PUBLICATION 600-106

Artemia Culture for Intensive Finfish

and Crustacean Larviculture
Brendan C. Delbos, Research Specialist, Virginia Seafood Agricultural Research and Extension Center
Michael H. Schwarz, Extension Aquaculture Specialist, Virginia Seafood Agricultural Research and Extension Center

Introduction the larviculture production system. As such, all Artemia

production and storage procedures must be conducted
In Virginia and throughout the United States, freshwater utilizing hygienic production protocols and proper hatch-
and saltwater finfish and shrimp aquaculture is expand- ery sanitation procedures. This document provides the
ing rapidly. During the cultivation of most marine finfish background, rationale, and detailed production protocols
and shrimp species – as well as some freshwater spe- for all stages of high-quality Artemia culture.
cies – live feeds are an essential component during the
larviculture stage. During larviculture, the rotifer is the
most commonly used live feed upon transition of the lar-
vae from endogenous (internal energy reserves) to exog-
Decapsulation of Artemia Cysts
enous (external) feeding. Upon completion of the rotifer
stage, the most commonly used live feed prior to conver-
Rationale
sion of the larva to a dry diet is Artemia (figure 1). Artemia represent one of the few live feeds that can be
cultured in sufficient numbers and are of appropriate
size for larva to transition to between rotifers and wean-
ing diets. During a portion of their life cycle, Artemia
hibernate as a desiccated cyst that is capable of with-
standing extreme environmental conditions for long
periods of time. Cysts are easily shipped and are thus
the form purchased by hatcheries.

However, Artemia cysts can cause problems during lar-

viculture because:

1. The shell of the cyst is indigestible and may cause

intestinal blockage when ingested by larva.

2. Cysts are a potential vector for pathogen introduction

Figure 1. Artemia: just hatched (left), enriched and 24 to the culture system.
hours old (right)
3. Artemia consume high levels of endogenous energy
As a food source for the larvae, it is imperative that reserves when hatching through the cyst shell.
Artemia is of high quality, as nutritionally complete as
possible, and maintained in this state until consumed 4. Cysts must be physically separated from the live
by the larvae. There are four distinct stages involved in Artemia after hatching.
Artemia culture. These stages are: (1) decapsulation, (2)
hatching, (3) enrichment, and (4) storage. Artemia also Decapsulation of Artemia cyst is a process whereby the
represent a potential vector for disease introduction into external shell or chorion is chemically removed from
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Virginia Polytechnic Institute and State University, 2009
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Issued in furtherance of Cooperative Extension work, Virginia Polytechnic Institute and State University, Virginia State University,
and the U.S. Department of Agriculture cooperating. Rick D. Rudd, Interim Director, Virginia Cooperative Extension, Virginia
Tech, Blacksburg; Wondi Mersie, Interim Administrator, 1890 Extension Program, Virginia State, Petersburg.
the cyst. This process addresses the concerns noted color from brown to grey, then to orange, and finally
above and has become standard practice by fish hatch- to bright orange. This bright orange color indicates that
eries looking to produce high quality Artemia. the process is complete. (Cyst buoyancy can also be
used as an endpoint indicator: when approximately 90
percent of cysts sink, the process is complete.)
Artemia Decapsulation Requirements
Artemia cysts: 1 kilogram (kg) The process should take from one to three minutes, but
time may differ due to temperature variations. Cysts
Decapsulation vessel: 20 liters (L) can easily be damaged by overexposure to the decapsu-
lation solution, adversely affecting the resulting hatch
Chlorine bleach (NaOCl; 5.5%): 8 L at 2-10 degrees rate. It is imperative to closely monitor the process and
Celsius (°C) standardize it for your particular conditions.

