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Novel additives to reduce the economic impact of disease on shrimp production


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Penaeid shrimp production is under continuous threat from bacterial and particularly viral infections which have caused disastrous collapses of the industry in all major shrimp producing countries. Disease problems in shrimp production are complex and often still poorly understood. Regulations, consumer demands and sustainable management strategies restrict the number of drugs available to treat pathogens. Vaccines are likely to be ineffective in crustaceans, which lack a specific immune system similar to that of vertebrates. Therefore, shrimp producers must consider the seed stock quality, husbandry procedures and healthy nutrition as the major tools to control disease. The current article reports on recent progress in the development of feed additives capable of reducing the impact of diseases on productivity and profitability in shrimp farming.

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Novel additives to reduce the economic impact of disease on shrimp production

  1. 1. I N C O R P O R AT I N G f i s h far m ing t e c h no l og y January | February 2013 Novel additives to reduce the economic impact of disease on shrimp production International Aquafeed is published six times a year by Perendale Publishers Ltd of the United Kingdom. All data is published in good faith, based on information received, and while every care is taken to prevent inaccuracies, the publishers accept no liability for any errors or omissions or for the consequences of action taken on the basis of information published. ©Copyright 2013 Perendale Publishers Ltd. All rights reserved. No part of this publication may be reproduced in any form or by any means without prior permission of the copyright owner. Printed by Perendale Publishers Ltd. ISSN: 1464-0058The International magazine for the aquaculture feed industry
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  3. 3. FEATURENovel additives toreduce the economicimpact of disease onshrimp productionby Peter Coutteau PhD, Business Unit ManagerAquaculture, and Tim Goossens PhD, R&D Engineer GutSupport, Nutriad International NV, BelgiumP enaeid shrimp production is under an average annual growth rate of 18 percent diseases consists of increasing the level of key continuous threat from bacte- over the period 1970-2008, which by far nutrients affecting the health and immunology rial and particularly viral infections exceeds growth for all other aquaculture spe- of shrimp, including vitamin C and E, phos- which have caused disastrous col- cies (FAO, 2010). World shrimp aquaculture pholipids, essential fatty acids, trace minerals lapses of the industry in all major shrimp is producing now well over four million mt of and carotenoids. These ‘booster feeds’ are producing countries. Disease problems shrimp (Valderrama, 2011). This rapid increase often supplemented with immunostimulants, in shrimp production are complex and in crustacean production largely often still poorly understood. Regulations, reflects the dramatic increase in white consumer demands and sustainable manage- leg shrimp culture in China, Thailand, ment strategies restrict the number of drugs Vietnam and Indonesia since 2000. available to treat pathogens. Vaccines are Despite this apparent success likely to be ineffective in crustaceans, which story in terms of production expan-lack a specific immune system similar to that sion, shrimp production in many of vertebrates. Therefore, shrimp produc- regions continues to suffer important ers must consider the seed stock quality, economic losses due to the impact husbandry procedures and healthy nutrition of a wide variety of diseases. Recent as the major tools to control disease. The events illustrate the impact of disease Figure 2: The hepatopancreas is the maincurrent article reports on recent progress outbreaks on shrimp production in organ of the shrimp’s digestive systemin the development of feed additives capable major producing countries. responsible for digestion, absorption and storage of nutrients. Esophagus (E),of reducing the impact of diseases on pro- The white spot syndrome virus gastric mill (GM), hepatopancreas (HP),ductivity and profitability in shrimp farming. (WSSV), one of the main causes mid gut (MG), hind gut (HG), and anus of stagnation in the shrimp indus- (A). try in the nineties, is significantly Diseases are number one threat affecting shrimp production in recent The production of crustaceans has shown years in Mexico and Brazil. Early Mortality mostly derived from the cell envelope of Syndrome (Acute micro-organisms, such as polysaccharides, Figure 1 - Table: Effect of booster feed on production parameters in a farm in NE Hepatopancreatic lipoproteins, and lipopolysaccharides. Brazil during episode of increased disease incidence due to a combination of Necrosis Syndrome, The continuous use of immunostimu-intensive rains and increased incidence of infectious myonecrosis virus (IMNV) AHPNS), for which the lants