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SRAC Publication No.

701

VI
October 2000 PR

Culture of Small Zooplankters for the


Feeding of Larval Fish
Granvil D. Treece1 and D. Allen Davis2

In nature, zooplankton is one of is a euryhaline species, small and tion occurs below 35 ppt. Most
the primary foods of larval fish. slow swimming, with good nutri- production facilities use 10 to 20
Two of the dominant zooplankton tional value. It is well suited to ppt salinity. Abrupt salinity
groups are Rotifera (rotifers) and mass culture because it is prolific changes of more than 5 ppt can
a sub-class of the Crustacea, and tolerates a wide variety of inhibit swimming or even cause
Copepoda (copepods). These two environmental conditions. death, so acclimation should be
groups are the preferred prey for Strain selection is important done slowly and carefully.
shrimp and fish and are the live because reproduction rate, size Temperature, salinity and feed
feeds used most often by cultur- and optimum culture conditions concentration all affect the growth
ists. The intensive larval culture of (temperature and salinity) can all rate of rotifers, but temperature is
most marine fish depends on a vary with different strains and the most critical factor. The opti-
large supply of zooplankton. species. Some freshwater rotifer mum temperature for most strains
Brachionus plicatilis (Fig.1a), is a variation can be seen in Figure 1b. is 28 to 32 oC (82.4 to 89.6 oF).
small rotifer first developed as lar- Two of the best known strains of Above 28 oC, the salinity and size
val fish food in Japan in the 1950s. brackishwater rotifers were of the strain are not very critical,
Since then, many methods of cul- thought to be morphotypes of B. but the density of feed is very
turing it have been developed. plicatilis, and were referred to as important. Below 26 to 28 oC
More than 60 species of marine the “large” (L) and “small” (S) (78.8 to 82.4 oF), the bigger strains
finfish are cultured using B. pli- types. Later it was found that tend to grow faster than the
catilis as live food. This publica- these are two different species smaller ones.
tion will concentrate on the cul- (L being B. plicatilis and S being Rotifers have broad nutritional
ture and feeding of rotifers, but B. rotundiformis). Mean dry requirements that must be met to
will include information on less weights are approximately 0.33 produce stable cultures. They are
used zooplankton such as clado- microgram/rotifer for the L type planktonic filter feeders, feeding
cerans (water fleas), copepods and and 0.22 microgram/rotifer for on organic particles brought to
tintinnid ciliates. An important the S type. The size of the S type their mouths by the movements of
larger zooplanktor used in aqua- is 126 to 172 micrometers accord- their coronas. The corona is a cili-
culture is the Artemia (brine ing to one source, and 100 to 340 ated organ on the head region that
shrimp), which is the subject of micrometers according to another. characterizes rotifers and is their
SRAC publication 702. The L type is 183 to 233 microme- means of locomotion. Rotifers
ters according to one source, and ingest many types of feed, includ-
Rotifers 130 to 340 micrometers according ing bacteria, as long as the size of
to another. Larval fish survive bet- the particle is appropriate, so a
B. plicatilis is the species used ter with L-type rotifers, probably
most commonly to feed larval fish variety of food sources can be
because the larvae use less energy used to rear rotifers. However,
in hatcheries around the world. It to feed on larger rotifers. rotifers cultured indoors often
1Texas A&M University, Sea Grant College Rotifers may tolerate 1 to 97 ppt require vitamin B12 and vitamin A
Program salinity, but optimum reproduc- supplements.
2Auburn University
Saltwater as “semi-continuous” or the com-
Freshwater
bined “batch/semi-continuous
a. b. technique.”
Nutrient sources for culturing
rotifers include baker’s yeast and
emulsified oils; algae (Isochrysis
galbana), yeast and emulsified oil;
algae alone; bacteria alone; and
outdoor culture using semi-pure
or wild strains of algae. The high-
est reproduction rate (21 offspring
per female every week) has
occurred when rotifers were fed a
pure diet of Isochrysis galbana
(Tahiti strain) and kept at a tem-
perature of 20 to 21 oC (68 to
Brachionus placatilis with eggs 69.8 oF). The optimum feeding
Adult 100 to 300 microns (0.003 to 0.01 rate is 105 to 107 cells of the algae
inches) in length Nannochloropsis oculata per indi-
vidual rotifer, or 106 to 107 cells of
baker’s yeast per individual
rotifer. The normal concentration
of rotifers is about 100 to 200 per
ml, but often reaches more than
1,000 per ml with an adequate