Sodium hydroxide (NaOH; 40%): 4 L at 2-10°C

Harvest
Sodium thiosulfate (Na2S2O3 ): 100 g When it is determined that the cysts are adequately
decapsulated, add 75 g of sodium thiosulfate to the
Harvest bag: 100 micrometer (µm) decapsulation vessel to neutralize the chlorine, then
immediately begin to drain cysts into the 100 µm har-
Artemia Decapsulation Procedure vest bag. During the harvest process (figure 2), rinse
with ample amounts of water (fresh or salt) while pro-
Hydration viding ample aeration via an air stone to keep decapsu-
lated cysts in suspension. When all decapsulated cysts
The first step in the decapsulation procedure is Artemia
have been collected, the remaining sodium thiosulfate
cyst hydration. Hydration of cysts allows for separa-
should be added to the harvest bag. Continue rinsing
tion of the nauplii from the chorion, facilitating the
the bag until water runs clear and no presence of chlo-
decapsulation process. For this step, Artemia cysts are rine can be detected.
placed in either fresh or saltwater at room temperature
for approximately one hour, using a concentration of 1
g of cysts per 15 milliliters (ml) of water. It is impor-
tant during this step to maintain sufficient mixing via
aeration to keep cysts well suspended. After one hour
of hydration, the water and hydrated cysts should be
drained through a 100 µm harvest bag; the concentrated
cysts are then placed back into the empty decapsulation
vessel.

Decapsulation
For decapsulation, pour the chilled sodium hydroxide
solution into the decapsulation vessel with hydrated
cysts, again making sure there is adequate aeration
within the vessel to keep cysts suspended. The chilled
bleach should then be added to the cysts to initiate the
decapsulation process. Because the chemical reac-
tion during decapsulation is exothermic, it is helpful
to begin with chemical solutions chilled to a tempera-
Figure 2. Harvesting decapsulated Artemia: note harvest
ture of 2°C to 10°C. These starting temperatures will
bag and rinse water leaving bucket
prevent the temperature of the chemical solution from
exceeding 35°C, which may damage the cysts.
Storage
As decapsulation progresses, the chorion is chemically Decapsulated cysts can be drained of excess water and
removed, resulting in the cysts gradually changing stored in an airtight container in a refrigerator for up

2
to two weeks. For longer-term storage (two months Lake City, Utah) can be used to help minimize growth
or more), cysts must be dehydrated by placing them of pathogenic bacteria in the hatching tank. The proper
in aerated brine (330 g of sodium chloride [NaCl] per stocking density for nondecapsulated cysts is approxi-
liter of water) at the concentration of 1 g of cysts per mately 2 g per liter.
20 ml of brine for 24 hours. They can then be drained
and placed into a suitable container, topped with fresh When using decapsulated cysts, approximately 5 g
brine, and placed in a refrigerator. per liter can be stocked. These numbers can be dou-
bled through the use of pure oxygen supplementation,
which is needed to maintain dissolved oxygen levels
Hatching of Artemia Cysts greater than 4 milligrams per liter. Attempting to hatch
at higher stocking densities can result in physical dam-
Rationale age to the nauplii and reduced quality.
While it is a straightforward process, proper hatching It is important to maintain sufficient aeration at the
and harvesting of Artemia nauplii is vital to maximiz- bottom of the cone to keep cysts suspended (figure 3).
ing quality. Standardization of protocols is important, When hatching large volumes of cysts, it is advanta-
as slight deviations in the process pro foundly affect the geous to use a food-grade antifoam product to mini-
hatching rate, nutritional makeup, and final size of the mize excessive foaming in the culture. Hatching times
harvested nauplii. Artemia cysts are expensive, making will vary based on strain and age of cysts, temperature
them one of the largest variable costs for a hatchery. and salinity of water, etc. Thus, it is important to mini-
As a result, every attempt must be made to maximize mize variation between hatches for consistency.
hatch rate and quality. Furthermore, because the risk
of pathogenic contamination is high, biosecurity mea- Generally, Artemia require 18 to 24 hours of incubation
sures should be in place to minimize this risk. to hatch. Decapsulated cysts, however, may be ready to
harvest after only 16 hours of incubation. When feed-
Artemia Hatching Requirements ing nauplii directly to fish, timing of the hatch is very
important. If nauplii remain in the hatching tank for
Temperature: 26-30°C too long, they will grow too large and their nutritional
quality will decrease. Determining the endpoint of the
pH: 8.0-9.0
hatch should be made through microscopic observation
Dissolved oxygen: > 4 mg/L of the relative numbers of hatched nauplii, prehatched
nauplii, and unhatched cysts.
Light level: ~2000 lux

Salinity: 25-35 parts per thousand (ppt)

Hatching density: ≤
 2 g dry cysts/L
(up to 5 g/L with supplemental O2)