is generally discouraged due to the and necrotising hepatopancreatitis (NHP). Booster feed based on enhancednutritional specifications and supplementation of an immunomodulator causative agent has not risks for over-stimulation of the immune (AQUASTIM S, Nutriad) versus standard feed. been identified so far defense system. Alternating on/off regimes (Flegel, 2012), is affect- for feed additives is often impractical in farm Control feed Boosted feed % change ing shrimp production operations. Encouraging results to improve in China, Vietnam, disease resistance have been obtained by the total pond area (ha) 25 25 Malaysia, and to a continuous use of health enhancing booster Culture period (days) 107 111 lesser extent, Thailand. feeds based on the selection of the appropri- ate immunostimulants in combination with a Survival (%) 77.1 80.7 +5% Traditional balanced nutritional supply of key nutrients Final weight (g) 12.77 14.01 +10% approaches to to support the enhancement of the immune Harvest yield (kg/ha) 1771 2034 +15% boost shrimp system (Table 1). FCr 1.86 1.85 -1% health through However, the efficacy of various commer- avg weekly growth (g) 0.84 0.88 +6% the feed cially available immunostimulants to improve A traditional stress and/or disease resistance of fish and relative production cost 100% 100% approach to reduce shrimp strongly depends on the type of the relative crop value 100% 119% +19% the impact of shrimp product and on the supply of adjuvant nutri- 44 | InternatIonal AquAFeed | January-February 2013
  4. 4. FEATURE could be effective ways of providing the • Discourage the use of non-sustainable • Nevertheless, the existence of small ‘research arm’ for partnering with ‘large sources of MBt librariesents that are essential to support the buildup both as a digestive gland companies willing to take part in pharma’of the immune system. Commercial context – investment, regu- as well as a storage depot biodiscovery de-risks this activity • Technology Centers for marine lation, IP for energy. Therefore, for big companies and justifies the Novel approaches (1): boosting biotechnology, with specialised infra- • The need for long-term stability means farmers routinely look at perceived need for entrepreneurial the nutritional status and lipid structure might provide ‘one-stop that VC and short-term investment squash preparates to evalu- companies to supply into bigger reserves of the hepatopancreas shops’ for chain connection strategies are not appropriate either ate the nutritional status of pharma (and equivalent ‘big’ compa-Science, technology and infrastructures:of the at set-up or for with Shrimps do not tolerate high levels hepatopancreas, longer survival of nies – food/nutraceutical, cosmetics/ new businesses; encouraging business dietary fat very well. is not being studies ample lipid reserves being • Marine biotech A number of well- cosmeceutical) served by lack of knowledge amongst angel groups and raising awareness show reduced growth at levels above 10 per- an indicator of better resist- • Better and more efficient recogni-cent of dietary lipid. Nevertheless, quality and ance to knowledge-levels amongst these technology-transfer offices and stress and disease tion, development and transfer of • More fora for meeting of scientists would be fruitfulquantity of dietary lipids play a primordial role challenges. academic IP in this area is needed and industrial players would generate in growth and health of shrimp. Shrimp have • Lipid digestion in shrimp Smaller players in innovation should • The attributes and benefits of MBt better understanding and sharing of consider more collaborations between no or very limited capacity to biosynthesize a occurs for a big proportion could be better communicated. In needs and possibilities them, and selling skills and knowledge, number of lipid molecules which are essential intracellular in the hepato- terms of giving MBt a different image, for normal growth, including cholesterol, high- pancreas epithelium from molecules at • A global source of ‘soft’ funding rather than pushing the higher hit rate could be a starting would promote the transition from ly unsaturated fatty acids and phospholipids. where it is transported to to go too far big pharma or trying point proof of concept are often the most the down the value-chain; it is usually too Fishmeal and fish oil to demonstration target organs via the Some of the points raised above were Figure 3. Effect of the supplementation of a and commercial-scale for innovations digestibility-enhancing further in Think Tank 5 in the difficult for small companies to handle important sources of cholesterol and HUFA haemolymphe under the discussed additive (Aquagest®S, • Specific incubator programmes marine form of lipoproteins (Fig. 2). of regulatory on context of of lipid vacuolization in and the in the diet. Increasing cost of these could the cost and stresses Nutriad) the degree Marine Biotechnology be recommended processesingredients has forced formulators to reduce The formation and absorp- the hepatopancreas of shrimp fed the different Environment. feeds during 30 days (van