tu r tiö ie g - e p u e j s o Q
:
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S H 3 G N V ld
ci J ( - '' '
S 'W W l A
(m
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y n A
A quaculture Research, 2007, 38, 1539-1553
doi:10,llll/j.l365-2109.2007.01814.x
Improved te c h n iq u e s for rearing m ud c ra b Scylla
param am osain (E stam pador 1949) la rv a e
Truong Trong N ghia1, M a th ieu W ille2, Tran C ong B in h \ H oang P h u o c T h an h 1, N guyen Van D a n h 1 &
P atrick S orgeloos2
'College of A quaculture and Fisheries, C anTho University, C anT ho City, Vietnam
la b o r a to r y of A quaculture & Artemia Reference Center, G hent University, Ghent, Belgium
C o rre s p o n d e n c e : T Trong Nghia, College of A quaculture and Fisheries, Can Tho University, 3 rd February avenue, Campus II. Can Tho
City,Vietnam. E-mail: ttnghia@ ctu.edu.vn
A b s tra c t
r a te fro m Z1 to c ra b 1 - 2 s ta g e s in la rg e r e a r in g t a n k s
A s e rie s o f r e a r in g tr ia ls in s m a ll 1 L c o n e s a n d la rg e
t a n k s o f 3 0 - 1 0 0 L w e r e c a r r ie d o u t to d ev elo p o p ti­
m a l r e a r in g te c h n iq u e s fo r m u d c ra b {Scylla p a ram a­
m osain) la rv a e . U sing w a te r e x c h a n g e ( d is c o n tin u o u s
p a r t ia l
w a te r r e n e w a l
o r c o n tin u o u s
tr e a t m e n t
o f 1 0 -1 5 % ( m a x im u m 30% ).
K eywords: S c y lla p a ra m a m o sa in , r e a r i n g te c h n i ­
q u es, w a te r e x c h a n g e , m ic r o -a lg a e , la r v a l d e n s ity ,
life food d en sity , p r o p h y la x is
t h r o u g h b io filtra tio n ) a n d m ic ro -a lg a e (Chlorella o r
Chaetoceros) s u p p le m e n ta tio n (d aily s u p p le m e n ta tio n
at
0 .1 -0 .2 m illio n cells m L - 1
1 - 2 m illio n s c ells m L - 1 ),
or
six
m a in te n a n c e
d iffe re n t
ty p e s
at
of
Introduction
r e a r in g sy s te m s w e r e trie d . T h e c o m b in a tio n o f a
A q u a c u ltu r e o f m u d c ra b s. Scylla spp., c o n tr ib u te s a
g r e e n - w a te r b a tc h s y s te m fo r e a rly sta g e s a n d a r e c ir ­
la rg e p r o p o rtio n to t h e w o rld p r o d u c tio n o f t h e g e n u s
c u la tin g s y s te m w ith m ic r o -a lg a e s u p p le m e n ta tio n
(FA O 1999). M oreover, m u d c ra b s r e p r e s e n t a v a lu a b le
fo r l a te r s ta g e s re s u lte d in th e b e s t o v e ra ll p e r fo r ­
c o m p o n e n t o f s m a ll-s c a le d c o a s ta l fish e rie s i n m a n y
m a n c e o f t h e c ra b la rv a e . N o c le a r effects o f crab
c o u n tr ie s in tro p ic a l a n d s u b tro p ic a l A sia, fo r w h ic h
s to c k in g d e n s ity
r o tif e r
th e r e h a s b e e n a g e n e r a l tr e n d o f in c r e a s e d e x p lo ita ­
(1 0 -
tio n in r e c e n t y e a r s (A n g eli 1992; K e e n a n 1999 a). I n
(5 0 -2 0 0 la rv a e L " ^
( 3 0 - 6 0 ro tife rs m L ~ 1)
A rte m ia
and
and
d e n s ity
of
V ietn am , th e m u d c r a b Scylla p a ra m a m o sa in is t h e
ro tife r
s e c o n d m o st i m p o r ta n t m a r in e s p e c ie s n e x t to
o f 3 0 - 4 5 m L ~ 1 fo r e a rly s ta g e s a n d A rte m ia fe e d in g
sh rim p , b e in g c u lt u r e d w id ely i n t h e c o a s ta l a re a .
a t 1 0 -1 5 n a u p l ii m L - 1 fo r Z 3 -Z 5 se e m e d to p r o d u c e
H ow ever, m u d c ra b fa rm in g c u r r e n tly re lie s e n tire ly
t h e b e s t p e r fo r m a n c e o f S. p a ra m a m o sa in la rv a e . O p­
o n th e w ild fo r se e d s to c k a n d th e m a in o b sta c le for
tim a l r a tio n s fo r c ra b la r v a e sh o u ld , h o w ev er, b e a d ­
e x p a n s io n is t h e u n a v a ila b ility o f h a tc h e r y - re a r e d
j u s te d d e p e n d in g o n t h e sp ecies, la r v a l sta g e , la r v a l
se e d (L iong 1992; R a tta n a c h o te & D a n g w a ta n a k u l
20 L -1 )
w e re
1 0 0 -1 5 0 zoea 1
o b se rv e d .
(Z 1 )L ~ \
A
s to c k in g
c o m b in e d
d e n s ity
w ith
s ta tu s , p re y size, r e a r in g s y s te m a n d te c h n iq u e s . A
1992; K e e n a n 19 9 9 a; S h elley & Field 1999; M a n n ,
p r a c tic a l fe e d in g s c h e d u le c o u ld b e to in c r e a s e live
A s a k a w a , P izzu to , K e e n a n & B rock 2001; X u a n
fo o d d e n s ity fro m 3 0 to 4 5 ro tife rs m L - 1 fro m Z1 to
2001).
Z2 a n d in c r e a s e t h e n u m b e r o f A rte m ia n a u p lii m L - 1
R e a rin g te c h n iq u e s , d is e a s e a n d n u tr itio n a r e th e
f ro m 10 t o 15 fro m Z3 to Z5. B a c te ria l d ise a se r e m a in s
t h r e e m a in a r e a s of r e s e a rc h t h a t h a v e s u p p o r te d
o n e o f t h e key f a c to rs u n d e r ly in g th e h ig h m o r ta lity
in t h e z o e a sta g e s. F u r t h e r r e s e a r c h to d ev e lo p sa fe
c o m m e rc ia l p ro d u c tio n o f m a r in e fish a n d c r u s t a ­
p ro p h y la c tic tr e a tm e n ts is th e r e fo r e w a r r a n te d . C o m ­
p e c ts a re to a larg e e x te n t in te r c o n n e c te d a n d
b in e d w ith p r o p e r live fo o d e n r ic h m e n t te c h n iq u e s ,
d ev e lo p in g h a tc h e r y te c h n iq u e s fo r a ‘n e w ’ sp e c ie s is
a p p lic a tio n o f th e s e fin d in g s h a s s u s ta in e d a s u rv iv a l
n o t p o ssib le u n le s s all t h e a r e a s a r e a d d re ss e d . T h e
© 2007 The Authors
Journal Compilation © 2007 Blackwell Publishing Ltd
c e a n la rv a e (Sorgeloos & L ég er 1992). T h e s e th r e e a s ­
1S39
Improved rearin g techniques for m ud crab larvae T T Nghia et al.
A quaculture Research, 2007, 38, 1539-1553
d e s ig n o f r e a r in g s y s te m s c o v e rs m o re t h a n p u rely
(30 g L - 1 ) a n d a m m o n ia a n d n i tr i te levels. E very
t e c h n ic a l a s p e c ts. S n b -o p tim a l r e a r in g c o n d itio n s
o t h e r day, t h e c ra b s w e r e p la c e d i n a 5 0 p L L - 1 for­
(e.g. p h y sic a l stre s s, la c k o f o x y g e n o r s u b -o p tim a l
m a lin b a t h for 1 h to r e d u c e o r p r e v e n t in f e s ta tio n of
w a te r q u a lity ) a ffe c t la r v a l h e a l t h a n d c a n c a u s e
t h e e g g s w i th f u n g i a n d b a c te r ia . D u r in g e g g in c u b a ­
m a s s m o r ta lity d u e to d is e a s e o u tb r e a k s . Sim ilarly,
tio n , t h e c ra b s w e r e n o t fed.
sy s te m d e s ig n in f lu e n c e s (live) food q u a lity a n d its
O n e to tw o d a y s b efo re h a tc h in g , t h e b e rrie d
fe m a le w a s m o v ed to a 5 0 0 L fib re g la s s ta n k . W h e n
av a ila b ility to t h e p r e d a to r la rv a e .
