U i6 tS
14-0
MARINE ECOLOGY - PROGRESS SERIES
Mar. Ecol. Prog. Ser.
V ol. 3: 3 0 3 -3 0 7 , 1980
P u b lish e d D e c e m b e r 15
International Study on Artemia*. XIV. Growth and
Survival of Artemia Larvae of Different
Geographical Origin in a Standard Culture Test
P. Vanhaecke and P. Sorgeloos**
A rtem ia Reference Center, State University of Ghent, J. Plateaustraat 22, B-9000 Ghent, Belgium
ABSTRACT: For characterization of strains of the brine shrim p A rtem ia of d ifferent geographical origin, a stan d ard culture test
has b e e n developed in order to com pare statistically grow th a n d survival of larv ae of different strains. 25 geographical strains
have been studied so far - inclu d in g , for 3 strains, analyses of cysts h arv ested at different times. Im portant differences in rates of
grow th and survival w ere observed b etw een strains b u t not am ong b atches of th e sam e strain. Best perform ances w ere noted for
strains from B ahia Salinas (Puerto Rico), Buenos Aires (A rgentina), C haplin Lake (C anada), G reat Salt Lake (Utah, USA), G alera
Zam ba and M anaure (Colombia).
INTRODUCTION
T he use of A rte m ia ** * as live food for larval fishes
and crustaceans has, th u s far, b e e n m ostly restricted to
nauplii (for details consult 'M arine Ecology', Volume
III: Kinne, 1977). A lthough it w as dem onstrated that, as
predators grow, b etter survival and grow th are
obtained w ith b rin e shrim p larvae (Walne, 1967; Sick
and Beaty, 1974; Purdom an d Preston, 1977), nauplii
are still p referred b ecau se of th e sim plicity of hatching
procedures, as opposed to m ore labour, eq u ip m en t and
food costs req u ired for raisin g larv ae (Brouillet, 1977 in
Girin, 1979). H ow ever, d u e to recen t developm ents in
A rtem ia cultivation on ch eap diets (Sorgeloos et al.,
1980; D obbeleir et al., 1980) in easy-to-operate ra ce ­
way system s (Bossuyt and Sorgeloos, 1980), th e use of
A rtem ia larv ae and p read u lts in aq u acu ltu re h atch ­
eries is g ain in g interest (Sorgeloos, 1980), not only
b ecau se of th e im proved o u tp u t of fish and crustacean
hatcheries, b u t also b ecau se of substan tial savings in
the quantity of A rtem ia cysts n e e d e d for hatchery oper­
ations (Fujimura, 1978).
W ithin the fram ew ork of th e ‘International Study on
Artem ia', w hose goal is to im prove th e use of brine
shrim p in aq u acu ltu re through characterization and
selection studies of various geographical strains of
A rtem ia (Sorgeloos, 1980), a com parative study of
grow th rates and conversion efficiencies is presently in
progress. A lthough there is evidence th a t grow th rates
can vary significantly from one strain of brine shrim p
to another (Gilchrist, I960; Sorgeloos, 1975; Tobias et
al., 1980) m ore inform ation is n eeded, preferably
obtained u n d e r standardized experim ental conditions.
This p ap e r reports on the rates of grow th and survi­
val of 25 different Artem ia strains (3 of w hich w ere
analyzed after differential harvesting in a standard
culture test.
’ International Interdisciplinary Study on A rtem ia strains,
coordinated by th e A rtem ia R eference C enter, State U ni­
versity of G hent, Belgium
' * 'B evoegdverklaard N avorser’ a t th e B elgian N ational Sci­
ence Foundation (N.F.W.O.)
* * * The bin o m en A rtem ia salina L. is taxonom ically no
lo n g er valid (see review s in Bowen and Sterling, 1978; Bowen
et al., 1980). In view of the im portant g en etical differences
b e tw een p arth en o g en etical strains of b rin e shrim p, (AbreuGrobois a n d Beardm ore, 1980) species definition in the genus
A rtem ia has becom e unclear. O ne of the m ain conclusions of
th e 'International Symposium on th e Brine Shrim p, A rtem ia
salina L.’ (Corpus Christi, Texas-USA, Aug. 20-23, 1979) was
that, unless th e exact sibling species can be identified (20
bisexual strains have been classified so far into 5 sibling
species by B ow en et al., 1978) a n d until sp éciation in brine
shrim p is b e tte r understood, only the genus designation
A rtem ia should be used (Persoone et al., 1980)
© by Inter-R esearch
0171-8630/80/0003/0303/$ 02.00
304
M ar. Ecol. Prog. Ser. 3: 303-307, 1980
MATERIALS AND METHODS
For each A rtem ia strain approxim ately 100 m g of
cysts w ere incubated u n d er optim um h atch in g condi­
tions (Sorgeloos, 1980). The culture experim ents w ere
carried out in test tubes filled w ith 25 m l of 0.2-/zm
filtered n atu ral seaw ater and k e p t in d arkness at 25 °C
± 0.5 C°. Each tube w as in o cu lated w ith 10 instar-I
nauplii.
