanism is called Quorum-Quenching (QQ), or the disruption of QS signaling or intercellular communication (Prazdnova et al., 2022). GPBs are thought to have developed this QQ strategy as an antagonistic mechanism against competitors, usually GNBs (Moradi & Hadi, 2021). There are several processes in which QQ can occur: (i) small molecules called Quorum-Sensing Inhibitors (QSIs) can inhibit or interfere with the synthesis of autoinducers or AHL (Tang & Zhang, 2014); (ii) QQ antibodies such as Cyclodextrin scavenge autoinducers (Park et al., 2007) or; (iii) extracellular hydrolysis of the autoinducers using QQ enzymes (Fetzner, 2015). All these QQ processes prevent autoinducers from achieving the maximum threshold value for detection (Moradi & Hadi, 2021; Weiland-Bräuer, 2021).

Quorum-quenching probiotics: A systematic review To verify the effectiveness of quorum-quenching potential of probiotics against aquaculture pathogens, a systematic review was conducted. The main objective is the synthesis of diverse relevant research findings and consolidate the current results in various published scientific journals.

An extensive literature search was conducted using the databases PubMed and ScienceDirect. The search query included a combination of the following keywords: ((quorum sensing) OR (quorum sensing inhibition) OR (quorum quenching)) AND (aquaculture). In total, 15 studies qualified for analysis (Table 1). Survival rate from the pathogen challenge test was the common criterion among the studies, thus, it was used to calculate the outcome effect in fish and in crustaceans following the methodology outlined by Neyeloff et al. (2012).

Given that the survival data in all studies were reported in absolute survival rate, the survival data from each qualified study was normalized using Formula (1) to allow comparison among the crosssectional studies.

Survival%N = Survival% –Survival% x100 (Formula 1) Survival% P

C

P

Where Survival%P is the survival rate of animals when treated with probiotics, Survival%C is the survival rate of

Figure 6. Identification, screening, and selection of relevant scientific journal articles to be included in the systematic review.

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RESEARCH Table 1. Summary of the fish studies included in the systemic study and the results.

Challenge test Duration

Study Animal host Probiotic Pathogen (1hr)

Santos et al., 2021

Danio rerio

Bacillus spp. (FI436)

Edwardsiella tarda

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Survival improvement (%) 61%

Torabi Delshad Onchorrynchus Bacillus thuringiensis Yersinia ruckeri 336 100% et al., 2018 mykiss

Chu et al., 2011

Carassius auratus gibelio

Bacillus spp. (QSI-1)

Aeromonas hydrophila

336

Other (% reduction)

Biofilm suppression: 22% QS inhibition: 100% Bacterial motility suppression: 68%

90%

Aeromonas Chu et al., 2014 Danio rerio Bacillus spp. (QSI-1) 336 84%

hydrophila (YJ-1)

Biofilm suppression: 80% Hemolytic activity reduction: 78%

Ghanei-Motlagh Lates calcarifer Bacillus cereus QQ2 Vibrio alginolyticus 288 63%

et al., 2021

Biofilm suppression: 45% Bacterial motility suppression: 50% Bacterial swarming suppression: 71%

Vinoj et al., 2015 Danio rerio

Pseudomonas aeruginosa PsDAHP1

Vibrio parahaemolyticus

480

75%

Chen et al., 2020 Danio rerio

Bacillus licheniformis (T-1)

Aeromonas hydrophila

72

71%

Vadassery & Pillai, Enterococcus Carassius auratus 2020 faecium (QQ12)

Aeromonas hydrophila

336

85%

Vadassery Haridas Carassius auratus et al., 2022

Lactobacillus plantarum (QQ8)

Aeromonas hydrophila

336

87%

Vadassery Haridas Carassius auratus et al., 2022

Lactobacillus casei (QQ10)

Aeromonas hydrophila

336

85%

Vadassery Haridas Carassius auratus et al., 2022

Enterococcus faecium (QQ12)

Aeromonas hydrophila

336

88%

Vadassery Haridas Carassius auratus et al., 2022

Bacillus thuringiensis (QQ17)

Aeromonas hydrophila

336

87%

Outcome (random effect model, value ± SE)

83 ± 1%

Biofilm suppression: 77%

Table 2. Summary of the shrimp studies included in the systemic study and the results.

