Professional Documents
Culture Documents
Amit Kumar Sinha - Biofloc Article
Amit Kumar Sinha - Biofloc Article
3 EDITORIAL
Finding the key to good biosecurity
4 RESEARCH FOCUS
Breaking inbreeding in domesticated shrimp (Litopenaeus stylirostris) in New
Caledonia
7 NEWS FEATURE
Venezuela passes new law in fisheries and aquaculture
8 RESEARCH FOCUS
Horizon scanning: the potential use of biofloc as an anti-infective strategy in
aquaculture – an overview
11 NEW PUBLICATION
Pearl Oyster Health Management manual on line
12 RESEARCH FOCUS
The BC Pacific Salmon Forum, Nanaimo, Canada
4 15 AQUAFEED
A news roundup courtesy of the Aquafeed website www.aquafeed.com
16 NEWS
Updates from around the globe
22 RESEARCH FOCUS
Selective breeding can reduce salmon lice problems
23 EVENTS
Forthcoming fish and shellfish events
24 COMMERCIAL FOCUS
Cytogenix announces agreement to develop DNA vaccines
28 LABORATORY FOCUS
The Central Fish-Health Laboratory in Israel
32 RESEARCH FOCUS
What’s new in pancreas diseases research?
34 BOOK REVIEW AQUACULTURE
Aquaculture biosecurity: prevention, control
HEALTH I N T E R
NIA BREAKS
36 ORNAMENTALS FOCUS S HRIMP INBREE
DING
A basic overview of multicellular parasitic
infections
38 VACCINE STATISTICS
Monthly Norwegian
vaccine statistics ON THE
provided by Pharmaq COVER ALIEN FISH IND
AQUATIC HEA ICATE
40 RESEARCH FOCUS A farmed shrimp BIOFLOC HA
S
LTH
TO FIGHT INF POTENTIAL
Spread and L stylirosis in New ECTION
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T
he team at Aquaculture Health International is delighted to bring they are “de-motivated” by the environment they are working in. This
you the thirteenth issue of the magazine, celebrating our third is perhaps a more serious issue than many fish health professionals are
full year of production. As always, we have endeavoured to bring aware of, and it’s to the credit of Scarfe, Lee and O’Bryen that they have
you a diverse range of articles, and so we have included contributions included a short chapter on this very topic.
from Israel, Ireland, Norway, India, New Caledonia, France, Venezuela, I once again find myself calling for a more holistic approach, as I
Belgium, the United States of America and Canada. often do in these columns, although this time on a different aspect of
This issue of Aquaculture Health International also contains a review aquaculture health management. It seems to me that there is a tendency
on David Scarfe, Cheng-Sheng Lee and Patricia O’Bryen’s excellent to compartmentalise the myriad functions involved in managing an
book entitled Aquaculture Biosecurity. Although this book was first aquaculture unit so that training in biosecurity and developing a highly
published in 2006, it is still the most authoritative and comprehensive motivated team are seen as having only a tenuous link at best.
publication on this subject that I’ve come across to date. The bottom line is that a properly motivated and valued workforce
Clearly, biosecurity is one of the most important issues for policy is much more likely to take on ownership of the biosecurity process,
makers, regulators, fish health professionals and aquaculturists alike. and this is surely to the advantage of all concerned. After all, sub-
You only have to follow the unfolding saga of the current ISA outbreak optimal biosecurity has the potential to seriously impact on public
in Chile on the news wires to get a sense of the centrality of this issue perception of the industry, in addition to a plethora of other negative
to the aquaculture industry. impacts of the biological and economic variety.
There is, however, one area that we tend to miss when discussing So I find myself musing on a number of issues. For example, how
biosecurity. Decision makers quite rightly have a tendency to focus can the latest advances in business communication theory be utilised
almost exclusively on setting and enforcing norms and standards in and developed to inculcate a sense of biosecurity awareness? What are
this arena, whether it be on an international, national or regional the motivational models that can best be applied to the aquaculture
platform. Where we often fall short, and this is an area that is touched environment to enable staff at all levels in the enterprise to promote
on in Scarfe, Lee and O’Bryen’s book, is the issue of the mindset of and foster “good practice?”
those at the “coal face” in the industry, namely the staff who work on It seems to me that these issues need to be investigated more
the farms. I’ve visited farms where the biosecurity procedures have thoroughly at both a theoretical and practical level. We need all the
been exemplary on paper but have been half-heartedly adhered to, not tools we can to foster and promote optimal biosecurity in whichever
because the staff are unaware of company policy, but rather because sector of the industry we work in. ■
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BREAKING INBREEDING
IN DOMESTICATED SHRIMP
(LITOPENAEUS STYLIROSTRIS)
IN NEW CALEDONIA
BY E GOYARD, C GOARANT, D ANSQUER, F BROUTOI, P BRUN, S DE DECKER, R DUFOUR, C GALINIE, J-R MAILLEZ,
JM PEIGNON, D PHAM, E VOUREY, J PATROIS AND Y HARACHE
IFREMER, DEPARTEMENT AQUACULTURE EN CALÉDONIE, NOUMÉA CEDEX, NEW CALEDONIA
I
n 1993, when temperatures dropped rapidly at the beginning of 2006, with particularly strong exports markets in France, Japan and
the cold water season, significant mortalities occurred among Australia.
the farmed shrimp (Litopenaeus stylirostris) population in New The domesticated strain, introduced into the country in the
Caledonia. The mortalities, due to Vibrio penaeicida, were locally late 1970s and reproduced in captivity since, shows good growth
referred to as “1993 syndrome” and within a few months they had performance profiles and an excellent virus status profile (resistant
spread to all the farms on the island. to IHHNV and free of all other known viruses).
Survival rates (pl’s to commercial size shrimps) declined However, we were convinced that the low number of founders had
repetitively down to 30-35 percent during the winter, making winter induced a high level of inbreeding with an important loss of genetic
production uneconomical. The industry concentrated on seasonal variability, possibly hampering its adaptability to modifications of
production during the warm months, with good results, although the rearing conditions, including resistance to new pathogens.
such an approach created bottlenecks for market availability and The results of the multi-disciplinary Ifremer DéSans programme
peak demand of post-larvae over short periods. pointed out that the outbreaks of seasonal diseases were linked to
At the end of 1997, another sanitary crisis appeared during the the instability of the pond ecosystem (sediment and water column)
warm season in the same farm initially struck with V penaeicida. during intensified production cycles, the virulence of two specific
This was due to a septicaemic summer vibriosis caused by Vibrio pathogens (Vibrio nigripulchritudo or V penaeicida, respectively) and
nigripulchritudo, which has occurred every year since on the the shrimps’ capacity to resist the stress induced by the interaction
contaminated site. V nigripulchritudo (both non-pathogenic and of these critical factors.
highly pathogenic strains) was also isolated in several other sites, but We assumed there was a genetic component to this global
was not always associated with mortalities. resistance capacity. The necessity to re-introduce “new blood” into the
While V penaeicida isolates show a low genetic variability, the strains broodstock line, even through a costly, complex and risky procedure,
of V nigripuchritudo appear highly variable, with striking differences appeared unavoidable. This provided the impetus for a range of
in their pathogenicity level. As New Caledonia’s production certified disease challenge studies using stocks of different genetic origins.
procedure totally prohibits the use of antibiotics in the pond-
growing phase, the presence of this pathogen was considered a major STOCKS TESTED
potential threat to industry expansion. A domesticated SPF strain from Hawaii (High Health Aquaculture
This motivated a specific monitoring policy, undertaken by Ltd) was purchased by the New Caledonian Industry (UPRAC
Ifremer in collaboration with DAVAR, the Nouvelle-Calédonie association), allowing it to test three discrete stocks for their
Veterinary Service. Significant summer mortalities, due to the same performance:
pathogen, appeared in 2003 and 2005 in two other farms. However, • CC. New Caledonian stock from Ifremer and private hatcheries. A
production continued to increase slowly, reaching 2430 tonnes in newly constituted CC control line (to reproduce every year) was
created with 16 bi-parental families and reproduced twice in 2005 dead shrimp found following rapidly dropping temperatures was
and 2006. typical of syndrome 93, suggesting a higher resistance by Hybrids-F1
• HH. Hawaiian stock introduced through a quarantine procedure to V penaeicida. The final weight, and the subsequent growth rate,
(UPRAC) was reproduced at Ifremer, constituting a new HH- were significantly higher for Hybrid F1 (0.22g and 0.24g/day) than
G9 and HH-G10 generations structured each in at least 16 bi- for both parent HH and CC lines (0.15g to 0.17g/day).
parental families from known genealogy, reducing inbreeding to
a minimum. From this G9, batches of juveniles for experimental Summer tests in floating cages (Private farm 2007)
evaluation of the Hawaii stock were constituted in 2005 and 2006. Because no summer mortalities caused by V nigripulchritudo
• Hybrid-F1. HH and CC animals were crossed each year to obtain had ever been observed in the Ifremer ponds, it was decided, in
two first-generation hybrid stocks named F1-2005 and F1-2006 coordination with the Farmers Association, to compare the results
for evaluation of performance, using several families for each of the three populations in a farm showing typical outbreaks of V
combination sex/origin. nigripulchritudo since 2003.
This test was conducted in small floating cages (4m2/3.2m3) installed
EVALUATION OF THE COMPARATIVE in a production pond affected by summer mortalities. Shrimps were
PERFORMANCES OF THE DIFFERENT stocked either as “pure line” cages or under “competition” (mixed
POPULATIONS: PRELIMINARY RESULTS population) during the recurrent outbreak period.
Winter tests in earthen ponds (Ifremer 2006 and 2007) The final survival after 77 days appeared significantly higher in
The different groups arising from 2005 (CC-2005, HH-2005 and Hybrids F1 (51 percent) than the parent lines, which expressed no
F1-2005) and 2006 reproduction scenarios (CC-2006, HH-2006 significant difference between CC and HH (41 and 44 percent) with
and F1-2006) were reared separately in earthen ponds at the Ifremer no significant difference between the conditions of testing (“pure
station. They were then branded with silicone injections (2.9g to 11g line” cages vs “competition” cages).
mean weight) and re-mixed in two 500m2 ponds seeded with 10,000 Haemolymph samples taken at day 29 showed a V nigripulchritudo
shrimp and reared for 130 days using normal production techniques. prevalence of 90 to 100 percent with no difference between groups,
The ponds were harvested after the beginning of the cold season, but a higher number of vibrios in CC samples (50 percent with more
with identification, counting and size sampling of all groups. than 300 vibrios in 10µl) while 50 percent of HH and F1 bore fewer
Under these “competitive” conditions, the average results showed, than 50 pathogens. Further samples did not indicate significant
for both years, a higher survival rate for Hybrid F1 (respectively trends in prevalence between groups, but confirmed a higher number
60.5 and 53.5 percent) compared to HH (60 and 32.5 percent) and of V nigripulchritudo CFUs found in CC haemolymph samples.
CC (51 and 34.5 percent). Although no moribund shrimp could be The average growth rates observed in Hybrids F1 were 40
observed, V penaeicida was not isolated, but the presence of a few percent higher than that of the parent lines (0.28g v 0.19g/day),
Production of Biomass in ponds - year 1 Production of Biomass in ponds - year 2 Production of Biomass in cages with a much better food conversion ratio than the CC line in the
Growth in ponds - year 1 Growth in ponds - year 2 Growth in cages
testing conditions, though this remains to be confirmed in other
Survival in ponds - year 1 Survival in ponds - year 2 Survival in cages
conditions.
240%
220%
A Experimental infections with Vibrio penaeicida (Ifremer 2007)
Six experimental infection tests were undertaken, three each in 2006
200% and 2007, in a bio-safe infection room at Ifremer. The experimental
180%
population comprising the three tagged populations (CC, HH and
Hybrid F1) was reared in an outdoor pond for three weeks. Groups
160% of each population were then introduced in the experimental facility
in 16 or 32 x 200-litre tanks. After acclimatisation for one week, the
140%
animals were infected by balneation or injection with a reference
120% strain of V penaeicida, following a standardised technique.