de Braak et al., 2012).dietary specifications for these essential lipids. tion of lipid micelles from Although these nutrient levels may not show the lumen of the hepato-significant differences on growth performance pancreas tubuli is therefore a limiting step to use fats as essential components and as in Companies attending this Think Tank: the lipid digestive process. Digestibility source of energy for growth and surviving feeding trials under controlled conditions, in Aqua Bio Technology ASA - A-Spark Good Ventures - Algal Bioenergy Special Interest Group - Bioalvo - BioNova - BioTech North - they may become critical for maintaining enhancers based on natural emulsifying episodes of stress or disease pressure. The Bretagne Developpement Innovation - CCMar - EMPA - European Marine Biological Resource Centre (EMBRC) - Government of Portugal health and immune defenses under disease agents, selected for their compatibility with enhancement of the lipid reserves in the - Grette Law - innoVactiv - Innovation Norway - JPI Oceans - The Research Council of Norway - Kiel Center for marine natural products challenges and fluctuating ambient conditions the shrimp’s digestive system, have shown to hepatopancreas of white shrimp Penaeus - Marealis - Marine Biotechnology Programme of Ireland - Max Planck Institute - National Research Council of Canada - Nautilus Biosciences Canada - Novagraaf Technologies - Novus International - Soliance - Marine Bio-Technologies Center of Innovation - National University of of encountered in production. be capable of complementing the process of indicus as a result of the supplementation Furthermore, the energy status of shrimp is emulsification and absorption of dietary fats in a digestibility-enhancing additive was demon- Ireland, Galway - OceanGate, Inc. - Oceanomics project, Roscoff - Polaris - Polytechnic Institute of Leiria - PwC - Roscoff Marine Biological largely determined by its lipid reserves depos- the hepatopancreas (Coutteau et al., 2012). strated recently by van de Braak et al. (2012). Station - Saint Malo Agglomeration - ScandiDerma AS - Univeristy College Cork - University of Aveiroited in the hepatopancreas which functions This in turn improves the efficiency of shrimp Histological analyses showed a three-fold AquaStar ® Fast growth in improved environment! ASIAN GATEWAY TO AN AQUATIC WORLD OF WONDER Probiotic strains support gut health. Biodegrading strains and enzymes stabilize water quality and pond bottom. co-located with • Im pr an oved gu d pe • Im rform t health prov ance • Co ed w n ater qual ba trol of pa ity The 4th International Pet cteri thog & Accessories Exhibition a enic For more information, please contact: Naturally ahead • Iman Tam January-February 2013 | InternatIonal AquAFeed | 33 January-February 2013 | InternatIonal AquAFeed | 45
  5. 5. FEATURE proteases, invasion factors cific bacterial biosensors and model organ- or other virulence factors isms. Synergistic blends of different natural (Defoirdt, et al., 2011). compounds resulted to be extremely efficient In recent years, research in QS quenching activity against signaling by focusing on ways to dis- Vibrio harveyi, an pathogen causing vibriosis in turb QS signaling (also penaeid shrimp (Figure 6). called quorum quenching) is therefore gaining par- Putting QS inhibition into Feeding manually from a boat and harvesting shrimp ticular interest (Figure 5). practice: effect of optimising at the CAMACO farm, Panama (with courtesy of Jorge This is especially true in gut health on productivity Cuéllar-Anjel) the field of human medi- and economics of semi- cine, where QS-inhibitors intensive shrimp farmingincrease of the percentage of shrimp with are investigated as potential alternatives to Shrimp are actively ‘grazing’ on the sub-a high degree of lipid vacuolization in the antibiotics in tackling pathogenic bacterial strate present in the pond bottom and hepatopancreas after supplementing the feed infections (Sintim et al., 2010). Interestingly, water column, and therefore highly exposed additive during one month (Figures 3, 4). The chances that bacteria build up resistance to exchanges of microflora between the results of a parallel pond study indicated two against QS disruptors are predicted to be low, environment and the digestive system. This percent higher average body weight (ABW), giving that the selective pressure against these increases the risk for the proliferation of four percent higher survival, and six percent in se non-lethal molecules is limited. This an unfavorable gut microflora or frequent higher biomass for the treatment ponds. stands in stark contrast with what is seen with destabilization of the microflora, which can affect the optimal functioning of the digestive system. Furthermore, the digestive system of shrimp is the main entry port for bacterial and viral infections, which remain a major risk for the profitability of shrimp production. Sustainable approaches to modulate the gut microflora in farmed animals include the use of selected bacteria to inoculate the gut (probiotics), specific nutrients promoting the development of selected