food supply. And if there is also a
pure oxygen supply instead of
aeration, the number will reach
Keratella spp. more than 10,000 individuals per
ml. Concentrated Chlorella sp. also
c. can be used for rotifer culture. No
one food source contains all the
nutrients required for the long-
term culture of a species. Several
food sources should be used for
Figure 1. Rotifers cultures that are to be maintained
for long periods of time.
The nutritional value of rotifers tinuous and feedback culture
for larval fish depends on the techniques evolved. Each system Larval culture with rotifers
rotifers’ food source. Researchers has advantages and disadvan-
have determined that highly tages. Batch culture is the most Rotifers usually are fed to fish lar-
unsaturated fatty acids (HUFAs) reliable but the least efficient. vae as soon as the larvae have
are essential for the survival and Semi-continuous is less reliable developed mouthparts. For larval
growth of marine finfish larvae. than batch but more efficient; red drum (Sciaenops ocellatus), this
Rotifer feeds containing DHA, however, it allows wastes to build will be on day 3 post-hatch.
22:6n-3, docosahexaenoic acid, up, which causes contamination. Rotifers are fed at a rate of three
and EPA, 20:5n-3, eicosapen- Continuous cultures are the most to five rotifers per ml until larval
taenoic acid, can be valuable, efficient and consistent but are fish can consume larger foods at
with DHA the more essential for maintained under strictly defined about day 11 post-hatch. Larval
marine fish larvae. Depending conditions and are almost always mullet (Mugil cephalus) require a
upon their food source, rotifers “closed” and indoors, which lim- food density of 10 rotifers per ml,
are about 52 to 59 percent protein, its the size and increases the cost when there are 25 to 50 larvae per
up to 13 percent fat, and 3.1 per- of the operation. The feedback liter, through day 40. Once rotifers
cent n-3 HUFA. system, developed in Japan, uses are harvested from the culture
wastes from rotifer culture (treat- system food is often limited, so
There are many methods of cul- the nutritional value of rotifers
turing rotifers. Some are low-den- ed by bacteria and the nutrients
retrieved) as fertilizer for algae decreases over time. It is best to
sity and some high-density. An feed them to fish at least twice a
early method involved daily cultured in a separate tank. The
Japanese consider this method the day, or replenish them whenever
transfers of rotifers to fresh tanks rotifer density drops below a des-
of the same size after most of the most efficient and reliable. The
culture technique described in this ignated number per ml. For exam-
algae were consumed. Following ple, in red drum larval culture,
this, batch, semi-continuous, con- publication is usually referred to
replenishment should occur when
rotifer density drops below 3 per gravity feed to rotifer tanks, umes may be harvested rou-
ml. Since one fish larva can eat as or the algae can be pumped tinely by dropping to the 50
many as 1,900 rotifers per day, to rotifer tanks. Gravity feed percent level. Even if the
from 13,300 to 57,000 rotifers are is preferred; it helps control rotifers are not needed in the
needed to feed one fish larva contamination of algae tanks hatchery, the volume in the
through this period (depending with rotifers. Rotifer tanks tank should be reduced, and
upon fish species and rotifer size). are usually the same size rotifers discarded.
Most producers estimate three (1,800 liters) as the algae 5) Drain-harvest rotifers for 1
times the amount of rotifers actu- tanks. Rotifer tanks must month unless a problem
ally eaten (1,900 X 3 = 5,700 also have drains and harvest occurs such as a “crash” or
rotifers per day) are fed per larva. baskets or nitex screen socks die-off. If this occurs, drain,
Therefore, as many as 39,900 (48- to 60-micrometer mesh) clean, disinfect and restart
rotifers (for a 7-day period) to to capture the rotifers. Both the tank. Restart the cultures
171,000 rotifers (for a 30-day peri- rotifer and algae tanks in clean tanks monthly.
od) may be required to feed one should have aeration and Starter cultures of rotifers
fish larva. Feeding too few rotifers illumination. should be maintained at low
often results in slow growth and 2) A few days after inocula- densities and in a separate
too much size variation; feeding tions the Tetrasalmis cultures facility. Densities of rotifers
too many rotifers can cause the will turn a darker green and at harvest will vary, but the
fish to ingest so much that assimi- cell densities will be about ranges to expect using this
lation becomes a problem. 132,000 cells per ml. Then technique are 100 to 150
For most marine finfish species gravity flow or pump algae per ml.