Natural contamination in 2007 with V nigripulchritudo prior
100%
30% 35% 40% 45% 50% 55% 60% to the V penaeicida experimental infections made the expression
Biomass production in ponds - year 1 Biomass production in ponds - year 2 Biomass production in cages
of results and the characterisation of a specific resistance to V
Growth rate in ponds - year 1 Growth rate in ponds - year 2 Growth rate in cages penaeicida difficult. However, in all experiments, the final survival
Survival in ponds - year 1 Survival in ponds - year 2 Survival in cages
of hybrids F1 was higher than that of CC and HH, with a significant
140% difference for 2006 experiments (57 percent v 50 and 47 percent
B respectively).
120%
100% CONCLUSION
80%
All the tests indicate that F1 hybrids are characterised by faster
growth, independently of the testing conditions: in earthen ponds,
60%
in cages, under competition with other strains or alone; while the
40% slower-growing pure strains CC and HH do not show significant
20%
differences.
Though the present results do not allow a conclusion to be drawn
0%
30% 35% 40% 45% 50% 55% 60% with respect to a specific resistance of the hybrids to the pathogenic
Vibrios, it was shown in all experiments that they provide higher
Average values for survival, growth rate and biomass, obtained in
ponds and cages, is expressed in % of the values observed in survival than any parent population in all cases (pond or cage
Caledonian shrimps (CC) for the same criteria. (Goyard et al., 2008) rearing, winter or summer period and global survival to experimental
handling) and infections. Moreover, indications of a lower pathogen
T
he government of Venezuela passed a new Fisheries and floating cages and pens
Aquaculture Law on March 14 which will have a considerable • algae, crustaceans, molluscs and finfish, and their larval stages
impact on the future development of its fisheries and where appropriate
aquaculture. • aspects of aquatic animal health, including quarantine matters
The new law, known as Ley de Pesca y Acuicultura, published as and genetics
Decree Nº 5930 in the Official Gazette, or Gaceta Oficial de la República and the levels of these activities (eg, artesanal, small-scale operations,
Bolivariana de Venezuela (GORBV), Nº 5877 (extraordinario), on larger commercial operations). Tariffs are established for issuing
March 14, is very extensive in its coverage and attributions, as much various types of permits, certificates, authorisations and other such
with reference to “fisheries” as to “aquaculture”. “documentation” on a sliding scale that favours artesanal producers
It specifically replaces and leaves without any legal effects the and becomes successively more “onerous” for the larger producers.
previous laws on fisheries and aquaculture, published in the GORBV In compliance with the terms of article 47 of Title VI (Communal
Nº 37323 of November 13, 2001, and in the GORBV Nº 37726 of Responsibility) of the law, for example, aquaculture producers at all
May 22, 2003, respectively. levels are now obliged to freely donate, directly and gratuitously to
The National Institute of Fisheries and Aquaculture (Instituto government and local community organisations a minimum of five
Nacional de Pesca y Acuicultura, or INAPESCA), opportunely percent of their production for distribution by those agencies to
created by these previous two laws as a dependency of the Ministry needy sectors of the population.
of Agriculture and Lands, or MAT, is now officially designated as the This article also states quite clearly that the competent authorities
Socialist Institute of Fisheries and Aquaculture (Instituto Socialista can increase this free contribution of five percent as and when
de Pesca y Acuicultura, or INSOPESCA), which continues to be a deemed necessary.
dependency of the MAT. In accordance with the general guidelines as detailed in the text
The printed version of this new law comprises some 120 pages of this new Fisheries and Aquaculture Law, the future development
of text, and its contents should be read very carefully by all those of aquaculture in Venezuela is to be oriented above all else toward
with an interest in aquaculture in Venezuela (preferably with some improving the “social welfare of the less economically favoured
professional input from the interested parties’ financial, legal and communities” within the country.
technical advisers, where convenient and/or necessary). The contents of this law should be carefully read and studied by
The law provides a series of quite clear definitions of the various any interested parties. Potential investors, in particular, would be
types of aquaculture activities to which it refers, eg strongly recommended to adopt a "caveat emptor" approach before
• freshwater and marine reaching any important decisions in principle. ■
• extensive, semi-intensive and intensive, including ponds, tanks,
▲
PRINCIPLE
I
ntensification of the aquaculture industry has in some cases been
accompanied by environmental pollution (high nitrogen toxicity) Bacteria and other micro-organisms require carbohydrates for their
and increasing severity of disease outbreaks. The increased level growth. As protein is the major component of new cell material,
of intensification requires more supplementary protein feed and in nitrogen is also required as an essential element. Thus, microbial
turn contributes to nitrogen pollution in the culture system. utilisation of carbohydrate or any other low nitrogen feed is
Decomposition of dead algae, uneaten feed, fish excreta and accompanied by the immobilisation of inorganic nitrogen. This is
other organic matter releases excess nitrogen in the form of the basic process carried out by the microbial community.
ammonia and nitrite and amplifies the nitrogen toxicity level.
Elevated concentrations of ammonia affect growth, moulting (in BIOFLOC AND WATER QUALITY
shellfish), oxygen consumption and even ammonia excretion, and IMPROVEMENT
can eventually cause mortality of fish/shellfish. Increased ambient Nitrogen control is stimulated by feeding bacteria with carbohydrates,
nitrite concentration negatively affects the growth performance and through the subsequent uptake of nitrogen from the water, by
and survival of fish/shellfish (Mallasen and Valenti 2006). Hitherto, the synthesis of microbial protein. The relationship between adding
many techniques such as photo-autotrophic algae-based systems carbohydrates, reducing ammonium and producing microbial
(green-water systems), frequent water exchange and the use of proteins depends on the microbial conversion coefficient, the C/N
biofilters have been used to remove excessive nitrogen. ratio in the microbial biomass and the carbon contents of the added
However, none of these options has proved to be technically material (Avnimelech 1999).
feasible and economically viable. Thus, the quest to develop an The C/N ratio has been widely used as an index of the rate at which
economical, eco-friendly and user-friendly strategy to remove or organic matter decomposes. If the organic matter is low in nitrogen
optimise the level of such toxic compounds in the pond water has content (ie a high C/N ratio), some of the nitrogen for microbial
become a priority. growth will be obtained from the water column and will become
immobilised as microbial protein. Previous studies have shown
that the immobilisation of inorganic nitrogen only occurs when
The breakdown of PHA inside the C/N ratio of the organic matter is higher than 10 (Lancelot and
Billen 1985). However, Avnimelech (1999) verified that to eliminate
the gastrointestinal tract can be inorganic nitrogen, the C/N ratio in pond should be 10.75, rising to
15.78 when fish are introduced into the holding pond.
carried out via enzymatic and
BIOFLOC AS FISH NUTRIENTS
chemical hydrolysis The bacterial protein and new cells (single-cell protein)
synthesised by the heterotrophic bacterial population are utilised
One such strategy for removing ammonium nitrogen from directly as a food source by the cultured organisms (carp,
water is through its assimilation into microbial proteins by adding tilapia, shrimp), thus lowering the demand for supplemental
carbonaceous material into the system. Nitrogenous compounds feed protein (Avnimelech 1999). Hari et al (2004) reported
present in the water combine covalently with carbon and produce that Penaeus monodon could effectively utilise the additional
flocs of protein molecules. These small protein molecules flocculated protein derived from the increased bacterial biomass as a result
together form a floating mass called bioflocs; these flocs are of carbohydrate addition. Burford et al (2004) suggested that
dominated by bacteria. An important aspect of this process is the “flocculated particles” rich in bacteria and phytoplankton could
potential utilisation of microbial protein as a source of protein contribute substantially to the nutrition of the Litopenaeus
for fish and shrimps. The success of this technique depends upon vannamei in intensive shrimp ponds.
optimising the amount of carbonaceous material to be added.
Recently, it was observed that excess addition of the carbon BIOFLOC AS AN ANTI-MICROBIAL AGENT
source enhances the development of polyhydroxyalkanoate (PHA) It was observed that the regular addition of carbon to the culture
accumulating micro-organisms. The properties of PHAs are very is known to select for polyhydroxyalkanoate (PHA) accumulating
similar to those of organic acid and have been proved to be effective bacteria (Salehizadeh and Von Loosdrecht 2004) such as Alcaligenes
bio-control agents, given that they beneficially affect the host’s eutrophus, Azotobacter vinelandii, Pseudomonas oleovorans and others
microbial balance in the gut. that synthesise PHA granules.
Such granules are synthesised under conditions of nutrient stress, exhibiting bacteriostatic and/or bacteriocidal properties, depending
that is, when an essential nutrient like nitrogen is limited in the on the physiological status of the host and the physiochemical
presence of an excess carbon source. (Avnimelech 1999). These characteristics of the external environment (Ricke 2003).
PHAs are polymers of β-hydroxy short chain fatty acids and if These indicate that biofloc can be a novel strategy for disease
degraded in the gut, they could have antibacterial activity similar management on a long-term basis, in contrast to conventional
to short chain fatty acids (SCFAs) or organic acids. The breakdown approaches such as antibiotic, probiotic and prebiotic application,
of PHA inside the gastrointestinal tract can be carried out via especially since in many cases the efficacy of the latter two remains
enzymatic and chemical hydrolysis (Yu et al 2005). to be demonstrated.
Enzyme hydrolysis is generally carried out by PHA depolymerases Short chain fatty acids have been known for a long time to
produced by various micro-organisms such as Aspergillus fumigatus,
Pseudomonas fluorescens, Comamonas sp and others (Khanna and
Srivastava 2004). Chemical hydrolysis can be carried out by treating
the PHA with NaOH (Yu et al 2005). Pre-treatment of PHA
polymer with NaOH can significantly accelerate the digestibility
of PHA polymer. For example, an increase of up to 85 percent was
demonstrated in pigs (Forni et al 1999).
The working mechanism of SCFA with respect to their antibacterial
activity is not well understood. It could, however, be related to the
reduction of pH, as well as their ability to dissociate, or the pH of
the surrounding milieu (Ricke et al 2003). The antibacterial activity
increases with decreasing pH value. SCFA in undissociated form is
able to pass through the cell membrane of bacteria (Cherrington et
al 1991) and once internalised into the neutral pH of the bacterial
cytoplasm, it dissociates into anions and protons. These ions
influence the stability of functional macro-molecules and inhibit
the action of important enzymes. Bacteria need to pump out excess
protons to maintain the neutral pH of the cytoplasm. Removal of
protons requires consumption of cellular ATP and may deplete the
cellular energy and thus growth is inhibited.
Another possibility is that SCFA may interfere with membrane
structure and membrane protein in such a way that electron
transport is uncoupled and ATP production is diminished. Russel
(1992) speculated that anion accumulation is the primary reason
for the antimicrobial effect of organic acid. Moreover, less direct
antimicrobial activities have also been attributed to SCFA, which
include interference with nutrient transport, cytoplasmic membrane
damage resulting in leakage, disruption of outer membrane
permeability and hindrance in macromolecule synthesis (Ricke
2003).
Apart from inhibiting the growth of pathogenic bacteria by
lowering the pH of surrounding milieu, SCFA have also been
shown to specifically down-regulate virulence factor expression
and positively influence the gut health of animals (Teitelbaum
and Walker 2002). Moreover, these compounds are capable of
inhibit the growth of pathogenic bacteria and they also have been P and Verstraete W 2007. The bacterial storage compound poly-
used in commercial diets of terrestrial animals to control pathogens b-hydroxybutyrate protects Artemia franciscana from pathogenic
such as Salmonella and E coli (Van Immerseel et al 2005) but their Vibrio campbellii. Environmental Microbiology 9. pp445-452
use in aquafeed is very limited (on a commercial scale). Forni D, Bee G, Kreuzer M and Wenk C 1999. Novel biodegradable
Recently, Defoirdt et al (2006) reported increased survival of plastics in sheep nutrition 2: Effect of NaOH pre-treatment of poly
Artemia nauplii when fed formic, acetic, propionic, butyric and (3-hydroxybutyrate-co-3-hydroxyvalerate) on in-vivo digestibility
valeric acid and challenged with a luminescence pathogenic Vibrio and in-vitro disappearance (Rusitec). Journal of Animal Physiology
campbellii strain. In another study, the same author (Defoirdt et and Animal Nutrition 81. pp41-50
al 2007) reported that commercial polyhydroxy butyrate (PHB)
Hari B, Kurup BM, Varghese JT, Schrama JH and Verdegem MCJ 2004.
particles or PHB accumulating bacteria offered a preventive and
Effects of carbohydrate addition on production in extensive shrimp
curative protection to Artemia against luminescent vibriosis.