T h e re h a s b e e n a g r e a t d e a l o f p ro g re s s in m a rin e
th e h a t c h i n g p ro c e s s w a s c o m p le te d , la r v a e w ere
la r v a l r e a r in g te c h n o lo g y s in c e its b e g in n in g in th e
se le c te d b a s e d o n t h e i r p h o to ta c tic b e h a v io u r. A e ra ­
1 9 6 0 s (S h e lb o u rn e 1964; H o w ell, Day, E llis & B aynes
tio n in t h e h a tc h in g t a n k w a s t u r n e d off f o r se v e ra l
1998). M a n y o f t h e s e te c h n ic a l im p ro v e m e n ts devel­
m in u te s a n d t h e a c tiv e la r v a e s w im m in g u p to th e
o p ed o v e r t h e p a s t d e c a d e s c o u ld be a p p lie d fo r m u d
s u rfa c e w e r e c o lle c te d b y g e n tle sc o o p in g .
c ra b w i t h so m e m o d ific a tio n s . A n o v e rv ie w o f th e
T h e la r v a e w e re t h e n tr a n s f e r r e d to t h e r e a r in g
r e a r in g sy s te m s c u r r e n tl y a p p lie d fo r l a r v ic u ltu r e of
c o n ta in e r s . A c c lim a tio n w a s p e r fo r m e d b y p la c in g
m u d c ra b s w a s p r e s e n te d b y D av is (2003). A lth o u g h
t h e la r v a e in a 5 0 L p la s tic m e s h b u c k e t a n d slow ly
k n o w le d g e h a s b e e n o b ta in e d fro m th e s e system s,
r in s in g t h e m w ith w a te r fro m t h e la r v a l r e a r in g c o n ­
th e r e is a n e e d to f u r t h e r o p tim iz e r e a r in g te c h n iq u e s
ta i n e r s f o r 2 0 - 3 0 m in b efo re re le a se .
i n o r d e r to m a x im iz e la r v a l s u r v iv a l a n d quality.
F u rth e rm o r e , te c h n iq u e s s h o u ld b e a d a p te d fo r ea c h
S cylla s p e c ie s (K een an , D a v ie & M a n n 1998; K e e n a n
Food a n d feed in g
1999b) i n r e la tio n to lo c a l c o n d itio n s (seaw a ter
so u rc e ,
s ta tu s
of
h a tc h e ry
m a n a g e m e n t,
lo cal
re so u rc e s). T h e a i m o f th i s r e s e a r c h is to a d a p t th e
M icro-algae c u ltu re
S ta r tin g c u ltu r e s o f th e m ic r o -a lg a e C haetoceros calci­
e x is tin g r e a r in g s y s te m s fo r l a r v ic u l tu r e to m u d crab
trans a n d Chlorella vulgaris w e r e m a i n ta i n e d in d o o rs
sp e cies (S. p a ra m a m o sa in b e in g t h e te s t case) a n d to
w ith W a ln e s o lu tio n i n s e a w a te r o f 3 0 g L “ 1 a t 25 °C.
im p ro v e o t h e r te c h n iq u e s i n o r d e r to m a x im iz e la rv a l
L a rg e -s c a le p r o d u c tio n w a s p e rfo rm e d in d o o r s in
s u rv iv a l a n d q uality.
5 0 0 L ta n k s u n d e r a tr a n s p a r e n t ro o f. A h a e m o c y to m e te r w a s u s e d to c o u n t m ic r o -a lg a l d e n s itie s .
M aterials a n d m e th o d s
R o tifer c u ltu re and e n rich m en t
Sou rce o f larvae
T h e s a m e B rachionus p lica tilis L -s tra in w i th a lo ric a
G ravid c ra b s w e re b o u g h t fro m lo c a l m a rk e ts a n d
l e n g th a n d w id th o f 1 6 4 ± 2 2 a n d 1 2 0 ± 2 2 p m , r e ­
tr a n s p o r te d to th e h a tc h e ry . B efo re s to c k in g in th e
spectively, w a s u s e d i n a ll t h e e x p e rim e n ts . R otifers
h atch ery , th e c ra b s w e r e su b je c te d to a b a th of
w e re c u l tu r e d in d o o rs in 1 0 0 L fib reg lass ta n k s o p e r­
10 0 p L L - 1 o f a 4 0 % fo rm a lin s o lu tio n fo r l h . T he
a te d in a b a tc h m ode, fo llo w in g t h e p r o c e d u r e d e ­
c ra b s w e re s to c k e d in d iv id u a lly i n 1 0 0 L c o m p a rt­
s c rib e d i n S o rg e lo o s a n d L av en s (1996). R o tife rs w e re
m e n ts o f a
in itia lly g r o w n o n b a k e r s y e a s t, b u t l a te r o n fed
eq u ip p e d
ro o fe d
w ith
a
2 x 2 x 0.5 m
biofilter.
c e m e n t tan k ,
R e a rin g
w a te r
of
30 ± 1 g L “ 1 s a lin ity w a s d ilu te d fro m b r in e (9 0 —
C u ltu re S e lc o ' (INVE A q u a c u ltu r e , D e n d e rm o n d e
B elg iu m ) b e fo re fe e d in g to t h e la rv a e . T e m p e ra tu re
110 g L " x) w ith ta p w a te r a n d c h lo r in a te d befo re use.
a n d s a lin ity w e re c o n tro lle d a t 25 °C a n d 2 5 g L _1
A m b ie n t
respectively. T h e y w e r e h a r v e s te d t h r o u g h a 6 0 p m
te m p e r a tu re
flu c tu a te d
slig h tly
aro u n d
28 °C. E v ery c ra b w a s fed a d a ily r a tio n of 1 0 -1 5 g of
s c re e n a n d r in s e d th o ro u g h ly .
fre sh m a r in e sq u id , b ivalve o r s h r im p m e a t alternately.
R otifers w e re e n r ic h e d w i t h m ic r o -a lg a e o r a rtifi­
A fte r 3 - 5 d a y s o f a c c lim a tio n , u n i la t e r a l e y e s ta lk
cial e n r i c h m e n t m e d ia b efo re b e in g fed to t h e crab
a b la tio n w a s a p p lie d to in d u c e s p a w n in g . A fter
la rv a e . E n r ic h m e n t w ith Chlorella w a s p e rfo rm e d at
s p a w n in g , b e r rie d c r a b s w e r e a g a i n su b je c te d to a
a d e n s ity o f 5 x IO6 cells m L ~ 1 fo r 3 h (D h e rt 1996).
1 0 0 pL L _ 1 f o r m a lin b a t h fo r 1 h a n d tr a n s f e r r e d to a
R o tifers w e r e a ls o e n r ic h e d w it h D ry I m m u n e S elcoa
7 0 L p la s tic t a n k c o n n e c te d to a b io filte r fo r eg g in c u ­
(DIS ", IN V E A q u a c u ltu re ), u s in g tw o s e p a r a te doses
b a tio n . D aily m a n a g e m e n t c o n s is te d o f s ip h o n in g o u t
o f 0.05 g L “ 1 a t a 3 - h in te rv a l. E n ric h m e n t w a s p e r­
w a s te m a te r ia ls a n d s h e d d e d e g g s fro m t h e t a n k b o t­
fo rm ed a t a d e n s ity o f 5 0 0 r o tife rs m L - 1 . T h e w a te r
to m a n d c o n tr o llin g t h e t e m p e r a t u r e (30 °C), s a lin ity
i n th e e n r ic h m e n t v e sse l w a s slo w ly h e a t e d to
1540
© 2007 The Authors
Journal Compilation © 2007 Blackwell Publishing Ltd, A quaculture Research, 38, 1539-1553
A quaculture Research, 2007, 38, 1539-1553
Improved rearing tech n iq u es for m ud crab larvae T T N g h ia et al.
T able 1 Overview of larval rearing systems applied in th is study based on th e m e th o d of w ater exchange and m icro-algae
supplementation
W ater e x c h a n g e
C o n tin u o u s w a ter
D is c o n tin u o u s m a n u a l
tr e a tm e n t th ro u g h th e
A lg a e su p p le m e n ta tio n
partial w a te r r e n e w a l
u s e o f a biofilter
No m icro-algae supplem ented (indoors)
C lear-B atch sy stem
C lear-R ecirc sy stem
M icro-algae supplem ented a t low levels to provide
A lgae-B atch sy stem
A lgae-R ecirc system
G reen-B atch sy stem
G reen-R ecirc system
extra food for live preys (Indoors or outdoors)
Micro-algae supplem ented a t a high concentration
a n d self-sustainable u n d er natural sunlight a s an
(Combination of G reen-B atch an d
extra food for live p rey and w ate r conditioning (outdoors)
A lgae-R ecirc sy stem a t early an d late
larval s ta g e s respectively)
2 9 - 3 0 °C to avoid e x p o s in g t h e ro tife rs to t h e r m a l
s h o c k w h e n th e y w e re a d d e d to th e la r v a l r e a r in g
ta n k s . B efore b e in g fed to t h e la rv a e , e n r ic h e d ro tifers
w e re rin s e d a n d r e - s u s p e n d e d in c le a n s e a w a te r a t
t h e sa m e te m p e r a tu r e a s t h e c r a b - re a r in g ta n k s .