being reared indoors, the weaning to the rotifer tank and The health of the rotifers will pri-
of larvae from live rotifers and replace the algae volume marily be determined by the
Artemia to dry food should begin with clean, sterilized seawa- availability of an adequate food
well in advance of the transfor- ter and nutrients. In the supply. Hence, algae should be
mation from larvae to juveniles. rotifer tank, place a stock supplied in slight excess. In gener-
This transition might be timed to culture of rotifers in the al, the cultures should not be
take 3 days or as long as 2 weeks, algae (at least 1 rotifer per cleared of algae in less than 24
but should be done gradually. ml). Note: The larger the hours (i.e., after replacing algae;
Food particles should be the stock culture, the faster the the culture should have a rich
largest that can be swallowed eas- desired numbers of rotifers color that will clear to a lighter
ily by the fish (one-fourth to one- will be reached. color in not less than 24 hours).
half of mouth width). Starter feeds 3) After several days the algae Rotifer numbers and health
should contain 50 to 60 percent numbers should be obvious- should be checked daily. Using a
high quality protein. As an exam- ly decreasing (water looks dissecting microscope, a sample of
ple, in the past red drum larvae clearer) and the rotifer num- the rotifers should be observed for
were generally fed rotifers from bers increasing. Start drain- swimming speed (fast is good,
day 3 post-hatch to day 11, harvesting the rotifer tank slow is bad), gut fill (well packed
Artemia nauplii from day 11 to 21, into the mesh sock or har- gut that is easy to see indicates
and then weaned onto dry feeds. vest screen until approxi- good feeding; little or no food
More recent protocols include the mately 30 to 50 percent of indicates poor food densities, an
co-feeding of micro-particulate the culture tank is drained. undesirable species of algae, or
larval diets starting at day 5. Replace the drained volume contamination), percentage of
Although live zooplankton are with algae culture. Initially, rotifers with eggs (the more eggs
still used, dependence on them as the collected rotifers can be the better the culture), and num-
the sole nutrient source has been placed back into the culture ber of egg sacks carried (one indi-
significantly reduced, and the container. However, once the cates an adequate culture, two or
need to wean the animals from desired density is reached more a very healthy culture).
live foods is eliminated. (about 100 to 150 per ml) Most problems with rotifer cul-
about half of the rotifers will tures are caused by an inadequate
Production examples have to be harvested each supply of algae because of poor
day. algae culture techniques, under-
Batch/semi-continuous culture
4) Continue harvesting or dis- sizing of algae production, or an
of rotifers fed algae inability of the culturist to match
carding rotifers and refilling
1) Culture Tetraselmis chuii in the rotifer culture tank with the rotifer populations with the
1.8-ton (1,800-liter or 475- new algae culture daily. algae supply. The latter is general-
gallon) circular, fiberglass Volumes harvested from the ly a result of not discarding excess
tanks. The elevated fiber- rotifer tank may vary rotifers.
glass tanks should be according to demands of the
equipped with drains that hatchery; or standard vol-
Batch/semi-continuous culture 4) Once rotifer densities are 200 seem to work well in a wide range
using mixed feeds per ml, drain-harvest by of densities, prepared feeds gener-
draining 30 to 50 percent of ally work best in super-intensive
Even though zooplankton are the tank volume daily and batch production systems (more
generally considered good food capturing the rotifers in a 48- than 100 rotifers per ml), which
sources, they can be deficient in to 60-micrometer mesh net. are harder to manage over long
several essential nutrients, espe- Repeat until rotifer density periods of time. Because they are
cially the n-3 highly unsaturated drops. This culture method not live products, they do not stay
fatty acids (n-3 HUFAs) required should maintain rotifer den- suspended without considerable
for good growth and development sities at 150 to 200/ml for aeration. It should be noted that
of marine fish larvae. This is one about 30 days. using batch cultures and intensive
of the primary disadvantages of feeding regimes, an initial starter
rotifers, especially if they are Emulsified oil is a mixture of sea-
water, fish oil, and egg yolk at a culture of 100 rotifers per ml can
grown on a food source that is not reach densities of 1,300 per ml in 6
rich in HUFAs. Because a variety ratio of 100 ml: 5 ml: 1g, with the
addition of vitamin mix at 0.5% to 7 days. Although such densities