They observed complete protection at a level of 1000mg/l of culture system. Aquaculture 241. pp179-194
commercial PHB or 107 cells/ml of PHB accumulating bacteria. Khanna S and Srivastava AK 2004. Recent advances in microbial
Although there are no reports of direct use of biofloc (PHA or polyhydroxyalkanoates. Process Biochemistry 40. pp607-619
SCFA) as antimicrobial neutraceuticals in fish feed, many projects Lancelot C and Billen G 1985. Carbon-nitrogen relationships in
have and are focussing on the characterisation and optimalisation nutrient metabolism of coastal marine ecosystems. In: Jannasch HW
of PHA production by these flocs and analysing their bio-control and Williams JJL (eds). Advances in Aquatic Microbiology, vol. 3.
efficacy in different host-microbe systems. Academic Press, New York, USA. pp263-321
Mallasen M and Valenti WC 2006. Effect of nitrite on larval
CONCLUSION development of the giant river prawn, Macrobrachium rosenbergii.
In essence, biofloc technology could be useful in improving the
Aquaculture 261. pp1292-1298
sustainability of fish/shellfish farming in both extensive and
modified extensive culture systems. There is clearly scope for Ricke SC 2003. Perspectives on the use of organic acids and short
further improvement of this management strategy, not only chain fatty acids as antimicrobials. Poultry Science 82. pp632-639
by optimising the quantity of carbohydrate addition at various Russel JB 1992. Another explanation for the toxicity of fermentation
intensities of culture, but also by comparing the potential of acids at low pH: anion accumulation versus uncoupling. Journal of
other carbohydrate sources. Additional research is required Applied Bacteriology 73. pp363-370
with respect to feed composition planning and feeding rate Salehizadeh H and Van Loosdrecht MCM 2004. Production of
determination. polyhydroxyalkanoates by mixed culture: recent trends and
biotechnological importance. Biotechnology Advances 22. pp261-
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Avnimelech Y 1999. C/N ratio as a control element in aquaculture Teitelbaum JE and Walker WA 2002. Nutritional impact of pre and
systems. Aquaculture 176. pp227-235 pro-biotics as protective gastrointestinal organisms. Annual Review
Burford MA, Thompson PJ, McIntosh PR, Bauman RH and Pearson on Nutrition 22. pp107-138
DC 2004. The contribution of flocculated material to shrimp, Van Immerseel F, Boyen F, Gantois I, Timbermont L, Bohez L, Pasmans
Litopenaeus vannamei nutrition in a high-intensity, zero-exchange F, Haesebrouck F and Ducatelle R 2005. Supplementation of coated
system. Aquaculture 232. pp525-537 butyric acid in the feed reduces colonisation and shedding of
Cherrington CA, Hinton M, Pearson GR and Chopra I 1991. Short Salmonella in poultry. Poultry Science 84. pp1851-1856
chain organic acids at pH5 kills Escherichia coli and Salmonella Yu J, Plackett D and Chen LXL 2005. Kinetics and mechanism of
spp without causing membrane perturbation. Journal of Applied the monomeric products from abiotic hydrolysis of poly [(R) -3-
Bacteriology 70. pp161-165 hydroxybutyrate] under acidic and alkaline conditions. Polymer
Defoirdt T, Halet D, Sorgeloos P, Bossier P and Verstraete W 2006. Degradation and Stability 89. pp289-299
Short-chain fatty acids protect gnotobiotic Artemia franciscana from
pathogenic Vibrio campbellii. Aquaculture 261. pp804-808
Defoirdt T, Halet D, Vervaeren H, Boon N, Wiele T, Sorgeloos P, Bossier CONTACT Kartik Baruah. Email baruahkartik4@rediffmail.com ■
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I
n 2007 the BC Pacific Salmon Forum, an independent citizen
body using science and stakeholder dialogue to advance sustainable
governance of BC Pacific salmon, launched a two-year collaborative
research programme to improve our understanding of the Broughton
Archipelago ecosystem.
The programme engages over 30 scientists and many others, with
a variety of disciplines and perspectives, in some 15 research projects.
THE BROUGHTON ARCHIPELAGO
They include defining oceanographic dynamics, monitoring salmon,
stickleback and sea lice population dynamics, investigating the impacts
of lice on salmon, and performing surveys of wild fish health. A number
of key findings have been identified from individual projects in 2007,
and although the data is preliminary and study will continue in 2008,
some key points of interest arose.
The following is an overview of the preliminary findings from the
2007 research period. (See end of article for contact details.)
ECOSYSTEM DYNAMICS
Oceanographic factors play a significant role in the dynamics of any
ecosystem, and in particular, in the distribution of organisms (both
fish and potential pathogens such as sea lice) within that system.
The Broughton Archipelago is a complicated system and to better
understand it, and to provide insights into the distributions of fish and
lice, several research groups have gathered oceanographic data.
MAP OF THE
The data indicates a considerable degree of wind-driven surface water BROUGHTON ARCHIPELAGO
circulation that may result in surface transport in directions counter to
tidal currents in the Broughton Archipelago. Although currents near
the surface move predominantly in a seaward direction, winds can
drive surface water (and potentially any particulates, including sea lice, although larvae were relatively rare, some patterns did emerge. Caligus
contained in these surface waters) up inlets contrary to the current copepodids were found in higher abundance in side inlets and low
flow. salinity areas, while Lepeophtheirus copepodids were most abundant
Oceanographic fluctuations have significant impacts on organisms near active farms.
living within an ecosystem. A delicate balance exists between a fish, Plankton tows in April were hampered by dense plankton blooms
the environment it lives in and potential pathogens. It is an accepted that congested nets and precluded further sampling. In 2008, new
fact that any change (environmental, nutritional or physiological) that equipment will allow the examination of vertical distributions of sea lice
pushes an organism beyond its ability to cope results in stress, which can larvae in the water column. Their salinity and temperature preferences
upset homeostasis and lead to increased pathogen susceptibility. and their diel (ie daily) migration patterns will also be investigated.
A comparison of data between 2003 and 2006 demonstrated that Several projects involved collecting wild juvenile pink and chum
surface water salinity and surface water temperatures were greater in salmon and evaluating their infestation rates. Although there was
2004 compared to other years, and this corresponded with an increased some degree of variation between research groups with respect to the
abundance of sea lice. Surface water temperatures increased (from 8.5˚C prevalence and intensity of lice observed on wild pink salmon captured
to 12.5˚C) and surface salinity decreased (from 26ppt to 19ppt) between in 2007, lice levels were low early in the season (March and April) and
April and June 2007. increased over time (April to June).
Overall, the prevalence of lice on juvenile pink salmon in 2007
SEA LICE STUDIES was described as being low, relative to previous years. In one study
Plankton tows were performed to identify the distribution patterns approximately 80 percent of all juvenile pink and chum salmon
of planktonic lice, both near to and distant from salmon farms. The captured had no lice (ie the prevalence was approximately 20 percent).
majority of early samples (February and March) contained no lice, but This represented the overall infection rate (prevalence) observed for
RIGHT: SAMPLING
ABOVE CENTRE:
JUVENILE SALMON UNDERGO A FITNESS TRIAL
with fish having no lice, there were no significant differences in their In some cases sticklebacks have been described as being, “feathery with
maximum swimming performance. Louse stage (at one louse per lice”. There is no evidence to date that lice are capable of carrying out
fish) had no apparent effect on maximum swimming performance. their entire life cycle on the stickleback, and the stickleback’s heavy
Copepodids artificially raised from egg strings were successfully armour is thought to protect it.
used to infect juvenile pink salmon in the laboratory, and while the In 2007 sticklebacks were captured in large numbers during juvenile
duration of survival of juvenile salmon did decrease with increasing salmon fry sampling and numbers were high relative to previous
lice loads, this effect was not statistically supported unless lice loads sampling years. In many cases sticklebacks were reportedly more heavily
were 10 times greater than levels reported in nature. Interestingly, infested by lice than juvenile pink and chum salmon captured in the
mortalities of pink salmon spiked when the lice entered a moulting same sets.
stage of their development. Interestingly, in a captive study, sticklebacks were observed
An examination of the impact of sea lice on the schooling behaviour cropping egg strings from gravid females on juvenile salmon and
of juvenile pink salmon in tanks in the field was carried out, and actively removing adult lice in some instances. It is not known what,
preliminary findings suggest that predation may occur selectively if any, role sticklebacks play in the transmission dynamics of lice in
on lice-infected fish. This data has not yet been fully analysed, and the Broughton.
modelling is necessary to substantiate the results and evaluate the
implications more thoroughly. CONCLUSION
Salmon populations can and do fluctuate for a great number of The 2007 research period has ended and much of the data is still
reasons, many of them environmental. Variations in the freshwater being analysed. It is difficult, if not impossible, to make well-founded
environment can impact early rearing stages of fry, environmental statements about the dynamics of organisms in a region without
conditions on marine entry can impact what is known to be a accurate assessments of populations and the environment over time.
physiologically challenging period, oceanic productivity can impact For this reason, the majority of the projects being funded by the
prey availability and parasite populations, and the high seas are a black Forum are being repeated for a second year to provide more data. The
box into which we have yet to accurately peer. 2008 research programme is now underway, and new data emerging
The overall catch of juvenile salmon in the Broughton Archipelago, over the coming year will add to our expanding knowledge of this
for sampling purposes in 2007, was described as being the lowest over complex system.
the previous five-year period of monitoring (despite an increase in the The Broughton Archipelago is very intricate, and its currents,
number of sampling sites) and there was greater variation in the size of winds, geography and ecology are very complicated. The Forum
fish than previously observed. has stated that, “Since it is clear we are dealing with a dynamic
Freshwater counts from a variety of systems and the migratory ecosystem that includes many factors, not simply sea lice, funding is
patterns of adult and juvenile salmon were tracked through the being allocated in 2008 to engage a range of researchers to develop
installation of video equipment, an acoustic counter, aerial surveys and an analytical framework that will incorporate all possible ecosystem
fry traps. factors in order to interpret the data that is emerging from this
The 2007 pink adult salmon returns (escapement), as a whole, research programme.”
in the mainland inlets of the Broughton Archipelago system are This analysis would be a first attempt to undertake a whole system
described as being similar or slightly improved, relative to the evaluation of many of the key environmental factors that affect salmon
brood return in 2005. The out-migration from the Glendale River in the Broughton Archipelago.
spawning channel was estimated at 7.7 million pink fry and 34,000 While research will not unravel all of the ecological mysteries of lice
chum salmon fry during 2007, but fry appear to have migrated out and salmon in the archipelago, perhaps some of the more pressing
later in 2007 than in past years. questions can be, at least in part, addressed through this scientific
The three spine stickleback fish has been of interest for several years, process.
following numerous reports of them being heavily infested with lice. See www.pacificsalmonforum.ca/research/index.php ■
SALMON GET TRAINING TIPS generation shrimp feed additive, IMS. SemBioSys is developing
FROM FOOTBALL a second-generation shrimp feed additive product in safflower,
What do juvenile salmon and the Spanish football team Barcelona called ImmunoSphere, which it says could significantly reduce the
have in common? Both use intensive training to improve their manufacturing costs of the feed additive.
form. The head coach is a Nofima scientist and the training is “As a result of the decision, SemBioSys Genetics intended to
healthy for fish, too. evaluate the impact it had on ImmunoSphere,” said the president
In order to make salmon more robust for transfer to sea water, an and chief executive, Andrew Baum. ImmunoSphereTM offered
interdisciplinary research group involving Nofima, the Norwegian a significant reduction to the cost of producing immune-
Institute for Water Research (NIVA), the Norwegian School of ostimulatory feed additives, Baum said. “We intend to evaluate
Veterinary Science, NTNU, Thelma AS and Aakvik Settefisk AS all the available options prior to committing to the next steps
decided to strengthen the salmon’s heart capacity and health with this product, which targets a large potential market that has
through intensive training. Benefits to humans through training a significant unmet need.”
include strengthening of the heart, muscles, skeleton and immune The global shrimp industry represents a US$18 billion market.
system and reduction of stress. One of the major challenges facing commercial shrimp producers
“We were concerned that the fish would develop lifestyle diseases,” is the outbreak of diseases, which cost the shrimp aquaculture
says senior scientist Harald Takle of Nofima Marin (formerly industry more than US$3 billion per year. Immuno-stimulatory feed
Akvaforsk), who headed the research group. The scientists have additives, like ImmunoSphere, are designed to provide prophylactic
found hearts in farmed salmon that differ from the heart form of protection to shrimp production.
wild salmon, so the thought that the fish needed training wasn’t far
off the mark. It was just a matter of starting to train 50g salmon. PROSOL CREATES NEW PRODUCTS
The scientists increased the tank water velocity to get the fish to BASED ON NUCLEOTIDES
swim faster. One of the challenges was to get tiny heart rate monitors The Italian company Prosol, which is a private producer of natural
made. These were inserted into the fish’s stomach with sensors nucleotides for dietary food applications, has launched a new range
attached to the heart. of nucleotide-based products for aquaculture. Based on well-known
The fish in the trial were divided into three groups: independent scientific literature, these products provide a specific
• a control group that lived the normal life of juvenile salmon source of Free 5' nucleotides (F5N). It has been proved that only
• one that received increased tank water velocity around the clock, nucleotides in free form enhance the immune system’s response to
and stress factors.