T he A rtem ia larvae w ere fed once daily w ith a
defined n um ber of algal cells (D unaliella viridis): Day
1, 12 X IO5 cells; Days 2, 3 and 4, 24 X IO5 cells; Days
5 and 6 , 36 X 1 0 s cells; Day 7 , 48 X 1 0 s cells. This
optim al feeding schedule has b e e n d eterm ined by
V an h aeck e and Coorem an (1979) in prelim inary cul­
tu rin g tests w ith nauplii h atch ed from San Francisco
Bay (California-USA) cysts (San Fancisco Bay Brand
Com pany, Batch 288-2596).
The alg ae w ere cultured according to De Pauw et al.
(1978), h arvested in th eir exponential grow th phase by
centrifugation, then resu sp en d ed in filtered seaw ater
and dilu ted to a final concentration of 12 X IO6 cells
m l-1. T he suspension was th en stored in d arkness at
4 °C d uring the w hole experim ental period. A lgal sus­
pen sio n w as distributed w ith an autom atic m icropipet
(100, 200, 300 or 400 /¿I tu b e-1 d-1) according to feeding
schedule. After each feeding th e test tub es w ere sh a­
k e n to assure good food distribution.
The n u m b er of surviving b rin e shrim p w ere counted
on Day 8, fixed w ith lugol's solution (100 ¡A p e r tube).
T hese w ere th e n transferred to m icroscopic slides
w h ere th e ir length (from the anterio r tip of th e head
along th e intestine to the base of th e furca) w as d e ter­
m ined by cam era-obscura draw in g at a m agnification
of 50x.
The m inim um n u m b er of rep licate test tubes neces­
sary for statistical analysis for each g eographical strain
w as d eterm in ed according to C ohran and Cox (in
Sokal and Rohlf, 1969) using d ata from prelim inary
experim ents (V anhaecke and C oorem an, 1979). Four
rep licate tubes assured 95 % probab ility for detecting
a 15 % difference from th e m ean a t the 0.05 level.
Since h ig h er m ortalities can be ex pected w ith some
experim en tal strains (the San Francisco Bay strain
gives an averag e of over 90 % survival) 5 replicate test
tu b es p e r strain w ere used. T est tu b es w ith a survival
b elow 70 % w ere not ta k e n into consideration for
grow th evaluation.
The culture tests are d esig n ed to en ab le statistical
in terp retatio n by m eans of a one w ay analysis of var­
iance, M odel I (Sokal and Rohlf, 1969). The survival
d a ta are norm alized through an arc sin V perceñt tran s­
form ation. The standard culture test w as re p eated for
th e strains from Shark Bay, M acau (harvested in M arch
1978), an d from A delaide and G reat Salt Lake (har­
v ested in 1977).
A com parison betw een all strains tested w as pos­
sible by using th e San Francisco Bay strain, Batch
288-2596 as in n er standard; th e grow th for each strain
(up to 5 strains w ere tested p e r experim ent) was ex­
pressed as p ercen tag e of the grow th recorded for the
reference strain.
RESULTS
For th e reference strain, average larval length after 7
days is 3.16 ± 0.17 /zm; the m e an percent survival was
94.2 ± 2.8 %. G row th and survival data for the 25
geographical strains tested are sum m arized in Table 1.
Significant differences in grow th rates occur be­
tw een th e geographical strains studied. Com pared to
th e reference strain, brine shrim p form L am aca Salt
Lake, Santa Pola, Salin du G iraud and an unknow n
locality in C hina grow significantly slower. In contrast,
hig h er grow th rates prevail for A rtem ia from A delaide,
M anaure, Bahia Salinas, G reat Salt Lake, Buenos
Aires, G alera Z am ba and C haplin Lake. For 14 other
strains no significant differences from th e reference
strain could be observed. The variations betw een diffe­
rent harvests from the sam e strain and b etw een the 4
re p eated experim ental runs are sm all and not signific­
ant. Furtherm ore, the differences in grow th rates b et­
w een th e San Francisco Bay strain an d the strains
originating from either the San Francisco Bay trans­
plantations in M acau and Barotac Nuevo (Sorgeloos et
al., 1979) or the Pj generation of San Francisco Bay
cysts produced u n d er laboratory conditions (Versichele and Sorgeloos, 1980) are statistically not sig­
nificant.
W ith regard to percen t survival, m ortality rates sig­
nificantly higher th a n in th e reference strain prevail for
A rtem ia from Bonaire, Salin du G iraud, G reat Salt
Lake 1966, L am aca Salt Lake, Lavalduc, Buenos Aires,
San Felix and S anta Pola. V ariations betw een different
harvests are sm all w ith th e exception of the G reat Salt
Lake sam ples 1966 and 1977. How ever, during th e last
decade this biotope has been subjected to considerable
ecological changes (Stephens an d G illespie, 1972).
DISCUSSION
T he standard culturing test, outlined above and
tested on 25 strains, appears to b e applicable for com­
paring A rtem ia strains on th e basis of larval growth
and m ortality. Since it has b ee n reported earlier that
th e optim al tem perature-salinity com bination can vary
from one A rtem ia strain to another (Sorgeloos et al.,
1976) it is possible that som e differences in grow th and
V anhaecke and Sorgeloos: In ternational study on Artem ia. XIV.