bacterial strains (prebiotics), and specific natural compounds Figure 4: Histological determination of the degree of lipid vacuolization of the hepatopancreas in shrimp Penaeus indicus, showing a high (left picture) and low (mostly derived from yeast and herbal extracts (right picture) level of lipid vacuolization (100x magnification; van de Braak et called ‘phytobiotics’) capable of modulating al., 2012) the microflora towards a favorable composi- tion, favoring the development of beneficial However, removal of outliers for survival from conventional antibiotics (Defoirdt et al., 2010). bacteria and inhibiting potentially pathogenic control and treatment set showed eight per- Initial studies of quorum sensing in aquacul- micro-organisms. The latter strategies have cent higher ABW, 12 percent higher survival, ture organisms are very limited but point out the advantage of being easily applicable at and 23 percent higher biomass. exciting results. Halogenated furanones isolated the feedmill on large volumes of feed and from red marine algae, for example, have been avoiding major adaptations of the production Novel approaches (2): Quorum demonstrated to reduce QS-regulated gene protocols at the farm. Sensing technology expression in Vibrio and to protect fish and A synergistic blend of botanical extracts Quorum Sensing (QS) is a form of bacteri- shrimp from vibriosis (Rasch et al., 2004; Defoirdt (Sanacore® GM, Nutriad) was originally selected al communication. Over the last decade, many et al., 2006). At the Nutriad Technology for its bacteriostatic and bactericidal properties bacterial species have been documented to Center, QS technology is being applied in against pathogenic and potentially pathogenic be able to produce and secrete small signal- a novel generation of natural feed additives bacteria in vitro using the disk diffusion method. ing molecules, such as acyl homoserine lac- capable of modulating gut tones or certain oligopeptides, which can be micro flora. Compounds are detected by adjacent bacteria of the same or tested for their capacity to of distinct species. When population density inhibit QS-signaling using an rises, these molecules will accumulate in the array of genetically modi-extracellular environment, thereby providing fied bacterial biosensors and a means for bacteria to quantitatively monitor QS-dependent infection the presence of other bacteria. These signal- protocols in simple model ing molecules will, upon reaching a certain organisms. Using these sensi-threshold concentration, initiate intrabacterial tive assays, potent QS modu-signaling that culminates in the activation of lators, able to shut down specific genes. QS communication is therefore QS signaling at concentra-used by bacteria to synchronize gene expres- tions far below the minimum sion alterations and coordinate biochemical inhibitory concentration, are responses within the entire population. being identified. Different In most pathogenic bacteria from which QS quenching activities are the QS system has been studied, QS has selected for agriculture and Figure 5: Quorum Sensing (QS), an innovativebeen associated with pathogenicity, such as aquaculture species based on mechanism to tackle pathogenicitybiofilm formation and the production of screening work using spe- 46 | InternatIonal AquAFeed | January-February 2013
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  7. 7. FEATURE Furthermore, this synergistic blend has proven to be a powerful interrupter of bacterial QS signaling at concentrations well below minimal inhibitory concentrations, allowing it to effec- tively modulate the gut flora towards a more favorable composition. The supplementation of Sanacore GM promoted growth significantly in healthy shrimp growing under controlled lab conditions; showing a remarkable 20 percent increase of weekly weight gain and four percent improvement on food conversion (Coutteau et al., 2010). The effect of this botanical product showing combined activities in QS inhibition and bactericidal action against a wide range of pathogenic bacteria was verified in a semi- Figure 6: Dose – response of a synergistic blend of botanical compounds (SANACORE GM, intensive shrimp farm in Panama.The second Nutriad) on Quorum Sensing signaling activity of Vibrio harveyi. Graphs show signaling activity in QS biosensor system Vibrio harveyi BB170, relative to control, exposed to for production season in Panama, stocked between different dilutions of the product extract (Nutriad Technology Center, in-house results). August-September, is characterised by unstable Table 2: Production results for P. vannamei in Panama during the second production cycle for control ponds and treatment ponds receiving a phytobiotic supplement after 141 days of culture (average and standard deviation of eight replicate ponds of three ha per treatment). average coefficient Weekly Crop Yield Feed (kg/pond of variation for treatment Survival Shrimp size FCr Growth(g/wk) (kg/ha) 3ha) parameters listed (CV%) Sanacore® GM 55.5 ± 7.1 a 16.6 ± 1.5 a 735 ± 78 a 4,170 ± 338 a 1.91 ± 0.23 a 0.825 ± 0.075 a 10% Control 44.6 ± 10.6 