Aquafeed vol 10 issue 3 2018

VOL 10 ISSUE 3 July 2018

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

Plant-derived compounds and natural products in aquaculture

The Maillard Reaction and amino acid availability Improving dryer productivity with intelligence Critical KPIs in factory performance Wood-based technology for aquafeed Hydroxy-Selenomethionine in fish broodstock, larvae and juveniles

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AQUAFEED

Volume 10, Issue 3

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

Contents •

Interview with Neil Auchterlonie

5

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News Review

9

•

The Maillard Reaction and amino acid availability in feeds

13

•

Improving dryer productivity with intelligence

17

•

Aquafeed Production: Critical KPIs for optimizing and sustaining factory performance

22

•

Using wood-based technology to bridge the aquafeed supply gap

27

•

The use of a functional diet to increase survival of white leg shrimp Litopenaeus vannamei (Boone, 1931) infected with acute hepatopancreatic necrosis disease (AHPND)

32

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Phytotherapy as an alternative to use of chemicals in aquaculture production and aquatic feeds

36

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Importance of plant-derived compounds and/or natural products in aquaculture

43

•

Dietary supplementation of Hydroxy-Selenomethionine enhances the reproductive performance and antioxidant status of fish broodstock, larvae and juveniles

48

•

Improvement of fish seed quality with plant based ingredients

52

•

Calendar of events

55

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

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AQUAFEED

Volume 10, Issue 3

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

Index to advertisers ADM

26

Adisseo

IFC

Aquaculture Without Frontiers

4

Aquafeed Workshop Mexico

8

Aquaculture 2019

47

Biomin

21

Extru-Tech

20

Famsun

BC

GFFC: 6th Global Feed and Food Congress

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Dr. Neil Auchterlonie is the Technical Director IFFO The Marine Ingredients Organisation. He joined IFFO in 2015, having worked previously in technical and production positions in aquaculture companies, consultancies, and government department and research agencies in the U.K. He has a strong track record in managing aquaculture and fisheries science programs in both public and private sector, specializing in commissioning, managing and reporting on science that addresses policy questions for both government and industry. He has held positions on a number of scientific and technical committees and has a strong interest in technology transfer within the seafood industry.

Neil holds a BSc in Marine and Freshwater Biology from Stirling University, a MSc in Applied Fish Biology from the University of Plymouth, and a PhD in Aquaculture (halibut physiology) from Stirling University.

Interview with Neil Auchterlonie AQUAFEED.COM Fishmeal and oil are frequently looked on as “unsustainable�, meaning in this instance, that they are a limited and diminishing protein and lipid source.

So perhaps we could start by understanding how much is there and is it going to run out?

NA IFFO collates annual data on fishmeal and fish oil, and has been doing so since in some shape or form since its inception in 1959. Over the last two decades the trend for production has been generally flat with approximately 5 million tonnes of fishmeal and just under 1 million

tonnes of fish oil. The steady nature of that production is testimony to the sustainable nature of the fishmeal industry, and the global businesses that produce these very important marine ingredients.

6 AQUAFEED.COM Is IFFO doing anything to actively encourage the use of fish waste at sea on fishing vessels and at the wholesalers for recycling into fishmeal?

NA IFFO recently funded a research project on the availability of fish byproduct for fishmeal and fish oil production. The investigation looked at both capture fisheries and aquaculture byproduct. That project was commissioned with the Institute of Aquaculture, University of Stirling, with Principal Investigator Dr. Richard Newton. Current production is based on utilization of approximately 20 million tonnes of raw material every year, of which 33% is byproduct. Dr. Newton estimated that according to FAO production data there is additional volume of byproduct available up to an additional 11 million tonnes (based on 2016 data).

IFFO has been raising awareness of this work in an effort to show that more production is possible, but there are some limitations around the capture, storage, and transport of this material. Interestingly, the growth in aquaculture carries some prospect for additional material as aquaculture processing tends to be centralized and collection of material may therefore be more straightforward.

AQUAFEED.COM Over the past months, IFFO has publicly rebutted moves to limit fishmeal and fish oil in aquafeeds. Would you give us an update on some of these? For example, you went head-on against the F3 (Fish Free Feed) Challenge and the way it was reported. Never-theless, the contest raised a good deal of

money and was successful and went on to launch the F3 Fish Oil Challenge. How do you view the contest now?

NA Our view on the alternatives is that they may be useful supplements to fishmeal and fish oil. Those materials are supplements only, and fishmeal and fish oil have always been and shall remain, the foundation of aquafeeds. Those alternatives should be responsibly-produced, nutritionally appropriate and safe. The aquaculture industry continues to grow, more feed is needed, and any significant additional volume of supply will not come from fishmeal and fish oil. The contest in general does not acknowledge the importance of fishmeal and fish oil, and distracts from those materials’ true contribution to global aquafeeds. There are often general misconceptions about the sustainability of the raw material supply to the fishmeal industry, which are included in the positioning of some of these alternatives in an effort to be seen as superior products in some way. IFFO will of course challenge those statements where we see them, and correct those inaccuracies where they occur. Interestingly, separate conversations with some of the producers of alternative ingredients indicate an acknowledgement of the position of these new products as supplements to fishmeal and fish oil rather than replacements, so it is likely that some of this narrative is perpetrated through the media rather than the companies themselves. The F3 Challenges are also part of that story.

AQUAFEED.COM Also, at the beginning of last year, you responded to the Aquaculture Stewardship Council (ASC)’s proposed amendment of the Salmon Standard Forage Fish Dependency Ratio (FFDR) to revise the level to 1.2 for fishmeal and 2.8 for fish oil, from the original levels of 1.35 for fishmeal and 2.95 for fish oil.

What was your argument and has the proposal been adopted?

NA This is an ongoing discussion, as it is with a number of stakeholders.

Our position on FFDR and the use of the similar term FIFO is that these are too crude as measurements to take into account the importance of the nutritional contribution of fishmeal and fish oil into aquafeeds. Fishmeal and fish oil provide essential nutrients to farmed fish species, as evidenced by the growing need for supplementation of some key nutritional elements when fishmeal and fish oil inclusion rates are reduced. Examples of that are specific amino acids, and vitamins such as vitamin D. in addition, conceptually these terms are flawed because they are largely based on a premise that it is somehow wrong to feed fish to fish. That position ignores some important points including the well-managed nature of the majority of the pelagic fisheries that produce the species used in fishmeal and fish oil production, as well as the lack of a direct consumption (i.e. food) market.

These materials are therefore transformed from a protein that has a very limited market into one which is readily accepted and supports global food security goals.

7 AQUAFEED.COM So much of your job seems to be in a defensive role, taking on the press: misreporting of a scientific paper on fishmeal and antibiotic resistance by the Economist, comes to mind. How do you think the industry as a whole could be more engaged in telling a positive story about seafood?

NA IFFO is very active in communicating the importance of the marine ingredients industry, but of course it is the headline stories that capture the attention, and it is correct to say that we do have an important role in defending this important industry.

We also provide an excellent information and data source on many different subjects, obviously as a service to our members in the first instance but also in our engagement with international organizations, national governments, NGOs and other industries. For the future, it may be interesting to note that IFFO is funding an increasing amount of research, and as this work reports much of this will be communicated to external audiences.

AQUAFEED.COM Common sense tells us that marine products are the natural and best food for marine species, so in terms of fish nutrition and end product quality, there is actually a very good story to tell.

What is IFFO doing to promote those benefits?

NA We of course see this as an essential component of IFFO’s work.

Common sense may tell us this, but so does the science. The nutritional contributions from fishmeal and fish

oil just cannot be found in any other single feed ingredient, and they are acknowledged to be particularly rich in some micronutrients that are important for the health and quality of farmed fish. These messages are a core element of IFFO’s general communications, whether in written format, or the many presentations we give to external audiences every year.

There are some very useful examples in the seafood industry that support this principle, such as the production of premium quality farmed salmon on high marine ingredient diets.

AQUAFEED.COM Turning to more proactive efforts, IFFO has been very active in fighting for antioxidants.

What is the position now on ethoxyquin and tocopherols, in the text of the International Maritime Organisation’s (IMO) IMDG Code and with EFSA? NA IFFO has been working across industries and regulators to support our principle of providing effective and safe antioxidants available to the fishmeal industry. This work is highly complex, and dependent on others taking up the market opportunities that exist (e.g. antioxidant manufacturers), but in our view it is important to have available options so that the industry may decide which antioxidant to use for an individual market. That leads to customization of product, which is important for the future of the sector.

AQUAFEED.COM Would you talk a little about the fishmeal sector in China?

NA China is an extremely important market for fishmeal, where it is as elsewhere in the world an important constituent of aquafeeds. China is the birthplace of aquaculture, and has an aquaculture industry that is very diverse in nature producing many different species of fish, and other aquatic animals such as turtles and frogs. Many of these species have complex nutritional requirements and fishmeal inclusions in some of the higher value species can often be high to ensure those needs are met.

Fishmeal is also important in providing a raw material base for pig feeds.

Where it is used in weaning diets, as the rich nutritional contribution supports the growth and health of piglets at a key point in the production cycle. It may only be used for feeds that are applied in a number of days, but they are critical, and fishmeal is regarded very highly by pig farmers.

AQUAFEED.COM What do you think IFFO will be saying about fishmeal and fish oil 10 years from now?

NA I expect that the key messages will be very similar. It is taking some time for the novel ingredients to reach commercial reality, and as mentioned above their position will be more of supplementation than replacement.

It is unlikely that any other ingredients will come along with the rich spectrum of nutrients seen in fishmeal and fish oil. Fishmeal and fish oil are certainly going to remain the foundations of aquafeeds, supporting the healthy production of farmed fish and in turn providing health benefits to the consumer.

AFΩ

feedconferences.com In association with

AQUAFEED WORKSHOP—MEXICO Guadalajara ▪ September 27th, 2018

An Aquafeed.com technical workshop

(in English and Spanish)

TECHNOLOGY

Robert Strathman

Dana Nelson

FORMULATION

Spencer Lawson

Extrusion Applications - Optimal Design and Quality Management of Aquafeed Robert Strathman, President, Famsun-USA Design and Engineering Develop and continually deliver exceptional products by understanding the four essential aquafeed design features: Buoyancy, Water Stability, Shelf Life, and Prevention of Water Pollution. In doing so, we hope to better enable the audience to develop and continually deliver exceptional products.

What You Should Know About Producing Both Floating and Sinking Feeds Dana Nelson, Market Development - Aquaculture specialist, Extru-Tech, Inc.

Albert Tacon (CHAIRMAN)

Gilberto Hernandez Gonzales

Bart Dunsford

Biosecure Shrimp Feeds and On-Farm Feeding Strategies Albert GJ Tacon, PhD., Aquaculture Nutrition & Feed Expert, Aquatic Farms Ltd. This presentation reviews the different feeds commonly used to produce farmed shrimp and discusses their potential risks, including the use of live hatchery and nursery feeds and the use of live and/or fresh food organisms for broodstock, and dry formulated shrimp feeds for shrimp grow-out operations.

The Use of Specifically Selected Probiotics in Shrimp Mariculture Bart R. Dunsford, Ph.D., PAS., Business Development Manager, Lallemand Animal Nutrition

Controlling the final bulk density of feed is not always as simple as it seems. Learn some of the lessons learned from the challenges of past and discuss equipment solutions directed at solving these problems.

Pre- and probiotics have often been a potential solution to meeting disease challenges. However, the frequent use of random products can lead to variable and inconsistent results.

Understanding the products is key to ensuring that the desired results are achieved.

Advanced Process Technologies for Micro Feed Production

Functional Feeds, Strategy for Reduction of Risks and Diseases in Aquaculture Under Challenging Times

Spencer Lawson, Process Technologist, Wenger Manufacturing Inc. While specifically focusing on micro aquatic feed, twin screw extruder technology exists that will allow for production of direct extruded feed as small as 0.5 mm. With proper preconditioning and extruder die technology, production rates that were once very difficult to achieve, are now possible.

Gilberto Hernandez-Gonzales, Aquaculture Manager N&CA, Nutriad A most important feeding strategy to tackle complex disease problems is feed management. Functional feeds are positioned as part of a preventive strategy to reduce risk and impact of disease in aquaculture.

9

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

Pilmico gains control of Gold Coin for $413 million AEV’s food subsidiary Pilmico International Pte Ltd has acquired a 75% equity interest from Golden Springs Group for Gold Coin Management Holdings Limited, one of Asia’s largest privately-owned agribusiness corporations, for US$413 million, as it expands its feeds business in the Asia Pacific region.

