 Lecture one.

 What is the (critical period)?

The transition from yolk sac larvae to exogenously feeding, which lead to high mortality in
A short time period of time.

 What are different factors affecting on mortality of fish larvae?

1) Advection "transport" : offshore by Currents

2) Diseases: especially during larvae and Juvenile Stage

3) Fishing: higher for fish that have recruited

4) Environment: currents, turbulence, temperature , Salinity , food abundance

 What are different types of malformation in fish larvae?

1) Skeletal deformation “mainly in round fish"

2) Default pigmentation "mainly in Flat fish”.

3) low Stress resistance

4) physical factors involved malformation in fish larvae

5) Congenital default inducing defective “embryonory development”

6) Nutritional factors involved in the quality of development

 What are different types of skeletal deformities?

1) Jaw malformation.
2) Scoliosis
3) Lordosis
4) Coiled Vertebral Column.
5) Missing, additional fin rays
 How can physical parameters affect fish larvae mortality?
1) Light: is a key environmental factor that synchronizes all life-stages of fish. as Fish
larvae consumed large quantities of food when they are under the lighting longer
periods.
2) Temperature: -High temperature during early ontogeny → disproportionate muscle and
bone development → lordosis in juvenile and fry → more severe by higher swimming
speed.
- Abnormal temperatures during early ontogeny → sensitivity to sex differentiation→
skewed Sex ratio and differential growth in adults.
3) Salinity: High salinity (40% or more) during early ontogeny→ poor swim bladder
inflation → skeletal deformities in juvenile and fry development.
 What are natural factors involved in quality of development?
3) phospholipids
4) highly unsaturated fatty acids
5) Peptides.
6) Amino Acids " tryptophan"
7) Retinoic Acid " Vitamin A"
8) Ascorbic Acid" vitamin C”
 What is the result for using (tube feeding technique)?
showed that free fatty acids would be absorbed much faster as protein and peptides and
would have a higher assimilation efficiency
Lecture two.
 What is the effect of HUFA on gene expression?
HUFA are known to modulate the transcription of genes involved in their metabolism, through
their nuclear receptors, These receptors form heterodimers with retinoid X receptors to regulate
the expression of more than 500 genes, involved in lipid metabolism, energy balance,
Morphogenesis, bone synthesis.
 Essentially of dietary phospholipid for fish larvae. Explain
1) for growth
2) For survival
3) For normal development.
 Effect of vitamin A on pigmentation.
1) Dietary incorporation of vitamin A, associated to PL and DHA is essential for reducing
pigmentation default in flatfish.
2) Color abnormality can be reduced by feeding flounder with rotifers enriched with high
vitamin A level .
 Why phospholipids are very important requirements in fish larvae? Explain?
1) Phospholipids constitute the cell membranes, and so are essential
For development of fish larvae
2) Dietary phospholipids affect lipid absorption and transport, as they have a specific role in the
synthesis and secretion of chylomicrons and VLDL.
3) Dietary phospholipid is important for growth & for survival& for normal development.
4) Phospholipid appeared to prevent skeletal malformations, lordosis, scoliosis and lower jaw
deformities.
Lecture three.
 In fish, the levels of digestive enzymes may be influenced by:
1) the age of the fish
2) Type of feeding.
3) Season and/or temperature of acclimatization.
 In larval fish diets and weaning diets, part of the protein fraction is usually supplied in
the form of hydrolysate. Explain.
- In most fish larvae the stomach is not differentiated at hatching Means that ingested protein
is neither exposed to the denaturing conditions imposed by the gastric acids nor pre-digested
by pepsin before entering the mid-gut.

- Fish larvae digestion relies mainly on pancreatic enzymes with an optimum of activity at
a neutral or alkaline pH, conditioning the range of proteins that fish larvae are able to
digest.

- This is a way to facilitate digestion and to increase the availability of peptides and amino
acids absorbed within the larval gut.
 The importance of adding pancreatic enzymes in miroparticles
Improving compound diet digestion by fish larvae. This exogenous enzyme supplementation
led to an enhanced diet assimilation and growth improvement in gilthead sea bream larvae but
was ineffective in sea bass larvae.

 Why larvae growth is depended on dietary lipids levels?

