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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3. FEATURE
Novel additives to
reduce the economic
impact of disease on
shrimp production
by Peter Coutteau PhD, Business Unit Manager
Aquaculture, and Tim Goossens PhD, R&D Engineer Gut
Support, Nutriad International NV, Belgium
P
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 main
current article reports on recent progress outbreaks on shrimp production in organ of the shrimp’s digestive system
in 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 enhanced
nutritional 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
5. 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 libraries
ents 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 fruitful
quantity 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 processes
ingredients 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 Aveiro
ited in the hepatopancreas which functions This in turn improves the efficiency of shrimp Histological analyses showed a three-fold
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January-February 2013 | InternatIonal AquAFeed | 33
January-February 2013 | InternatIonal AquAFeed | 45
6. 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 farming
increase 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 innovative
been associated with pathogenicity, such as aquaculture species based on mechanism to tackle pathogenicity
biofilm formation and the production of screening work using spe-
46 | InternatIonal AquAFeed | January-February 2013
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8. 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 ---
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January-February 2013 | InternatIonal AquAFeed | 47
9. 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.com
observed during shrimp farming in both treatments Website: www.nutriad.com Unit Aquaculture.
under similar frequency and severity; WSSV virus
48 | InternatIonal AquAFeed | January-February 2013
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