Larval rea rin g e x p erim en ts: o b jectiv es and
ex p er im e n ta l d e sig n
In e x p e r im e n ts 1 ,2 a n d 3, t h e e ffe ct o f d ifferen t w a te r
e x c h a n g e s c h e m e s a n d t h e a d d itio n o f m ic ro -a lg a e
o n la r v a l s u r v iv a l a n d d e v e lo p m e n t w e re e v a lu a te d .
In e x p e r im e n ts 4 - 8 , o th e r c u lt u r e a s p e c ts s u c h a s
A rte m ia c u ltu re a nd e n rich m en t
Z1 s to c k in g d e n s ity , live food d e n s ity a n d th e effect of
A rte m ia n a u p lii (V in h C h a u s tr a in ) w e re h a tc h e d a s
d e s c rib e d by V an S ta p p e n (1996). B o th n e w ly h a tc h e d
o r e n r ic h e d A rte m ia n a u p lii w e r e u s e d in t h e e x p e ri­
m e n ts o f t h i s stu d y . A rte m ia w e re e n r ic h e d w ith
Chaetoceros i n t h e s a m e m ic r o -a lg a l d e n s ity a s fo r r o ­
tife r e n r ic h m e n t. T h e n a u p lii w e r e a lso e n ric h e d w ith
DIS “ (u sin g tw o s e p a r a te d o s e s o f 0.3 g m L “ 1 a t a 6 -h
in te rv a l). W a te r t e m p e r a t u r e a n d s a lin ity w e re m a in ­
ta in e d a t 3 0 °C a n d 3 0 g L ~ \ respectively, d u r in g
A rte m ia e n r ic h m e n t. T h e d e n s ity o f A rte m ia d u r in g
e n r ic h m e n t w a s 2 0 0 m L - 1 . B efore fe e d in g to th e
c ra b la rv a e , t h e A r te m ia w e r e r in s e d w ith d isin fe c te d
s e a w a te r a n d s u s p e n d e d a t a k n o w n d e n s ity in
se a w a te r.
d iffe re n t p r o p h y la c tic t r e a tm e n t s w e r e in v e stig a te d .
T h e w a te r q u a lity m a n a g e m e n t s c h e m e s te s te d i n ex­
p e r im e n ts 1 - 3 a r e s u m m a r iz e d in T able 1. A n o v e r­
v ie w
of
th e
e x p e r im e n ta l
d e s ig n
and
c u ltu r e
c o n d itio n s o f a ll t h e e x p e r im e n ts is p r e s e n te d in
T able 2. T h e s m a ll- s c a le e x p e r im e n ts ( 1 - 3 0 L) w e re
c a r r ie d
out
in
a
te m p e r a tu re - c o n tr o lle d
ro o m
( 2 8 - 3 0 °C). T h e e x p e r im e n ts in 1 0 0 L t a n k s w ere
p e rfo rm e d o u td o o r s a t a m b ie n t te m p e r a tu r e (2 7 31 °C). T h e s o u r c e a n d th e d is in f e c tio n p r o c e d u r e of
th e s e a w a te r fo r la r v a l r e a r in g w e r e s im ila r t o th o s e
u s e d for b ro o d s to c k r e a r in g . F o rm a lin a t a c o n c e n tr a ­
tio n o f 2 0 pL L ~ 1 w a s a p p lie d e v e ry o th e r d a y a s a
p ro p h y la c tic t r e a t m e n t i n e x p e r im e n ts 1 - 6 .
E x p e rim e n t 1
Feeding
L a rv a l s u r v iv a l a n d g r o w th in a c le a r w a te r s y s te m
R o tifers w e re fed to t h e c r a b la r v a e fro m 0 to 6 days
w it h d a ily p a r t ia l w a te r e x c h a n g e (C lear-B atch ) w a s
a f te r h a tc h (DAH 0 - 6 ) [ ro u g h ly c o rre s p o n d in g to
c o m p a re d w i t h th o s e i n a c le a r w a te r r e c irc u la tin g
z o e a 1 (Z l)-Z 2 sta g es]. N e w ly h a tc h e d A rtem ia o r
s y s te m (C lear-R ecirc). I n t h e first r e a r in g sy stem ,
A rte m ia m e ta - n a u p lii w e r e o ffered fro m DAH 6 (Z3
3 0 - 5 0 % o f th e w a te r w a s r e p la c e d daily. I n t h e r e c ir ­
sta g e) o n w a rd s. R o tifers a n d A rte m ia w e re a d d e d
c u la tin g s y s te m , a ll r e a r in g t a n k s w e re c o n n e c te d to
d a ily a t 3 0 - 4 5 a n d 5 - 1 0 m L - 1 to th e r e a r in g ta n k
a c e n tr a l biofilter. W ater w a s r e c irc u la te d a t a ra te o f
re sp e c tiv e ly ( e x p e r im e n ts 1, 2, 3, 4, 7 a n d 8). For
a p p ro x im a te ly 100% o f th e t a n k v o lu m e ev ery 3 - 4 h.
e x p e r im e n ts 5 a n d
Live food a n d c ra b la rv a e w e re r e ta in e d i n th e r e a r in g
re q u ire d
p re y
6, live feed w e re fed a t th e
d e n s itie s
based
on
th e
p la n n e d
t a n k s w ith b y a m e s h s c re e n o f 7 0 a n d 3 0 0 p m d u r ­
tr e a tm e n ts . W h e n e v e r th e c r a b la rv a e w e re fed e n ­
in g t h e r o tife r a n d A rtem ia fe e d in g s ta g e respectiv ely .
r ic h e d live feed, a lg a e - o r DIS - e n ric h e d live feed
L a rg e r m e s h s c re e n s (250 a n d 5 0 0 - 1 0 0 0 p m fo r ro ti­
w e re u s e d o n a lt e r n a te days.
fer a n d A rte m ia s ta g e resp ec tiv ely ) a n d h ig h e r flow
© 2007 The Authors
Journal Compilation © 2007 Blackwell Publishing Ltd, Aquaculture Research, 38, 1539-1553
1541
A quaculture R esearch. 2007, 38, 1539-1553
Improved rearing techniques for mud crab larvae T T N g h ia et al.
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© 2007 The Authors
Journal Compilation © 2007 Blackwell Publishing Ltd, Aquaculture Research, 38, 1539-1553
A quaculture Research, 2007, 38, 1539-1553
Improved rearing te c h n iq u e s for m ud crab larvae T T N g h ia et al.
r a te s w e r e u s e d u p o n d aily flu s h in g o u t o f u n e a te n
( e x p e rim e n t 4) o r a C lea r-R ecirc s y s te m ( e x p e rim e n ts
live food a n d w a ste .
5 a n d 6) a s d e s c r ib e d above.
E x p e rim e n t 2
E x p e rim e n t 7
A C lea r-R ecirc s y s te m w a s c o m p a re d w it h
tw o
I n e x p e r im e n t 7, t h e effect o f p ro p h y la c tic c h e m ic a ls
s y s te m s w h e r e m ic r o -a lg a e w e re ad d ed . R e a rin g c o n ­
o n t h e s u rv iv a l o f t h e la rv a e w a s in v e s tig a te d . T h re e
d itio n s fo r th e C lea r-R ecirc s y s te m w e re s im ila r to
tr e a tm e n ts , c o n s i s t in g o f a c o n tr o l (no c h e m ic a ls
t h o s e d e s c rib e d in e x p e r im e n t 1. I n t h e A lg a e -R e c irc
u sed ), d a ily a d d i ti o n o f fo rm a lin a t 2 0 pL L ~ 1 a n d d a i­
sy s te m , m ic r o -a lg a e w e re a d d e d d a ily to t h e r e c irc u ­
ly a d d itio n o f O x y te tra c y c lin e a t 10 m g L “ \ w e re r u n
la tin g s y s te m a t a lo w c o n c e n tr a tio n r a n g in g fro m 0.1
in 1 L p la s tic c o n e s . A ll c o n e s w e re p la c e d in a w a te r
t o 0.2 m illio n cells m L ~ \ T h e o p e ra tio n o f t h e r e a r ­
b a th i n o r d e r to m a i n t a i n t h e r e a r in g te m p e r a t u r e a t
in g ta n k s w a s s im ila r to th e C lear-R ecirc tre a tm e n t.
3 0 °C. W ater w a s r e p la c e d a lm o s t c o m p le te ly daily.