• one also received continual “jogging”, a daily spell of
high-intensity training.
The findings showed that the fish that trained grew
considerably quicker, but that it did not influence feed
The new knowledge about juvenile
utilisation. salmon could lead to salmon farmers
“We are now conducting tests at the VESO Vikan
fish farm to see if the fish with headbands and heart putting more jets in fish tanks and
rate monitors have greater powers of resistance to a
deadly virus. The findings are extremely promising. regulating the tank water velocity
Moreover, the fish tackle stress better when they are
in better form. Less stress means the fish have greater
energy reserves to tackle the challenges of everyday life,” says Takle. Nucleo 5 Prime, the latest in the Prosol range, is developed from
The new knowledge about juvenile salmon could lead to salmon the company's deep knowledge in nucleotide extraction technology.
farmers putting more jets in fish tanks and regulating the tank water In response to feed producers’ requirements, Nucleo 5 Prime is said
velocity. “In the long term, we believe that this can make the fish even to offer:
more robust. It’s just like with us humans, healthier fish thrive better, • expertise in production
and this will in turn increase profitability for the salmon farmers,” • a constant ratio among free 5' nucleotides
Takle said. • a precise amount of free Single nucleotides
• determination of free 5' nucleotides in each batch
AQUA BOUNTY SCALES BACK COMMITMENT • GMP+ certification, and
SemBioSys Genetics Inc., a biotechnology company developing a • optimal ratio quality/price.
portfolio of therapeutic proteins for metabolic and cardiovascular Nucleo 5 Prime is obtained in its free form by enzymatic hydrolysis
diseases, says its joint development partner on the ImmunoSphere of ribonucleic acid in yeast. Nucleo 5 Prime is then spray dried and
shrimp feed additive programme, Aqua Bounty Technologies Inc, provided in two different concentrations: six percent F5N and 40
intends to scale back marketing and registration efforts on its first- percent F5N content. ■
CHILE
New general manager
identify the source of the infection. Importation of fish from
the affected source will be prohibited pending the conclusion
of these investigations.
The clinical signs of SVC can include darkening of the skin,
swollen eyes, abdominal swelling, pale gills, trailing faecal casts
and protrusion of the anus. Infected fish may be lethargic and
for Pharmaq Chile show areas of bleeding in the gills and skin.
Dr Bernd Wrege has been appointed general manager of Anyone noting deaths in carp or any other species susceptible
Pharmaq Chile Ltda, situated in Puerto Montt. He will to SVC, with signs of disease similar to those above, should
continue his responsibilities as technical director. Dr Wrege immediately contact the Cefas Fish Health Inspectorate at the
has held key positions within the company since 2004, and has Weymouth laboratory on 01305 206673/4.
extensive experience within the aquaculture industry in Chile. Anyone who imports, keeps or retails carp or other
Pharmaq Chile Ltda is as a subsidiary of the Norwegian susceptible fish should take strict precautions to prevent the
company Pharmaq AS, and started its activities in Chile in spread of SVC and follow the advice set out in Defra’s booklet
2001. The company says it has experienced a strong growth in Keep Fish Disease Out, available from the Cefas Fish Health
the Chilean market and occupies the number one position in Inspectorate or through www.efishbusiness.com
vaccines and treatment against Caligus.
The aquaculture industry in Chile is facing several challenges Salmon industry to invest £400k in
related to diseases, and the company says it is committed to research and development
helping the industry by introducing new products and Scottish salmon farmers are to benefit from a £400,000
customer services. investment in research and development projects over the
next three years to help secure the long-term socio-economic
Pharmaq introduces vaccine and environmental sustainability of the industry.
in a new product line “This project will be highly significant in developing new
Pharmaq Chile Ltda has been granted a licence by Servicio farming practices in the Scottish industry,” said the chief
Agricola y Ganadero de Chile (SAG) for the new vaccine executive of the Scottish Salmon Producers’ Organisation,
Alpha Ject® micro 3. This is a three-component vaccine for Sid Patten. “It is hoped that the information generated will
salmon in a micro-dose (0.05ml/dose) to protect against enhance sustainability, while improving both environmental
infectious pancreatic necrosis, vibriosis caused by Vibrio performance and reducing the cost of production.
ordalii, and SRS caused by Piscirickettsia salmonis. “To be successful, it must report good quality, robust
“With all the efforts and resources put into this project scientific evidence. By working closely with stakeholders, the
we are confident that we have developed an effective and initiatives address many of the current challenges and they will
safe vaccine for our customers,” said the company’s general help the industry to achieve its full potential,” Patten said.
manager in Chile, Dr Bernd Wrege. A range of initiatives has been identified, including a
Alpha Ject® micro 3 represents the first micro-dose SRS scientific environmental modelling project and the adoption
vaccine delivered from Pharmaq. “The documentation of of new technology and husbandry practices on farms. An
this new-generation micro-dose product from Pharmaq enhanced monitoring programme has been commissioned to
is comprehensive. We had to develop new technologies to develop the accuracy of predicting environmental performance
ensure the right concentrations of antigens for optimum of marine farms. A second initiative is to look at economic
protection of this product.” modelling of production to determine optimum sustainable
As well as ensuring the efficacy of fish vaccines from farming conditions.
Pharmaq, all the products go through laboratory and field Projects addressing alternative sea lice management
tests to establish safety documentation as well as safeguard the strategies, the application of new technology in freshwater
maximum growth of the fish. farms and further development of acoustic devices to deter
The introduction of the new vaccine follows the company’s predators are under consideration.
launch one year ago of the first SRS vaccine, Alpha Ject® 4-1. The initiative is being managed and partly funded by
Contact Bernd Wrege, general manager Pharmaq Chile Ltda. the producers’ organisation under the auspices of the new
Phone (+56) 65 483091, or email bernd.wrege@pharmaq.cl Demonstration Project, with financial and other support from
the Scottish government, salmon farmers, industry suppliers,
UNITED KINGDOM Highlands and Islands Enterprise, The Crown Estate, Scottish
Isolation of spring viraemia of carp virus Aquaculture Research Forum and regulators.
Recent routine import checks have identified spring viraemia
of carp, or SVC virus in goldfish imported from a supplier in Scientists gather for workshop
Hong Kong, the Tung Hoi Aquarium Company. Marine scientists from around the world took part in a
SVC has no implications for human health. It is nonetheless, special workshop in Weymouth on infections and diseases
a serious viral disease affecting common and ornamental carp, in marine life during May. The 27 delegates took part in a
as well as tench, roach, rudd, goldfish, pike and wels catfish. histopathology workshop at the Centre for Environment,
The virus was found in a random sample taken directly from Fisheries and Aquaculture Science Laboratory.
imported boxes of fish. The Fish Health Inspectorate at Cefas The three-day workshop featured presentations from
has placed movement controls on, and taken samples from, all experts, laboratory tours and breakout sessions, and placed
sites receiving fish from the SVC-positive consignment. The significant emphasis on “hands-on” practical training and the
FHI has also provided advice to other businesses that may diagnosis of diseases and pathological changes. Participants
have recently imported fish from the same source to increase studied tissue samples through microscopes and examined
awareness of any potential SVC risk. tissue sections under high-magnifying monitors.
The authorities in the exporting country have been informed Species examined included salmonids, cyprinids, molluscs
of the isolation of SVC and are conducting investigations to and crustacea. One of the organisers, pathologist John
IRELAND
Improved sea lice strategy
the Intervet Dieter Lütticken for salmon farms
Award at World Aquaculture The Minister for Agriculture, Fisheries and Food, Mary
2008, held in Busan, Korea from Coughlan TD, and Minister of State John Browne TD,
May 19 to 23. The award, which recently launched a new strategy to improve pest control on
carried a prize of €20,000, was Irish salmon farms.
for his innovative statistical The sea louse is the common enemy of both the salmon
and epidemiological research farmer and wild fisheries. Monitoring of sea lice on salmon
methods in sea lice treatments in farms was initiated in 1991 and involves inspecting and
salmon farming. The jury found sampling at all fish farms 14 times per year. Treatment is
his research methods were an required where sea lice levels are above target levels.
PROFESSOR
excellent example of commitment The strategy intends to build on the existing monitoring
GEORGE
to the advancement of reducing, GETTINBY and treatment regime through intensifying and revitalising
refining and replacing the use the single-bay management approach and making it central
of animals in research, development and production of to national policy for sea lice management. The new approach
veterinary medicines. will incorporate “real time” management of sea lice infestations
Professor Gettinby’s research focused on applying statistical on a case-by-case basis.
and epidemiological methods in sea lice treatments in salmon The strategy also calls for more work to be done to identify
farming in order to minimise the number of animals required in further treatment options, and optimising the use of available
test groups. The research also provides an alternative to assessing licensed salmon farm sites from a sea lice management
the effectiveness of veterinary medicines in fish, which supports perspective.
the replacement principle. This alternative is the adoption A national implementation group comprising representatives
of mathematical models, which can simulate the effects of from the Minister’s Department, the industry, BIM and the
treatments on lice populations. Marine Institute, is to be established to review progress made
Dr Dieter Lütticken, who presented the award, said there to reduce sea lice levels and to report within six months with
had been a long and intimate relationship between life sciences recommendations on further steps, if any, required to redress
and statistics. “Professor Gettinby’s innovative statistical and the situation.
epidemiological research methods are an excellent example of
refining and reducing the use of animals in sea lice treatments CANADA
in salmon farming.” Public updated on salmon migration
The jury panel praised Professor Gettingby’s innovating British Columbia’s largest aquaculture company, Marine
mathematical model approach for investigating the optimal Harvest Canada, placed full-page advertisements in the North
use of treatments that involve the minimal use of animals. ”In Island Gazette and a later edition of the Victoria Times-
addition, these methods also minimise the use of veterinary Colonist on April 3 to update the public on its success in
medicines in the environment.” minimising the number of sea lice on its farmed fish in
Professor Gettinby said he was honoured to receive the the Broughton Archipelago area during the current out-
Dieter Lütticken Award. “It is pleasing to know that the migration of wild juvenile salmon.
award has come as a result of the increasing role of statistics The advertisement said that that four of its Broughton
and mathematical modelling in the pursuit of medicines for farms were now empty of fish. Six other farms showed sea lice
animals and the improved health of species in aquaculture.” levels of no more than 0.2 lice per farmed fish, well under the
He acknowledged the collaboration, support and work provincial government’s threshold limit of three lice per fish,
of Marine Harvest Ltd, Scottish Quality Salmon, the UK at which time fish must be treated to eliminate the lice. The
Department for Environment, Food and Rural Affairs and latest monitoring was completed in early March.
the Epi-informatics research group at the University of “This data has been available on our website since 2004,” said
Strathclyde. the Director of Environmental Compliance and Community
The Dieter Lütticken award, established in 2004, aims to Relations, Clare Backman. “We monitor for sea lice levels diligently
encourage research into the use of alternative models for and manage our operations closely to ensure that our fish do not
animal testing with significant impact on the development or add to the many other threats that young wild salmon face.”
production of new animal health products. It is named after The ad was the second placed by Marine Harvest Canada to
Dr Dieter Lütticken, a committed researcher in microbiology update the public on its sea lice management plan since the
and virology. He led Intervet’s research and development for out-migration began in March. “It is part of our commitment
more than 25 years, and retired in 2003 as vice-president and to being open with those interested in our business about
head of research and development. what we are doing and what results we are attaining. We do
The award covers in-vitro models used in research and not pretend to have all the answers, but we are committed to
development which replace animal testing for licensing reviewing and supporting science and improving our practices
purposes, as well as studies avoiding the use of animals so that we minimise risk to our salmon and wild stocks.”
in efficacy, safety and quality testing in the production of Marine Harvest Canada produces 45,000 tonnes of salmon
biologicals and pharmaceuticals for animals. year-round in farms off Vancouver Island and the central coast.