Established in Singapore in 1953, Gold Coin employs 2,600 people across 19 production facilities, in 9 countries.

With a milling capacity of approximately 2.5 million tonnes per year, Gold Coin offers products for both

livestock and the aquaculture, including young animals and hatchery feed, premixes, concentrates and compound feed. Gold Coin’s aquaculture business is reported to be worth $90 million in sales.

Pilmico International’s first ASEAN venture was in Vietnam in 2014 when it bought a 70% stake in Vinh Hoan Feeds (VHF), one of the country’s major aquafeed manufacturer. In 2017, it bought an additional 15% stake in VHF, effectively increasing its stake to 85%.

Conserving krill Conservation groups, seafood and aquaculture industries - including krill fishing companies and aquafeed firm BioMar, have agreed to support the creation of marine protected areas, including large no-fishing zones in the Antarctic.

The agreement was made at a Greenpeace-led retailer roundtable event in Cambridge, UK. The fishing conservation commitment was initiated by Aker BioMarine, a supplier of MSC certified Antarctic krill.

Nutreco invests in ViAqua to combat disease in the shrimp market Nutreco announced in July that it has invested in Israeli startup ViAqua as part of its focus on combatting disease in the aquaculture industry.

ViAqua is developing the first orallyadministered treatment for shrimp that improves resistance to viral diseases, including White Spot Syndrome Virus (WSSV), and prevents viral epidemics. Nutreco has taken a meaningful minority share that will condition-

ally grow over time.

ViAqua’s platform for improving resistance to viral diseases in shellfish and fish utilizes a proprietary RNAi particle formulation that triggers a cell reaction to disable the viral infection.

The coated particles are delivered via feed or immersion to produce maximum effect and to reduce costs. In the future, ViAqua intends to expand to include more species, like other

shellfish such as lobster and crab, and small fish.

Nutreco’s aquaculture division Skretting will work directly with ViAqua to effectively deliver the solution to the market.

Other shareholders in the company include The Trendlines and the Technion – Israel Institute of Technology, both based in Israel.

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BioMar expanding capacity of Alimentsa in Ecuador Less than a year after acquiring the majority stake in Alimentsa, BioMar is accelerating development of the Ecuadorian shrimp feed producer with the commissioning of a fifth production line and reorganization of personnel to strengthen the integration of the two companies.

"We have just commissioned a fifth production line in order to secure sufficient capacity next year," said Danny Velez, who took over the position as General Manager shortly after the acquisition.”

"It is a relatively small investment as the new line fits into our present facilities, and thereby we expect it to be ready already early next year. Our new feed trial and development facility announced in November is also progressing according to schedule and will be operational later this year."

Mr. Velez underlined that further investments are planned for the near future in order to fulfil the strategic ambitions of the company. He also outlined how BioMar's investment program in Ecuador is accompanied by a strengthening of the organization in Alimentsa.

From left to right: Danny Velez, General Manager Alimentsa; Andres Rivadulla, Finance Manager; John Tinsley, Technical Director and Laurence Maussaut, R&D Manager.

"We want our customers to see the tangible benefits of becoming a part of BioMar, so especially product development and technical customer support are being reinforced, however, also other areas of the company are being strengthened while we integrate Alimentsa into BioMar," he explained.

The Technical Director for BioMar Central America, John Tinsley, relocat-

ed to Guayaquil in May to join Alimentsa's R&D Manager, Laurence Massaut, in a first step to build a team for feed trials and product development around the new Aquaculture Technology Center. Andres Rivadulla has joined Alimentsa as Finance Manager from the multinational plastic giant BIC, and a number of new positions are being filled.

ADM announces plans to buy Neovia In a move that would greatly increase ADM’s aquaculture footprint, Archer Daniels Midland Company (ADM) has agreed to terms granting exclusivity in talks to purchase Neovia. The 100 percent cash deal has an approximate value of €1.535 billion.

Founded in France in 1954, Neovia manufactures and sells a wide range of nutrition solutions for the feed industry, operating in business lines including premix and value-added services, additives and ingredients, aquaculture and complete feed.

Through acquisition, the current

portfolio includes Epicore and BernAqua hatchery feeds and specialty feed firm, Pancosma. The company, which is currently majority owned by leading French agricultural cooperative group InVivo, has about 8,200 employees.

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Fish bones color salmon muscle red

Researchers at Nofima have discovered that a mineral-rich ingredient extracted from fish bones can affect the color of salmon muscles.

In experiments where salmon were fed diets containing the ingredient, the color of the salmon muscles increased considerably. The effect, which was first observed in an experiment with salmon smolt, was visible to the naked eye and confirmed by chemical analyses.

The color came from astaxanthin.

Only a limited portion, usually less than 10 per cent, of the astaxanthin in salmon feed is absorbed in the muscle of farmed salmon. This can be caused by limitations in absorption and transport via blood and liver, or limited absorption and pigmentation in muscle tissue.

“It is somewhat unexpected that a mineral ingredient can affect pigment utilization," said Sissel Albrektsen, senior researcher at Nofima (pictured above). "But at the same time, it is very positive to see that nutrients liberated from fish bones can considerably increase the utilization of astaxanthin in salmon feed."

Feed with the phosphorus-rich min-

eral ingredient was also tested on slightly larger salmon during the growth period from 1.7 to 2.5 kilograms, and compared with salmon that received the same feed to which a regular source of commercial phosphorus was added. In muscle, the researchers found 35 per cent more color, measured as milligrams of astaxanthin per kg of fish growth. The salmon’s ability to digest astaxanthin increased by nearly 20 per cent in fish fed the mineral ingredient, while the pigment levels in both the blood and liver also increased.

“We believe that the main explanation of why the muscle becomes redder is that the salmon digest more of the astaxanthin with the mineral ingredient present in the feed,” said Albrektsen.

It has been repeatedly proven in salmon that the mineral ingredient stimulates increased growth, and in some cases this is explained by increased digestibility of nutrients.

In the initial experiment with smaller salmon, it was found that the astaxanthin levels in blood, liver and whole fish were 55, 29 and 22 per cent higher, respectively, compared with fish that were fed an ordinary source of commercial phosphorus.

“In an experiment now taking place, we will take a closer look at how minerals and other liberated components in fish bones can affect processes including intestinal function,” said Albrektsen.

Low protein Novacq can increase prawn biomass Ridley has shared the results of its commercial feed trial with three prawn farms. Two steam-pelleted prawn diets, nutritionally identical with the exception that one diet contained 5% Novacq and one did not, were compared on each farm.

The trials in commercial sized ponds began in January 2018, and harvested in late April/May. Overall the trials demonstrated a positive response to Novacq. Some operators reported record biomass harvest per hectare, and there was a high degree of variation in the survivability of prawns on the various farms in both treatments.

In addition, a low protein Novacq diet was successfully trialed at one farm, and results found it can potentially enable Australian farmers to increase biomass by up to 25% if they are at or reaching maximum nitrogen farming limits set by the EPA. “We tried a reduced protein version of the Novacq diet and the results look very promising, as we saw healthier and faster growing prawns on the Novacq diets,” stated Matt West, General Manager of Australian Prawn Farms, one of the farms participating in the study.

“We see the benefits from the last 2 years across survival, biomass productivity and nutrient reduction.”

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Aquaculture still growing, says new FAO SOFIA report FAO has released its latest official world fishery and aquaculture statistics report, the State of World Fisheries and Aquaculture (SOFIA) 2018.

meal and fish oil) are excluded.

According to the report, with capture fishery production relatively static since the late 1980s, aquaculture has been responsible for the continuing impressive growth in the supply of fish for human consumption.

The contribution of aquaculture to the global production of capture fisheries and aquaculture combined has risen continuously, reaching 46.8 percent in 2016, up from 25.7 percent in 2000.

With 5.8 percent annual growth rate during the period 2001–2016, aquaculture continues to grow faster than other major food production sectors.

Global fish production peaked at about 171 million tonnes in 2016, with aquaculture representing 47 percent of the total and 53 percent, if nonfood uses (including reduction to fish-

The report also revealed that the growth of farming of fed aquatic animal species has outpaced the farming of unfed species in world aquaculture.

The share of unfed species in total

Aquaculture Feed Extrusion, Nutrition and Feed Management short course at Texas A&M A one-week Practical Short Course on Aquaculture Feed Extrusion, Nutrition and Feed Management will be presented on August 26-31, 2018 at Texas A&M University by staff, industry representatives and consultants.

The program will cover information on designing new feed mills and selecting conveying, drying, grinding, conditioning and feed mixing equipment.

Current practices for preparing full-fat soy meal processing; recycling fisheries by-products, raw material, extrusion of floating, sinking, and high fat feeds; spraying and coating fats, digests and preservatives; use of encapsulated ingredients and preparation of premixes, nutritional requirements of warn water fish and shrimp, feed managements and least cost formulation will be reviewed.

There will also be a practical demonstration of sinking, floating, and high fat aqua feed produced on four major types of extruders - dry, interrupted flights, single and twin screw - using various shaping dies. Other demonstrations include vacuum coating and lab analysis of the raw material for extrusion. https://perdc.tamu.edu/extrusion/

aquatic animal production decreased gradually from 2000 to 2016, shrinking by 10 percentage points to 30.5 percent.

Aquafeed Extrusion Course Chile Australian company FoodStream, working with extrusion expert Dennis Forte, is expanding its extrusion courses to Chile. One participant from the Philippines commented “The Extrusion Course is unique in that it is able to present the blend of the fundamental principles of extrusion technology with its practical applications. The speakers transcend the hurdles of mathematical and engineering concepts by using appropriate learning and citing their vast production experiences”.

The 3-day program covers the principles of extrusion, the design of extrusion processes, and the formulation of extruded aquafeeds. Topics cover the basics of extruders and their configuration, through what is happening inside the extruder barrel, to an understanding of extruder dies and extruder instability. Examples in product formulation and the design of extrusion processes demonstrate application of the theory. Principles learned will be applied during the practical demonstration in the University of Temuco pilot plant.

The course will be presented in English, but attendees from the University will be available to help translate questions www.foodstream.com.au/events

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The Maillard Reaction and amino acid availability in feeds

By Colin Mair, Feed Consultant, U.K.

It has been stated by feed nutritionists for years now that any reaction between proteins and reducing sugars binds the amino acids in very stable compounds that make the amino acids unavailable to the animal eating them. It has recently been suggested that a mix of molasses and amino acids can be reacted at ‘low’ temperatures (110 degrees Celsius) and this will partially bind the amino acids, the result that these amino acids are absorbed more slowly than if they were imbibed in their crystalline state. This is important. If all the amino acids required by a tissue reach that tissue at the same time then protein manufacture is optimized. If crystalline amino acids are in the feed they pass through the gut wall very quickly, creating a situation where the more slowly digested amino acids in the proteins of the feed reach the target tissue later. This results in poor conversion ratios and increased excretion of nitrogen into the water (for aqueous species). How then can this partial binding of the amino acids be achieved?

The Maillard Reaction This is a highly complex reaction. It is simplistically explained as where the carbonyl group of the sugar reacts with the amino group of the amino acid, producing N-substituted glycosylamine and water. After this the glycosylamine undergoes rearrangement, forming ketosamines.

These can then react further in several ways, producing reductones, diacetyl, aspirin, pyruvaldehyde and other short chain hydrolytic fission products and then brown nitrogenous polymers and melanoidins. These latter chemicals are highly stable and there is no way that the digestive system can release the amino acids.

The full course of the reaction is not inevitable. There are some amino

acids that are more susceptible than others to this reaction. These are lysine, arginine, tryptophan, tyrosine, cystine, methionine and threonine.

They are more basic and generally have a secondary amine group. These amino acids are labelled ‘high

browning’. Note that this group includes lysine, methionine and threonine, the three most important essential amino acids in fish feeds.

Different compounds are formed at different temperatures, as shown in table 1 below.

Table 1. High browning amino acids

Amino Acid

100 deg C

120 deg C

180 deg C

Arginine

Buttery Note

None

Burnt Sugar

Histidine

Buttery Note

No significant aroma

Cornbread, buttery note

Lysine

Baked sweet potato

Weak

Bread-like

Methionine

Overcooked sweet potato

Strong baked potato

Potato

Threonine

Chocolate, maple

Weak

Burnt

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The temperatures are not exact, the reaction being more pronounced under alkaline or neutral conditions and being suppressed under acid conditions (Vitamin C is a good suppressor of the reaction). Also the type of sugar is important. Pentoses react more readily and brown more readily than hexoses (fruit sugar, fructose, is a pentose. Glucose and lactose are hexoses.) Note that reducing sugars are required. None reducing sugars such as sucrose (table sugar) first need to be cleaved using alkaline, acid or heat conditions.