I n t h e G re e n -B a tc h tre a tm e n t, a c la s sic a l ‘g r e e n -
U p o n w a te r e x c h a n g e , th e s u r v iv a l w a s d e te r m in e d .
w a t e r ’sy ste m , m ic r o -a lg a e c o n c e n tr a tio n s i n t h e c u l­
t u r e ta n k s w e r e k e p t a t a te n fo ld h ig h e r level o f
E x p e rim e n t 8
1 - 2 m illio n c e lls m L - 1 . I n t h i s system , t h e c u ltu r e
t a n k s w e r e in itia lly o n ly filled to 50% o f t h e i r c a p a ­
c ity a n d g r a d u a lly in c r e a s e d to 100% by t h e e n d o f
t h e Z2 s ta g e b y a d d in g w a te r a n d a lg a e daily. L a te r
o n , 1 0 - 3 0 % o f th e r e a r in g w a te r w as re p la c e d d a ily
b y c le a n s e a w a te r a n d /o r alg ae, d e p e n d in g o n th e
d e n s ity o f m ic r o -a lg a e r e m a in in g in th e r e a r in g
ta n k s . U p o n w a te r e x c h a n g e , u n e a te n live food w a s
a ls o f lu s h e d o u t t h r o u g h a m e s h s c re e n (m e sh siz es
a s d e s c rib e d i n e x p e r im e n t 1). T h e s a m e a m o u n t of
live fo o d ( 3 0 -4 5 r o tife rs m L “ 1 a n d
5 - 1 0 A rtem ia
n a u p lii m L ~ L) w a s fed i n a ll t h e tr e a tm e n ts . I n t h e
s y s te m s u s i n g a lg a e , Chlorella w a s u s e d fo r Z 1 -Z 3
To avoid t h e u s e o f d r u g s a s a p ro p h y la x is, d ir e c t o z o ­
n a tio n o f th e c u l t u r e ta n k s w a s te s te d i n a e r a te d 1 L
p la s tic co n es. O z o n e ( 0 3) w a s in je c te d d ire c tly v ia a n
a ir s to n e in to e v e ry la r v a l r e a r in g e o n e u p o n c h a n ­
g in g w a te r a n d fe e d daily. S ix tr e a tm e n ts w it h th r e e
re p lic a te s w e re a r r a n g e d c o n s is tin g o f a c o n tro l
( w ith o u t 0 } a p p lic a tio n ) a n d 0 3 in je c tio n fo r 2, 4, 6,
8 a n d 10 m in ( e q u iv a le n t to a re s id u a l 0 3 level i n th e
w a te r o f 0 ,0 6 ,0 ,1 2 ,0 .1 5 ,0 .1 7 a n d 0.19 m g L " 1 a s m e a ­
s u r e d b y a te s t k it u p o n fin is h in g t h e in je c tio n ). O th e r
r e a r in g c o n d itio n s a n d d a ily m a n a g e m e n t w e re sim i­
l a r to th o s e d e s c rib e d f o r e x p e r im e n t 7.
s ta g e s (w h ic h is u n s u ita b le a s a food s o u r c e for
A rtem ia); fro m Z 4 o n w a rd s . Chlorella w a s g r a d u a lly
r e p la c e d w i th Chaetoceros.
E valu ation cr iter ia
T h e s u r v iv a l r a te s i n th e e x p e r im e n ts u s in g la rg e
E x p e r im e n t 3
( 3 0 - 1 0 0 L) c o n ta in e r s (e x p e rim e n ts 1 - 6 ) w e re e s ti­
I n th is e x p e r im e n t, a G re e n -B a tc h a n d a G reen -R ecirc
m a te d by v o lu m e tric s a m p lin g . D e p e n d in g o n t h e
s y s te m w e r e s e t u p in o r d e r to f u r th e r e v a lu a te th e
t a n k v o lu m e a n d t h e d e n s ity o f th e s u r v iv in g la rv a e ,
a p p lic a tio n o f m ic r o -a lg a e o n th e p e r fo r m a n c e o f
c ra b la rv a e . T h e firs t r e a r in g s y s te m w a s a b a tc h sy s­
te m w i t h a d d itio n o f h ig h c o n c e n tr a tio n s o f a lg a e a s
d e s c rib e d i n e x p e r im e n t 2. T h e s e c o n d s y s te m c o n ­
s is te d o f a c o m b in a tio n o f t h e G re e n -B a tc h sy s te m
fo r e a r ly c ra b s ta g e s (Z 1 -Z 2 ) a n d a A lg ae-R ec irc sy s­
t e m fo r l a t e r s ta g e s (Z3 o n w ard s).
trip lic a te 3 0 0 - 1 0 0 0 m L s a m p le s w e re ta k e n fro m
e a c h ta n k . M e g a lo p a e (M) (DAH 1 5 -1 8 ) a n d first
c r a b s (C l) (DAH 22) w e r e c o u n te d in d iv id u ally . I n ex ­
p e r im e n ts 7 a n d 8 (u s in g s m a ll co n es), th e a v e ra g e
s u r v iv a l ra te w a s c a lc u la te d by in d iv id u a lly c o u n tin g
a ll s u r v iv in g la rv a e in e a c h rep licate.
L a rv a l d e v e lo p m e n t w a s m o n ito re d e v e ry 3 d ay s
b u t d a ily in e x p e r im e n ts 7 a n d 8 b y id e n tify in g th e
E x p e rim e n ts 4 - 6
a v e ra g e z o e a l i n s ta r s ta g e o f a s a m p le o f l a r v a e a n d
a s s ig n in g i t a valu e: Z1 = 1 , Z 2 = 2, etc. M eg alo p a
I n th e s e e x p e r im e n ts , t h e effe ct o f Z1 s to c k in g d e n s ity
s ta g e w a s a s s ig n e d a v a l u é o f 6. To c o m p a re t h e la r v a l
(50, 1 00, 150 a n d 2 0 0 L ~ \ e x p e r im e n t 4), ro tife r
d e v e lo p m e n t in e a c h t r e a tm e n t, a n a v e ra g e la rv a l
fe e d in g d e n s itie s a t 30, 4 5 a n d 6 0 m L - 1 fo r Z 1 -Z 2
sta g e in d e x (LSI) w as c a lc u la te d fro m t h e a v e ra g e
( e x p e r im e n t 5) a n d A rte m ia d e n s itie s a t 10, 15 a n d
LSI v a lu e o f a ll re p lic a te c o n ta in e r s in th e s a m e tr e a t­
2 0 m L “ 1 fo r Z3 o n w a r d s (e x p e rim e n t 6) w a s e v a lu ­
m e n t. F or larg e c o n ta in e r s (e x p e rim e n ts 1 -6 ), five o r
a te d . T h e s e e x p e r im e n ts w e r e r u n in a G re e n -B a tc h
10 la r v a e (in 3 0 a n d 100 L ta n k s resp ec tiv ely ) w e re
© 2007 The Authors
Journal Compilation © 2007 Blackwell Publishing Ltd, Aquaculture Research, 38, 1539-1553
1543
Improved rearin g techniques for m ud crab larvae T T N g h ia et al.
A quaculture Research. 2007, 38, 1539-1553
T ab le 3 E xperim ent 1: survival rates and larval stage index (LSI) values of Scylla paramamosain larv ae cultured in two differ­
ent rearin g system s
D ays afte r h a tc h
3
6
Clear-Batch
85 ± 6a
7 9 ± 9a
C lear-Recirc
84 ± 4a
7 8 ± 8a
C lear-Batch
1.5 ± 0.2a
Clear-Recirc
1.5 ± 0,2a
T reatm ent
9
12
15
18
70 ± 6a
6 4 ± 7a
4 2 ± 6b
32 ± 5b
72 ± 5 a
70 ± 5a
63 ± 9 a
47 ± 6a
2.7 ± 0.1a
3.5 ± 0.4a
4.0 ± 0.0a
4.6 ± 0.2a
ND
2.7 ± 0.1a
3.6 ± 0.3a
4.2 ± 0.3a
4.8 ± 0.1a
ND
Survival rate (%)*
LSI’
’ Survival rates or LSI values in the sam e colum n followed by the same superscript letter are not statistically different (P > 0.05).
For treatm ent descriptions, refer to Table 1.
ND, not determined.
sa m p le d fro m e a c h t a n k to c a lc u la te th e a v e ra g e LSI.
B a tc h s y s te m ( b o th a t P < 0.01) (T able 3). A lth o u g h
T h e sa m p le d la r v a e w e re s ta g e d u n d e r a d is s e c tin g
slig h tly h i g h e r in t h e r e c ir c u la tin g sy stem , LSI w a s
m ic ro sc o p e . I n e x p e r im e n t 8, u s in g s m a ll c o n ta in e rs ,
n o t sig n ific a n tly d iffe re n t b e tw e e n tr e a tm e n ts . T h e
a ll la rv a e w e r e s ta g e d v is u a lly u p o n c o u n tin g d aily
b e tte r la r v a l p e r fo r m a n c e i n t h e C lea r-R ecirc s y s te m
s u rv iv a l.
w a s a c c o m p a n ie d b y s ig n ific a n tly lo w e r a v e ra g e
I n th is r e s e a rc h , six la r v a l r e a r in g sy s te m s w e re
a p p lie d fo r t h e e x p e rim e n ts . E a c h r e a r in g sy s te m
a m m o n ia levels (P c O .O l) a n d s lig h tly lo w e r n itr ite
levels (see T able 2).
h a d its o w n fe a tu re s, i.e. w a te r q u a lity a n d ‘e a s e of
o p e r a tio n ’. T h e re fo re , th e s e f e a tu r e s in c o m b in a tio n
m a d e u p a tr e a t m e n t a s a ty p e o f r e a r in g sy ste m . T h e y
w e re n o t c o n s id e re d a s v ariab les.