Intervet said it welcomed submissions from all life-science The company employs 500 people from Duncan to Klemtu.
research institutions for the next award. This year’s deadline is See www.marineharvestcanada.com/documents/MHC_
November 15. Commercial organisations are excluded. sealice_update_April_08. pdf ■
S
ome salmon of the same species are more susceptible
to lice infestation than others. Utilising this knowledge
in designing selective breeding programmes not only
has the potential to save millions in lost revenue, but also
reduce the infestation pressure of salmon lice among wild
salmonid populations.
These are the findings of a recent Nofima research
project, and follows on from work published by Akvaforsk
researchers in 2005 (Kolstad et al 2005). This initial work
recorded susceptibility to the salmon louse Lepeophtheirus
salmonis in three year-classes of Atlantic salmon with 300
(year-classes 2000 and 2001) and 50 (year-class 2001) full- BJARNE GJERDE MARINE SALMON FARM
sib families.
The genetic correlations between body weight and lice numbers of salmon that are more resistant to lice infestation.
lice were of moderate size, showing that it was possible to improve “The aquaculture industry and authorities should join forces
both body weight and resistance to L salmonis simultaneously to breed a salmon with greater powers of resistance to salmon
through selection. Moreover, the genetic correlation between the lice,” says Nofima (formerly Akvaforsk) senior scientist Bjarne
numbers of lice recorded in a challenge test and during a natural Gjerde. “Breeding for resistance to salmon lice should be included
infection was very high (See Figure 1). as a supplementary measure in the industry’s and authorities’
It was concluded in this initial study that the potential for action plan against lice, in the same way as there is today an
improving resistance to sea lice in Atlantic salmon by selective order to delouse as soon as a stipulated number of lice per fish is
breeding was good. In addition, as natural infection is highly variable exceeded.”
in time and magnitude, the study recommended that challenge tests
should be used in selective breeding to increase the resistance of INFESTATION OF WILD SALMON
salmon to L salmonis. Although the issue is a complex one, salmon lice from farmed
Recent Norwegian media coverage has indicated that salmon salmon is one of the threats facing wild salmonids, given that wild
lice have become resistant to anti-lice medication. However, one salmon may be infested by lice spread by coastal currents. “The
measure to combat this particular problem is by breeding farmed responsibility to take good care of wild salmonids and the risk that
the lice develop resistance to medication means that it is important
to increase the farmed salmon’s powers of resistance to salmon
SEA LICE lice,” says Gjerde. “Purposeful breeding for a farmed salmon that is
infested by salmon lice to a lesser extent than today will also reduce
the infestation pressure on wild salmonids, as there would be fewer
salmon lice along the coast.”
FIGURE 1: Breeding values for the number of lice per fish of 50 Atlantic
salmon full-sib families under controlled (challenge test with post smolts)
and natural infection (Gjerde et al 2008, data from Kolstad et al 2005)
1.6
1.5
Field test, no. of lice per fish
1.4
1.3
rg = 0.88 ± 0.26
1.2
1.1
1.0
0.9
0.8
45 50 55 60 65 70 75 80 85 90 95
Challenge test, no. of lice per fish
ECONOMICS
Economic losses due to sea lice infestation are substantial in the 150 families and financed by the Aquaculture Industry Research
salmon industries of the major producing countries. Selective Fund (FHF) and Salmo Breed AS will be available later this
breeding for increased resistance to salmon lice can potentially (northern) summer.
provide significant economic gains in two ways. Firstly, farmed Contact Bjarne Gjerde, phone (+47) 93 06 15 41 or email bjarne.
salmon will require fewer treatments against salmon lice and gjerde@nofima.no
secondly, the infestation pressure on wild salmonids will
decrease. REFERENCES
The first is of major economic value to the salmon farming
industry, while the latter is of major value to society. “But it is Kolstad K, Heuch PA, Gjerde B, Gjedrem T and Salte R (2005). Genetic
important to emphasise that this, as with other traits selected for variation in the resistance of Atlantic salmon (Salmo salar) to the
in breeding programmes, is a long-term measure,” says Gjerde. “It salmon louse Lepeophtheirus salmonis. Aquaculture 247. pp145-151
will be some years before we reap the big gains, so it is important to Gjerde B, Sonesson A, Storset A and Rye M 2008. Selective Breeding
commence this work as quickly as possible.” and Genetics – Atlantic salmon. In: Aquaculture – Production of
Aquatic Organisms (2000-2005). Aquaculture Research: From Cage
ACKNOWLEDGEMENTS to Consumption (eds) M Thomassen, R Gudding, B Norberg and
The project is financed by the Research Council of Norway and L Jørgensen. The Research Council of Norway, ISBN 978-82-12-
Salmo Breed AS. The findings from a fresh project involving 02409-0 (pdf version). ■
CYTOGENIX ANNOUNCES
AGREEMENT TO DEVELOP
DNA VACCINES
SCOTT PEDDIE, AQUACULTURE HEALTH INTERNATIONAL, UNITED KINGDOM
T
he US-listed company CytoGenix Inc announced on March
5 that it has completed an agreement with Taiwan Cobia
Inc to develop DNA vaccines against fish viruses. Under this
agreement, CytoGenix will develop DNA vaccines against selected
viruses, and TCI will vaccinate the fish and test immunological
responses.
The major diseases and target species of interest include viral
nervous necrosis disease (VNN; Betanodavirus, Nodaviridae) affecting
marine fishes such as parrot fish, groupers, flounders, sea bass,
turbot, and striped jack. DNA vaccines against pathogenic Iridovirus
in grouper (Lymphocystivirus, Megalocytivirus or Ranavirus) are also
being investigated.
“Fish is man’s most important single source of high-quality THE CYTOGENIX TECHNOLOGY
protein and provides ~16 percent of the animal protein consumed The CytoGenix team has developed a method for manufacturing
by the world’s population (United Nations Food and Agriculture large amounts of high quality therapeutic nucleic acid constructs
Organization, 1997),” says the president and chief executive officer (synDNATM) which is conducive for the rapid development of
of TCI, Cory Huang. compounds with new sequences of interest. The process offers
According to the FAO, aquaculture is one of the fastest growing significant improvements over traditional bacteria-based
food-producing sectors of the world, and it continues to grow more fermentation for plasmid DNA production and polymerase chain
rapidly than all other animal food-producing sectors. Asia and reaction (PCR)-based methods.
the Pacific region contribute over 91.5 percent of the total global It incorporates a cell-free method for producing therapeutic
production and 82 percent of the value, Huang said. quality DNA for use as drug substances in both humans and
“However, aquaculture faces many challenges, especially when it animals. According to CytoGenix, such an approach offers a number
comes to health management of fish stocks. CytoGenix’ synDNATM of advantages, including:
technology has the capability to provide the large quantities of • Minimal hazardous contaminants: Cell-free amplification of
affordable DNA vaccines necessary to address the need for simpler therapeutic nucleic acid has many important benefits, beginning
and more effective aquaculture vaccination regimens than currently with a smaller size and less complex version of the plasmid. Under
available.” this system, there is no need for bacterial replication sequences or
CytoGenix’s chief scientific officer, Dr Yin Chen, says DNA selection markers such as antibiotic resistant genes found in the
vaccines have already been shown to provide protection against plasmid DNA backbone. In most cases, this will reduce the size
pathogenic challenges in several animal models, including fish. and weight of the therapeutic product by at least 3000 base pairs.
“The vaccines carry only a small portion of the infective agent, and In addition, the absence of bacteria and growth medium greatly
therefore carry almost no risk of inadvertent infection, particularly simplifies the methodology required to purify the manufactured
when manufactured using CytoGenix’s proprietary cell-free method product. This process employs a simplified chromatography-based
(synDNATM),” Dr Chen said. purification procedure developed at CytoGenix. Depending on
“Additionally, the prophylactic potential of genetic vaccination the intended application, nucleic acids of various grades of purity
in fish culture is being commercialised, due to the advantages can be produced ranging from 70 percent (ie, no purification)
offered by genetic vaccines over conventional preparations ie, ease to about 95 percent pure. This reduces or eliminates the need
of production, stability, cost and production of vaccines against for mechanical or chemical purification methods to remove
organisms which are difficult or dangerous to culture in the contaminants such as bacteria cellular debris, endotoxins, RNA or
laboratory.” genomic DNA molecules.
The president and chief executive officer of Cytogenix, Malcolm • Robust biological activity: According to CytoGenix, the results
Skolnick, says the synDNATM vaccine platform is highly adaptable. of their experiments have shown that the biological response to
“These vaccines are based on an expression cassette that may be used this material (devoid of vector backbone) is similar to traditional
against a large number of pathogens by substituting appropriate plasmids. Moreover, experiments conducted in several animal
sequences to express antigenic proteins for a given pathogen. models have shown that linear DNA prepared with CytoGenix
“This agreement represents a beneficial joint effort where synDNATM process triggers a robust response (immune or
CytoGenix develops a vaccine and our strategic partner tests it and physiological, depending on the application) in treated groups
has the ability to commercialise the results. This agreement with compared to placebo or plasmid DNA-treated animals.
Taiwan Cobia further exemplifies the versatility of our technology and • Low risk, competitive cost, universal accessibility and fast cycle
provides an avenue for expansion into the worldwide marketplace,” time: CytoGenix scientists claim that the entire process is bench-
Skolnick said. scale and requires little equipment, space or human intervention
in comparison to bio-process or bacterial fermentation Prior to joining the School of Public Health, Dr Skolnick managed
manufacturing facilities. the Health Science Centre’s Office of Technology Management and
This process easily lends itself to liquid-handling automation, oversaw the university's activities in protecting and licensing its
and a skilled technician can synthesise multi-gram quantities of patent portfolio of technology.
this material within a few weeks, while working in a compact, Dr Skolnick held several clinical trials while he was at the Health
room-sized facility. Unlike PCR, this process requires only basic Science Centre in Houston in pain management, smoking cessation
laboratory equipment and is therefore accessible to many facilities and reduction of withdrawal symptoms in drug addiction. He is
around the world, especially rural settings in the field, where a registered patent attorney, a patented inventor and is licensed to
access to specialised equipment is limited and often prohibitive. practice law in the state of Texas. Dr Skolnick has been active in
• Improved regulatory profile: A major benefit of using this patent prosecution and licensing for selected clients, and has served
cell-free DNA manufacturing technology, again according to as an expert witness in intellectual property, product liability and
CytoGenix representatives, relates to the regulatory agency review accident reconstruction matters.
and compliance perspective. CytoGenix claims that product
cGMP manufacturing procedures detailing methods for cell Greg S Taylor, CPA. Vice-president of finance and administration
collection, processing and cell culture conditions are no longer and chief financial officer.
necessary, and therefore reduces the level of risk, the amount of Greg Taylor has almost 20 years of investment banking, venture
documentation and the amount of required space, as well as QA/ capital, corporate advisory and entrepreneurial experience and
QC and compliance costs.