Obviously free amino acids react more readily than amino acids bound in protein chains, so added crystalline amino acids are more at risk.

The Maillard reaction is a multi stage reaction that goes to completion at about 180 degrees Celsius, the first stage of the reaction starting at about 100 degrees Celsius. This first stage produces a relatively unstable compound, the initial condensation reaction (where two molecules are combined, driving off a water molecule) forms a sugar-amine. The compound formed from this is known as a Schiff’s Base. Shortly after this further addition reactions occur where cyclic compounds are formed to form nitrogen-substituted glycosylamines. These are quite stable and from this point on the amino acids become increasingly unavailable to the digestive system, with increasing levels of polymerization

and increasingly unavailable amino acids.

How do we make amino acids slowly available without locking them up?

The Maillard Reaction can be slowed down or stopped by several means.

1.

Limit the amount of reducing sugars available. This is hard. Even in wood there are sugars that are almost impossible to leach out by even quite intensive pressure washing. Kiln browning is a maillard product that is quite a problem.

2.

Make the conditions more acidic. This can be done by using

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... the use of additives will not only allow more efficient absorption of added amino acids but will also allow maximum digestive absorption of amino acids in proteins which will not be bound in undigestible Maillard complexes.

SMS will do the job. The SMS is a process aid and disappears during the process. There could be a ‘metallic’ flavor off-note, but this can be masked using available masking agents. The result of this is that the amino acids are boded only partially in the first stage of the Maillard Reaction. These amino acids are still available to the animal, but are released more slowly than the original crystalline amino acids. The pH, temperature, available water, pressure, amount of SMS and amount of amino acids should all be balanced to optimize the reaction.

References:

Note the use of additives will not only allow more efficient absorption of added amino acids but will also allow maximum digestive absorption of amino acids in proteins which will not be bound in undigestible Maillard complexes.

https://onlinelibrary.wiley.com/doi/ pdf/10.1111/j.13652621.2000.tb15949.x

[Reference links live in online issues.]

https://www.wattagnet.com/ articles/22489-the-maillard-reaction-s -impact-on-animal-nutrition Nutrition Research Reviews 26(2):119 · August 2013 https://www.allaboutfeed.net/Home/ General/2001/3/The-Maillardreaction-in-feed-manufacturingAAF011008W/ https://conservancy.umn.edu/ handle/11299/146912

https://pubs.acs.org/doi/ abs/10.1021/jf502497r

To set up a feed with added crystalline amino acids add reducing sugars. The cheapest source of these in the feed industry is molasses. Add SMS at say 0.1%. ascorbic acid (vitamin C) or lactic acid.

These acids only have a limiting effect.

The ascorbic acid is changed to dehydroascorbate as it reacts, making it ineffective.

3.

Use sulphur containing compounds. Sometimes sulphur dioxide is used, but more often sodium metabisulphite (SMS). The advantage of this is that the first part of the Maillard Reaction tends to take place but further polymerization is prevented. The level of SMS needed is related to the type of amino acid being protected, but typically 0.1-0.4%

Make feeds and test by feeding. Note that the chemical standard test system prescribed by the AACC and AOM will not test for the availability of the amino acids. When companies report on free amino acids these have been separated by hydrolysis, and the results do not relate to in vivo (live animal) tests.

It would be possible to add molasses as a separate ingredient and blend the amino acids with the molasses, making the production control much easier. The mix can then be pumped into the production process as a separate stream.

AFΩ

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Colin Mair, Feed Consultant E: colinmair@btinternet.com

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17

Improving dryer productivity with intelligence

By Paul McKeithan, Head of Digital Services at BĂźhler Aeroglide

Customer journeys into IoT: discovering sustainable value with cloud-based data management.

The buzz words IoT, IIoT, smart devices and connectivity fill the news these days, and with good reason.

Thanks to a surge in device availability, coupled applications, combined with opportunities for constant connections, a smarter world has emerged around us. Homes with smart meters for lighting and thermostats are giving us control over our energy bills, urban infrastructures are beginning to manage waste collection, public transportation and energy usage, just as Alexa and Google are learning personal preferences that make our lives easier. For years the industrial processing sector has lagged behind the consumer market in cutting edge computing technology; however, this is changing as the expectations we

have in our personal lives collide with the work place.

By 2050, the needs of approximately nine billion people will require approximately three times our current resources. At this time, challenges will accelerate for the deficiencies of resources and the enormous production of current waste. Planning for the future, BĂźhler has a goal to reduce both energy consumption and waste at customer sites by 30%, by the year 2020. Sustainable business models will help customers get there, focused on safe and efficient feed processing that saves energy, and digitalization that provides integrated operational connectivity. Sustainable processing must meet present needs without compromising future viability.

It’s not just a corporate responsibility,

sustainability directly relates to efficiency, minimizing waste, finding cost-savings and new profitability.

Discovering efficiency, making use of volumes of data One of the biggest challenges with connectivity is understanding the vast volumes of data collected and being able to make use of it. This includes realizing the potential for sustainability to deliver a competitive advantage.

Over drying, for example, is a common occurrence, and one single processing deviation can dramatically affect results and yield. When feed is over dried, too much water is evaporated which has to be replaced

18 with raw ingredients. This can result in dramatic product losses over time.

A connection to proven algorithms for moisture control Access to full time, real-time monitoring with hardware, software, and ongoing process expertise, has enabled one BĂźhler customer to consistently reach a moisture target within +/- 0.5% of target.

Traditionally, the operator collected samples off the discharge end of the dryer and took them to a lab to measure target moisture, returning to adjust the dryer controls as needed.

The process was slow, with a retention time of 25-30 minutes, plus time in the lab to analyze. After one reading, it could be an hour before results were available, all the while product and time was wasted.

Without continuously automated data monitoring, the operator had to keep taking measurements and making adjustments. But when the operator turned on Moisture Pro--a product in a suite of cloud-based tools that reduces time-based moisture deviation based on decades of drying expertise--a reading began to be captured every second. As an inline moisture sensor took a reading, an algorithm written to control the dryer controlled temperature settings to consistently achieve the desired output. This drying expertise has been available for decades, introduced by field engineers during site evaluations.

But now, the connectivity to this expertise could be continuous. For this customer, connectivity not only increased bottom line profits, more efficient processing meant waste was minimized.

Real-time trending reports take data to the point of analysis. Insights confirm newly discovered efficiencies are maintaining, and customers appreciate the facts.

With Moisture Pro turned on, a report for the same customer showed an average actual moisture was 5%--with a moisture target of 6.8%--and the standard deviation from the target was 1.37%. It also showed that in one particular month, Moisture Pro was only active 39% of the time, which was a 3% improvement from the previous month. With Moisture Pro activated, the customer was able to achieve moisture control within 1% of the target, 59% better than manual operations, while during manual operation, capabilities to control within 1% were achievable only 13% of the time.

19

Turning Data into Insights for Overall Equipment Effectiveness Overall Equipment Effectiveness (OEE) is the gold standard for measuring productivity, and cloud-based data management does this best by reporting average energy consumption, along with a treasure trove of insights for sustainable value. But generating data through IoT connected technologies is only valuable when manufacturers have the capacity to process it in a meaningful way. The Internet of Things connected to technology is only one part of the process. When data can be fed into a data management system, the manufacturer can begin to see longterm benefits.

Discovering an energized dryer with no product Changes over time, in recipes and operating parameters, as a result of operator preferences, turnover, or

constraints, can lead to significant losses in energy efficiency.

A customer using our cloud-based management discovered that a dryer was energized 20% percent of the time, with no product. Data tracked over time showed that on Tuesdays, the dryer was energized 8%. On Wednesdays, it was 30%. From this, the question became why is third shift energizing the dryer this way? This led to a review that recognized third shift processed a different product. From there, management was able to pinpoint and review a process that reduced energized time and prevented this waste of energy.

Making Improvements Sustainable with IoT From the Bühler cloud, all data collected is displayed on web-based dashboards, enabling users to pinpoint at-risk parts and components before they fail. IoT data management Interconnects information to help

A trusted human-connected digital solution will go a long way toward managing the deficiencies in resources and the enormous production of current waste. .

customers realize new business models, optimize internal processes and achieve major cost savings. It allows customers to take advantage of process knowledge, full time, and make it sustainable. After all, it doesn’t make sense to collect and monitor energy usage data if the data cannot be put to good use—to drive sustainable efficiency. One Bühler customer discovered this recently, after repeating a processing evaluation.

When upstream operations change, product going into the dryer can have a higher moisture. As a result, more energy can be required to dry according to specifications. Bühler engineers solved this problem by maximizing bed depth for an even airflow that could thoroughly dry the wet product core, adjusting the processing depths for longer retention, at lower temperatures. The changes saved the customer 12% in

20 energy costs; however, they didn’t last. Eight months later, the customer requested another evaluation. When field engineers returned, they recommended the exact same changes. We explained that the same adjustments had been made during the previous evaluation. This became a turning point for the customer as we described how cloud-based management works. He started to see the value of being connected to processing expertise in real time, 24 hours a day, seven days a week. By joining the digital world and trusting the data, those energy savings will never disappear again.

Sustainable processing is the result of continuous improvement. By using subscription-based data management, connected to processing expertise 24/7, customers will be able to reduce

energy consumption and waste by the year 2020. Manufacturers already have sensors that collect volumes of data, but because of traditional technology restraints, and different departments having different data needs, information has historically been stored in silos. The centralizing and synthesizing of data, for analysis that yields meaningful insights will become more and more valuable as we prepare for the world’s growing

populations. A trusted humanconnected digital solution will go a long way toward managing the deficiencies in resources and the enormous production of current waste.

More information Paul McKeithan, Head of Digital Services, Bühler Aeroglide E: paul.mckeithan@buhlergroup.com/919-851-2000 www.Bühlercloud.com

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22

AQUAFEED PRODUCTION Critical KPIs for optimizing and sustaining factory performance

By Rob Strathman, President, Famsun-USA Design and Engineering

A defining characteristic of any manufacturing operation, because of its direct impact on the cost of goods sold, is the overall technical and fiscal performance of the factory.

Sustaining acceptable results is the pursuit of many plant management teams. The root of their challenge is in finding and correcting productivity losses through continuous improvement programs that ensure reoccurring issues are put to rest for good or at least delaying their reoccurrence.

For the efficient production of aquafeed two KPIs are essential: 1. Yield Efficiency (YE) and 2. Overall Equipment Effectiveness (OEE) Both YE and OEE metrics fall under the strategic tenets of Lean Manufacturing, which are intended to remove waste and inefficiency from production. Since raw material represents 60 to 70% of the total cost of production, the importance of monitoring YE cannot be overstated. OEE tracks wasted runtime to gauge the reliability of the line.

Fig. 1. Basic raw material loss model

Yield efficiency Shown in Figure 1, is a basic Material Loss Model of the aquafeed manufacturing process. It illustrates how none of the cost centers of the operation are immune to experiencing some level of material loss. More importantly, the diagram shows how the numerous forms and sources of lost materials can all be broken down into two categories: Controllable and Uncontrollable.

Controllable Losses are all sources and forms of material loss that can be mitigated through equipment design

23 or operational practices. Alleviating these losses should be the central focus of factory management.

Uncontrollable Losses are those that the business must incur. More specifically this is the portion of the raw material in the form of water that is given up during the drying operation to create a mold resistant, shelf stable, product.

In a scenario where the average raw material moisture is 10%, and the needed moisture of the finished product to prevent mold growth is 9%, a 1% material loss will unavoidably be experienced. In the production of aquafeed, the opposite response is also possible; such as when the average raw material moisture is less than the target moisture of the finished product. In this case, the operation receives a material gain.

Material gains are great for obvious reasons but provide a disservice to factory managers who are searching for improvement opportunities, as gains inflate the actual yield and easily hide the real material loss opportunity.

Figure 2, illustrates the two types of material losses that occur during a production run and provides the Yield Efficiency calculations. YE measures how well the controllable portion of the raw material was successfully converted into a finished product. A 100% YE means there were no controllable losses, while a 97% YE would indicate a 3% loss of controllable materials.

One of the shortfalls of the aquafeed production process is the routine (fines) and intermittent (startup and

Fig. 2. Yield Efficiency calculations.