Because maximal capacity of lipase synthesis was reached for 15% triglycerides in diet.
Therefore, regulation of lipolytic enzyme synthesis seems to be efficient in young larvae,
explaining why larval growth is so dependent on dietary lipid levels.
Lecture four.
 Definitions of (nutritional requirements).
1) requirement for maximal growth and/or survival
2) fish-diet-feeding has an important effect in the determination of the
3) requirement for body maintenance as the minimum rate of nutrient expenditure needed
4) requirement for least cost production
5) Requirement for fish health
 What is The major differences in terms of larval nutritional physiology compared to
larger fish?
Poorer capacity to digest and/or absorb complex proteins and much higher AA requirements.
 The optimum protein level is higher in larvae and juveniles than in adults for the same
species. Explain.
This difference was attributed to the high growth rate and high utilization of protein as energy
in larvae.
 Amino acid requirements of fish larvae are not easily determined. Explain.
Due to difficulties in the use of formulated diets and on the manipulation of the live feed protein
profile.
 The bottlenecks for the determination of quantitative PL requirements.
1) Pelleted diets are not suitable for most marine fish larvae, and the traditional diets (live feeds)
are unsuitable as they contain PL and further enrichment can be difficult.
2) both pelleted feeds or microdiets can have problems of low palatability as the formulations
must be either fish meal-free or use defatted fish meal
3) Although commercial PL preparations (lecithins) are available, they are not pure products.
4) the alternative use of pure PL species (classes) has limited availability and is costly.
5) replacement of one lipid class with another and comparing different PL preparations will
invariably alter the fatty acid composition of the diets and this can be difficult to control
fully.
 In fish larvae nutritional requirements, several gaps remain to be covered by PL
studies?

1) to define the quantitative requirements for most species, considering larval age and gut
development as well as source and its content of essential fatty acids

2) To determine the qualitative requirements in terms of the type of PL and their optimum
dietary ratios.

 Effect of The n-3 highly unsaturated fatty acids (HUFA)

They have an effect on swimming, feeding and escaping behavior and water reabsorption
in red sea bream and gilthead seabream larvae on skeleton development and on flatfish
pigmentation.
 Mineral requirement studies are also complicated. Explain
By the presence of mineral in seawater and by the fact that different forms of minerals may have
different bioavailabilities.
Lecture five.
 What are two types of larvae Fish biologist's categories?

1) Precocial larvae are those that, when the yolk sac is exhausted, appear as mini-adults

2) Altricial larvae are those that, when the yolk sac is exhausted, remain in a relatively
undeveloped state.

 Why it is difficult to maintain the fish larvae?

1) Is the small size of larvae (about 1 mm) and their need to feed junior much smaller.
2) Their need for natural feed in the early stages of her life.
3) Weakness of their mobility.
4) Weakness ability to devour feed.
5) The emergence of long-chain unconventional dietary factors such as unsaturated fatty acids
(HUFA).
6) Digestive system is not developed.
 What is the Conditions to be met in larval feed?
(A)Needs of the farmer:
1) To be available.
2) Be easy to produce all year round.
3) Time to be produced and Regrowth Short.
4) Be economically produced.
5) Available to the local environment and fit with the surrounding environmental conditions to the
hatchery.
(B) Needs of the larva:
1) To meet the needs of larva feed of essential fatty acids along with other elements.
2) Make it easier for the larva that can be found around.
3) To be commensurate with the size of the slot mouth of larva.
4) To be easy to digest.
5) To be free of contaminants so that diseases are not transferred to the ponds
 The importance of use live feed for marine fish larvae.
high vital value,because it contains essential amino acids and essential fatty acids and the most
important of these organisms are rotifers.

 What is the specification for the production of micro diets that may be replaced the live
feed must meet?
1) Float in the water for a period of not less than three minutes, without loss of contents and
acceptable taste.
2) Contain all necessary for fish larvae food needs.
3) Easy digestion and absorption.
4) Up to the size of rotifers after absorbing water.
5) Do not cause any disease of the larvae.
 Microencapsulated can be divided into three different types (explain)?
1) Micro- encapsulated diets (MED).
Feed containing thin membrane coated on the inside nutrients in a Suspense picture.
2) Micro- binding diets (MBD).
Feed made up of the nutrients associated with materials bind together.
3) Micro- coated diets (MCD).
 What is the advantage of using method of larval rearing without use of algae and rotifers?
1) The availability of untapped area for the cultivation of algae and rotifers.
2) The availability of labor and energy consumed.
 What is The aim of breeding algae in fish hatcheries?
1) Feeding rotifers.
2) Feeding the fish with the improvement of the properties of water used for fish larvae and
added to the water in which larvae works to improve the water district as well as the
survival of rotifers in the environment of fish that feed on the fish.
3) Stimulate the immune larvae.
4) Limit the bacterial load of water.
5) Reducing the concentration of nitrogen and phosphate rate because they depend on them
in their growth.
 What is the types of microalgae that fit farmed marine fish hatcheries in green and brown
algae?
1) Green algae: -
a. Tetraselmis chuii algae
b. Chlorella spp:
c. Nannochloropis oculta:
2) Brown algae: -
a. Isochrrysis spp:
 What are Rotifers?
- Very small cells of multi-range animals wandering objects (metazoa) uses a whirling motion
in the so-called rotifers.
- Play an important role in feeding larvae.
- It is a very small crustacean follows the animal genus Brachionus sp.