In terms of production, CytoGenix states that once all the
production parameters are optimised for a given construct, INFORMATION BOX
manufacturing of gram quantities of the desired product can usually Taiwan Cobia Inc, a private company founded in 2004,
be accomplished within five to 10 working days. Quality control tests was the first company to introduce the concept of organic
are incorporated for quality assurance and within another week, the aqua-farming in Taiwan. The company is currently adding
DNA can be released for use. Additional developments may reduce production sites in China and South East Asia. TCI also
the lag period between custom optimisation identification and DNA has collaborative agreements with research institutes and
release. universities, including Academia Sinica, University of
CytoGenix has entered into an exclusive supply agreement with California at San Diego, National PengHu University, and
GE Healthcare Bio-Sciences Corp to purchase custom kits and National Taiwan University, as well as various government
reagents for making large amounts of nucleic acid in accordance agencies in Asia and Africa.
with our proprietary technology. The agreement gives CytoGenix CytoGenix Inc is a Houston-based bio-pharmaceutical
the exclusive, worldwide right to purchase the necessary reagents company pioneering the new field of gene-based medicine.
from GE Healthcare for use in producing therapeutic synDNATM. It is focused on developing innovative vaccines and
therapeutic products using its three proprietary technology
ABOUT CTYOGENIX platforms:
CYTOGENIX MANAGEMENT TEAM • synDNATM enzymatic, non-bacterial production
Malcolm Skolnick, methodology
PhD, JD. Chairman/president and chief executive officer • single stranded DNA (ssDNA) expression, and
Malcolm Skolnick received his PhD in physics from Cornell • OligogenixTM, an oligonucleotide based anti-bacterial
University and his JD from the University of Houston Law Centre. technology.
Prior to joining CytoGenix, Dr Skolnick held academic positions in CytoGenix currently holds 13 granted patents, two
the Graduate School of Biomedical Sciences and the School of Public allowances and 55 additional international or US patent
Health at the University of Texas Health Science Centre at Houston. applications claiming methods and materials in connection
He is currently an adjunct professor in the School of Public Health, with these platform technologies.
where he formerly served as Professor of Technology and Health See www.cytogenix.com
Law.
THE CYGX SCIENTIFIC TEAM. FROM LEFT: CHIEF SCIENTIFIC OFFICER YIN CHEN,
PHD, SENIOR LABORATORY DIRECTOR MS HARILYN MCMICKEN, BS, MT,
FREDERIC AND HARILYN NEXT TO TC HOOD (FUNCTIONAL ANALYSIS IN SENIOR SCIENTIST FRÉDÉRIC KENDIRGI, PHD AND SENIOR RESEARCH SCIENTIST
CULTURED CELLS OF THERAPEUTIC DNA PREPARED WITH SYNDNATM PLATFORM) XIN-XING TAN, PHD
has completed over $2BB in transactions. He has also served in Prior to working for Cytogenix, Ms Schertz has had over 15 years
management positions as both a chief executive officer and chief of experience in accounting in various industries, including a casket
financial officer. manufacturer, an independent power company and an architecture
He was most recently managing director of Monterey Capital firm. Management positions have included serving as controller for
Partners, a private equity investment firm, for over 10 years. a high-end clothier and a civil engineering firm.
Previously, he served as a vice-president with the investment banking
division of Sumitomo Trust and Banking and with Bunker Hill Cindee Ewell, PhD, JD.
Associates, a merchant banking firm. Greg Taylor began his career Vice-president of legal affairs and secretary
with Ernst and Whinney, a big eight accounting firm in Houston, Dr Ewell has been with CytoGenix since 2004 and is the company’s
Texas as a certified public accountant. resident corporate and patent counsel. She received her law degree
He has served on the board of directors of several private from the South Texas College of Law, her bachelor’s degree in
companies and charitable organisations. He has a Bachelor of microbiology from Cornell University, and her doctorate in
Business Administration in accounting and finance from Baylor biochemistry and molecular biology from the University of Texas
University and a Master of Taxation from Baylor University. Health Science Centre at Houston.
She is a registered patent attorney and is licensed to practice law in
Yin Chen, PhD. Chief scientific officer and vice-president of the state of Texas. Her experience includes nearly eight years of research
research and development at Baylor College of Medicine, about four years of patent prosecution
Dr Chen earned his PhD in molecular biology and biochemistry and freedom to operate experience in a large law firm environment,
from the University of Maine. Subsequently, he was a post-doctoral and several years of contract and corporate law experience.
fellow at the Beth Israel Deaconess Medical Centre, a teaching Ms Ewell entered the legal field in 1999 as a biotechnology
hospital of the Harvard Medical School. He was briefly employed by technical advisor with a focus on intellectual property. She has since
InGene, Inc in Kansas City, MO as a senior research scientist. attained her law degree and has established herself as a patent and
Dr Chen joined CytoGenix In 2000 as chief research scientist. He corporate attorney with a specialty in biotechnology.
was promoted to vice-president of research and development in
2001. Dr Chen oversees the research and development team at the Key CytoGenix staff
company and is the leading inventor of the company’s proprietary Xin-Xing Tan, PhD. Senior research scientist
technologies. Dr Tan earned his PhD in biochemistry from the Chinese Academy
of Science, Shanghai, China. He was an NIH-sponsored post-
Pam Schertz, CPA, Controller doctoral fellow at Rice University, Houston. He has authored or co-
Pam Schertz has been appointed as interim chief financial officer. authored 20 scientific published papers. Dr Tan joined CytoGenix in
Ms Schertz has served the company as controller since 2003. She 2002 and is co-inventor of an anti-bacterial technology developed in
received her BBA in accounting from the University of Houston the company’s laboratory. He leads the company’s research efforts in
and is a licensed CPA in the state of Texas. After establishing her developing novel antimicrobial therapeutics.
own practice offering consultant services as a CPA in October 2003,
Cytogenix became her first client. Frederic Kendirgi, PhD. Senior scientist
T
he Central Fish-Health Laboratory is located in the Jordan the CFHL, Shmuel Sarig, who was one of the originators of the
River basin, Beit-Shean Valley, within a rural area where almost European fish pathology associations (COPRAQ, EAFP).
70 percent of Israeli fish farming is centred. CFHL belongs CFHL’s research priorities are set by the current needs of the fish
to the Department of Fisheries and Aquaculture, Ministry of industry in Israel, and these needs have changed over time. The
Agriculture and Rural Development, Israel. It specialises in fish and severe shortage of water and land in Israel has lead to intensification
shellfish animal health and environmental quality, provides disease of fish culture from 30kg to 200kg per hectare, and a move to use
surveillance and diagnostic services, export certifications and offers relatively brackish water, thus providing suitable conditions for the
advice and guidance on health management and treatment for edible occurrence of Prymnesium. These conditions raised the awareness
and ornamental fish growers. of the importance of water quality examinations introduced by the
The Central Fish-Health Laboratory addresses the basic CFHL as a routine test performed within the field as well as in the
requirements of a functioning fish-diseases laboratory in terms of lab.
fish and water sampling methodology and expertise, and experience Israel’s location within the main routes of the mass migration
in parasitical, viral and bacterial diseases of aquatic animals, as of birds between Eurasia and Africa introduced a vast number of
well as in histopathology, molecular biology, immunology, vaccine pathogens into aquaculture. Consequently, the CFHL was involved
development and pathogenicity studies. in introducing many new treatments and nutrition programmes for
The CFHL has been the only institute in Israel specialising in Israeli aquaculture, while remaining cognisant of ecological impact
aquatic animal health for many years and is involved in a wide in accordance with internationally accepted standards.
range of research projects concerning fish health and culture. The CFHL’s research focused on parameters of stress, mostly in
laboratory works with individuals, companies, organisations and tilapia, the main fish raised in Israel. This research, led by the
academic institutions to develop and provide health management CFHL’s supervisor for 30 years, Dr Itzhak Bejerano, elucidated the
services, ensuring high standards of veterinary care and attention. relationship between farm management, water quality and pathogen
The CFHL was established in 1941 when aquaculture in Israel occurrence.
began, and its main objective was research of the toxin-producing A major research priority of CFHL research was the urgent need to
alga Prymnesium parvum. One of its founders, Professor Moshe find a substitute treatment to malachite green, the use of which was
Shilo and his team from the Hebrew University of Jerusalem, prohibited a number of years ago, for saprolegniasis. Saprolegniasis
then discovered the mode of action of P parvum ichthyotoxin and infections cause vast economic losses to Israeli aquaculture as a
developed a bioassay for its quantification and verification. consequence of the immunological stress experienced by tilapia
This research was done in collaboration with the supervisor of during the winter. In the course of this study the whitening agent
We think globally but have the right products for local use.
Our quality products are led by the Norvax® range.
PD VIRUS
P
ancreas disease (PD) remains one of the most serious
and economically important infectious diseases of farmed PD virus is classified as a salmonid alphavirus (SAV), and
Atlantic salmon (Salmo salar L) in Europe. Over the last molecular analysis now indicates that there are at least six
few years our knowledge and understanding of the PD virus and known sub-types, with very interesting geographical and species
the disease has been expanded considerably, due to the efforts and distribution. See Table 1.
collaboration of excellent research scientists, veterinarians and SAV 3 is the only sub-type found to date in Norway and can
the salmon industry. This effort has been coordinated and greatly cause disease in both Atlantic salmon and sea-reared rainbow
assisted by the formation of the Tri Nation PD Research Group. trout (Oncorhynchus mykiss [Walbaum]). SAV 2 has primarily
The aim of this article is to summarise the new and important been associated with disease in freshwater rainbow trout and
information in pancreas disease research. For a recent review article was only recently introduced to the United Kingdom K. SAV 2,
see Alphavirus infections in salmonids – a review. MF McLoughlin as distinct from freshwater strains, has recently been recognised
and DA Graham, Journal of Fish Diseases, September 2007, Volume in marine Atlantic salmon in Scotland. SAV 1, 4 and 5 are present
30 Issue 9, Special Alphavirus issue. pp509-572 in Ireland and Scotland.
Further work to compare the pathogenicity of these sub-types is
TABLE 1: Geographical and Species Distribution of SAV Isolated in Europe planned in the near future. Interestingly, there is regional separation
of sub-types in Ireland with SAV 1 in Connemara (west) and SAV
SAV SUBTYPE SPECIES COUNTRY
4 in Donegal (north). Given that smolts from one region have
SAV 1 Atlantic salmon Ireland (Mainly Connemara) regularly been transferred to the other, it suggests that vertical
Scotland
transmission has not been a major source of infection between
SAV 2 Rainbow trout France
England & Scotland
these regions (Fringuelli et al 2008 submitted).
Italy
Spain
Germany* presumed SAV2
SAV SURVIVAL
Atlantic salmon Scotland A major gap in the knowledge of SAV was its survival outside
the fish and response to temperature, pH and disinfection,
SAV 3 Atlantic salmon Norway Marine only
Rainbow trout Norway which are very important in the overall control of the disease
SAV 4 Atlantic salmon Scotland
spread. SAV was found to be rapidly inactivated in the presence
Ireland of high levels of organic matter at 60˚C at pH7.2. It is also
SAV 5 Atlantic salmon Scotland
inactivated at pH4 (acid) and pH12 (alkali) at 4˚C, suggesting
that composting, ensiling and alkaline hydrolysis would all be
SAV 6 Atlantic salmon Ireland
effective at inactivating the virus in fish mortalities and waste.
MAP 1:
DISTRIBUTION OF MAP 2: DISTRIBUTION OF
CONFIRMED PD CONFIRMED PD OUTBREAKS
OUTBREAKS IN 2004 UP TO 01/12/2007. SOURCE
MATTILSYNET
2004
AQUACULTURE BIOSECURITY:
PREVENTION, CONTROL AND
ERADICATION OF AQUATIC
ANIMAL DISEASE
EDITED BY A DAVID SCARFE, CHENG-SHENG LEE AND PATRICIA J O’BRYEN. BLACKWELL PUBLISHING, 2006. PP182. ISBN: 978-0-
8138-0539-9
D
isease in aquaculture is responsible Fish and Wildlife Service’s aquatic animal
for widespread and often health policy is discussed, with particular
economically debilitating losses emphasis on the innovative approaches
and has implications for stock welfare, to disease management contained therein.
international trade and consumer A detailed account is presented of the
perception and purchasing patterns. As a plan’s risk assessment procedures, which
consequence, aquaculturists, veterinarians, were developed to both standardise and
policy-makers, scientists and a range of optimise the decision-making process
other stakeholders have become increasingly pertaining to aquatic animal relocation.
aware of the need to develop a coherent The situation in the USA makes for an
and integrated approach to aquatic animal interesting discussion point and is explored
disease prevention and management. It is in detail in this book. That many states
against such a backdrop that the theory have no regulations, while others address
and practice of biosecurity has gained only salmonids, provides an entry point to
increasing prominence in recent years. showcase Wisconsin’s veterinary approach
During the World Aquaculture Society’s to fish health, the topic of chapter six.
annual conference in Hawaii in March 2004, Wisconsin’s approach is an interesting one,
the Oceanic Institute and the American given that it includes non-salmonids and
Veterinary Medical Association brought deals with fish farm registration, certification
together a diverse group of stakeholders of veterinarians, health standards for fish
for a three-day session entitled Aquaculture introduced into public waters and the
Biosecurity 2004. The aim of this session issuing of fish import permits. Moreover,
was to present and assess the range of the programme evolved as a result of an
approaches being taken globally, and effective working relationship between the
then to identify the key steps necessary to effectively implement state government, aquaculturists and veterinarians, and is therefore
biosecurity programmes in the production environment. Aquaculture an important example of multi-stakeholder co-operation.