Fig. 3. Material losses points of the aquafeed production process.

shutdown) loss of materials from the process flow. These diverted materials are one of the most significant sources of yield loss for the entire factory. Thus, the first tactic used by most factory managers is to “re-mill� as many of these diverted materials as possible by adding them back in at the batching system as depicted in Figure 3.

The hot, wet, and gummy materials diverted from both the preconditioner and the extruder are the most difficult to re-mill and are typically at the greatest risk of being lost. In most cases, significant labor is required to collect and dry these materials before they can be metered back in at the batching system.

24 The benefit of measuring YE on a plant level is realizing the significance of the factory’s true material loss opportunity. Calculating YE on a production run and diet level can create a much better understanding of system reliability, product design, and the sources of material loss. More importantly, the data enables strategic planning and re-engineering of the recipe, equipment, and the process, allowing sustained long-term efficiency improvements.

equipment breakdowns. However, because there are less intuitive causes of runtime losses, such as the time wasted producing defective products, it can be a challenge for many managers to define, identify, and improve their situation.

Overall equipment effectiveness Critical to factory performance is achieving productivity (tons per day) targets. The most obvious hindrance to meeting productivity goals is

Fig. 4. OEE calculations.

OEE’s strength as a performance indicator is in tracking four different forms of runtime losses that are illustrated in Figure 4. OEE data enables managers to discover their truly effective runtime and to clearly see where to focus their improvement resources.

25 Are we going digital?

In part 2 of this series, we will explore next generation aquafeed factories and take a look at what digital manufacturing actually is, the evolution of factory management systems and changes in extrusion technology.

Fig. 5. Pareto chart of runtime losses

An OEE of 100% represents perfect production; meaning no defects were created while running at the target rate, and without stopping. According to OEE implementation experts, it is not uncommon for companies just beginning to use this improvement strategy to find themselves operating at or below 60% OEE. These same individuals consider an OEE of 85% or higher a World Class Operation.

An OEE best practice is to record the reasons for unplanned downtime.

This data provides excellent visibility into the reliability of the operation.

The Pareto Chart seen in Figure 5, lists the events that stopped an extrusion line along with both the frequency and the duration of each.

The OEE metrics and their associated data make downtime losses more transparent and highlight areas of needed improvement. Along with an integrated Continuous Improvement Program, OEE data can drive significant factory efficiency gains.

The future use of KPIs in aquafeed manufacturing Lean Manufacturing has a proven track record of improving and sustaining manufacturing performance targets including:

More information

• Maximizing throughput • Reducing quality defects and re-mill • Optimizing first pass quality

• Limiting material losses • Maximizing asset utilization • Lowering inventory holding costs Historically, however, most businesses have declined to use metrics like YE and OEE due to the complexities of gathering the needed data; but not anymore. Fortunately, with today’s automation and IT capability, the hurdles to data collection and rapid KPI generation have been lowered significantly. These same technologies are promising to usher in a more efficient, data-driven, means of manufacturing, which is being touted as the “Digital Factory”.

AFΩ

Rob Strathman, President, FamsunUSA Design and Engineering E: rob.strathman@famsun-usa.com This article is based on a presentation at Aquafeed.com’s 11th Aquafeed Horizons Asia conference, which took place March 27th, 2018 in Bangkok.

Visit feedconferences.com for upcoming Aquafeed technical conferences and workshops.

27

Using wood-based technology to bridge the aquafeed supply gap

By Ricardo Ekmay, PhD, Director of Animal Nutrition, Arbiom Challenges facing the feed supplydemand gap today have aquafeed producers wondering where the next big breakthrough will come from

animal-derived protein demand projected to double by 2050, to more than 465 million tons of meat and more than one billion tons of milk.1

A growing global population, combined with factors such as changing socio-demographics, will place increased pressure to provide not only more but also different types of food. The increased demand for protein is particularly concerning, with

Fish protein currently contributes 17 percent of the global human population’s intake of animal protein and, already today, aquaculture producers are experiencing protein sourcing challenges, with rapidly increasing production and greater

need for high-quality protein sources.2 As demand continues to expand, there are growing concerns about the availability of resources—including fish meal and fish oil—to safely and sustainably feed the world.

As we begin to approach the limits of conventional agricultural production systems, alternative proteins are gaining growing interest and could expand to encompass up to a third of

28 the market by 2054.3 In this article, we will explore more about the advancement of alternative protein sources to complement fish meal with a focus on single cell proteins (SCPs) derived from an unlikely source.

The Search for a Solution In the search for new resources to help bridge the supply-demand gap, much of the focus has centered around a two-fold question: how to increase protein production while also addressing sustainability challenges?

Or, alternatively, how to produce more using less resources and reducing environmental impact of production?

SCPs include microbes such as yeast, fungi, bacteria and microalgae, which can be produced via fermentation— thus requiring less land, water and fertilizer than agricultural crops or livestock. SCP biomass can serve as a high-quality ingredient in aquafeed as it is comprised of protein and essential amino acids, along with micronutrients.

One SCP in particular, Torula yeast,

provides a number of benefits as a protein source. A nutritious alternative to conventional plant and animal protein sources, Torula yeast is a highquality protein source with an excellent amino acid profile, providing valuable building blocks that improve nutrition and performance.4

Torula yeast also contains functional fibers, such as beta glucans and mannan-oligosaccharides, which are known to impact gut health as immunomodulators and pathogen binders – thereby assisting an animal’s ability to fight disease. A particularly important quality for producers to consider in light of regulatory and consumer pressure to reduce antibiotic use.

Additionally, Torula yeast is devoid of the endogenous allergens that are found in milk, egg, wheat and soy products, making them an ideal protein source for susceptible populations. And unlike other classes of SCPs, Torula yeast has regulatory approval for use in feed and food applications, a history of safe use and more consumer appeal than many insect, bacteria or fungi-based feed sources.

Wood to Food Technology What has limited the potential of SCP as a feed protein source? Until today, it has been a challenge to produce a high-protein SCP product in commercially relevant volumes in a process that is economical at commercial scale, as well as sustainable and safe in terms of inputs and source material required. But a team of researchers at Arbiom are now looking in a place many have never even considered: wood. Wood is a renewable, readily available, natural and organic carbon source that has not historically been a part of the food supply chain, even though it contains sugars (C5 & C6 hydrolysates). Despite wood’s promising attributes, bioconversion processing technology to extract its nutritional components has not been effective or efficient. With technology being pioneered by Arbiom, wood has significant potential to help address global protein supply challenges in a way that is both sustainable and scalable.

Arbiom’s scientists have developed bioprocessing technology to maximize the value of woody biomass by

Arbion’s Integrated Technology Platform

29 extracting its fermentable components and optimizing conditions for SCP production. The proprietary technology integrates pre-treatment and processing with state-of-the-art fermentation of an enhanced strain of Torula yeast (Candida utilis), enabling production of a highly-digestible, highprotein ingredient from wood that can be produced in industrial-scale quantities.

Wood offers important advantages over the use other feedstock material used to produce SCP, such as waste and wastewater, methane, and glucose from food crops: • Wood is sustainable, abundant, and environmentally-friendly • No irrigation or fertilizer required • Traceability through strong mature industrial supply chains • Advancements in global silviculture practices ensure forest health,

productivity and biodiversity are maintained / enhanced • Additive to food supply chain; does not compete with food crops • Available year-round For feed formulators, Arbiom’s torula yeast protein product is differentiated from any other alternative protein available today as it is produced using wood hydrolysates as the substrate.

Arbiom’s torula yeast (brand name: SylPro®) is an ideal high-quality alternative to soy protein concentrate, delivering nutritional, economical, traceable and sustainable protein source for multiple species. Beyond aquaculture, other market segments face similar production challenges due to reduced use of growth promoting antibiotics, for example, and are exploring these types of alternatives to safely and sustainably rear young animals.

In addition to the benefits that woodbased SCP offers the agricultural industry, recent developments in bioconversion technology also bring potential new opportunities to wood companies. Mills, for example, typically experience 30-34 percent mass losses on site in the form of slabs, edgings, sawdust, fines and bark.5 By aligning themselves with a partner in bioconversion technology development, these companies may be able to find a more profitable use of their current wood wastes and/or new market segments to explore.

The Road Ahead Technology is poised to play an increasingly important role in meeting protein supply-demand needs as aquafeed market requirements continue to evolve and expand. New SCP-based protein solutions offer

30 promising ingredients for feed formulators and ultimately end consumers, bridging the gap between agronomy and industry and opening up the door to a new era of renewable resources that can be utilized to meet market demands head-on.

Innovators across the food supply chain recognize that to meet society’s growing nutritional needs, we must produce food that is more ecologically -sustainable and biologically healthy.

Arbiom’s bioconversion technology is a strong example of how innovation can help address global food supply needs by identifying ways historically non-food material, such as wood, can expand global food production potential in a more resource-efficient process to produce protein. It’s a winwin for the industry and a milestone for helping drive healthier, more sustainable solutions to meet global nutritional demands.

Strategies and Factors Influencing a Sustainable Equilibrium. Foods 2017, 6, 53.

2. United Nations Food and Agriculture Organization (FAO) (2014).

Economic Analysis of Supply and Demand for Food up to 2030 – Special Focus on Fish and Fishery Products.

FAO Fisheries and Aquaculture, 1089, p. 22.

With technology being pioneered by Arbiom, wood has significant potential to help address global protein supply challenges in a way that is both sustainable and scalable.

3. Lux Research (2014). WhooPea: Plant Sources Are Changing the Protein Landscape. State of the Market Report.

.

4. VTT Technical Research Centre of Finland Ltd. (2017). Single Cell Protein—State-of-the-Art, Industrial Landscape and Patents 2001–2016.

Frontiers in Microbiology, 6, p. 1.

More information

5 Ramage et al. (2017). The wood from the trees: The use of timber in construction. Renewable and Sustainable Energy Reviews, 68, Part 1.

References 1. US National Library of Medicine National Institutes of Health (2017).

Future Protein Supply and Demand:

Dr. Ricardo Ekmay,Director of Animal Nutrition, Arbiom E: rekmay@arbiom.com AFΩ

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32

The use of a functional diet to increase survival of white leg shrimp Litopenaeus vannamei (Boone, 1931) infected with acute hepatopancreatic necrosis disease (AHPND)

By Matthew A.G. Owen Ph.D, Charles McGurk Ph.D, Polyana F. Silva Ph.D, and Cesar Molina-Poveda Ph.D, Skretting Sustainable aquaculture development requires disease prevention strategies. Within the last twenty years, the global shrimp industry has suffered from great economic losses due to viral, bacterial and fungal diseases caused by pathogens such as white spot disease virus (WSSV), Taura syndrome virus (TSV), Vibrio parahaemolyticus and Vibrio harveyi (Qiao et al., 2011). Understanding the interaction of shrimp immunity, diet and environment is of critical to the successful implementation of strategies to ameliorate the effect of these pathogens.

Beginning in approximately 2009, a new, emerging disease called ‘early mortality syndrome’ or ‘EMS’ [more descriptively called acute hepatopancreatic necrosis disease or AHPND; Lightner et al. (2012)] began to cause significant production losses in southern China. As has been the case with other epizootic shrimp diseases (Stentiford et al., 2012), EMS was causing serious production losses in affected areas and is also impacting employment, social welfare, and international markets (BondadReantaso et al., 2012; Mooney, 2012).

Fig. 1. Images from L. vannamei hepatopancreas showing morphological tissue variations characteristic from AHPND pathology. (A) Histomorphology from non- infected shrimp showing normal hepatopancreas (HP) tubules with distinct R- (resorptive), B- (blisterlike), F- (fibrillar) and E-cells (embryonic). (B) Histopathology of HP, showing typical lesions of AHPND: progressive degeneration of the HP tubule epithelial cells, accompanied by a decrease of R, B and F-cells followed by a marked reduction of mitotic activity in E-cells and inter-tubular hemocytic infiltration (arrows); (C) necrotic sloughing (SL) of the HP tubule epithelial cells into the lumens and the remnants of HP tubules surrounded by hemocytic infiltrates. This last with no R, B or F-cells and some epithelial cells presenting prominent karyomegaly (enlarged nuclei). Histological sections were stained with H&E. Scale bars = 150 μm.

An example of the typical pathology of the hepatopancreas of AHPND infected shrimp can be seen in Fig. 1.

Skretting Aquaculture Research Centre (ARC) undertook a series of experiments to investigate if a functional diet could reduce the mortality associated with the disease.