E xp erim en t 2
O n DAH 9, la r v a l s u r v iv a l in t h e C lea r-R ecirc s y s te m
w a s s ig n ific a n tly lo w e r (P < 0.05) t h a n in b o th tr e a t­
m e n ts w it h m ic r o -a lg a e s u p p le m e n ta tio n (A lgae-R e­
S ta tistica l a n a ly sis
c irc a n d G re e n -B a tc h s y s te m s) (Table 4). O n D AH 12,
O n e -w a y a n a ly s is o f v a r ia n c e (a n o v a ) w a s u s e d to
s u rv iv a l in t h e C lea r-R ecirc tr e a tm e n t w a s lo w e r
c o m p a r e d a ta . H o m o g e n e ity o f v a r ia n c e w a s te s te d
(P < 0.05) t h a n in th e A lg a e -R e c irc sy stem , w h e r e a s
w ith t h e L ev en e s ta tis tic (P o r a v a lu e w a s s e t a t
t h e G re e n -B a tc h s y s te m h a d in te r m e d ia te re su lts.
0.05). If n o s ig n if ic a n t d iffe re n c e s w e r e d e te c te d b e ­
T h e LSI v a lu e s o n DAH 15 s h o w a s im ila r tre n d ,
tw e e n th e v a r ia n c e s , t h e d a ta w e r e s u b m itte d to a
a lth o u g h n o t s ig n ific a n tly d ifferen t.
o n e -w a y a n o v a . T u k ey 's h o n e s tly s ig n if ic a n t differ­
T h e a v e ra g e levels o f a m m o n ia a n d n i tr i te in t h e
e n c e p o s t h o c a n a ly s is w a s u s e d to d e te c t d iffe re n c e s
C lea r-R ecirc a n d A lg a e -R e c irc sy s te m s w e r e signifi­
b e tw e e n m e a n s a n d to in d ic a te a r e a s o f s ig n ific a n t
c a n tly lo w e r (P < 0.01) t h a n th o s e in t h e G re e n -B a tc h
d ifferen ce. I f s ig n if ic a n t d iffe re n c e s w e re d e te c te d
s y s te m (see T able 2). I n th e G re e n -B a tc h sy stem ,
b e tw e e n v a r ia n c e s , d a ta w e re tr a n s f o r m e d u s in g t h e
p e a k s o f a m m o n ia a n d n itr ite c o n c e n tr a tio n s o f 3
a r c s in e - s q u a r e r o o t (for p e r c e n ta g e d a ta , i.e. s u r v iv a l
a n d 1 m g L " \ respectively, w e re re c o rd e d a t th e e n d
ra te ) o r lo g a r ith m ic tr a n s f o r m a tio n s (for LSI v alu e)
o f th e e x p e r im e n t.
(Sokal & R o h lf 1995). A ll a n a ly s e s w e r e p e rfo rm e d
u s in g t h e s ta tis tic a l p r o g ra m s t a t i s t i c a 6.0.
E xp erim en t 3
T able 5 p r e s e n ts th e la rv a l p e r fo r m a n c e o f th e crab
R esu lts
la rv a e c u ltu r e d in tw o r e a r in g sy stem s. T h e s u rv iv a l
r a te s a n d LSI v a lu e s o f b o th r e a r in g sy ste m s w e re n o t
E xp erim ent 1
sig n ific a n tly d ifferen t. H ow ever, t h e s u rv iv a l ra te s o n
S u rv iv a l i n t h e C le a r-R e c irc s y s te m a t Z 4 -Z 5 sta g e s
l a te r d ays (fro m DAH 1 2 - 2 2 ) i n t h e G reen -R ecirc sy s­
o n DAH 15 a n d in th e m e g a lo p a s ta g e o n D AH 18
te m te n d e d to be h ig h e r t h a n th o se in t h e G reen -
w a s s ig n ific a n tly h i g h e r t h a n t h o s e i n th e C lea r-
B a tc h sy ste m . T h e b io filter h a d a positive im p a c t o n
1544
© 2007 The Authors
Journal Compilation © 2007 Blackwell Publishing Ltd. Aquaculture Research, 38, 1539-1553
A quaculture Research, 2007, 38, 1539-1553
Improved rearing te c h n iq u e s for m ud crab larvae T T N g h ia et al.
T ab le 4 Experim ent 2: survival rates and larval stage index (LSI) values of Scylla param amosain larvae cultured in three
different rearing systems
D ays a fte r hatch
T reatm ent
3
6
9
12
15
Survival rate (%)*
Clear-Recirc
74 ± 12a
63 ± 7a
44 ± 6 b
26 ± 11b
Algae-Recirc
74 ± 12a
63 ± 9a
61 ± 7 a
43 ± 7 a
15 ± 8a
G reen-Batch
74 ± 11a
67 ± 9 a
58 ± 9 a
35 ± 9 ab
13 ± 6 a
8 ± 7a
LSI*
Clear-Recirc
1.9 ± 0.1a
2 ,7 ± 0 .2 a
3.9 ± 0.1a
5.0 ± 0.1a
5.1 ± 0.1b
Algae-Recirc
2.0 ± 0.1a
2 .8 ± 0.3a
4 .0 ± 0.1a
5.0 ± 0.1a
5 .6 ± 0.2a
G reen-Batch
2.0 ± 0.1a
2 .8 ± 0 .2 a
4 .0 ± 0 .1 a
5 .0 ± 0.1a
5.1 ± 0.1ab
‘ Survival rates or LSI values in the same column followed by the same superscript letter are n o t statistically different (P > 0.05).
For treatment descriptions, refer to Table 1.
T ab le 5 Experim ent 3: survival rates and larval stage index (LSI) values of Scylla paramamosain larvae cultured in two dif­
ferent rearing systems
D ays afte r h a tc h
TVeatment
3
6
12
9
15
22
Survival rate (%)*
G reen-Batch
94 ± 6a
88 x 9 a
80 ± 3 a
6 6 ± 15a
44 ± 20a
9 ± 1a
G reen-R ecirc
94 ± 6a
89 ± 8 a
8 0 ± 5a
6 8 ± 11a
56 ± 11a
12 ± 3 a
LSI*
G reen-Batch
1.4 ± 0.3a
2.7 ± 0 .1 a
3 .8 ± 0.4a
5 .0 ± 0 .0 a
5 .2 ± 0 .2 a
ND
G reen-R ecirc
1.4 ± 0.2a
2.6 ± 0 .2 a
3 .9 ± 0 .3 a
5 .0 ± 0 .0 a
5 .3 ± 0 .1 a
ND
‘ Survival rates or LSI values in the same column followed by the same superscript letter are not statistically different (P > 0.05).
For treatment descriptions, refer to Table 1.
ND, not determined.
w a te r q u a lity in t h e s e c o n d p a r t o f t h e e x p e rim e n t,
th a t
w ith s ig n ific a n tly re d u c e d a m m o n ia ( P < 0 .0 5 ) a n d
3 0 r o tife rs m L ” 1 a n d
LSI w a s
a lw a y s
n it r it e (P c O .O l) c o n c e n tr a tio n s a s a c o n s e q u e n c e
6 0 r o tife rs m L " \
th e
th e
lo w e s t
in
tr e a tm e n t
h ig h e s t
in
tr e a tm e n t
(seeT ab le 2).
E xp erim en t 6
E xp erim en t 4
T able 6 s h o w s t h e s u r v iv a l a n d d e v e lo p m e n t r a te of
T ab le 8 p r e s e n ts th e s u r v iv a l a n d d e v e lo p m e n t r a te o f
c ra b la rv a e fed A rte m ia n a u p lii a t th r e e d e n s itie s (10,
c ra b la r v a e sto c k e d a t f o u r d iffe re n t Z1 d e n s itie s (50,
15 a n d 2 0 m L “ :) fro m t h e Z3 s ta g e o n w a rd s. No sig­
100, 150 a n d 2 0 0 L - *). T h e s u r v iv a l ra te s w e r e n o t
n if ic a n t d ifferen ces w e r e o b s e rv e d b e tw e e n t h e t r e a t ­
s ig n ific a n tly d iffe re n t a m o n g tr e a tm e n ts . O n ly o n
m e n ts. T h e re se e m e d , h o w e v e r, to b e a w e a k tr e n d
DAH 6 w a s a n e g a tiv e c o r r e la tio n b e tw e e n LSI a n d
to w a r d s h ig h e r s u r v iv a l a n d LSI w ith in c r e a s in g
la r v a l d e n s ity o b se rv e d .
