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Aquatic Biotechnology, Aquaculture Modern Biotechnology
Captive fishery vs
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Hatchery phase
a
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Fertilize eggs to develop to form embryos (fry), which are reared indoor in tanks until they reach
a fingerling size
2
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Fingerling is usually transferred outdoors to raceways and ponds
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Salmon are usually transferred to net pens or cages to
reach final size for market sale
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Ongrowing/grow out phase
a
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Fish and fishery products remain some of the most traded food commodities in the world
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FIsh with the highest production
Salmonidae: Some common species cultured
- The Atlantic Salmon: Salmo salar
- Chum Salmon: Oncorrhynchus keta
- Sockeye Salmon: O
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Grass carp: herbivorous on vegetation
2
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Silver carp: phytoplankton feeder
4
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Black carp: mainly feeling on molluscs
Different fishes including indian carps could have very different biology
Gill rakers
- Prevent potential loss of prey and small food particles through the gills
Pharyngeal teeth of Grass Carp
- The grass carp can conspire its wright in begeration each day and reduce plant matter to 2-mm^3
particles with its pharyngeal teeth
- The movement of one set of teeth against the other and against a hory pad on the roof of the bucal
cavity
The different states of almost
- The eggs hatch and the salmon emerge as sac fry or alevins
- When the yolk is absorbed its called fry
- As they grow, vertical striped develop on their sides, called parr
- The parr eventually go through a physiological change in preparation to live in seawater → called
smoltification
- The result is juveniles salmon called smolt

Traditional Practices
1
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Co-aquaculture
3
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Isolation of wild seeds
(fingerlings)
Grass Carp
- Artificial reproduction technology
developed in 1958
- A single species accounting for
20% of total freshwater
aquaculture production in China
Hatchery Phase
Many fish and shellfish species have
- Large number of gametes, from tens to million eggs
- External fertilisation
- Many reach sexual maturity by age 1 or younger

To introduce spawning:
- A variety of hormone preparations such as human gonadotropin (HCG), luteinizing hormone-releasing
hormone analog (LHRHa), or extracts from pituitary glands over several fishes like salmon, carp,
snapper, catfish and others

-

Maturation → spawning

Feed Formulation
- Composed of individual feedstuffs (fishmeal, blood meal, SCP, wheatmeal)
- In order to estimate the exact percentage of individual component to be pun in the final compound, we
need to know the dietary requirements of the target fish species
- A general approval is the percentage of proteins
- 40% protein (most expensive), 40% carbohydrates, 10% lipids
- And the overall nutrient compositions on the ingredients

The brain-pituitary-gonad axis
- Gonadotropin releasing hormone (GnRH)
- The reproductive axis in fish
Nutritional Biotechnology
- Balance cost and growth rates
- Utilization of unwanted feedstuff
- Formulation of compound (formulated) feeds
Fish meal: drying and grinding or otherwise treating fish or fish waste
Nutritional management
Feeds Conversion Ratio (FCP)
- Weight of fish to produce weight of fish
- The number of kilos of feeds eaten to produce one kilo of fish
Wish list
- Enhance growth rate
- Improve diseases resistance
- Enhance environmental tolerance
- Reduced reproductive differentiation
- Additional nutritional values
Major application points of biotechnology in aquaculture
1
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Strains improvement, new species
2
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Induced breeding, larval culture in the hatchery
3
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Balance cost and growth rates
4
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Vaccination
5
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Growth rates
6
...
Depending on the culture systems
Genetic manipulations are easier and cheaper in fish and shellfish species than in terrestrial livestock such as
chickens or pigs
...
There was no evidence of
mutational effects due to insertion
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Properties and benefits stem from the regulated expression of its specific gene construct
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Faster to market
2
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Reduced pollution
4
...
carp
and catfish
- Homologous recombination, random insertion
Transgenesis incorporation nuclear transfer
- Nucleus from donor cell must be inserted into an egg
- Egg is prepared with enucleation
- Pipette suctions out of nucleus
- Nucleus from donor cells transferred into enucleated egg cell
- Embryo is transferred to a surrogate mother for gestation
GMO
- FDA approval for food fishes drastically differ from genetic engineered aquarium fish
- Present cost for production are too high for many and increased production = increased environmental
problems, coastal use, effluent, escaped, disease
- Faster growth rate → shorter time to market
- Improved food conversion ratio
- Lower cost of production
- Less susceptibility to disease

Population control in aquaculture
1
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Manual sexing
3
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High density effects
5
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Triplody - small bood size
7
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Sustained protection (during the propagation period to market size_
2
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Efficacious for a board number of fish species
4
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Multiple pathogens
Aquaculture
- Conversion of fish mean to sellable products
- Salmon farming, grouper farming
- Production of low cost proteins
- Carp farming, from feather meal, from plant materials
- For food safety
- Conservation
- Production of bioactive compounds
- CRISPR : genome-location targeted
- Possible to do genetic deletion without leaving a recombinant constructs
Destruction of coastal areas
- The destruction of major habitats (mangroves, wetland) are involved in the construction of many coastal
aquaculture
- Many of these areas are important as nursery grounds for local fishery
Introduction of diseases
- Furunculosis, the major salmanoid disease of scotland, is through to have introduced through the
importation of brown trout
Introduction of Invasive species
- From piranhas to crocodiles
- Besides predation or competition with local fauna and
flora, eg
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dsDNA viruses (poxvirus)
2
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dsRNA viruses (reovirus)
4
...
(-)ssRNA viruses RNA (rhabdovirus)
6
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dsDNA-RT viruses DNA with RNA intermediate in life-cycle
Positive-sense RNA (Translation ready)
Nucleo plasmid Protein: coat the genome helps the formation of viral particles
Membrane Proteins: Embedded in the envelope of the visu with a tail inside the virus interact with the N
protein
Spike Protein: virus attachment to host cell surface receptor
...
Target of
neutralizing antibodies
...