These experiments included in vitro, in vivo and also field trials. Candidate functional ingredients were screened in vitro for anti- bacterial activity against V. parahaemolyticus and V.

harveyi. After potential ingredients were identified, both the singular ingredients and their combinations were tested in vivo at the University of Arizona Aquaculture Pathology Lab to a standardized protocol. Triplicate tanks of 1 g shrimp were either fed control based diets (not including functional ingredients) or the functional diets for a period of 21 days prior to challenge with causing AHPND - causing V. parahaemolyticus. The challenge was either per os, by the soaking of feed, or immersion where

33

Fig. 2. Survival time course of L. vannamei fed one of test (Lorica) and control diets and challenged with V. parahaemolyticus at the University of Arizona.

Fig. 3. Survival time course of L. vannamei shrimp fed either the functional diet (Lorica) or a control base diet, while challenged with V. parahaemolyticus at AquaMekong ShrimpVet Lab.

by a bacterial suspension was added directly to the tank containing the shrimp.

were infected which was confirmed by histological scoring of the hepatopancreas (HP).

The results showed a mortality of 96% in the control fed group, whereas in the group fed Lorica’s functional ingredients the mortality was approximately 50%. This represents a relative percent survival increase of 48% (Fig. 2). Note that there was mortality in the Lorica fed group, so it is clear that all animals in all tanks

In order to confirm if the results were reproducible, an independent test at AquaMekong ShrimpVet Lab in Vietnam under similar testing conditions (21 days feeding prechallenge; control and functional diet: 4 replicate tanks per treatment group) was conducted.

Results showed the mean mortality

for Lorica peaked at 46%, whereas the control reached a mean mortality of 70%, at eight days post infection. This represents a relative percent survival increase of 34% (Fig. 3). While the effect was not as large as that seen in the University of Arizona, it did nevertheless reduce mortality, and again the survival curve displays some mortality that plateaus so we knew the diet was being effective in stopping the disease. The infection was confirmed in this experiment by the use of histology; these results also demonstrated that the animals fed Lorica were able to recover quickly after the infection. In short Lorica not only shielded the animals but also supported them during the infection.

Skretting ARC then confirmed the appropriate feeding protocol, both feeding period prior to challenge and the optimum feeding frequency. As shrimp do not have an immunological memory and the fact that the diet is both bacteriostatic and reduces toxin production means that the diet must be present within the animal to be active. Thus the feeding frequency, a minimum of 4 meals per day, is required to maintain efficacy of the diet. During field trials optimum growth was seen to be at 6 meals per day or more. Thus Skretting recommend that the feed is fed at least 6 times per day. Field trial results from Skretting Ecuador showed the beneficial effect of diets on survival and, subsequently, pond productivity.

At the end of the production cycle, in an intensive system (32 / m2), a difference of more than 25% survival was found among the Lorica treatment and the control (Fig. 4).

34 When the yield is analyzed in terms of pounds per hectare per day (Fig. 5), an increase greater than 60% is observed as a consequence of the greater survival and the higher harvest weight. The growth rate was higher for the group of shrimp fed with Lorica, regarding the control.

Conclusions Skretting has demonstrated the efficacy of the functional diet Lorica in laboratory trials as well as field trials.

The functional ingredients are proven to reduce Vibrio bacteria in vitro, and promote survival during in vivo challenge trials at two independent research facilities. The laboratory results were further corroborated by field trials that showed Lorica improved both survival and ultimately pond productivity. The use of Lorica is a valid alternative to the use of antibiotics, particularly considering that these compounds do not require time of withdrawal before the harvest; nor do they cause resistance in the pathogenic bacteria present in the culture system.

References Bondad-Reantaso, M. G., R. P. Subasinghe, H. Josupeit, J. Cai and X. Zhou (2012). "The role of crustacean fisheries and aquaculture in global food security: Past, present and future." Journal of invertebrate pathology 110: 158- 165.

Lightner, D. V., R. Redman, C. Pantoja, B. Noble and L. Tran (2012). "Early mortality syndrome affects shrimp in Asia." Global Aquaculture Advocate Magazine 40.

Mooney, A. (2012). An emerging shrimp

Fig. 4. Survival of L. vannamei fed a control diet or test (Lorica) diet in ponds under commercial conditions in Guayas province, Ecuador (October 2016).

Fig. 5. Pond productivity (lb/ha/day) of L. vannamei fed a control diet or test (Lorica) diet in ponds under commercial conditions in Guayas province, Ecuador (October 2016).

disease in Vietnam, microsporidiosis or liver disease.

Qiao, J., Z. Du, Y. Zhang, H. Du, L. Guo, M. Zhong, J. Cao and X. Wang (2011).

"Proteomic identification of the related immune-enhancing proteins in shrimp Litopenaeus vannamei stimulated with vitamin C and Chinese herbs." Fish & Shellfish Immunology 31: 736-745.

Stentiford, G. D., D. M. Neil, E. J. Peeler, J. D. Shields, H. J. Small, T. W. Flegel, J. M. Vlak, B. Jones, F. Morado, S. Moss, J. Lotz, L. Bartholomay, D. C. Behringer, C.

Hauton and D. V. Lightner (2012). "Disease will limit future food supply from the global crustacean fishery and aquaculture sectors." Journal of invertebrate pathology 110: 141-157.

AFâ„Ś More information Matthew A.G. Owen, Ph.D Skretting ARC, Norway E. matthew.owen@skretting.com

HE 6TH GLOBAL EED AND FOOD CONG RESS 2019

THE 6TH GLOBAL FEED AND FOOD CONG RESS

NGKOK, THAILAND. 11—13 MARCH

THE FUTURE OF FEED & FOOD ARE WE READY? Date & Venue

Theme & Topics

The 6th Global Feed & Food Congress (GFFC), organized by the International Feed Industry Federation (IFIF) with technical support provided by the Food and Agriculture Organization of the United Nations (FAO) and in collaboration with VIV worldwide will be held at the exclusive Shangri-La Hotel in Bangkok, Thailand, on 11-13 March 2019.

The 6th GFFC theme ‘The future of Feed & Food – are we ready?’ links to the global challenge to provide safe, affordable, nutritious and sustainable animal protein sources through innovative solutions to feed 9 billion people by 2050 and reflects our shared vision to achieve this for a growing world population now and for the future.

Join us

The 6th GFFC will look ahead at key topics for the feed & food chain, including:

The 6th GFFC is expected to attract executive level delegates from Asia, Europe, Africa and the Americas.

Join us and you will: — Experience exceptional speakers who will share their insights on the future of feed and food.

— Network and discuss strategy with business leaders, senior government officials, experts and policy makers from the feed & food value chain.

— Engage with leading animal nutrition and food companies, food chain partners, international organisations, national authorities and international civil society.

Organiser

Technical Support

— Digital Revolution — Sustainability — Feed & Food Safety — Nutritional Innovation — Global Regulations & Policy — Markets & Trade — Future of Farming Systems

How to register?

For more information please visit www.gffc2019.com

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The 6th Global Feed & Food Congress 2019. 11 – 13 March 2019, Shangri-La Hotel, Bangkok, Thailand. Learn more at www.gffc2019.com

36

Phytotherapy as an alternative to use of chemicals in aquaculture production and aquatic feeds

By Gianluigi Negroni use of antibiotics, especially those with a wide spectrum, goes hand in hand with resistance to the same pathogens.

Dangerous pathologies are a serious risk to aquaculture producers, and the resistance of bacterial agents to antibiotics is one of the major problems facing the industry. In the case of pathological events, there are currently no other solutions to antibiotics and other chemotherapeutic substances, especially for intensive and hyper-intensive breeding.

Chemotherapeutic agents, although designed to combat diseases of fish and other aquatic animals, also leave harmful contaminants in fish and unwanted residues in the water and on treated surfaces. These chemical agents address the problems of fish diseases and disinfection, but also do more harm than good to aquaculture facilities as well as to the environment as a whole. It is then more beneficial to invest in “greener� solutions that can both be good for fish products and the environment. In this article, phyto (from plants and algae) therapies and their most common

products will be discussed as better alternatives for aquaculture treatments and processing.

Inappropriate use of antibiotics in the aquaculture supply chain Unfortunately, there is often a redundant use of antibiotics, disinfectants and synthetic chemotherapeutic substances in both human and veterinary medicine and plant defense. The massive and periodical

Natural therapies The use of herbs, algae and plants could replace the present chemotherapeutic agents as agents against pathogens, promoters of growth and use in increasing feed productivity as in biosafety. The above needs appropriate studies to better define the characteristics and effects in farms; until now little has been done.

Naturopathy (also called phytogenic therapy, herbal medicine or phytotherapy) is the use of natural plants

37 and herbal extracts. Numerous herbs and plants have been used as home remedies in many cultures around the world for thousands of years to treat humans, farmed animals and plants.

Some of these remedies have strong characteristics against pathological events and diseases with antibacterial, antiviral and antifungal effects, so they are also dangerous if used inappropriately (e.g., rotenone can be highly toxic in large doses).

Other properties are recognized for principles such as anti-stress, growth promoters, appetizers, tonics and immune stimulants.

Other favorable features and benefits of the active ingredients contained in plants and algae, as an alternative to the industrial principles currently used, are: more environmentally friendly, bio-degradability and biocompatibility, and low toxicity (depending on concentration). Up to now, no immunological plant pathogens have been discovered. In fact, the plants used usually have a combination of several active ingredients from which pathogens struggle to defend themselves.

Despite the wide global knowledge about the benefits of medicinal herbs and algae, only a few natural plant/ algal principles are commercially available for industrial aquaculture.

Phytotherapy utilization in aquaculture The strong anti-pathogenic action characteristics of some plant and algal species are well known and documented in the aquaculture sector. The

action of flavonoids, phenols, polysaccharides, terpenes, and proteoglycans can considerably prevent and treat disease and all types of pathogen in aquaculture production such as bacteria, viruses, and parasites. Vibrio presence in the species Penaeus monodon could be reduced by a third when herbs (Solanum trilobatum, Andrographis paniculata, and Psoralea corylifolia) are given as feed to Artemia during the larval phases of P. monodon as feed (Citarasu et al. 2002 and 2009).

Some Chinese researchers have reported remarkable antimicrobial activities from extracts from five herbaceous species (Stellaria aquatica, Impatiens biflora, Oenothera biennis, Artemisia vulgaris, and Lonicera japonica) against 13 species of bacteria and two viral pathogens of fish species (Shangliang 1990).

Aeromonas salmonicida and Edwardsiella ictaluri were the major bacteria that were sensitive to herbal extracts.

S. aquatica was the most effective extract in terms of the highest number of inhibited bacteria and the degree of inhibition. L. japonica showed the highest degree of inhibition against infectious pancreatic necrosis (IPN) and infectious hematopoietic necrosis (IHN) viruses while A. vulgaris and S. aquatica inhibited only the IHN virus.

Numerous species of Indian herbs and plants such as A. marmelos, C. dactylon, L. camara, M. charantia and P. amarus have shown strong antiviral activity towards white spot syndrome virus (WSSV) when extracted (Kolkovski 2013). Numerous studies have also been published demon-

Despite the wide global knowledge about the benefits of medicinal herbs and algae, only a few natural plant/algal principles are commercially available for industrial aquaculture.

strating the antibacterial and antiviral effects of plant extracts with different species of fish and crustaceans. Plant extracts are also known for antifungal and antiparasitic properties.

One study found that a mixture of plant extracts including Benedenia seriolae added to fish feed, helped to decrease fish skin and gill parasites.

P. kurroa was used in experiments to eliminate the free radicals and to reduce the stress in P. monodon as with other herbs (Citarasu et al.

2006). The use of garlic and onion extracts, included in the formula for the production of pellets in the daily ration of farmed shrimp, contributes to the prevention of bacterial infections (Direkbusarakom 2004). A scientific study also highlighted the antibacterial activity of guava (Psidium guajava) against the pathogenic bacteria of crustaceans (Direkbusarakom et al. 2004).

Minimum inhibitory concentrations were tested on Vibrio and A.

38 hydrophila at 1.25 and 0.625 mg/ml respectively. Still others found how guava (Psidium guajava) and neem (Azadirachta indica) extracts eliminated luminescent bacteria from the black tiger shrimp (P. monodon) more effectively than tetracycline (Mahfuzul et al. 2007).

Hatchery specialists that practice maturation diets in crustaceans (P. monodon) use natural commercial products (e.g., Nutrabrood). Such products use the hormone production properties of some plant extracts (W. somnifera, M. pruita, F. asafoetida, and P. longum). In particular, they are used for the production of off-season gametes or with species with problems of fertilization and gonadal development, low sperm mobility and low spermatic volumes (Babu 1999).