A rte m ia d e n s ity to w a r d s t h e e n d o f th e tria l.
E xp erim en t 5
E xp erim en t 7
T able 7 s h o w s t h e s u r v iv a l r a te s a n d t h e LSI v a lu e s o f
T able 9 s h o w s th e s u r v iv a l a n d d e v e lo p m e n t r a te to
c ra b l a r v a e fed th r e e d iffe re n t r o tif e r d e n s itie s i n th e
t h e m e g a lo p a s ta g e (DAH 22) o f la rv a e re c e iv in g dif­
Z 1 -Z 2 sta g e s. N o s ig n if ic a n t d ifferen ces w e re fo u n d
fe re n t p ro p h y la c tic tr e a tm e n ts . F ro m D AH 6 o n ­
for a n y o f th e p a r a m e te r s . I t c a n , h o w ev er, be n o tic e d
w a rd s, t h e s u r v iv a l r a te o f la r v a e i n t h e tr e a tm e n t
© 2007 The Authors
Journal Compilation (£) 2007 Blackwell Publishing Ltd, Aquaculture Research, 38, 1539-1553
1545
Improved rearin g techniques for m ud crab larvae T T N g h ia et al.
A quaculture Research, 2007, 38, 1539-1553
T ab le 6 E xperim ent 4: survival rates and larval stage index (LSI) values of Scylla paramamosain la rv a e stocked at four differ­
en t 21 densities (21L ~ *)
D ays a fte r hatch
T re atm e n t
3
6
9
12
15
22
4 ± 6a
Survival ra te (%)*
50
79 ± 9a
56 ± 19a
42 ± 16a
31 ± 17a
2 8 ± 12a
100
80 ± T
74 ± 6a
71 ± 10a
5 6 ± 11a
4 5 ± 8a
5 ± 4a
150
79 ± 2a
57 ± 12a
45 ± 9a
31 ± 12a
2 8 ± 10a
5 ± 1a
200
85 ± 5a
53 ± 17a
42 ± 8 a
34 ± 3a
30 ± 5a
5 ± 1a
LSI’
50
1.7 ± 0.23
3.0 ± 0.1a
3 .9 ± 0.1a
5 .0 ± 0.1a
ND
ND
100
1.8 ± 0.1a
3.0 ± 0.1ab
4 .0 ± 0.0a
5 .0 ± 0.1a
ND
ND
150
1.8 i 0.2a
3.0 ± 0.1ab
3 .9 ± 0.2a
5 .0 ± 0.1a
ND
ND
200
1.8 ± 0.2a
2.7 ± 0.1b
3 .7 ± 0.2a
4 .8 ± 0.1a
ND
ND
’ Survival rates or LSI values in the same column followed by the same superscript letter are not statistically different (P > 0.05).
ND, n o t determined.
T ab le 7 E xperim ent 5: survival rates and larval stage index (LSI) values o f Scylla paramamosain larvae-fed th ree different
rotifer densities (rotifers mL ~ ) from day 0 to day 6 after hatch
D ays afte r hatch
T reatm en t
3
6
9
12
15
Survival rate (%)*
30
89 ± I a
53 ± 10a
30 ± 8a
13 ± 7 a
10 ± 7 a
45
87 ± 6a
58 ± 9a
35 ± 7a
18 ± 9 a
14 ± 8 a
60
87 ± 5a
55 ± 7a
32 ± 6 a
16 ± 11a
11 ± 10a
30
1.8 ± 0.2a
2.7 ± 0.1a
3 .6 ± 0 .2 a
3.8 ± 0.2a
4.0 ± 0.4a
45
1.8 ± 0.2a
2.8 ± 0.2a
3 .8 ± 0 .1 a
3 .9 ± 0.1a
4.3 ± 0.5a
60
1.8 ± 0.2a
2.8 ± 0.2a
3 .8 ± 0 .2 a
4 .0 ± 0.1a
4 .4 ± 0 ,5 a
LSI*
’ Survival rates or LSI values in the same colum n followed by the same superscript letter are not statistically different (P > 0.05).
u s in g a n tib io tic s w a s s ig n ific a n tly h i g h e r t h a n th o se
t h e s u r v iv a l o f tr e a tm e n t O zo n 2 b e c a m e th e h ig h e s t
i n th e r e m a i n i n g tr e a tm e n ts . T h e s u r v iv a l r a te s o f th e
(52% ); h o w e v e r, th is w a s n o t s ta tis tic a lly d iffe re n t
c o n tr o l a n d f o rm a lin tr e a tm e n ts w e re s im ila r on
fro m th e c o n tr o l (25%). O n DAH9, t h e r e w e re n o sig­
m o s t d ay s. F ro m D A H 6 o n w a rd s , t h e LSI v a lu e s of
n if ic a n t d iffe re n c e s in s u rv iv a l in a ll tr e a tm e n ts .
th e f o rm a lin tr e a t m e n t w e re g e n e ra lly h ig h e r t h a n
fo r th e o t h e r t r e a tm e n ts (n o t a lw a y s sig n ifican t). On
DAH 15 a n d 18, t h e a n tib io tic tr e a t m e n t re s u lte d in
lo w e r LSI v a lu e s (P < 0.01).
D iscu ssio n
R earin g sy stem
R ecirculation
E xp erim en t 8
W a te r r e c ir c u la tio n th r o u g h a b io filter i n th e C lear-
T able 10 p r e s e n ts t h e s u r v iv a l a t t h e Z2, Z3 a n d Z4
R e circ s y s te m p o sitiv ely affe cted la r v a l p e rfo rm a n c e
sta g e s o f la r v a e t h a t w e r e d a ily e x p o s e d to differen t
c o m p a r e d w i t h m a n u a l p a r tia l w a te r r e p la c e m e n t in
lev els o f 0 3. O n D A H 3 (Z2 stag e), s u r v iv a l in th e c o n ­
t h e C le a r-B a tc h s y s te m (e x p e rim e n t 1). T h e a d v a n ­
tr o l a n d t h e tr e a t m e n t w i t h 0 3 in je c tio n for 2 m in
ta g e s o f r e c ir c u la tin g sy s te m s in c o m m e rc ia l fish
(O zon2) w a s h ig h e r ( 6 7 -7 8 % ) c o m p a r e d w ith t h e
a n d c r u s t a c e a n la r v a l p r o d u c tio n h av e b e e n p ro v e n
o t h e r t r e a t m e n ts (2 4 -4 5 % ). O n D A H 6 (Z3 stage),
b efo re fo r o t h e r sp e cies. R e s e a rc h in to r e c irc u la tin g
1546
© 2007 The Authors
Journal Compilation © 2007 Blackwell Publishing Ltd, Aquaculture Research. 38, 1539-1553
Improved rearing tech n iq u es for m ud crab larvae T T N g h ia et al.
A quaculture Research, 2007, 38, 1539-1553
T ab le 8 E xperim ent 6: survival rates and larval stage index (LSI) values of Scylla paramamosain larvae-fed th ree different
instar-1 Artemia densities (Artemia mL ^ 1) from 6 days after hatch
D ays afte r hatch
12
9
T re atm e n t
15
Survival rate (%)*
10
2 6 ± 10a
12 ± 5 a
8 ± 3“
15
3 0 ± 6a
13 ± 7 a
10 ± 6 a
20
32 ± 8a
19 ± 9a
18 ± 9 a
10
3.1 ± 0.2°
3 .7 ± 0.4a
4.3 ± 0 .5 a
15
3.1 ± 0.1a
3.7 ± 0.2a
4.3 ± 0 .5 a
20
3 .2 ± 0.1a
3 .8 ± 0.3a
4.6 ± 0.3a
LSI*
*Survival rates or LSI values in the same column followed by the same superscript letter are not statistically different (P > 0.05).
T a b le 9 E xperim ent 7: survival rates and larval stage index (LSI) values of Scylla paramamosain larvae treated daily w ith
prophylactic chem icals
D ays a fte r h atch
T reatm en t
3
6
9
12
15
18
22
Survival ra te s (%)*
Control
85 ± 3a
64 ± 1°
48 ± 8b
3 4 + 13b
28 ± 11b
17 ± 7b
9 ± 5b
Formalin
84 ± 7a
66 ± 8 b
47 ± 7b
3 4 ± 12b
26 ± 8 b
13 ± 10b
11 ± 8 ab
Antibiotics
91 ± 4a
80 ± 2 a
74 ± 4a
66 ± 8 a
52 ± 6 a
34 ± 3a
21 ± 5a
Control
1.8 ± 0.1°
2 .6 ± 0 .3 a
3.4 ± 0 .2 ab
4 .5 ± 0.3a
5.1 ± 0.1a
5.5 ± 0 .1 b
5 .8 ± 0 .2 a
Formalin
1.8 ± 0.1a
2 .8 ± 0 .1 a
3,7 ± 0 .1 a
4 .3 ± 0.3a
5.1 ± 0.1a
5.8 ± 0 .1 a
6 .0 ± 0.1a
Antibiotics
1.9 ± 0.0a
2 .7 ± 0 .2 a
3.3 ± 0 .2 b
4 .3 ± 0.1a
4 .9 ± 0 .1 b
5.3 ± 0 .2 b
5 .6 ± 0.2a
LSI*
"■Survival rates or LSI values in the same column followed by the same superscript letter arc not statistically different (P > 0.05).