of factors influence the nutritional are desirable, the cultures are
quality of the rotifer, most produc- weight/volume of oil mixture.
Vitamin E is also added at 0.1% much harder to manage and
tion systems now use several food require careful attention to water
sources to enhance the nutritional weight/volume of oil mixture.
This mixture usually is a cod liver quality and feeding regimes.
content of live feeds.
or menhaden oil, raw chicken egg
Follow these steps to culture yolk, vitamin E (Tocopherol), and Copepods, Cladocerans,
rotifers on algae, baker’s yeast a vitamin mix (AIN Vitamin and Tintinnid ciliates as
and oil emulsion: Mixture 76®). The mixture is live feed
1) Follow the steps above for blended for 2 minutes in a blender
growing rotifers with algae. and then stored in a refrigerator Copepods are common zooplank-
Hold a rotifer starter/back- up to 1 week. The oil adds essen- ton both in freshwater and in
up culture at lower densities tial fatty acids and vitamins not brackishwater. They are natural
(100 per ml) in green water found in yeast. The eggs can be feeds for larvae and juveniles of
and use it to initiate the purchased at a grocery store. The many finfish and crustaceans
cultures as previously oil, vitamin E and vitamin mix- (Figs. 2a and 2b). In the wild,
described. ture can be purchased from ICN most marine larvae feed on cope-
Nutritional Biochemicals, pod eggs and nauplii during the
2) After algae is depleted for
Cleveland, Ohio. The menhaden first few weeks of life. Because
the first time in the rotifer
oil is produced by Zapata-Haynie some species of copepods have
tank, stop feeding algae.
Corp., Reedville, Virginia. Dry very small larvae (a necessity for
Instead, add the following
baker’s yeast can be obtained some larval fish species) and can
two products daily: baker’s
from wholesale grocery compa- have very high levels of HUFAs
yeast at 0.5 g/10 liters and
nies or most grocery stores. Some and other essential nutrients, they
oil emulsion (see makeup
researchers and commercial pro- are an excellent food source for
below) at 1 to 2 ml/10 liters.
ducers choose not to mix their first-feeding larvae. In fact, a
The remaining volume can
own oils, but prefer to purchase number of marine larval fish can-
be replaced with clean sea-
commercial enrichment products. not be reared using rotifers as the
water or de-chlorinated tap
There are also a variety of pre- first feed but have been reared on
water. Lowering the salinity
pared rotifer feeds that can be either laboratory reared or wild
to 16 to 18 ppt in the rotifer
used as a replacement for the caught copepod nauplii. Research
tank can be beneficial and
yeast. with several species, such as the
may improve growth after
turbot and red snapper, has
the rotifers are no longer Commercial enrichments and
shown that when offered mixed
being fed algae. Discard rotifer feeds are available from
plankton diets, young larvae con-
water when the rotifers are companies such as Aquafauna
sume more copepod nauplii than
harvested so they can adjust Biomarine Inc. ( California),
rotifers and prefer copepod nau-
rapidly to the higher salinity Sander’s Brine shrimp Co. (Utah)
plii because of the differences in
in the fish larvae environ- and Inve Aquaculture, Inc. (Utah).
size and swimming patterns of
ment. Algae pastes or concentrates are
the two prey types. Consequently,
3) Once rotifer population den- also available (Reed Mariculture
there is considerable interest in
sity reaches 100 per ml, Inc., California). The algae is
the use of copepods as feed
increase this daily yeast and grown under controlled condi-
sources for small marine larval
oil emulsion level to 0.7 to tions, concentrated using a cream-
fish.
1.0 g yeast per million separator, then preserved and
packaged. These products can be Copepods are cylindrical with a
rotifers and 2 to 3 ml oil
refrigerated for 1 month or frozen trunk comprised of 10 segments,
emulsion per million rotifers.
for more than a year. Although consisting of head, thorax and
the concentrated algae products abdomen. Adult copepods range
from 0.5 to 5.0 mm. The larval (Tisbe and Tigriopus spp.), and which gives them an advantage
stages consist of six naupliar and cyclopoids (see Fig. 2a for shape over the rotifers. Copepods can
six copepodite stages. The main differences). also eat detritus. They differ from
suborders of copepods found in Herbivorous copepods are pri- Artemia (brine shrimp) and
brackishwater are calanoids marily filter feeders and typically rotifers in that they do not repro-
(Acartia, Calanus and feed on very small particles. But duce asexually. Copepods mate
Pseudocalanus spp.), harpacticoids they can feed on larger particles, after maturing and the female