 Rotifers sizes vary depending on the different morphotypes into two main types depending
on the length of the lorica in which. Explain
1) First-class (large type) Brachionus plicatilis rotifers and known as (L-Type) ranges along the
lorica between (200-300 microns).
2) The second class (small type) Brachionus rotundiformis rotifers and known as. (S-Type) ranges
along the lorica between (100-200 microns).

 The importance of using rotifers.

1) Rotifers is small size (less than 400 microns in length) to suit the size of the mouth of the fish
and crustacean larvae in the first days after hatching.
2) Can breeding rotifers in an easy trading system depend on the algae from 3 to 17 microns to get
the bumper production of rotifers.
3) Small size and slow swimming (float) velocity make them a suitable prey for fish larvae that
have just resorbed their yolk sac but cannot yet ingest the larger Artemia nauplii.
4) Rotifers contain certain enzymes that help the digestive process in the digestive system of fish
larvae.
5) Rotifers contain easily digestible proteins.
6) Available from several types, some of which can be breeding in fresh water while others in salt
water.
7) Tolerance to a wide range of environmental conditions.
8) High reproduction rate.
 What is it’s Optimum conditions for growth?
- Temp- 25-30 C.
- PH- 7.5-8.5.
- Salinity- 20-30 ppt.
- Ammonia- <1mg/L.
- D. O> 2.2 mg/L.
 What is The optimal conditions for hatching Artemia?

1) Temperature above 25 C, with 28 C being optimum.

2) Salinity to be maintained between 15-35 ppt.

3) Aeration –10-20 liter air per minute. Oxygen level nearer to saturation.

4) Constant illumination (example: two 40- watt fluorescent bulbs for a series of four 1-liter
hatching cones).

5) A pH 8-8.5. (1 gm. Na bicarb in One L of water.).

6) Density – one gm cyst in one L.

 There are many advantages to replacing live prey with microparticulate diets. explain
- First and foremost, microdiets have a uniform nutritional composition, which can be
tailored to match the needs of a given species or adjusted for research purposes.
- Second, microdiets are consistently available year - round without having to dedicate culture
space and time to maintain stocks .
- third, even though feed costs remain a significant fraction of the total cost of production, once
larvae can be weaned to artificial diets, the feed costs per individual decline dramatically
 What is the physical properties of microparticulate.
1) Size and color:
- Particle size is particularly important in first - feeding larvae and needs to be adjusted
for different ontogenetic stages of the target species.
- Diet color can vary depending on diet formulation, the water and light conditions, and the
background color of the rearing system.
2) Movement or behavior:
- Live prey swim in the water column with movements that capture the attention of visually
feeding predators such as fish larvae.
- microdiets may need to remain in the water and have motion that is fast enough to draw
attention but slow enough so the larva can catch it. This is accomplished through adjusting
the density of the particle, and water and/or air movement in the rearing system.
3) Chemical attractiveness :
- Fish live in an enriched chemical environment with highly developed chemosensory
systems and are especially attracted to amino acids, organic acids, nucleotides, and bile
salts .
- Live prey, especially copepods, contain relatively large amounts of free amino acids
(FAA) including glycine, arginine, and betaine shown to be strong inducers of feeding
behavior in larval fish.
4) Leaching:
- Another difference between live and microparticulate diets is in the degree of leaching of
small - molecular - weight water – soluble (SMWS) nutrients. There is good evidence that
these types of compounds could be very important in the nutrition of fish larvae.
5) Sinking rate:
- Along with leaching, the sinking rate determines the actual composition of what the
larvae eat once the diet is introduced to the tank.
- Slower sinking diets have the advantage of being more available to the larvae for longer
periods of time, are presumably easier to catch, and result in less buildup on the tank
bottom.