Biosecurity is a compilation of twelve of the 30 presentations given Chapter seven makes a coherent and succinctly presented case
at the stakeholder session. for a harmonised system of accreditation to ensure diagnostic data
Chapters one and two provide an overview of the biosecurity and test result validity in aquatic animal health diagnostic systems.
approaches undertaken at the international level by focussing This chapter forcefully promotes that valid data and test results
on the World Organisation for Animal Health, the World Trade are a critical pre-requisite for effective biosecurity planning and
Organisation and the Codex Alimentarius Commission and their implementation.
respective standards aimed at eliminating trans-boundary disease Chapter eight provides an excellent and in-depth review of
transmission. the types of disinfectants applicable to aquaculture, as well as
Chapter three presents an interesting and informative example stressing the need for disinfection as an integral component of
of a regional approach to aquatic animal health management. In effective biosecurity at all levels. Information on the efficacy and
this chapter the scope and utility of characteristic of commonly used disinfectants, fish pathogen
programmes designed by the Network classification and transmission/susceptibility characteristics of
of Aquaculture Centres in Asia- important fish pathogens is presented clearly and helpfully in
Pacific is presented as an example of tabular format and makes an excellent point of reference for the
effective regional co-operation. This is interested reader. Moreover, the “example disinfection protocols”
followed by a comprehensive and well- are another informative reference point and a welcome addition to
written account of the development a well-rounded chapter.
and functioning of Canada’s national The ninth chapter highlights the importance of aquatic animal
Aquatic Animal Health Programme, health surveillance in effective policy making, not only for disease
using relevant case studies to highlight control but also for quarantine and health certification.
the need for an overarching and all- Chapter 10 deals with the key issue of biosecurity at farm level
encompassing approach to biosecurity. and how to create the optimum environment and “state of mind”
The national dimension is continued among farm workers and managers to significantly minimise risk
A. DAVID SCARFE
in chapter five, where the United States in aquaculture facilities. This chapter succeeds in emphasising that
the absence of mortality may be related to chronic sub-clinical TEST SERUM HEART KIDNEY GILL PANCREAS MUSCLE
infection (Graham et al 2006, Diseases of Aquatic Organisms 72. Virus R R R R
pp193-196).
Recent epidemiological results from Ireland revealed that SAV RTPCR R R
mean PD-related mortality was 18.8 percent in 2003 and 14.8 SAV
Antibody R
percent in 2004, with an estimated loss of growth of 11.8 percent
over the two years. The risk factors associated with PD outbreaks Histology R R R R R
included positive farms in the area, a history of PD on the site,
more than 250,000 fish on the site and high lice burdens (Rodger
and Mitchell 2007 30. pp157-167). Serology and RT-PCR have
PD LABORATORY TESTS also been successfully used
Various laboratory tests have been developed and tested in the
past few years and will aid the confirmation of a PD outbreak. as surveillance tools enabling
Serology and RT-PCR have also been successfully used as
surveillance tools enabling the identification of acute and
the identification of acute and
chronic infections. Some companies now carry out routine
serology testing in advance of the risk period(s) for PD as part
chronic infections.
of their overall health plan, and have found the early warning
system where viraemia is detected in pre-clinical cases to be very H and Todd D 2005. Longitudinal serological surveys of Atlantic
useful in managing PD outbreaks. salmon Salmo salar L using a rapid immunoperoxidase-based
Antibodies develop two to three weeks post infection at 12- neutralisation assay for salmonid alphavirus. Journal of Fish
14˚C and have been found to persist up to harvest and are a Diseases 28. pp373-379
good retrospective test for PD infection. Real-time PCR has Graham DA, Jewhurst H, McLoughlin MF, Sourd P, Rowley HM,
also proven to be a very sensitive tool for detecting viral nucleic Taylor C and Todd D 2006a. Sub-clinical infection of farmed
acid, but this signal does not persist as long as the presence Atlantic salmon Salmo salar L with salmonid alphavirus – a
of antibody (Graham et al 2005, Journal of Fish Diseases 28. prospective longitudinal study. Diseases of Aquatic Organisms
pp373-379). Recent work indicates that pooled sera can be used 72. pp193-199
effectively for detecting circulating SAV activity as individual Graham DA, Cherry K, Wilson CJ and Rowley HM 2007.
samples, thus making population screening more cost-effective. Susceptibility of salmonid alphaviruses to a range of chemical
The work on PD survival has also indicated that transporting disinfectants. Journal of Fish Diseases 30. pp269-277
sera for virus isolation should be at 4˚C to prevent false negative Graham DA, Staples C, Wilson CJ, Jewhurst H, Cherry K, Gordon
results. A and Rowley HM 2007. Biophysical properties of salmonid
Table 3 summarises the test and samples required to confirm a alphaviruses; influence of temperature and pH on virus survival.
diagnosis of SAV infection and to differentiate them from similar Journal of Fish Diseases 30. pp533-277
pathologies.
Rodger H and Mitchell S 2007. Epidemiological observations
REFERENCES of pancreas disease of farmed Atlantic salmon Salmo salar L in
Ireland. Journal of Fish Diseases 30. pp157-167
Alphavirus infections in salmonids – a review MF McLoughlin
Taksdal T, Olsen AB, Bjerkås I, Hjortaas MJ, Dannevig BH,
and DA Graham. Journal of Fish Diseases, September 2007,
Graham DA and McLoughlin MF 2007. Pathology of pancreas
Volume 30 Issue 9, Special Alphavirus issue. pp509-531
disease (PD) in farmed Atlantic salmon Salmo salar L and
Fringuelli E, Rowley HM, Wilson JC, Hunter H and Graham rainbow trout Oncorhynchus mykiss W in Norway. Journal of
DA 2008. Phylogenetic analyses of partial E2 and ns P3 gene Fish Diseases 30. pp545-558
nucleotide sequences and molecular epidemiology of European
salmonid alphaviruses (SAV). Submitted Special note: Regular updates on pancreas disease will be posted on
Graham DA, Jewhurst VA, Rowley HM, McLoughlin MF, Rodger a new, dedicated PD section at www.aqua.intervet.com ■
A BASIC OVERVIEW OF
MULTICELLULAR PARASITIC
INFECTIONS
BY GERALD BASSLEER, BASSLEER BIOFISH, BELGIUM
This article is based on material contained in The New illustrated Guide to Fish Diseases by Gerald Bassleer
ISBN 90-807831-2-9. See www.bassleer.com
CARASSIUS AURATUS SARASSA. PARASITIC AND CARASSIUS RED ORANDA. PARASITIC INFECTION. RED CARASSIUS RED ORANDA. PARASITIC INFECTION.
BACTERIAL INFECTION. SARASSA WITH NUMEROUS SKIN ORANDA WITH SMALL, WHITE SPOTS ON THE HEAD CAUSED RED ORANDA WITH A CRATER IN THE HEAD
FLUKES AND BACTERIAL INFECTION ON SKIN AND TAIL BY SKIN FLUKES (GYRODACTYLUS) CAUSED BY SKIN FLUKE (GYRODACTYLUS)
DAMAGE
T
he most common multicellular parasitic infections in ornamental, however, that there are strains of skin flukes that have already
tropical and pond fish are skin and gill flukes. External infections become resistant to certain medications.
are caused by skin and gill flukes (Monogenea or monogenean
trematodes) Gyrodactylus species and Dactylogyrus species are DACTYLOGYRUS (GILL FLUKE)
especially common in aquarium fish. However, various other species, This parasite mainly occurs on the gills, but it can also occur on the
similar to these two, also occur. head and skin. Reproduction occurs by means of eggs that develop
in the substrate (or in the filter), so that the entire holding pond (and
GYRODACTYLUS (SKIN FLUKE) its filter) will have to be treated. If the water temperature is low, the
This parasitic fluke quickly reproduces itself on the skin and because larvae do not emerge from the eggs until several weeks or months
it is live-bearing, the young flukes (up to three at a time) are released have passed. This should be taken into account in the spring when
from the adults on the skin. It may also be found in the gills and the water warms up and the gill flukes may appear. These parasites
can readily be identified in a skin scraping under the microscope. (usually 1mm) can be easily identified by examining a skin or gill
The lower the temperature the slower skin flukes will develop and scraping under the microscope.
reproduce. Small numbers usually do not cause any harm, except in In terms of clinical presentation, the fish scrape over the substrate
small or juvenile fish. or against objects, they are emaciated, breathe rapidly and have flared
In terms of clinical presentation, there is usually excess open opercula. A small number of parasites may be present on the
slime formation on the skin, the fish scrape over substrate fish without causing any damage; relatively large numbers of them
or against objects, red patches or lesions can be observed, are required to cause a clinically significant infection. Infection can
with secondary bacterial infection after damage by the skin cause physical damage, which in turn leads to secondary bacterial
flukes. If the parasites have also infected the gills, the opercula infections that can kill affected fish.
will be flared open and the fish will be breathing rapidly. As Praziquantel (250mg/100 l for three days) is one of the most
skin flukes are usually accompanied by bleeding patches on effective medicines currently available. It is recommended that
the skin caused by physical damage or bacterial infection, the treatment be repeated for three weeks to ensure that all larvae
parasitic infection is often “masked”. Moreover, morbid fish may have been killed; the filter must also be treated as it may harbour
be affected by other parasitic infections, such as Ichthyobodo or larvae. Moreover, all of the sick fish can be removed from the
Spironucleus. aquarium or pond and treated in a separate tank. Over this
Treatment with Fenbendazol (200mg/100 l) is often effective, as period the flukes in the aquarium/pond will die in the absence
is Trichlorfon, Flubendazol, Formalin, Mebendazol, Praziquantel or of a host. However, at low temperatures the larvae take a very
salt. In the case of severe infections, an anti-bacterial (and perhaps long time to hatch (at <5˚C this may even take months), or
another anti-parasitic) medicine will have to be used to combat they emerge from the eggs as soon as the temperature starts to
secondary infections. A second treatment will also be necessary rise. This should be taken into account when treating ponds,
within seven days of administration of the first treatment. Note, especially in the spring. ■
PHARMAQstatistics
Fish vaccines in Norway - March 2008
Oil based vaccines for salmon and trout Record breaking vaccination of salmon in March
March 2008 March 2007 Last 12 months
Doses Share Doses Share Doses Share In March 12.1 million salmon were vaccinated. In comparison
2 diseases 0 0,0 % 110 000 1,6 % 3 590 000 1,5 % to March 2007 the similar number of vaccinated salmon was
3 diseases 1 000 000 7,9 % 380 000 5,5 % 3 585 000 1,5 % 6.5 million. Over the first quarter 2008 as much as 52.5 million
4 diseases 1 040 000 8,3 % 380 000 5,5 % 18 810 000 7,9 % salmon were vaccinated. This constitutes an increase of 24 %
5 diseases 10 545 000 83,8 % 5 980 000 87,3 % 213 295 000 89,1 %
compared to the same period last year. The rolling long-term
Sum 12 585 000 6 850 000 239 280 000
Salmon 12 085 000 96,0 % 6 450 000 94,2 % 223 775 000 93,5 %
trend over the last 12 month period now shows us that nearly
Trout 500 000 4,0 % 400 000 5,8 % 15 505 000 6,5 % 240 million salmon and trout are being vaccinated.
PD vaccines 730 000 420 000 23 165 000
In March 2.4 million vaccine doses were used for farmed cod.
This is at the same level as in March 2007. On the other hand,
Oil based vaccines for salmon and trout this month more that 2 million cod were vaccinated with an oil-
100,0 %
based injection vaccine. Since August 2007 there has been a
90,0 % nice development in the long-term trend of cod vaccination.