Copepods – saltwater Cladocerans (water fleas) – freshwater

a. c.

HARPACTICOID

Egged female

Daphnia

CALANOID
Daphnia
CYCLOPOID

Copepods – freshwater
b.

Macrothrix

Cyclops Diaptomus
Figure 2. Copepods and Cladocerans
produces 250 to 750 fertilized Tigriopus and Acartia for rearing viduals per ml could be main-
eggs (rotifers produce 15 to 25 per fish larvae approximately 7mm in tained on Tetraselmis chui after
female). The copepod lifespan is length. U.S. researchers com- maximum density was attained
40 to 50 days (5 to 12 days for pared the growth and biochemical (for general culture). In Thailand,
rotifers), and it has a longer gen- composition of mahi-mahi culturists are growing
eration time (1 to 3 days for the (Coryphaena hippurus) larvae that Diaphanosoma on Chlorella sp. In
rotifer and 7 to 12 days for the were fed brine shrimp, rotifers 1998, researchers at SEAFDEC in
copepod). and the copepod Euterpina acu- the Philippines successfully used
Unlike the rotifer, copepods are tifrons, cultured in 700-liter tanks. Diaphanosoma as an Artemia sub-
more difficult to culture on a Larvae fed copepods survived stitute for Barramundi larvae
commercial basis. Only a few better under stressful conditions. (Lates calcarifer).
species of copepods, such as A system for the mass culture of a Other cladocerans considered
Tigriopus japonicus, have been benthic marine harpacticoid cope- promising species are Evandne
mass cultured successfully. Even pod, described by Sun and tergestina, Penilia avirostris and
this technique employs the com- Fleeger (1995), should be useful Podon polyphemoides. The cladocer-
bination of rotifer culture and the for aquaculture. an Moina macrocopa has been used
use of baker’s yeast or omega-3 Other copepods considered to be in Southeast Asia as feed for sea
yeast as feed. Unfortunately, the promising species for mass cul- bass fry immediately after wean-
amount of yeast used to produce ture are Acartia clausi, A. lon- ing from Artemia and prior to
the copepod and rotifer combina- giremis, Eurytemora pacifica, feeding minced fish flesh. During
tion outdoors is fairly high. There Euterpina acutifrons, Oithona brevi- this period, sea bass, being a
are outdoor production systems cornis, O. similis, Pseudodiaptomus catadromous species (moving into
that can produce large numbers inopinus, P. marinus, Microsetella freshwater for a portion of its life
of copepods; however, these sys- norvegica and Sinocalanus tenellus. cycle), may be reared at lower
tems are very inefficient in terms salinities and fed freshwater zoo-
Cladocerans or water fleas (Fig.
of number of copepods per liter plankton. This practice is not
2c), such as Daphnia magna, have
of culture water. Considerable commonly used or proven to be
been cultured as live food using
work needs to be done on culture viable on a commercial scale. A
techniques similar to those
and harvest techniques before related cladocera, Moina salina,
described for rotifers. Many labo-
copepods become as widely used has been used in finfish culture in
ratories use Daphnia as the inver-
as rotifers. Spain.
tebrate of choice to conduct toxici-
One interesting advantage of ty tests because it is easy to cul- Tintinnid ciliates are consumed by
copepods is that under appropri- ture and maintain in the laborato- larval fish and crustaceans in the
ate conditions some species will ry. Cladocerans are mainly fresh- wild and are considered promis-
produce a resting egg similar to water zooplankters; most do not ing candidates for mass produc-
that of Artemia. So once commer- tolerate salinities higher than 3 tion. However, since the technolo-
cial techniques are developed, ppt., and are generally not found gy for mass production of rotifers
copepod eggs could be collected in brackishwater. One exception is is well established and micropar-
in large numbers and stored for Diaphanosoma celebensis ticulated diets are being co-fed
months, like Artemia (brine (=aspinosum). In Asia there is a with rotifers or have been devel-
shrimp) and rotifer cysts. growing use of this species. This oped to partially substitute for
Photoperiod and temperature is a saline-tolerant (1- to 42-ppt) live food, the role of copepods,