305
T able 1. A rtem ia. P ercent grow th and survival of d ifferent g eo g rap h ical strains of brine shrim p u n d e r stan d ard culture conditions
Survival at Day 7
(%)
G eographical strain
L am aca Salt Lake (Cyprus)
S anta Pola (Spain)
S alin d u G iraud (France)
C h in a (unknow n locality)
S hark Bay (Australia)
S an Francisco Bay (California/USA; cysts produced in th e laboratory)
San Francisco Bay (California/USA; Batch No 236-2016)
M acau (Brazil; harv ested in M arch 1978)
B arotac N uevo (Philippines)
T ien tsin (China)
San Francisco Bay (California/USA; Batch No 933-235)
A iguës M ortes (France)
Izm ir (Turkey)
M arg herita di Savoia (Italy)
San Felix (Spain)
M acau (Brazil; harv ested in M ay 1978)
B onaire (N etherlands Antilles)
San Pablo Bay (California/U SA; Batch No 1628)
Port A raya (V enezuela)
E ilat (Israel)
Lavalduc (France)
A delaide (Australia)
M anaure (Colombia)
B ahia Salinas (Puerto Rico)
G reat Salt Lake (Utah/USA; h arvested in 1977)
B uenos A ires (Argentina)
G alera Z am ba (Colombia)
G reat Salt Lake (Utah/USA; harvested in 1966)
C h ap lin Lake (C anada)
70*
76*
66*
84
90
92
98
84
86
96
94
90
96
96
74*
94
66*
90
90
92*
70*
88
90
88
94
72*
98
66*
88
84®
96a
88a
94 a
G row th expressed as %
recorded for A rtem ia
referen ce strain,
San Francisco Bay,
Batch 288-2596
88
88
90
91
95
96
96
96
97
98
99
99
101
102
102
103
104
105
107
107
109
113*
115
122
119
126
126
127
130
—
96a
98a
113a
125a
—
a result of replicate test in tim e
* significantly different from the reference strain at th e 0.05 level
• * significantly different from th e reference strain at th e 0.01 level
survival b e tw e e n th e strains studied have been
m ask ed by our p resen t culture technology (25 °C;
n a tu ra l seaw ater of ca 35 ppt). In any case, the order of
grow th perform ance rep o rted by Sorgeloos (1975) for
th ree strains (San Francisco Bay, G reat Salt Lake and
China) is id en tical to th e sequence obtain ed h ere for
the sam e strains.
In this study, all A rtem ia strains th at grew signifi­
cantly faster th a n th e reference strain w ere bisexual.
H ow ever, G ilchrist (1960) reported th at a parth en ogentic stra in from La Palm e (France) grew faster th a n a
bisex u al A rtem ia from San D iego (California, USA). It
ap p ears from our results th a t this observation cannot
b e ascribed to a difference in grow th rate re la te d to th e
m ode of reproduction. O ur study provides fu rth er ev i­
dence th a t grow th rates of A rtem ia larvae are straind ependent. T h ere are indications th at this p h e n o m e ­
non m ight b e g en etically controlled: the strains p ro ­
duced from S an Francisco Bay inoculations in Brazil
an d th e Philippines, or from a San Francisco Bay po pu­
lation m ain tain ed under laboratory conditions ex h i­
bited grow th perform ances sim ilar to the parental
strains.
W hile b rin e shrim p strains b etw een w hich the g e n ­
etic distance is know n to be large (e. g. Buenos Aires
an d C haplin Lake; A breu-G robois and Beardm ore,
1980) can hav e sim ilar grow th rates, different rates
m ay b e observed in strains of th e sam e sibling species,
e. g. from C haplin Lake and San Francisco Bay, w hich
are both A rtem ia franciscana (Bowen et al., 1978). It is
furtherm ore in teresting to note th a t no correlation
exists betw een grow th rates an d biom etrical charac­
teristics of nau p lii from corresponding strains (Van­
h a eck e an d Sorgeloos, 1980).
W ith reg ard to th e practical use of A rtem ia in
aquaculture, th e results rep o rted here provide a
gu id elin e for the selection of specific geographical
strains. W hile data on growth rates are useful, a d d i­
Mar. Ecol. Prog. Ser. 3: 303-307, 1980
306
tional criteria should be tak en into consideration.
A ccordingly, our com parative analyses have now b een
ex ten d ed to include biom ass production, food conver­
sion efficiency and tem peratu re-salin ity preferences.
A ckn o w led g em en ts. W e are greatly in d eb ted to th e m any
people w ho provided us w ith cyst sam ples from different
geographical b rin e shrim p strains. This study w as supported
b y the B elgian N ational Science Foundation (N. F. W. O.)
through G rant F. K. F. O. - 2.0010.78.
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This p ap er w as p resen ted by Professor G. Persoone; it w as accep ted for p rin tin g on A ugust 8, 1980