80,0 %
According to our information, approx. 700.000 doses of a
70,0 %
single PD vaccine (against pancreas disease), were
60,0 % administered to salmon in March. Re-vaccination with another
vaccine is needed to add protection against other pathogens.
50,0 %
40,0 %
30,0 %
20,0 %
10,0 %
0,0 %
March 2008 March 2007 Last 12 months
Doses of oil based vaccines for salmon and trout Doses of vaccines for
against different diseases - 12 months rolling salmon and trout - 12 months rolling
250 250
Millions
Millions
200 200
150 150
Doses
Doses
100 100
50 50
0 0
2 diseases 3 diseases 4 diseases 5 diseases Salmon Trout - oil based Trout - water based
Doses* of oil and water based vaccined for cod 2 diseases = furuculosis and vibriosis
12 months rolling 3 diseases = 2 diseases + coldwater vibriosis
4 diseases = 3 diseases + winter sore
5 diseases = 4 diseases + IPN
24
Millions
20
Last 12 months value is the sum of doses used the previous 12 months.
18 The 12 months rolling graphs shows the trend by using the 12 months values
Doses
over a period - here we follow the trend for the last two years
16
* One dose administered to cod is defined as 0.1 ml of vaccine.
It is important to be aware that the calculated number of vaccinated fish
14
shows an over estimate because immersion vaccines are included in the
calculation.
12
10
40,0 %
30,0 %
20,0 %
10,0 %
0,0 %
April 2008 April 2007 Last 12 months
Doses of oil based vaccines for salmon and trout Doses of vaccines for
against different diseases - 12 months rolling salmon and trout - 12 months rolling
250 250
Millions
Millions
200 200
150 150
Doses
Doses
100 100
50 50
0 0
2 diseases 3 diseases 4 diseases 5 diseases Salmon Trout - oil based Trout - water based
Doses* of oil and water based vaccined for cod 2 diseases = furuculosis and vibriosis
12 months rolling 3 diseases = 2 diseases + coldwater vibriosis
4 diseases = 3 diseases + winter sore
5 diseases = 4 diseases + IPN
24
Millions
20
Last 12 months value is the sum of doses used the previous 12 months.
18 The 12 months rolling graphs shows the trend by using the 12 months values
Doses
over a period - here we follow the trend for the last two years
16
* One dose administered to cod is defined as 0.1 ml of vaccine.
It is important to be aware that the calculated number of vaccinated fish
14
shows an over estimate because immersion vaccines are included in the
calculation.
12
10
© PHARMAQ AS 2008
RESEARCH FOCUS
N
ew strains and varieties of alien fish species have been diversification efforts and are likely to be susceptible to invade
introduced in several countries, including India (DAIS 2004, new environments and ecosystems. Naturally populating alien fish
Sena De Silva et al 2006, Singh and Lakra 2006), to compete species have therefore been considered as an indicator of degraded
on the world market. However, environmental, socio-economic and stream conditions or the poor health of the aquatic ecosystem.
biodiversity issues are important considerations for regulating the Extensive studies have demonstrated that five to 45 percent exotic
importation of alien fishes in India or in other countries. fish were present in river stretches and streams of Ganga and Yamuna
The introduction of alien fish raises questions about how best to in Uttar Pradesh, India. The maximum contribution of commercial
meet food demands and preserve environmental quality. Four broad catch was constituted by alien tilapia (Oreochromis niloticus) and
categories exist for ecological impacts of alien fish species. They are: common carp (Cyprinus carpio), while there was stray occurrence of
• basic species interactions such as predation and competition other alien fish species like grass carp, silver carp and bighead carp
(DIAS 2004, Singh and Lakra 2006) or even African catfish Clarias gariepinus.
• genetic impacts (Senanan et al 2004, Mabuchi et al 2008) Tilapia (Oreochromis niloticus) and common carp (Cyprinus
• disease impacts (Arthur 2005) and carpio) have formed breeding populations and contributed a large
• habitat alteration (IUCN/SSC 2000). percentage of the exploited stock in the river. These alien fish species
Alien species are fundamentally indicators of biological integrity have started competing for food and space in riverine stretches,
in two aspects. Firstly, the spread of alien fish species in places showing diet overlap with native and endemic fish species and thus
other than where it was originally introduced (ie, post-introduction environmental changes have been perceived (Table 2).
expansion). Secondly, alien fish species have been associated with the Thus it may be expected that riverine ecosystems consisting of
decline in or extirpation of native fish in a range of systems because various natural foods in different niches of the river is important in
of predation, competition and/or transmission of disease. diversified zones that could be homogenised by a feral population
In recent years, many alien fish species have been introduced of alien fish species. Unethical over-fishing, the use of chemicals,
into India illegally or otherwise (Table 1). These alien species have poisons and dynamite, a wide array of prohibited fishing methods
escaped into streams and rivers by human activity and aquaculture and habitat destruction of natural spawning and breeding grounds
COMMON NAME SPECIES YEAR SOURCE OF REASON OF SPREAD INTO NATURAL WATER BODIES
INTRODUCTION INTRODUCTION
Mozambique Orechromis 1952 Indonesia Aquaculture Reservoirs of southern part, lakes, wetlands of West
tilapia mossambicus 1962 Bangkok Bengal and Assam, back waters of Kerala, river
1985 Sri Lanka stretches of northern plain.
Bangladesh
Nepal
Nile tilapia Oreochromis niloticus 1987 Thailand Aquaculture and sewage River stretches of Ganga and Yamuna in Uttar Pradesh
Israel fed fisheries
Common carp Cyprinus carpio 1939 Sri Lanka Aquaculture (Composite Most of the river stretches, reservoirs and lakes.
(Scale carp) communis 1957 Bangkok Fish Culture) Recently formed commercial catch in Yamuna and
Ganga in UP
Mirror carp Cyprinus carpio 1939 Sri Lanka, Aquaculture in high Yamuna, Jhelum, Beas, Satluj
specularis 1957 Bangkok altitudes
Grass carp Ctenopha- 1959 Japan, Aquaculture Reservoirs and few river stretches
ryngodon Hongkong (Composite Fish Culture)
idella
Silver carp Hypophthalmichthys 1959 Japan Aquaculture (Composite Reservoirs, lakes and few river stretches
molitrix Hongkong Fish Culture)
Bighead Aristichthys nobils 1987 Possibly Aquaculture Reservoirs, lakes and few river stretches
Bangladesh &
Nepal
African catfish Clarias gariepinus Not known Neighbouring Aquaculture Few reservoirs, Yamuna and occasionally in
countries Godawari river
Sutch catfish Pangasius sutchi Not known Neighbouring Aquaculture River stretches in West Bengal
countries
OREOCHROMIS NILOTICUS
of the fish through various human activities in the river has FROM YAMUNA RIVER
contributed to the population decline of the freshwater fish. The
recent invasion of alien species in riverine stretches has further
threatened the existence of local and endemic fishes (Singh and
Lakra, 2006).
The eroding health of the riverine ecosystem due to the invasion
of alien fish species is attributable to the following facts:
1 The ability of many introduced fish species to thrive in degraded
aquatic habitats and their potential to impact on aquatic ecosystem
structures and functions. Thus, introduced fish may represent
both a symptom and a cause of decline in river health and the
integrity of native aquatic communities.
2 The varying sensitivities of many commonly introduced fish 4 Alien fish species found to be widespread in distribution and
species (Cyprinus carpio, tilapia and Aristichthys nobilis, Clarias often abundance in rivers and streams could be considered to
gariepinus and Pangasius sutchi) to degraded stream conditions be relatively tolerant to river degradation, making them good
to become invasive because of the environmental and biological candidate indicators of river health.
characteristics of the receiving water body, are all confounding 5 Potential confounding factors that may influence the likelihood of
factors that may obscure the patterns of introduced fish species successful establishment of an alien species have the potential to
distribution and abundance and therefore their reliability as disrupt the biological integrity of natural aquatic ecosystems.
indicators of river health. It is therefore important to assess the associated risks and benefits
3 The relationships of alien species distributions and indices of any alien fish introduction. If found appropriate, develop and
of abundance and biomass with the natural environmental implement a plan for their responsible use. The development of
features, the biotic characteristics of the local native fish codes of practice is the mechanism to assist in the responsible use of
assemblages and indicators of anthropogenic disturbance introduced species.
are subject to varying intensities of human interventions on In India, the National Bureau of Fish Genetic Resources has also
alien species introduction for aquaculture without adopting developed plans and guidelines to regulate the introduction of exotic
biosecurity and bio-safety measures. fish to safeguard fish biodiversity and biological integrity. The
Oreochromis mossambicus Displaced Gangatic carps, Puntius dubius and Labeo kontius and Singh and Lakra, 2006
now posing threat to Etroplus suratensis in back waters of Kerala
Aristichthys nobilis Displacement of Catla and silver carp, hybridization with silver Singh and Ponniah 2001; Mia et al 2005;Taylor 2005;
carp and Catla Singh and Lakra, 2006
Cyprinus carpio Displacement of local spp. Schizothorax, Osteobrama belangiri, Tor Singh and Das 2006; Singh and Lakra 2006
putitora etc.
O. niloticus Reduced catches of indigenous fish species Sugunan 2002 ; Singh and Lakra 2006
Clarias gariepinus Environmental problem posing threat to biodiversity. Thakur 1998; Singh and Ponniah 2001 Singh and
Risks of hybridisation with native fishes, loss to local culturable fishes. Mishra 2001; Sugunan 2002; Singh and Lakra 2006
Hypophthalmichthys molitrix Naturalised in some reservoirs and displacement of Catla Pandey 1997; Singh 2004
Pangasius sutchi Evironmental problems are realised but yet to be documented. Under study by NBFGR
aquaculturists and farmers are advised to comply with the available and McAndrew BJ 2005. Detection of hybridisation between Chinese
regulatory mechanisms for alien fish introductions, along with strict carp species (Hypophthalmichthys molitrix and Aristichthys nobilis)
conditions of sanitary and hygienic standards. in hatchery brood stock in Bangladesh, using DNA microsatellite
loci. Aquaculture 247. pp267-273
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Available for purchase from the Office International des symposium on fish health management and sustainable aquaculture.
Épizooties (OIE), 7 Rue de Prony, 75017 Paris, France, or for College of Fisheries, GB Pant University of Agriculture and
consultation on-line at www.oie.int Technology, Nov 1-2, 2000 Pantnagar. (eds) Singh UP, Chauhan RS
and Sharma AP. pp93-96
• Aquatic Animal Health Code, 11th edition 2008. English
Sugunan VV 2002. Clarias gariepinus (African catfish) gravitates
language version. 170pp
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• Code sanitaire pour les animaux aquatiques, 11th edition
Chimes 22. pp50-52
2008. French language version. pp160
• Código Sanitario para los Animales Acuáticos, 11th edition Taylor RM and Pegg MA 2005. Management and ecological note
2008. Spanish language version. pp160 response of bighead carp to a bio-acoustic behavioural fish guidance
• Manual of Diagnostic Tests for Aquatic Animals, 5th edition system. Fisheries Management & Ecology 12. pp283-286
2006. English language version. pp469 Thakur NK 1998. A biological profile of African catfish, Clarias
• Manual de Pruebas de Diagnóstico para los Animales gariepinus and the impact of its introduction into Asia. In: Fish
Acuáticos. 5th edition 2008. Spanish language version. Genetics and Biodiversity conservation (eds) Ponniah AG, Das P, and
pp480 Verma SR. Natcon Publication, Muzaffarnagar (UP), India. pp275-292
• Changing trends in managing aquatic animal emergencies
(ed) Eva-Maria Bernoth. Scientific and Technical Review CONTACT DETAILS
27 (1): pp300 (trilingual English-French-Spanish). April
AK Singh and WS Lakra. National Bureau of Fish Genetic Resources,
2008. (Editor’s note: This publication will be reviewed in a
forthcoming issue of Aquaculture Health International.) Canal Ring Road, PO Dilkusha, Lucknow-226002 (UP), India. Email
• Risk analysis in aquatic animal health (ed) CJ Rodgers. aksingh56@rediffmail.com
Thematic Publications 2001. pp346 Arvind Mishra. UP State Fisheries Department, Varanasi-221002,
Uttar Pradesh, India ■