Asparagus racemosus used in India as Ayurvedic principle (ancient Indian medicine system) is also used as a growth promoter and has similar effects even on larvae of Labeo rohita (Kavita and Sharma 1999).

Finally, scientific experiments were carried out with specific advanced acidifiers in a species that does not produce acids in its digestive system such as L. vannamei, against the presence of Vibrio spp (a key pathogen for the species). The results were excellent with a positive linear effect on the survival of the shrimp at 60 days between the control and the various treatments with the specific acidifier. The same group of researchers has carried out tests with flavonoids on carp to test the possibilities of intestinal health,

palatability of the ration, improvement of digestive processes, and improved absorption of nutrients as results of the antimicrobial and antioxidant effects of phytogenic preparations. The results of the carp growth were marked after six weeks of breeding, with differences of more than 20% in daily increase (Tzuc et al.

2014). Research at the aquatic animal health laboratory of the Institute of Desert Research, which carried out experiments with rosemary extracts with respect to various pathogens, including S. iniae and S. agalactiae, gave excellent results in vitro and in vivo for infected tilapia (Zilberg et al.

2010). The use of antibiotics (oxytetracycline) showed bacterial resistances that were not shown in the rosemary extract.

39

Healthcare concept of the digestive tract The combined efforts of an appropriate formulation of rations, breeding management, biosafety, and the use of additives can optimize the achievement of “gut health” or “gut welfare” (or so called biodome welfare according with the latest studies), which is defined as “the ability to execute normal physiological functions, to maintain homeostasis that support the ability of the bred animal to overcome without consequences the stresses of infectious and non-infectious diseases” (PlusVet Animal Health 2013). During the life cycle on the farm, having a healthy digestive tract is important to extract maximum productivity without the use of antibiotics. The herbal principles can replace antibiotics for promoting digestive tract health of animals raised in aquaculture.

Antibiotic growth promoters currently used can be replaced by additives with bactericidal or bacteriostatic activities, such as: • Zinc oxide and copper sulphate: have good antibacterial properties but are harmful to the environment when used in high doses; used with limitations in some countries.

• Not all organic acids are effective anti-microbials; effectiveness depends on their hydrophobicity and dissociation capacity.

• Many plant and algal extracts in vitro demonstrate an antibacterial activity, many of which confirm the ability to cope with in vivo experi-

ments. Plant extracts and organic acids have a synergistic effect. With in vivo experiments, synergistic effects have been confirmed (PlusVet Animal Health 2013).

Additives to nutrition can also promote antimicrobial action and indirectly positively influence the health of the digestive tract, such as: • Enzymes, which reduce the level of polysaccharides (anti-nutritional factors) • Probiotics and prebiotics, which increase the number of beneficial bacteria • Immune boosters to help prevent infections

The possible role of plant and algal extracts in aquaculture Plants contain an abundance of chemical compounds that allow them to resist attacks by micro-organisms and insects. These compounds defined as “secondary” or “phytochemical” can potentially provide growth promoting activities in aquaculture species, as mentioned in the following points: • Antioxidant properties: Antioxidants help the body to reduce stress caused by free radical damage. The flavonoids and phenolic acids, widely present in higher plants, are effective against the destructive effect of ROS (Reactive Oxygen Species, the so-called free radicals). Some compounds of basil (Ocimum basilicum), cinnamon and

other plants possess a strong antioxidant activity (Chanu et al.

2012).

• Microcide direct effects: Some components have the ability to adhere to the cell wall, thus forming breaks and killing the microorganism.

Canella and marjoram are examples of plants with micro biocidal characteristics of their active ingredients, as well as guava (Psidium guajava), basil, Indian almond (Terminalia catappa), cannabis (C. sativa), oregano (Origanum vulgare with more than 30 antibacterial principles), and many more.

For many years, the undersigned has been using natural extracts of citrus seeds to prevent bactericidal and fungicidal diseases in frogs during the reproduction, egg and larval stages of frogs (known for their sensitivity to synthetic antimicrobials), trout, marine fishes and crustaceans. The benefits are summarized below: • Antiviral activities: Numerous active ingredients present in plants have a potent viricidal activity in crustaceans and fish.

• Antifungal activity: Herbal extracts can cause the lysis of the cell

40 membranes of myocytes to alter their metabolism and even bring them to death. Note that fungal infections are controlled also by basil (Ocimum basilicum), tamarind (Tamarix dioica) and Rhazya stricta.

• The protective effects of herbs against subacute aflatoxicosis in Oreochromis niloticus were solved with extracts of rosemary and parsley, both of which were able to protect fish from aflatoxynosis even at low administration volumes (El-Barbary & Mehrim 2011). Other studies used macro algal extract mixed with clay as an anti-mycotoxin agent.

• Anti-stress activity: Numerous herbs (Withania somnifera, Emblica officanalis, Asparagus racemosus, Ocimum sanctum, Tribulus terrestris and Piper longum) are believed to have adaptogenic, anabolic effects, increasing the vital energy of animals.

Picrorhiza kurroa is used as an antistress factor in shrimp farming (Citarasu 1998) like the bioflavonoids extracted from Toona sinensis, which acted as antioxidant and anti-stress factors in Litopenaeus vannamei infected by Vibrio alginolyticus (Hsieh 2008).

• Hormonal activities: P. monodon fed maturation diets containing extracts of Withania somnifera, Mucuna prurita, Ferula asafoetida and Piper longum, have increased fertility and the weight of the gonads, reducing the period between the mutes compared to the control shrimp groups (Babu 1999). Similar results were obtained with semi-herbal commercial diets based on herbs with P. vannamei.

• Probiotic effects: Some plants rich in oligosaccharides, such as chicory, are capable of stimulating the growth of beneficial bacteria such as lactobacilli and ibifidus bacteria without promoting the growth of pathogenic bacteria.

• Prebiotic effects: instead of stimulating the growth of beneficial bacteria, they also increase the production of short chain fatty acids (SCVFA) with lactobacilli and Bifidobacterium. SCVFAs inhibit the growth of numerous proteolytic putrescent bacteria.

• Some plant extracts are not able to stimulate the growth of lactobacilli, so they are not real prebiotics, but instead can promote the growth of SCVFA with lactobacilli present in the intestine. Cinnamon and garlic are some of these promoters of the production of SCVFA. • Blocking the adhesion of bacteria on the surface of the digestive tract: Some phytochemical compounds, such as those contained in the carob, can block the adhesion of pathogens in the intestinal mucous layers, thus preventing the development of infection.

• Immune-stimulatory effects: Interesting immune stimulant properties of plants have been recognized in extracts from Emblica officinalis, Cynodon dactylon, Adhatoda vasica, Ocimum sanctum, Withania somnifera, Myristica fragrans, Aegle marmelos, T. cardiofila, Picrorhiza kurooa and E. alba against bacteria (Vibrio harveyi, A. Hydrophila) and viruses (e.g., WSSV in prawns). Some algal extract also

work as the above plants.

• Lymphoid tissues associated with other digestive tract health GALT (gutassociated lymphoid tissue) play a key role in the digestive immunity of animals raised in aquaculture.

Prebiotic compounds, such as chicory oligosaccharides, can stimulate and induce beneficial effects on gastric health by positively influencing local digestive immunity through the prompt response of GALT. Other plant substances such as fungal polysaccharides or garlic allicin can be used in general (on the whole animal) as an immuno stimulant because they activate both innate and adaptive immunity.

• Digestive enzymes secretion and appetite stimulation: some plant components, such as those contained in the canella (with cinnamaldehyde), pepper (with piperine and other substances) and cardamom are able to stimulate the production of enzyme secretions in the digestive tract of the pancreas and intestinal mucosa, inducing a better feed efficiency and an increase in zootechnical performances.

Conclusion The development of pathogens resistant to the medication and disinfectant spectrum currently applied in zootechnical, and particularly aquaculture industries, originates from all areas of animal husbandry.

Plants, algae, herbs and the extracts of their various parts are a great resource and a great alternative to the use of chemotherapeutic agents,

41 disinfectants and other banned substances that could improve the treatment of humans and animals reared in the aquaculture industry, as well as the environment where these substances would spread. The legislative bodies must seriously consider formulating and implementing regulations to allow the appropriate use of herbs, plants and algae in animal production, particularly in aquaculture and processing.

Furthermore, the certification of herbal products to be used in the breeding sector is essential for breeders who will use them, especially for those that are dealing with organic production aquaculture.

It must be noted that phytotherapy cannot quickly replace chemotherapy, although it may mitigate its use, especially in the following applications: adjustment of diets, appropriate use of good animal husbandry, appropriate use of the principles of known medicinal plants, and increase in water quality and consequently of the environment in which the aquatic animals are kept.

References Babu, M. (1999). Developing bioencapsulated ayurvedic product for maturation and quality larval production in P. Vannamei (doctoral dissertation).

Manonmaniam Sundaranar University, Tirunelveli, India.

Citarasu, T., Immanuel, G., & Marian, M. P. (1998). Effects of feeding Artemia enriched with stresstol and cod liver on growth and stress resistance in the Indian white shrimp Penaeus indicus postlarvae. Asian Fisheries Science 12(1998), 65-75.

Citarasu et al., 2002 and 2009 as cited in Kolkovski, S. (2013). Herbal medicine in aquaculture. International Aquafeed, 1306, 34-37.

Citarasu et al., 2006 as cited in Kolkovski, S. (2013). Herbal medicine in aquaculture.

International Aquafeed, 1306, 34-37.

Chanu, T. I., Sharma, A., Roy, S. D., Chaudhuri, A. K., & Biswas, C. (2012).

Herbal medicine—an alternative to synthetic chemicals in aquaculture feed.

World Aquaculture, 45(2012), 14-16.

Direkbusarakom, S. (2004). Application of medicinal herbs to aquaculture in Asia.

Walailak Journal of Science and Technology, 1(1), 7-14.

Direkbusarakom, S., Ezura, Y., Yoshimizu, M., & Herunsalee, A. (1998). Efficacy of Thai traditional herb extracts against fish and shrimp pathogenic bacteria. Fish Pathology, 33(4), 437-441.

El-Barbary, M. I., & Mehrim, A. I. (2011).

Protective effect of antioxidant medicinal herbs, rosemary and parsley, on subacute aflatoxicosis in Oreochromis niloticus.

Journal of Fisheries and Aquatic Sciences, 4 (4), 178-190.

Hsieh, T. J., Wang, J. C., Hu, C. Y., Li, C. T., Kuo, C. M., & Hsieh, S. L. (2008). Effects of Rutin from Toona sinensis on the immune and physiological responses of white shrimp (Litopenaeus vannamei) under Vibrio alginolyticus challenge. Fish and Shellfish Immunology, 25(5), 581-588.

Mahfuzul Hoque, M. D., Bari, M. L., Inatsu, Y., Juneja, V. K., & Kawamoto, S. (2007).

Antibacterial activity of guava (Psidium guajava L.) and neem (Azadirachta indica A. Juss.) extracts against foodborne pathogens and spoilage bacteria.

Foodborne Pathogens and Disease, 4(4), 481-488.

PlusVet Animal Health. (2013). Can plant extracts replace antibiotic growth promoters?

Shangliang, 1990 as cited in Citarasu, T. Herbal biomedicines: A new opportunity for aquaculture industry. Aquaculture International, 18(3), 403-414.

Tzuc, J. T., Escalante, D. R., Rojas Herrera, R., Gaxiola Cortés, G., & Ortiz, M. L. (2014).

Microbiota from Litopenaeus vannamei: Digestive tract microbial community of Pacific white shrimp (Litopenaeus vannamei). SpringerPlus, 3(280), 1-10.

Zilberg, D., Tal, A., Froyman, N., Abutbul, S., Dudai, N. & Golan-Goldhirsh, A. (2010).

Dried leaves of Rosmarinus officinalis as a treatment for streptococcosis in tilapia.

Journal of Fish Diseases, 33(4), 361-369.

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More information

Kavita, K., & Sharma, L. L. (1999). Use of herb (Asparagus racemous wild) supplemented diet for promoting growth in the fry of Labeo rohita. In M. Mohan Joseph, N. R. Menon, & N. Unnikrishnan Nair (Eds.), The Fourth Indian Fisheries Forum Proceedings (pp. 241-243). Kochi, India.

Kolkovski, S. (2013). Herbal medicine in aquaculture. International Aquafeed, 1306, 34-37.