Resortment: mechanism of antigenic shift
- Possible new virus from genetic reassortment
Influenza viruses are constantly evolving by mutation or by reassortment
Mutation can cause small changed hemagglutinin and neuraminidase antigens on the surface of the virus
A new vaccine is required every year because influenza virus has the ability to undergo antigenic drift
Post Translational Modification
1
...
Glycosylation: attached a sugar, usually to an N and O amino acid side chain
Bacteria DO NOT have post translational modification, cannot use bacteria for vaccines so had to develop
other eukaryotic translational systems (more expensive)
...
Parasites which were spread by anopheles mosquitoes
- In 2015 there were 214 million malaria causes and 438,000 deaths
- A vaccine
Artemisinin
Stunted growth in transgenic plants
- 12 genes
- Suspected the problem was the whole pathway diverting too much precursor from the chloroplast
- Expressed 6 genes in the tobacco plant
Vaccine for Malaria
- RTS,S/AS01 → first vaccine to be tested in phase 3 clinical trials and the first to be assessed in routine
immunization programs in malaria-endemic areas
- Results show children ages 5-17 month who received 5 doses, vaccine efficacy of malaria was 36%
over 4 years
- $5 each dose
Current treatment
- Artemisinin-based combination therapy (ACT)
- But a certain parasite in becoming resilient
- As it had become resistant to other drugs, chroquire and sulfadoxine-pyrimethamine

Apart from malaria
- Mosquito-transmitted viruses
- Yellow fever virus
- Dengue virus
- 60-100 million cases each year
- + strand RNA virus
- Most are transmitted by arthropods (mosquitos or ticks)
- Chikungunya virus
- West nile fever virus
- Zika virus
Sterile mosquito
- Transgenic male breed with wild-type female mosquito results in;
- sterile or mosquito offspring die before reaching adulthood
- Sterile insect technique (SIT1) is a method of pest control with a strong record of success against a rant
of agricultural pest insects
- Genetically engineered mosquitoes
Malaria resistant mosquito?

Marine Toxins Shellfish Poisoning Syndromes
- Diarrhetic shellfish poisoning, DSP
- Neurotoxic shellfish poisoning, NSP
- Paralytic shellfish poisoning, PSP
- Azaspiracid shellfish poisoning, ASP
- Ciguatera fish poisoning, NSP
- Amnesic shellfish poisoning, ASP
How is lobster red?
- The orange-red astaxanthin is released when heated, turning the shell of crustaceans bright red
Astaxanthin
- A naturally occurring carotenoids
- Found in microalgae, yeast, salmont, krill, shrimp, crayfish and the feathers of some birds
- Proved the red colour of salmon meat and cooked shellfish
- A carotenoid in larger family of terpenes
- Is added to fish feed to ensure a good flesh colour

Potential market of astaxanthin
1
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>9
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Fight inflammatory diseases through oral, injection and inhalation
c
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Photo-protective product market
a
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Prevent direct sunlight on skin through physical and chemical means
c
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It has been
assumed that astaxanthin biosynthesis proceeds along both routes
...
Synthesis of Astaxanthin by wittig reaction
2
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Oxidation of zeaxanthin
Comparison on natural and synthetic Astaxanthin
Natural

Synthetic

Related products: BioAstin

Related products: AstaSana

>95% Esterifies

All are free form

Natural fatty acids attached to either end of the
molecule
More stable and better bioavailability with its SS
stereochemistry shape

Less stable and poorer bioavailability with various
stereochemistry shape (RR, SS, RS, SR)
Only around 25% are in SS form

Vitamin A
- Group of unsaturated nutritional organic compounds that includes retinol, retinal and several provitamin
A carotenoid
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Food security
a
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Food demand and food production
c
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Energy and water security + land utilization
a
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Carbon footprint
3
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Climate change
5
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Biosafety and zoobiotics

Photosynthesis and Carbon Fixation
- Rubisco, the key enzyme of the CO2 fixation
- Light independent reaction→ RuBP + Co2 → glycerate-3-phosphate
- PSII and PSI
- Photosynthesis occurs in organelle chloroplasts
- 50% from photosynthesis in the sea → chloroplast, endosymbiosis
Bacteria
- Plants are multicellular, eukaryotic cells
- E
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drought resistance)
- N
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many traits (eg
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Keep it in the fridge to keep it solid for a few more days
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Disease Resistance
- Bacillus thuringiensis (bt) is a bacterium that kills harmful insects through production of a Bt protein
- It has been used as a bioinsecticide

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