b 15.7± 2.9 a 543 ± 90 b 3,464 ± 396 b 2.17 ± 0.39 a 0.776 ± 0.137 a 18% % change Sanacore +24.4% +5.8% +35.2% +20.4% -12.1% +6.3% -41% vs Control P Value 0.0304 0.4395 0.0004 0.0018 0.7130 0.3876 --- THE BEST WAY TO PREDICT THE FUTURE IS TO CREATE IT. —Peter F. Drucker Why retire a workhorse that’s still doing the job? Simply put, your old dryer may be costing you a bundle. In fact, today’s Wenger dryer could save you enough in operating efficiency alone to cover the replacement of your old dryer. Additionally, our new advanced dryer designs give you less potential for cross-contamination and bacteria build-up; feature new direct drive spreaders for level product bed and uniformity of final prod- uct moisture; and afford quicker, easier inspection and cleaning. Contact us now. With new concepts and fresh initiatives, we’re ready to help you develop the product possibilities of the future. Turning ideas into opportunities. PROGRESSIVE AQUAFEED PROCESSING What will tomorrow bring BElGIUm TAIWAN BRASIl CHINA TURkEY INDIAWenger12_AQ_210x147mm.indd 1 8/8/12 12:01 PM January-February 2013 | InternatIonal AquAFeed | 47
  8. 8. FEATURE About the authors Tim Goossens got a Masters degree in Biotechnology at Ghent University, and subsequently worked as an aca- demic researcher in was confirmed by immu- the Laboratory of no-chromatography and Molecular Biotechnology at the University nested-PCR tests. The of Antwerp. After working for four years presence of a synergis- on the characterisation of a gene family tic blend of phytobiot- involved in bone development, he moved ics provided an array of to the KU Leuven, where he took up a antimicrobial activities, doctoral project on neurodevelopment in including quorum sens- the Laboratory of Developmental Genetics. ing inhibition capabilities, After obtaining his PhD. in Biomedical in the shrimp’s digestive Sciences, he joined Nutriad to work as an system. This offers addi- R&D Engineer, focusing on the develop- tional protection against ment and technical support of the Gut co-infections with oppor- Support range of feed additives. Figure 7: Survival percentage at harvest for tunistic bacteria such as control ponds and treatment ponds receiving the vibriosis, often the major Peter Coutteau, cur- phytobiotic supplement in two production cycles in semi-intensive production of white shrimp L. cause of mortality in rently Business Unit vannamei (average and standard deviation of WSSV-infected shrimp Manager Aquaculture eight and five replicate ponds of three ha per (Phuoc et al., 2009). for Nutriad, obtained treatment, respectively; data from Vaca et al., The evaluation in in 1992 a PhD. in 2010, 2011). the second cycle on Biological Sciences at eight replicates per the Laboratory of climatological conditions, resulting in strong tem- treatment allowed a good evaluation of Aquaculture & Artemia Reference Center, perature fluctuations which in turn affect shrimp variability among ponds for the different University of Gent on the filter feeding biol-growth and increase the impact of outbreaks of production parameters. The addition of ogy of Artemia and bivalves. He continued white spot virus (WSSV). The first production the phytobiotic reduced drastically the academic research at postdoc level till 1997 cycle, seeded between January-April, provides variability of production results among on lipid nutrition of bivalves, fish and shrimp, more suitable growth conditions and generally ponds fed the same feed (average coef- publishing over 40 refereed papers in scien-results in better survival and productivity. ficient of variation between ponds for tific journals. In 1997, joined the INVE group, Two treatments were compared which the six production parameters: control as head of research and product develop-only differed with regard to the supplementa- 18 percent versus Sanacore group 10 ment in the aquaculture division. During tion or not of the phytobiotic growth pro- percent; Table 2). This further indicated 2002-2008, he was responsible for global moter (Sanacore® GM) to the standard feed the importance of increased control of product development and customer service used at the farm. The supplementation of the gut microflora on the reproducibility of for feed concentrates and additives as prod-botanical feed additive drastically improved production in semi-intensive pond envi- uct manager farm nutrition for INVE’s survival, amounting to a relative increase with ronments. Business Unit Aquaculture. Following 24 percent and 18 percent compared to the References available on request restructuration of the INVE group in January control group for the second and first cycle, 2009, the support team, research activities respectively (Figure 7). and product lines for aquaculture additives More InforMatIon: Natural White Spot Disease outbreaks were were reorganised under Nutriad’s Business Email: p.coutteau@nutriad.comobserved during shrimp farming in both treatments Website: Unit Aquaculture.under similar frequency and severity; WSSV virus 48 | InternatIonal AquAFeed | January-February 2013
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