Gianluigi Negroni E: gigineg@gmail.com

43

Importance of plant-derived compounds and/or natural products in aquaculture

By Vikash Kumar and Peter Bossier, Department of Animal Production, Ghent University

Aquaculture, comprising all forms of culture of aquatic animals and plants in freshwater, brackish water or marine environment, is the fastest growing food producing sector in the world with an average growth rate of 7.1 % per year (FAO 2013). However, low growth rate of fish species, stressful condition due to water quality and disease outbreak is considered to be a significant constraint to the development of this sector, causing huge economic losses per year worldwide. Conventional approach such as antibiotics and disinfectants had limited success in the mitigation stressful conditions in aquaculture sector. Moreover, their usage is under severe scientific and public scrutiny due to development of (multiple) resistance. Because of such concerns, FAO has concluded that there is an urgent need for the development of alternative control technique in aquaculture, with the emphasis on prevention, which is likely to be more cost-effective than cure (Defoirdt et al. 2011). In fact, plant-derived compounds and/or natural products have been reported to improve water quality, enhance

growth performance and feed utilization, reduce stress and confer protection and/or enhance immune reactivity to pathogens in aquatic animals. In addition, they can be administered as a whole plant or parts (leaf, root or seeds) or extract compound, via water routine or feed additive, either singly or as a combination of extract compound or even as a mixture with prebiotics or immunostimulants (Table 1). Hence, the natural compounds/molecules can be a promising alternative control technique to avoid stressful condition and prevent aquatic disease in aquaculture system (Baruah et al.

2015).

the cost of production (Gatune et al.

2014). In fact, with increased growth rate of aquaculture there is an increasing demand of high quality fish feed and the high price of fish meal (important source of protein) made it necessary to evaluate alternative protein source of plant origin in fish diets as partial or total replacement

The increasing incidence of drug-resistance pathogens from the use of chemical compounds have drawn the

Plant-based compounds as growth promotors Plant-based compounds have the potential to provide diet containing all the necessary nutrients in suitable proportion to increase the feed intake by improving the palatability of diet, enhances the digestive enzyme activity and optimize the growth performance of fish, while reducing

attention the pharmaceutical and scientific communities towards the use of plant based compounds as a potential antimicrobial agent

44 Table 1. Role of plant-based compounds in fish and shellfish health

Class

Sub-class

Example

Role in aquatic species

Phenolics

Quinones,

Allium sps. (Allium cepa, Allium sativum, Immunostimulant, antioxidant,

flavonoids,

Allium tuberosum), Cynodon dactylon,

antimicrobial, growth promotor, anti-

flavones, tannins,

Viscum album, etc.

helminthic, antiviral

Camellia sinensis, Nicotiana tabacum,

Immunostimulant, antioxidant,

Aconitum napellus, Atropa belladonna,

antimicrobial, growth promotor, anti-

Conium maculatum, etc.

helminthic, antiviral

Terpenoids and

Pistacia terebinthus, Lavandula

Immunostimulant, antimicrobial,

essential oils

angustifolia, Mentha piperita,

antioxidant, anti-helminthic, growth

Melaleuca alternifolia, etc.

promotor

flavonols Alkaloids

Lectins and

Glycine max, Arachis hypogaea, Triticum Antioxidant, antiviral,

polypeptides

aestivum, Cocos nucifera, etc.

immunostimulant

Polyacetylenes

Anethum graveolens, Carum, carvi,

Immunostimulant, antimicrobial,

Daucus carota, etc.

antioxidant

for fish meal. There are several potential plant-based compounds are used e.g., essential oils, alkaloids, flavonoids, saponins, polyphenols, etc. that shows heath benefiting effect in aquaculture species.

However, the presence of antinutritional factors must be considered and evaluated in plant products before the application in fish feeds as it can adversely affect the utilization of food by fish.

Plant-based compounds as immunostimulant The use of natural or plant based immunostimulants growing interest for boosting the defense mechanism (innate or non-specific) and conferring protection of animals from infectious diseases. Several plants based

compounds e.g., berberine, gelselegine, piperine (alkaloids); Zligustilide, tetramethylpyra-zine (essential oils), luteolin, apigenin (flavones), phloroglucinol, quinones, etc. are reported as immunostimulatory characteristics and have been used in aquaculture enhance the immune system and improve the fish health (Kumar et al. 2018). Plant based products have been also demonstrated to facilitate the function of phagocytic cells (e.g., macrophages, monocytes, granulocytes), increased their bactericidal activity, and stimulates the natural killer cells, complement, lysozyme activity and antibody responses in fish and shellfish. In fact, the role of plant based immunostimulants in shellfish aquaculture is more significant as they mostly rely on innate immune system to combat infectious pathogens.

There are several reports, which suggest that treatment of crustacean species (like brine shrimp, Macrobrachium sps) with polyphenols, significantly enhances the innate immune response and provide protection to abiotic (salinity, heat) and biotic (pathogenic bacterial infection) stressors (Figure 1, 2) (Baruah et al. 2014; Kumar et al.

2018).

Plant-based compounds as anti-microbial agent The increasing incidence of drugresistance pathogens from the use of chemical compounds have drawn the attention the pharmaceutical and scientific communities towards the use of plant based compounds as a potential antimicrobial agent. Plants

45

Fig 1. Phloroglucinol at an optimum concentration confers protection to Artemia larvae against Vibrio parahaemolyticus.

are rich source of bioactive compounds like alkaloids and glycosides, and synthesize aromatic compounds mostly phenols or their oxygen substituted derivates. Medicinal plants have been reported to contain phenolic rings (quinones, flavones,

flavonoids, tannins, coumarins), isoprene structures (terpenoids and essential oils), heterocyclic nitrogen compounds (alkaloids), or some heterogenous structures and their use in traditional medicine as antimicrobial agent has been known for

thousands of years around the world (Wunderlich et al. 2017). As an alternative to the conventional methods of fish disease treatment, like antibiotics and other chemical compounds, plant based products, e.g., essential oils and phenolic compounds have been tested and used as an efficient and alternative treatment against microbial infection in aquaculture. The important function of plant based compounds as antimicrobial includes binding to substrate or metal ions and making unavailable for microbial pathogens, microbial cell membrane disruption, binding to bacterial cell adhesins or other proteins and inhibiting the binding of bacteria to cell membranes, inactivating the microbial enzymes, blocking the viral cell fusion or adsorption in host cell, etc. Moreover,

46 FAO Fisheries and Aquaculture. (2013) Report of the FAO/MARD technical workshop on early mortality syndrome (EMS) or acute hepatopancreatic necrosis syndrome (AHPNS) of cultured shrimp.

Report n° 1053. Hanoi, Vietnam.

Gatune, C., Vanreusel, A., Ruwa, R., Bossier, P. and De Troch, M., 2014.

Growth and survival of post-larval giant tiger shrimp Penaeus monodon feeding on mangrove leaf litter biofilms. Marine Ecology Progress Series, 511, pp.117-128.

Fig 2. Induction of Hsp70 protein in Artemia larvae either or not pretreated with

Fig 3. Residual effect of plant based or natural compounds and conventional compounds in aquaculture species and environment (Adapted from Wunderlich et al. 2017).

the natural or plant based products are preferred because of their biodegradability in the environment (Figure 3), i.e., the residues from plant derived compound treatment tend to be biodegradable in the water whereas, from antibiotics or other chemical treatment might have effect on non-target organism (e.g. other non-target culture fish and shellfish species).

References Baruah K, Norouzitallab P, Linayati L, Sorgeloos P, Bossier P. Reactive oxygen species generated by a heat shock protein

(Hsp) inducing product contributes to Hsp70 production and Hsp70-mediated protective immunity in Artemia franciscana against pathogenic vibrios. Dev Comp Immunol. Elsevier Ltd; 2014;46: 470–479. doi:10.1016/j.dci.2014.06.004 Baruah K., H.P.P.D. Phong, P. Norouzitallab, T. Defoirdt, and P. Bossier. 2015. The gnotobiotic brine shrimp (Artemia franciscana) model system reveals that the phenolic compound pyrogallol protects against infection through its prooxidant activity. Free Radical Biology and Medicine 89:593-601.

Defoirdt T., P. Sorgeloos, and P. Bossier.

2011. Alternatives to antibiotics for the control of bacterial disease in aquaculture.

Current Opinion in Microbiology 14:251258.

Kumar, V., Baruah, K., Nguyen, D.V., Smagghe, G., Vossen, E., Bossier, P., 2018.

Phloroglucinol mediated Hsp70 production in crustaceans : protection against Vibrio parahaemolyticus in Artemia franciscana and Macrobrachium rosenbergii. Front. Immunol. 9. doi:10.3389/fimmu.2018.01091 Wunderlich, A.C., Zica, É.D.O.P., dos Santos Ayres, V.F., Guimarães, A.C. and Takeara, R., 2017. Plant-Derived Compounds as an Alternative Treatment Against Parasites in Fish Farming: A Review. In Natural Remedies in the Fight Against Parasites. InTech.

More information

Vikash Kumar E: Vikash.Kumar@ugent.be

Peter Bossier

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48

Dietary supplementation of Hydroxy-Selenomethionine enhances the reproductive performance and antioxidant status of fish broodstock, larvae and juveniles

By Martin Guerin, Aquaculture Nutrition Consultant, Kuala Lumpur, Malaysia In nature, fish produce large numbers of small-sized fry that face difficult and multiple challenges to survive in their aquatic environment. This small size, complicated by important metabolic and morphological changes, means that fish fry survival rates can be very low before they reach adult size. Hatcheries and nurseries strive to offer better environmental and nutritional conditions in order to improve those survivals. However, despite these efforts, for many species, survivals less than 50% from hatched egg to juvenile sold to commercial farms are not uncommon.

Salmonid hatchery-produced fry have relatively good survival chances thanks to large-size eggs containing a large yolk sac that delivers important nutrients to the swim-up fry for several days until it is capable of

feeding itself efficiently. This biological cost of producing large size eggs and fry is compensated by a number of eggs that is often much

smaller in salmonids than in marine fishes such as gilthead seabreams.

Indeed seabreams produce instead larger quantities of smaller eggs that

49 will need to go through several larval stages after they hatch before they reach the juvenile stage. Irrespective of size and number of eggs, in all species, reproductive performance is often measured by the number of spawns, number of eggs produced, hatching rate and fry survival rate.

Oxidative stress from environment changes, fast-paced metabolism, internal metabolic and morphological changes, pathogens, and interventions by hatchery operators, can rapidly weaken the immunity of fish fry and aggravate mortalities during these early days. Selenium (Se) is a key component of the Seleno-Cysteine, an amino acid located at the reactive site of the selenoproteins such as Glutathione peroxidases (GPx), a family of antioxidant enzymes critical to oxidative stress resistance and immunity. As a result, Selenium is a key dietary component of starter feeds to improve the antioxidant status of fry.

Organic Selenium is now widely acknowledged as having superior biological value over inorganic Selenium such as Sodium Selenite (NaSe) and is the preferred form of

Selenium added in fish hatchery and starter diets. Hydroxy-SelenoMethionine (OH-SeMet, commercialized as SELISSEOÂŽ) represents the latest generation of organic Selenium, that offers not only 100% purity and superior biological value but also high stability under aquafeed processing conditions.

Effects on larvae A recent study at the University of Las Palmas, Gran Canaria, Spain, demonstrated that high levels of OH-

SeMet [6 ppm Se] fed to gilthead seabream larvae improved their growth, survival under stress challenge, and gene expression of GPx , vs a control diet with 4 ppm Se from raw materials or diets with the same levels of Se from an inorganic source: NaSe (Figure 1). In addition, it has been demonstrated that only OHSeMet was able to boost the gene expression of GPx and Superoxide Dismutase antioxidant enzymes in these larvae, unlike Selenium Yeast or NaSe (Figure 2). In further testing, juvenile fish (6.16g ABW) were fed a

Fig. 1. Effect of OH-SeMet supplementation on growth of Gilthead Seabream larvae.

Fig. 2. Effect of OH-SeMet on gene expression of key antioxidant enzymes Glutathione Peroxidase and Superoxide Dismutase.

50

Females fed OH-SeMet accumulated more Se in the muscle and liver than the two other diets with the unsupplemented trout showing

Control diet without added Se (Control diet at 0.8 ppm Se) or supplemented with 0.2 ppm and 0.5 ppm Se from Sodium Selenite or OHSe-Met. Fish fed OH-SeMet had the highest growth rate. The study also showed that OH-SeMet supplementation helped Se deposition in the muscle and liver and reduced lipid peroxidation and MDA concentrations in those tissues.

the lowest levels.