T a b le 10 E xperim ent 8: survival rates of Scylla paramamosain larvae treated daily by ozone for different durations of tim e
(min)
T re atm e n t
DAH 3 (Z2)
DAH 6 (Z3)
DAH 9 (Z4)
Control
7 8 ± 6a
2 5 ± 9ab
9 ± 6a
O zon2
6 7 ± 6ab
5 2 ± 14a
11 ± 10a
O zon4
4 0 ± 17abc
3 3 ± 11ab
5 ± 2a
O zon6
4 5 ± 25abc
2 7 ± 15ab
5 ± 5a
O zon8
31 ± 2bc
19 ± 3 b
0 ± 0a
O zonlO
2 4 ± 15c
12 ± 9 b
0 ± 1a
Survival rates in the same column followed by the same superscript letter are not statistically different (P > 0.05).
DAH, days after hatch; Z, zoea; Control, without ozonation; Ozon2,4, 6,8 and 10, duration of ozone injection from 2 to 10 min, which is
equivalent to 0.06,0.12, 0.15, 0.17 and 0.19 mg L " 1 of the residual ozone respectively.
s y s te m s h a s a ls o b e e n id e n tifie d a s a p rio rity for
t a in a h ig h b io lo g ical c a r r y in g c a p a c ity in relativ ely
s h r im p c u l tu r e (L a w re n c e & L ee 1997). I n th e s e sy s­
little s p a c e (Q u ille re, M arie, R o u x , G osse & M o ro t-
te m s, w a te r e x c h a n g e is m in im iz e d th r o u g h t h e u s e
g a u d r y 1993; T w a ro w sk a , W e s te rm a n & L o so rd o
o f b io lo g ical, c h e m ic a l a n d / o r m e c h a n ic a l filtra tio n
1997). F o r c ra b la rv ic u ltu re , re c irc u la tin g sy s te m s
to m a i n ta i n g o o d w a te r q u a lity c o n tin u o u sly . A s th e y
a lso a p p e a r to w a r r a n t f u r t h e r in v e s tig a tio n in o rd e r
p ro v id e le s s s tr e s s a n d c o n f e r c o n s ta n t goo d w a te r
to d e c r e a s e la b o u r r e q u ire m e n ts a n d s e a w a te r c o n ­
q u a lity to t h e la rv a e , th e s e sy s te m s a r e able to m a in ­
s u m p tio n , p ro v id in g a m o re s ta b le c u ltu r e m e d iu m
© 2007 The Authors
Journal Compilation © 2007 Blackwell Publishing Ltd, Aquaculture Research, 38, 1539-1553
1547
Im proved rearing techniques for m ud crab larvae T T N g h ia et al.
A quaculture R esearch. 2007, 38, 1539-1553
a n d t h u s re d u c in g la rv a l stre s s. If t h e s y s te m d e sig n
d u a lly fill u p t h e t a n k s w ith f r e s h s e a w a te r, a lg a e a n d
is k e p t sim p le, re c irc u la tin g sy s te m s c o u ld a ls o b e s u i­
ro tife rs t h a n flu s h in g o u t old r o t i f e r s i n th e r e c ir c u la ­
ta b le fo r la rg e -s c a le p ro d u c tio n .
tio n sy stem . I n th e r e c ir c u la tin g sy ste m , th e y o u n g
la rv a e m a y be p ro n e to p h y s ic a l d a m a g e a n d m a y
R ole o f su p p le m en ted m icro-algae
s p e n d c o n s id e ra b le e n e r g y t r y i n g t o sw im u p a g a in s t
T h e a d d itio n o f m ic ro -a lg a e to t h e r e c irc u la tio n
s y s te m s r e s u lte d in b o th h ig h e r s u r v iv a l a n d fa ste r
d e v e lo p m e n t in th is stu d y . M ic ro -a lg a e h a v e b e e n
p r o v e n to b e b en eficial by v a r io u s m o d e s o f a c tio n .
T h e y c o u ld h e lp m a in ta in th e q u a lity o f live food. As
in t h e c u l t u r e o f m a r in e fis h la rv a e , u n c o n s u m e d
r o tife rs m a y re sid e in t h e ta n k s fo r s e v e ra l d a y s a n d
t h e i r n u t r it i o n a l v a lu e m a y b e c o m e s e v e re ly re d u c e d
(M a k rid is & O lse n 1999). F u rth e rm o r e , a c c o r d in g to
th e s e a u th o r s , p o o rly fed ro tife rs w e re m o r e s e n sitiv e
to s ta r v a tio n t h a n w ell-fed ro tifers, a s t h e i r n itro g e n
c o n t e n t d e c r e a s e d a t a h i g h e r rate.
M ic ro -a lg a e a ls o play a n im p o r ta n t ro le i n sta b iliz ­
i n g w a t e r q u a lity v ia e ith e r a m m o n ia u p ta k e o r oxy­
g e n p r o d u c tio n (Tseng, H u a n g & L iao 1991). B e c a u se
t h e C le a r-R e c irc sy ste m a lr e a d y p ro v id e d o p tim a l
w a te r q u a lity , i t is u n lik e ly t h a t t h e s ta b iliz in g effect
o n w a t e r q u a lity is re sp o n s ib le fo r t h e im p ro v e d p e r­
f o r m a n c e i n t h e a lg a e -s u p p le m e n te d
sy s te m . In
b a tc h c u l t u r e sy stem s, th is effe ct w o u ld p ro b a b ly be
m uch
m o re
p ro n o u n c e d .
A
d ir e c t
c o m p a r is o n
b e tw e e n a g r e e n a n d c le a r w a te r b a tc h s y s te m w as,
h o w e v e r, n o t m a d e in th is study.
I n a s tu d y o n t h e effect o f Chlorella o n th e p o p u la ­
t i o n o f lu m in o u s b a c te ria Vibrio harveyi, n o lu m in o u s
b a c te r ia w e re re c o v e re d o n d a y s 2 a n d 3 i n flasks
w i t h Chlorella, w h ile th o s e w i th o u t t h e m ic r o -a lg a e
still h a r b o u r e d lu m in o u s b a c te ria a t d a y 3 (T endencia
& d e la P e n a 2003). Also, th e d ia to m Chaetoceros h a s
b e e n s h o w n to p ro d u c e n a t u r a l a n tib io tic s a n d h ig h
c o n c e n tr a tio n s o f th is m a r in e d ia to m w ill e lim in a te
Vibrio vu ln ificus a n d o th e r p a th o g e n ic b a c te ria , w h ic h
c o n tr ib u te to th e p ro p a g a tio n o f v iru s e s in t h e s h rim p
p r o d u c tio n e n v ir o n m e n t (W an g 2003).
th e c u r r e n t. E a rly c ra b la r v a e a r e d e lic a te d u e to t h e i r
s m a ll size a n d th e th r e e lo n g s p i n e s o n t h e c a r a p a c e
t h a t a r e e a s ily d a m a g e d iv h e n t h e y a r e e n tr a p p e d o n
th e m e s h s c re e n d u r in g f lu s h in g o u t o f u n e a t e n feed
i n t h e r e c irc u la tio n s y s te m (D a v is 2003). T h e n u t r i ­
tio n a l effect of m ic ro -a lg a e is p r o b a b ly a lso m o r e p r o ­
n o u n c e d d u r in g th e r o tif e r f e e d in g s ta g e t h a n d u r in g
t h e A rtem ia fe ed in g stage. F u r th e r m o r e , it is n o t n e ­
c e s s a r y to re c irc u la te w a te r d u r i n g th e s e first days,
a s t h e c o n c e n tr a tio n s o f a m m o n i a a n d n it r it e a r e s till
low. U sing t h e A lg ae-R ec irc s y s te m in l a te r s ta g e s is
m o re fa v o u ra b le fo r re d u c in g t h e in c r e a s in g a m m o ­
n ia a n d n itr ite c o n c e n tr a tio n s a s m o r e w a s te m a te r i­
a l is p ro d u c e d by t h e c ra b la r v a e . M oreover, a s t h e
la rv a e develop in to m o re effic ie n t p r e d a to rs , feed is
c o n s u m e d faster, a n d m a i n t e n a n c e o f o p tim a l feed
q u a lity is less o f a n issu e. M a n y s tu d i e s su c c e s s fu lly
a p p lie d a s im ila r c o m b in e d r e a r in g te c h n iq u e d u e to
its b e n e fit for th e la rv a e a n d c o n v e n ie n c e fo r m a n ­
a g e m e n t. p a r tic u la r ly fo r la rg e r e a r i n g c o n ta in e r s .