Sodium bicarbonate (NaHCO3): 0.5 g/L

Antifoam (silicone based): 1 ml/100 L

Artemia Hatching Procedure

Fill a clean, cone-bottomed hatching tank with warm,
filtered seawater. If warm seawater is not available,
allow enough lead time for water to be warmed to 26°C Figure 3. Artemia hatching cone: note pure oxygen
to 30°C in the hatching tank via submersible heaters. injection regulators on wall and wire from submersible
Add 0.5 g of sodium bicarbonate per liter of water in heater on front edge of tank
order to maintain the pH between 8.0 and 9.0 through-
out the entire hatching process. The use of antimicrobial
products such as INVE’s Hatch Controller (INVE, Salt

3
Artemia Harvesting Artemia Counting
The harvesting procedure varies depending upon Counting of harvested Artemia is necessary to deter-
whether decapsulated or nondecapsulated cysts were mine accurate dosage rates for feeding and enriching,
hatched. and as quality control for the hatch. Hatch rates will
vary depending on strain and age of cysts, but gener-
When harvesting previously decapsulated cysts, sim- ally speaking, experienced users should see hatch rates
ply drain the entire water column into a 125 harvest with GSL cysts of 200,000 or more nauplii per gram
bag. An air stone should be placed in the bag to main- of nondecapsulated cysts. While there are a number of
tain oxygen levels while keeping nauplii in suspension. methods used for counting hatched Artemia (two are
After all nauplii have been collected, rinse them in the presented below), it is important to choose the best one
bag with clean water for at least five minutes. for your situation and stick with it to develop standard-
ized counting protocols for your facility in order to
After rinsing, attempt to separate the hatching mem- minimize variation.
branes (which remain intact after the decapsulation
process) from the nauplii. To do this, place Artemia into Method 1:
a cooler, enrichment tank, or other clean container at a After harvesting and rinsing Artemia, store in a clean,
density less than 5 million per liter. Using a micropore well-aerated container at a density no greater than 5
oxygen diffuser (Point Four Systems, British Colum- million per liter. To count Artemia, a small subsample
bia, Canada), oxygen should be injected into the cooler. should be collected from the well-mixed storage con-
The tiny oxygen bubbles will adhere to the membranes tainer and diluted 10-fold. Load a Sedgwick-Rafter
and they will begin to float after a few minutes, where slide (figure 4) with 1 ml of the diluted sample and
they can be skimmed from the top. After removing add one to two drops of formalin or Lugol’s solution to
membranes, nauplii are ready to be fed to the larva, immobilize the Artemia. Count under low magnifica-
transferred to subsequent enrichment, or placed into tion and record the number of intact, healthy-looking
cold storage. nauplii. Counts should be conducted two to three times
to determine an average. Multiply the average by 10
If harvesting nondecapsulated cysts, turn off the (rate of dilution) to determine the number of Artemia
air for 10 to 15 minutes. This will allow nauplii – as per milliliter in the storage container.
well as any unhatched cysts – to settle to the bottom
of the cone, while hatched cysts will float to the sur-
face. Keeping the air off for more than 15 minutes may
result in Artemia suffocation. Artemia move toward
light, so covering the tank and/or placing a light source
at the bottom of the cone will aid in separation. After
settling, slowly open the bottom drain and purge off the
unhatched cysts that will come out first.

Conversely, a standpipe can be placed in the tank prior

to settling to allow the bottom 1 inch of settled material
to remain undisturbed. Newly hatched nauplii should
then be collected in the harvest bag and rinsed for at
least five minutes. If nauplii have settled properly, only
75 percent of the water column will need to be drained. Figure 4. Newly hatched Artemia on Sedgwick rafter
While harvesting, check on the relative ratio of nau- counting slide; round object is the cyst the Artemia just
plii to cysts by transferring a sample to a glass beaker. hatched from
This will help determine when the harvesting process
is finished or if more time is needed to allow Artemia Method 2:
to settle. The nauplii are now ready to be fed to your Concentrate the rinsed Artemia in 10 L of saltwater
fish, transferred to subsequent enrichment, or placed using vigorous aeration. Collect 10 ml of Artemia and
into cold storage. add to 990 ml of saltwater. Collect 1 ml of sample with