Challenge test Duration Study Animal host Probiotic Pathogen

(1hr)

Survival improvement (%)

Shaheer et al., 2021 Penaeus monodon Bacillus spp. (MFB7) Vibrio harveyi MFBP5 120 91%

Vibrio coralliilyticus VibC-Oc-193

72

37%

Stenotrophomonas Reina et al., 2019 Artemia salina

maltophilia (M9-54-2)

Vibrio coralliilyticus VibC-Oc-193

72

37%

Pande et al., 2015

Macrobrachium rosenbergii

Bacillus cereus LNE7 (AM397642.1)

Vibrio campbellii

144

37%

Widanarni et al., 2024

Litopenaeus vannamei

Bioflocs (no bacterial identification)

Vibrio parahaemolyticus

120

98%

Reina et al., 2021 Artemia salina

Psychrobacter sp.

(M9-54-1)

Outcome (random effect model, value ± SE)

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Other (% reduction) Biofilm suppression: 64% QS inhibition: 72% QS inhibition: 100%

Biofilm suppression: 32%

61 ± 12%

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Figure 7. One of the methods to indirectly detect the production of autoinducers in bacteria is to employ a second bacteria, also called "reporter" bacteria, which can detect these autoinducers and elicit an observable response, or in this photo, fluorescent light.

The V. anguillarium and Y. ruckeri strains produced autoinducers which caused this specific E.coli strain to glow.

animals without probiotics treatment, and Survival%N is the normalized survival rate (percentage improvement from the control).

The outcome of the systematic review showed that probiotics with quorum-quenching capabilities, when tested in vivo, improved the survival rate of pathogen-challenged animals by 83% for fish and by 61% for crustaceans. Some of the qualified studies, by in vitro confirmation, demonstrated that several virulence factors were suppressed. This virulence suppression may partially (or fully) explain the significant improvement in survival rate during pathogen infection.

Conclusion The quorum-quenching ability of some probiotics is a promising approach to directly address the devastating effects of aquaculture crop losses during pathogenic bacteria outbreaks. This systematic review revealed in vitro suppression of some virulence factors and in vivo significant improvement in the survival rate of the animals. Although more research is necessary to further elucidate the QQ mechanisms, it is quite clear that some probiotic strains can be used as an environmentally friendly substitute for antibiotics in the aquaculture sector thanks to their QQ-mechanism.

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It must also be noted that these results are all based on carefully-controlled conditions. In actual environments, the situation may be more complex due to the interactions between bacteria, their host organisms, as well as the surrounding environment which can further complicate the outcomes.

References available on request.

More information: Edeth Novia Tubal Master student, Student Intern VDS, N.V. e: viasogorg@gmail.com

Brett Laudato R&D Aquaculture VDS, N.V. E: Brett@vdspremix.be

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RESEARCH

Innovative sustainable recirculating aquaculture feeds: The InSuRAFeed project Dr Nicola Pontefract, Hannah Mance, Dr Jack James, Pontus Research Ltd.

Over 300 species of fish, crustaceans, mollusks, and other aquatic organisms are now produced by the aquaculture industry, and over half of the seafood consumed globally is produced through aquaculture (FAO, 2022), making it a vital sector in meeting the world's demand for fish. Aquafeeds account for approximately 50-70% of total industry production costs (Barreiro, 2023) and must be formulated to meet the specific nutritional requirements of each individual species (Glencross, 2025). At present, ingredients for aquafeeds are sourced from across the world, reflecting the global nature of the industry (OlivaTeles et al., 2025), but also impose large associated “feed miles”, greenhouse gas emissions and carbon footprints upon the finished aquafeed, subsequently diminishing sustainability.

InSuRAFeed project Considering the increasing global demand for sustainable and resource-efficient protein sources, the need to reduce the environmental footprint of the aquaculture industry, and the commitment of the UK government to support the growth of the aquaculture sector, Pontus Research launched the InSuRAFeed project in 2023 with generous financial support from the Welsh government and the Co-Op Carbon Innovation Fund. This project investigates the feasibility and potential of producing aquafeeds utilizing exclusively UK-sourced ingredients for two commercially valuable UK species, mirror carp (Cyprinus carpio) and rainbow trout (Oncorhynchus mykiss), and demonstrates their performance in vivo in comparison to a commercially available control diet.

The conditions determining an ingredient's suitability for inclusion within the project were simple:

• Non-soy, including derivatives or products with a soyderived product added during finishing • Locally sourced and processed (i.e. within the UK) • Sustainably sourced (i.e. where possible, not captured from wild fisheries, impacting ecosystem provisions, with the exception of fishmeal) • Attractive nutrient profile (i.e. favorable protein/lipid content) A wide range of ingredient suppliers was approached during the scouting phase for prospective aquafeed ingredients: from the well-established ingredient

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RESEARCH suppliers to a variety of UK startups, in addition to university-based research teams producing novel ingredients via alternative manufacture routes, local Welsh farmers, two independent UK mills, and government-funded research institutions.