Fig. 3. Effect of OH-SeMet supplementation on number of trout spawning (Se1: no Se, Se2: NaSe, Se 3: OH-SeMet)

Enzyme activity of total GPX

Effects on broodstock More recently a study conducted at the INRA research station in St Pee-sur -Nivelle, France, looked at the effect of dietary Selenium on rainbow trout broodstock physiology, reproductive performance, quality of eggs and progeny. The study compared the effect of feeding broodstock a basal diet with 0.3 ppm Se from raw materials, vs the same diet supplemented with 0.3 ppm additional Se from inorganic source: Na-Selenite or organic source: OH-SeMet for a period of 6 months. Females fed OH-SeMet had the highest number of spawns (24) vs those fed selenite (21) and control diet (17 only, which was significantly lower than Se supplemented diets) (Figure 3). OH-SeMet fed trouts also spawned much earlier than un-supplemented or NaSe supplemented ones. Moreover female fed OH-SeMet had a significantly higher hatching rate than those fed NaSe. Females fed OH-SeMet accumulated more Se in the muscle and liver than the two other diets with the un-supplemented trout showing the lowest levels. Similarly, oocyte Se

GPX1 gene expression

Fig. 4. Efficacy of OH-SeMet supplementation on total GPx activity and gene expression of GPX1 in swim-up fry (Se1: no Se, Se2: NaSe, Se 3: OH-SeMet)

51 was higher in females fed OH-SeMet vs those fed selenite or unsupplemented diets. Finally, feeding OH-SeMet to the parents led to higher levels of GPx activity and GPx gene expression in the swim-up fry, i.e. before they have a chance to be fed extra levels of antioxidants (Figure 4).

Trout egg hatching rates vs inorganic Se supplementation.

In conclusion, these studies highlight the importance of dietary organic Selenium, preferably supplied as pure source of organic Se such as OHSeMet, to fish broodstock, larvae and juveniles. Indeed, OH-SeMet supplementation resulted in improved:

Sources: Mechlaoui et al. 2018, ISFNF, Las Palmas, Wischhusen et al. 2018, ISFNF, 2018

As a result, OH-SeMet is a highly recommended Selenium supplement for hatchery diets aimed at broodstock, larvae, fry or juveniles.

... these studies highlight the importance of dietary AFΩ

preferably supplied as pure

Growth and survival under stress in larvae and juvenile Deposition of Se in the liver and muscle of seabream juveniles and trout broodstock as well as oocytes -

organic Selenium,

source of organic Se such as OH-SeMet, to fish More information

broodstock, larvae and juveniles.

Resistance to oxidative stress:

o Higher gene expression of antioxidant enzyme GPx in seabream larvae and trout swim-up fry

.

o Reduced liver and muscle lipid peroxidation and MDA levels Spawning frequency, and parental transfer of Se to oocytes and improved antioxidant status of swimup fry

Martin Guerin, Consultant, Kuala Lumpur, Malaysia E: martin.guerin.aqua@gmail.com

Hatchery suppliers!

Starting in September HatcheryFeed Magazine is launching a classified market section in each issue.

Contact Michael at sales@aquafeed.com to learn how to bring your products and services to our 7,000+ (and growing) readers with affordable 2”x 2” mini-ads.

52

Improvement of fish seed quality with plant based ingredients

By Alex Diana, Tilman Wilke and Dzenan Hozic, Dr. Eckel Animal Nutrition GmbH & Co. KG

Fish consumption The total food fish supply and hence global consumption has been growing at an average annual rate of 4.3 percent in the period 1961-2009 and of 6.0 percent in the period 19902009, while the world’s population is expanding today at a rate of 1.09% per year. The proteins derived from fish, crustaceans and molluscs account for between 13.8% and 17.0% of the animal protein intake of the human population. World per capita fish consumption increased from an average of 9.9 kg in the 1960s to 20.2 kg in 2013-15 (Figure. 1) and it is projected to reach 21.8 kg by 2025.

The per capita availability of fish and fishery products has therefore nearly doubled in 40 years, outpacing population growth.

Moreover, fish provided more than 3.1 billion people with almost 20 percent of their average per capita intake of animal protein. In addition to being a rich source of easily digested, high-quality proteins containing all essential amino acids, fish provides essential fats (e.g. longchain omega-3 fatty acids), vitamins (D, A and B) and minerals (including calcium, iodine, zinc, iron and selenium), particularly if eaten whole.

Even small quantities of fish can have

Fig. 1. Per capita fish consumption. Source OECD/FAO (2016), “OECD-FAO Agricultural Outlook”

a significant positive nutritional impact on plant-based diets, and this is the case in many LIFDCs and leastdeveloped countries. Fish is usually high in unsaturated fats and provides health benefits in protection against cardiovascular diseases for example.

Availability of freshwater fish seed It has recently been recognized that availability of freshwater fish seed of suitable quality is a major constraint on the development of aquaculture in Asia. Fish culture systems, from intensive, commercial operations to extensive ‘backyard’ ponds require eggs and/or fry of sufficient quality and quantity to meet their production requirements. Stress during nursing,

especially at the sex reversal stage, and transport, which may be either a result of management practices or environmental factors, can reduce performance in grow-out. Normally acute stress results in death, sublethal effects include the reduction in growth, limited reproduction, and immune deficiency, and thus stress is a major issue in the question of fish seed quality.

Sex Reversal and plant based solutions Taking into account the wide spectrum of economic and rearing benefits, sex reversal techniques are paid greater attention over time in aquacultured fish systems. Achievement of high growth rate, prevention

53 of large energy dissipation into reproduction and courtship behavior, reducing aggressiveness, uniformity in size, avoiding undesirable effects of sexual maturation on appearance and meat quality, and reducing undesirable environmental impacts are the main contributory factors in raising interest about monosex fish cultures.

One of the most practiced sex reversal techniques is hormonal induction especially using methyltestosterone that has been tested in more than 25 fish species. However, this technique faces several limitations and disadvantages like time and cost consumption, low survival of sex reversed male or female, delayed sexual maturity, sterility at high dose, paradoxical sex reversal, as well as carcinogenicity of hormonal residues and other health hazards for humans and especially the operators at the farm.

The sex reversal in tilapia farming is such a challenging period, during which farmers are especially concerned about survival of fry.

Nevertheless, it is common practice as the use of exclusively male tilapia in the grow-out stages leads to a better output in terms of volume and thus to a higher profitability. The reason is that male tilapia are more energy efficient and grow bigger and faster compared to their female counterparts. Numerous studies have shown that polyphenol based feed additives, especially those rich in flavonoids, have beneficial effects on animals and humans. These plant extracts are known to have anti-bacterial properties and to improve the immune response of the animal as

Table 1. Performance results. Means in a row with different letters were significantly different (P<0.05)

Control Diet

Anta®Ox

22,712a

30,000b

Survival (%)

76a

100b

FCR

2.85

2.49

SGR (%bw/day)

9.6

10.2

No. Harvested

well as their health and performance.

In critical phases of the production cycle, such as sex reversal, these positive effects are of particularly high importance.

Field A phytogenic feed additive (Anta®Ox

by Dr. Eckel Animal Nutrition, Germany) was supplied to red tilapia during sex reversal in hapas in PElined ponds. The objective was to test whether this polyphenol and flavonoid based additive leads to beneficial effects on growth and survival of fry. Six hapas (5 m2 each) were set up in a double PE-lined pond and stocked with 30,000 red tilapia swim-up fry. The animals were fed a fishmeal-based diet containing 60 ppm 17-α methyltestosterone (MT) over 21 days. The feed allocated per day was divided into five feeds and fed at 8:00, 9:30, 11:30, 13:30 and 16:30 by broadcasting the powdered feed over the water surface. The test diet was prepared by mixing the product Anta®Ox (dosage 0.6 kg/Mt) into the fishmeal followed by addition of an ethanol/MT solution and thorough mixing in a blender-type mixer. Aeration was provided in each

pond by 12 homemade diffusers consisting of PVC pipe and material.

After 15 days the 5 m2 hapas were replaced with 10 m2 ones to supply more space for growth.

The addition of Anta®Ox resulted in a significantly higher survival rate (Table 1). Total harvest of fish per hapa was enhanced by almost 1/3 from 22,712 individuals in the control to 30,000 in the treatment group. The outcome clearly shows the potential of this additive for the improvement of animal health and profitability (Figure 2).

In order to better understand the mode of action of Anta®Ox Aqua, a parallel study aimed at quantifying the minimum inhibition concentration (MIC) that could completely stop the growth of common pathogens found in aquaculture. This is considered an alternative strategy to control pathogenic bacteria. The results of the MIC test show the remarkable antimicrobial properties of Anta®Ox Aqua and its powerful bacteriostatic activity against aquatic pathogens.

These were selected in the group of Vibrio spp, including Vibrio parahaemolyticus that are well-known as a major cause of acute hepatopancreatic necrosis disease (AHPND).

54 The positive outcome (Figures 3 & 4) suggests that the use of AntaÂŽOx Aqua is beneficial for the control of AHPND in the field.

The natural feed additive with its preventive mode of action could be a valid and much more sustainable alternative to the frequent use of therapeutics products such as antibiotics which have resulted in the development of resistant aquaculture pathogens in the past.

References on request.

AFâ&#x201E;Ś

Fig. 2. Trial results with flavonoid-based AntaOx fed during sex reversal of tilapia

Fig. 3. Mic results on petri dish

More information Alex Diana Dr. Eckel Animal Nutrition GmbH & Co. KG E: a.diana@dr-eckel.de Fig. 4. Graph is order by MIC percentage for Vibrio parahaemolyticus since data was available for all raw materials mentioned.

55

EVENTS 2018 & 2019 August

For details of industry events: visit the Aquafeed.com Calendar

15—16 TARS 2018 Chiang Mai, Thailand

17– 19: 9th OMC: High Energy Mariculture 2018 Corfu, Greece

http://www.tarsaquaculture.com

http://www.offshoremariculture.com

20 - 22: Food & Feed Extrusion Technology Short Course Werribee, VIC, Australia https://fie.com.au/events/extrusionaust

25- 29: AQUA 2018 Montpellier, France http://www.was.org

26- 31: 25th Annual Practical Short Course on Aquaculture Feed Extrusion, Nutrition and Feed Management College Station, Texas, USA https://perdc.tamu.edu/event/ aquaculture-feed-extrusion-nutrition-andfeed-management-short-course/

September 25—27: GOAL Guayaquil, Ecuador https://www.aquaculturealliance.org/ goal/

26 - 28: 13th International Aquaculture Forum Guadalajara, Mexico http://www.fiacui.com

27: Aquafeed Workshop Guadalajara, Mexico http://feedconferences.com

October 15– 17: IFFO Annual Conference Rome, Italy http://www.iffoevents.com

19: 7th International Conference on Aquaculture and Fisheries Rome, Italy http://www.worldoceansday.org/ events2017/7th-international-conferenceon-aquaculture-amp-fisheries

22—24: Aquaculture Extrusion Technology short course (South America) Temuco, Chile www.fie.com.au/aquafeedsouthamerica

23 – 26: Latin American & Caribbean Aquaculture 2018 Bogotá, Colombia https://www.was.org/meetings

22 – 23: 12th World Aqua Congress New Delhi, India http://worldaquacongress.org/ about_wac.html

March 7 -11: Aquaculture 2019 New Orleans, Louisiana, USA http://www.marevent.com/ AC19_NEWORLEANS.html

11 -13: 6th Global Feed & Food Congress 2019 Bangkok, Thailand

25 - 27: The 8th International Conference of Aquaculture Indonesia (ICAI 2018) Yogyakarta, Indonesia

http://gffc2019.com

http://icai.aquaculture-mai.org/

12 - 14: VICTAM International Cologne, Germany

29 – November 1: Algae Biomass Summit Salt Lake City, Utah, USA

https://victaminternational.com

http://www.algaebiomasssummit.org/

November 7 – 9: AFIA Equipment Manufacturers Conference San Antonio, Texas, USA

June

12 : 12th Aquafeed Horizons Cologne, Germany https://feedconferences.com

18- 20: Asian Pacific Aquaculture 2019 Chennai, India www.marevent.com

http://www.afia.org/ ev_calendar_day.asp?date=11/7/2018% 208:00:00%20AM&eventid=122

Submit your event details to: editor@aquafeed.com