4
a pipette and count the number of live Artemia within (see Artemia hatching protocol) and a second day for
the pipette. Return the sample to the container, stir, and the enrichment process. Having a second, dedicated
repeat the process five to 10 times; then determine an enrichment tank is necessary to facilitate this process.
average. The average of these counts conducted in this As with hatching, a cone-bottomed tank is ideal for
fashion multiplied by 1 million (rate of dilution) equals enrichment and helps to ensure adequate mixing and
the total number of Artemia. complete draining during harvest. Prior to stocking,
the enrichment tank should be filled with a suitable
amount of water, and water-quality parameters (salin-
Enrichment of Artemia ity, temperature, and pH) must be adjusted to match the
requirements listed above.
Rationale
Before being fed to larvae, Artemia nauplii are usually It is important to begin the enrichment process with
fed a specialized diet in order to increase their size and healthy, high-quality nauplii. Nauplii that are damaged
nutritional profile. While freshly hatched Artemia nau- or sluggish prior to enrichment will result in suboptimal
plii are rich in protein and can serve as a bridge between nutrient uptake. Care should be taken to remove hatched
rotifers and enriched Artemia for many species, they cysts (nondecapsulated cysts) or hatching membranes
are largely void of the beneficial fatty acids required (from decapsulated cysts) as described in the Artemia
for proper growth and development of most larvae. For hatching section. Artemia nauplii should also be rinsed
the purpose of the following Artemia enrichment pro- well prior to stocking into the enrichment tank. This
cedure, the protocol developed for the use of the INVE is especially important when using an additive such as
product, DC DHA SELCO, will be utilized. INVE’s Hatch Controller or antifoam during the hatch-
ing process, as ingredients in these products can inter-
fere with enrichment uptake.
Artemia Enrichment Requirements
Temperature: 25°C During enrichment, vigorous aeration should be applied
through the bottom of the enrichment vessel, and dis-
pH: 8.0-8.5 solved oxygen levels should be closely monitored
throughout the process (figure 5). The use of supple-
Dissolved oxygen: > 4 mg/L mental oxygen during this stage will likely be necessary
to maintain oxygen levels above 4 milligrams per liter.
Salinity: 20-30 ppt Temperature must also be maintained at 25°C through
Density: ≤ 300 nauplii/ml the use of submersible heaters or ice packs, as dictated
by ambient conditions.
DC DHA dosage: 0.6 g/L

Enrichment duration: 20-24 hours

Artemia Enrichment Procedure

There are a number of commercially available Artemia
enrichment products. Because these products have dif-
ferent ingredients, nutritional profiles, and enrichment
protocols, it is up to hatchery managers to decide which
product is most suitable for their conditions and species.
Once an enrichment product is chosen, it is important
that standardized protocols be developed and strictly
followed. Slight changes in temperature or enrichment
time, for example, can have significant effects upon the
size and nutritional quality of the final product. Figure 5. Multiple Artemia enrichment cones: note heavy
aeration
Preparation of enriched Artemia requires a two-day
lead time: one day is required for hatching of Artemia

5
Harvest at 2°C to 10°C, with adequate aeration to prevent set-
At the end of the enrichment process, the entire volume tling (figure 6). Under these conditions, Artemia can be
of water should be drained into a 125 µm harvest bag concentrated as high as 5,000 per milliliter and stored
for up to 24 hours.
with sufficient aeration to keep enriched Artemia in sus-
pension. Oxygen levels should be closely monitored in
the harvest bag. The bag containing the Artemia should
be rinsed well for five minutes or until the water runs
clear. Thereafter, Artemia should be transferred into a
container containing clean water of a known volume,
aerated vigorously, and enumerated as discussed above.
If Artemia will not be fed to larvae immediately, it should
be placed directly into cold storage, as described below.

Cold Storage
Artemia not fed to larvae or enriched immediately
needs to be stored under cold conditions. Cold stor-
age of Artemia dramatically decreases its metabolism,
which directly reduces further growth and metabolism Figure 6. Cold-banked Artemia: ice jugs for temperature
of their protein and lipid stores. Artemia should be control and air line for aeration to keep Artemia
transferred to a cooler or suitable container and stored suspended

This is a joint publication of Virginia Cooperative Extension and Virginia Sea Grant. (VSG-09-05)

Disclaimer: Commercial products are named in this publication for informational purposes only. Virginia Cooperative
Extension does not endorse these products and does not intend discrimination against other products which also may
be suitable.