largely determine the production water flea in the 400- to 800- cladocerans and tintinnid ciliates
of copepod resting eggs. Labora- micrometer range that has been is not as important.
tory production of these eggs is successfully cultured in backyard
possible, but has not yet proved hatcheries. Biomasses of up to 1 Overview
to be economically feasible. It is kg in 1 cubic meter of water every
hoped that using copepods as a 3 days have been reached. To be Finfish producers are concerned
food source can improve the cul- effective as a replacement, the with improving the quality, quan-
ture of a variety of species, such organism must be enriched before tity and cost effectiveness of their
as the red drum, by reducing the it is fed. This enrichment is live feed production facilities.
size variability and mortality. accomplished with a source of Many of them now supplement
DHA, but usually not one with cultures with omega yeast, vita-
The use of copepods, especially mins (E, D, C and B12), marine
the harpacticoids (Fig. 2a), is well an oil emulsion base because of
gill and water fouling problems. oils or other HUFA sources, and
documented in marine fish cul- vitamin B12-producing bacteria to
ture. Researchers have reared Schizochytrium (a spray-dried or
drum-dried algae developed by improve feed quality. Today, live
copepods in vessels of 100 liters feeds for fish larvae are being
(26 gallons) and 450 liters (118 Omega-Tec, Inc.), is the most
common enrichment agent used improved by adjusting their bio-
gallons) and reported that the chemistry through controlling
system provides 250,000 nauplii in Thailand for Diaphanosoma.
Researchers have found that their diet and supplementing the
per day. The Japanese have rou- cultures with microencapsulated
tinely cultured the copepods mean densities of 72 to 100 indi-
feeds or emulsified oils. While Japan. In the past it was thought Suggested reading
algae and rotifers are the most that fish larvae have low concen-
widely used live food items, their trations of digestive enzymes until Davis, C. 1955. The Marine and
use is not without problems and they reach approximately 6 mm; it Freshwater Plankton. Michigan
limitations. Rotifer and copepod was also thought that they are State University Press, East
cultures are subject to collapse or unable to digest inert feeds. Lansing, Michigan.
“crash.” Producers are finding Evidence is accumulating to sup- Hyman, L.H. 1951. The
new species of live food organ- port the idea that properly formu- Invertebrates: Acanthocephala,
isms better suited for specific cul- lated diets are digested and pro- Aschelminthes and Ectoprocts,
ture situations. Larvae of some vide a controlled way of deliver- Vol. III. McGraw-Hill, New York,
species (e.g., angelfish, butterfly- ing nutrients to larvae. For many New York.
fish, damselfish, parrotfish) have commercial species, the co-feeding Fulks, W. and Main, K.L. 1991.
small mouths and might require of live and artificial feeds during Rotifer and Microalgae Culture
prey smaller than rotifers. the larval stages is recommended. Systems. The Oceanic Institute,
Dinoflagellates such as Microencapsulated diets do have Honolulu, Hawaii.
Gymnodinium sp., ciliates such as one very positive attribute—they
Euplotes sp., and the nauplii of are an alternative way to adminis- Sun, B. and Fleeger, J.W. 1995.
many copepods are in the size ter vaccines and therapeutic Sustained mass culture of
range suitable for those fish lar- agents to larvae. Even though the Amphiascoides atopus, a marine
vae. Larvae of oysters, barnacles large-scale, intensive production harpacticoid copepod, in a recir-
and sea urchins have also been of microalgae and rotifers is culating system. Aquaculture 136:
used, but are not as reliable in expensive and often unreliable, 313-321.
quantity and quality. Advances the production of live food organ-
have also been made in the area of isms continues to be a very impor-
formulated feeds, especially in tant first step in aquaculture.
The work reported in this publication was supported in part by the Southern Regional Aquaculture Center through Grant No. 97-38500-4124 from
the United States Department of Agriculture, Cooperative States Research, Education, and Extension Service.

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