A fundamental aim of this research was to design experimental aquafeeds that would be comparable to commercial controls, not only in terms of the health and growth of the fish but also in terms of price. Therefore, the wholesale cost of a metric tonne of each individual ingredient was provided by the respective suppliers, allowing for the determination of the cost of the experimental aquafeed during the formulation phase.

As expected, there was a wide variation in price for the different ingredients, ranging from the most expensive novel protein sources such as hemp and algae (£7,000-£8,000/tonne) to insect proteins and novel animal byproducts such as mussel and crab meals (£2,000-£4,000/tonne), to the alternative plant-based proteins such as faba beans, lupins, peas and rapeseed (£300-£550/tonne).

Trials in mirror carp and rainbow trout From the 22 suitable ingredients identified, and with reference to the current price of an appropriate commercial control diet, “expensive”, “moderate”, and “economical” experimental aquafeeds were formulated for each species: iso-nitrogenous, isocalorific, and iso-lipidic aquafeeds that meet the known nutritional requirements of mirror carp and rainbow trout respectively, aligning with the proximate

composition of an appropriate commercial control as closely as possible.

The experimental aquafeeds were manufactured

by Pontus Research’s in-house feed mill. Following confirmation of their suitability for administration, and further internal quality control checks, feeding trials in both species commenced during the summer of 2024.

The mirror carp trial ran for a total of 93 days.

As presented in Figure 1, the “expensive” and “moderate” experimental diets performed very well, demonstrating significantly improved Specific Growth Ratios (SGR) and Feed Conversion Ratios (FCR) in comparison to the fish administered the commercial control and “economic” experimental feed.

After 42 days of administration, rainbow trout fed the “expensive” and “economical” experimental aquafeeds demonstrated significantly improved SGR and FCR values in comparison to those fed the commercial

Figure 1. Mirror carps trial results

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Figure 2. Rainbow trout trial results

control diet (Fig. 2). Additionally, all three experimental trout aquafeeds exhibited optimal Apparent Digestibility Coefficients (ADC) for protein (91-94%), lipid (89-92%), and energy (80-86%), suggesting the diets are optimally balanced, promoting excellent fish performance, and are suitably sustainable without causing a nutritional deficit or nutrient waste.

Further trials Exhibiting a significantly greater weight gain (data not shown) in comparison to all other treatments and considering its strong performance in comparison to the commercial control, iterations of the “moderate” experimental mirror carp aquafeed will be investigated further. Similarly, following the encouraging results from the “expensive” and “economical” experimental aquafeeds for rainbow trout, these diets will also be further refined and tested in vivo in the spring of 2025.

This research remains ongoing. Additional analyses (not presented here) are still underway, including histological analysis of the gills, skin and distal intestine that will provide further insights into the mucosal health of the fish following administration; a range of hematological parameters; and assessment of epidermal mucus quality. Life Cycle Analysis of the different ingredients is also underway which will provide valuable information regarding the environmental impacts of the experimental diets.

A final and notable characteristic of the designed aquafeeds is that they are specifically intended for Recirculating Aquaculture Systems (RAS). Waste management is a crucial aspect of successful and

efficient RAS operation, as particulate waste from feed pellets and animals can contribute to system dysfunction, impacting water quality and animal health (Ramil et al., 2020). We are therefore additionally performing waste characterization evaluations in the form of particle size analysis, to determine the effect of the experimental aquafeeds on the efficiency and functionality of the system.

Historically, the UK has struggled with the longterm success of RAS facilities, often due to increased operating costs, suitable and cost-effective aquafeed sources, and facility issues. By offering locally sourced, RAS-specific, competitively priced aquafeeds and limiting the impact of poor feed choices on facility infrastructure, the InSuRAFeed project will support the development of a sustainable, profitable, and viable land-based aquaculture sector in the UK. Furthermore, this project will help the UK meet its commitments to increasing aquaculture production and promote the adoption of sustainable practices in the industry. It is our ambition to continue the InSuRAFeed project for other high-value species in the future.

References available on request.

More information: Nicola Pontefract Research Scientist Pontus Research Ltd. E: nmp@pontusresearch.com

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