U n d e r g r e e n - w a te r c u lt u r e c o n d itio n s , w a te r is n o t
e x c h a n g e d fo r th e first 3 days. T h e re a fte r , w a te r ex­
c h a n g e is slo w ly in c r e a s e d fro m 1 0 - 2 0 % d a y “ 1 for
Z 2 -Z 3 to b e tw e e n 4 0 % a n d 50% d a y ~ 1 a t t h e e n d o f
t h e r e a r in g cycle (Z 4 -M ) (M a n n , A s a k a w a & P iz z u to
1999; Q u in itio . P a ra d o -E ste p a , M illa m e n a . R o d rig u e z
& B o rlo n g a n 2001). I n J a p a n , a m e s o c o s m s y s te m is
u s e d for c u ltu r in g la rv a e in la r g e r t a n k s ( > 10 m 3).
T h e t a n k s a r e p a r tia lly filled w i th g r e e n w a te r a t Z1
( 2 0 -2 5 % volum e), t a n k s a r e t h e n fille d u p w it h c le a n
s e a w a te r d u r in g th e c o u r s e o f t h e Z 2 -Z 3 s ta g e s a n d
d u r in g t h e Z4 a n d M s ta g e s w a te r is e x c h a n g e d o n a
flo w -to -w aste b asis (H am asa k i, S u p ra y u d i & T a k e u c h i
2002).
I n c o n c lu s io n , m ic ro -a lg a e in m u d c ra b la r v a l r e a r ­
in g m a y p la y a ro le i n im p ro v in g a n d m a in t a in i n g live
fo o d q u a lity a n d c o n tr o llin g b a c te r ia levels.
O ther rearing te c h n iq u e s
ZI stocking d en sity
Choice o f sy s te m
No sig n ific a n t effect o f la r v a l d e n s ity w a s o b s e rv e d
I n e x p e r im e n t 3. t h e G reen -R ecirc s y s te m (w h ic h is a
fro m 50 to 2 0 0 Z 1 L - 1 . T h is w o u ld s u g g e s t t h a t th e
c o m b in a tio n o f a G re e n -B a tc h s y s te m d u r i n g t h e r o ­
l a r v a e c a n be g r o w n a t 2 0 0 Z l L ~ \ V a ria tio n i n t h e
tife r fe e d in g s ta g e a n d a A lg a e -R e c irc s y s te m t h e r e ­
fin al s u r v iv a l b e tw e e n re p lic a te t a n k s a lso s e e m e d to
a fte r) s e e m e d to be b e tte r t h a n t h e G re e n -B a tc h
d e c re a s e a t h ig h e r d e n s itie s . F or S. p a ra m a m o sa in ,
s y s te m . T h e G re e n -B a tc h s y s te m s e e m s to b e m o re
D ju n a id a h , M ardjono, W ille. K o n ta r a a n d S o rg e lo o s
a p p r o p r ia te for e a rly s ta g e s o f c ra b la r v a e (Z 1 -Z 2 ) a s
(2001) f o u n d a te n d e n c y o f in c r e a s e d s u r v iv a l to Z5
it is le s s s tr e s s f u l fo r th e e a r ly z o e a e a n d e a s ie r t o g ra ­
a s a f u n c tio n o f t h e Z l s to c k in g d e n s ity (i.e. s u r v iv a l
1548
© 2007 The Authors
Journal Compilation © 2007 Blackwell Publishing Ltd, Aquaculture Research, 38, 1539-1553
A quaculture Research. 2007, 38, 1539-1553
Improved rearing te c h n iq u e s for m ud crab larvae T T N g h ia et al.
r a te s o f 2 7 % , 39% a n d 63% b e in g o b ta in e d a t d e n s i­
tio n b y la r v a l s t a g e s in th is w a y c o m p e n s a te s fo r
tie s o f 50 ,7 5 a n d 100 Z l L “ 1 respectively). B a y lo n a n d
th e i n c r e a s e d i n g e s ti o n o f c ra b la r v a e a s t h e y g ro w
F a ila m a n (1999) a lso r e p o rte d h ig h e r s u rv iv a l a n d
(B aylon, B ravo & M a n ig o 2 0 0 4 ). F o r e a rly la rv a e ,
m e ta m o r p h o s is o f Scylla serra ta a t 5 0 Z 1 L - 1 c o m ­
h o w ev er, food a m o u n t c a n n o t b e r e d u c e d to th e ir
p a r e d w it h lo w e r d e n s itie s o f 10 a n d 25 Z l L ~ x. I n ­
m a x im u m i n g e s ti o n p o te n tia l a s t h e y a r e q u ite ineffi­
c re a s e d s u r v iv a l a t h ig h e r la r v a l d e n s itie s s o m e h o w
c ie n t p r e d a to rs a n d th e re fo re m ig h t r e q u ire a m in i­
s e e m s c o n tra d ic to ry . H ow ever, in d ire ctly , food r a tio n
m a l d e n s ity t o m a x im iz e e n c o u n te r .
m ig h t be resp o n sib le. E x cess food in tr e a tm e n ts w i t h
S im ila r to o u r s t u d y m o s t s tu d ie s in v e s tig a tin g t h e
lo w la r v a l d e n s itie s m a y p o llu te th e w a te r a n d m a y
effect o f r o tif e r d e n s i t y a d d e d t h e live food i n o n e s in ­
t h u s c a u s e m o rta lity . I n o u r stu d y , w e n o te d h ig h e r
gle ra tio n . U n d e r t h e s e c ir c u m s ta n c e s , th e o r e tic d e n ­
c o n c e n tr a tio n s o f a m m o n ia a n d n itrite in t h e tr e a t­
sities a r e o n ly a t t a i n e d u p o n fe e d in g a n d g r a d u a lly
m e n t h a v in g 5 0 Z 1 L - 1 (seeT ab le 2). F or t h e h ig h e s t
d e c re a s e a s la r v a e c o n s u m e th e prey. O p tim a l live
s to c k in g d e n s itie s te s te d in o u r s tu d y (2 0 0 Z l L ~ x),
food q u a n titie s c a n n o t , h o w ev er, be s e p a r a te d fro m
t h e la r v a l d e v e lo p m e n t ra te s e e m e d slig h tly im p a ire d .
fe e d in g freq u en cy . B e c a u s e z o e a la r v a e c a n c o n s u m e
T h is h i g h s to c k in g d e n s ity m a y h a v e c a u s e d c o m p e ti­
th e i r o p tim a l r a t io n w i t h i n l h , G en o d ep a , S o u th g a te
tio n fo r feed, r e s u ltin g in slo w e r d ev elo p m en t. T h e re ­
a n d Z eng (2 0 0 4 ) s u g g e s te d t h a t th e y c a n be fed o n c e
fore,
Zl
s to c k in g
d e n s itie s
in
th e
ran g e
of
1 0 0 - 1 5 0 Z l L “ 1 m ig h t b e o p tim a l.
a day. B e c a u se o f t h e se v e re re d u c tio n in t h e n u t r i ­
tio n a l v a lu e o f r o tif e r s w i th lo n g e r r e te n tio n tim e s in
r e a r in g c o n ta in e r s (M a k rid is & O lsen 1999) a n d th e
R o tife r d e n s ity f o r feed in g early larval stages (Z 1 -Z 2
stages)
f a c t th e r e is a m i n i m u m p re y d e n s ity n e e d e d fo r th e
passiv e fe e d in g b e h a v io u r o f z o e a la rv a e ( H e a s m a n &
F ie ld e r 1983; Z e n g & Li 1999), t h e in te r a c tio n b e tw e e n
A lth o u g h th e r e w a s a tr e n d to w a r d s in c r e a s e d s u r v i­
t h e o p tim a l ra tio n a n d fe e d in g fre q u e n c y s h o u ld be
v a l a n d g r o w th w ith in c r e a s in g ro tife r d en sity , n o sig­
f u r t h e r in v e stig a te d .
n if ic a n t d iffe re n c e s in la r v a l s u r v iv a l o r g r o w th w e re
f o u n d b e tw e e n th e d iffe re n t r o tif e r d e n s itie s teste d .
A lth o u g h n o t sig n ific a n t, t h e h ig h e s t s u rv iv a l w a s
g e n e r a lly o b se rv e d a t 4 5 r o tife rs m L - \ w h ile a d e n ­
A rte m ia f o r feed in g la te r la rv a l stages (from Z 3
onw ards)
s ity o f 6 0 r o tife rs m L ~ 1 re s u lte d in th e fa s te s t la rv a l
W e fo u n d n o d iffe re n c e b e tw e e n fe e d in g Z3 a d a ily
d e v e lo p m e n t. T h e d iffe re n c e s w ere, how ever, n o t v e ry
feed r a tio n o f 10, 15 o r 2 0 A rte m ia n a u p lii m L - 1 .
m a rk e d , a n d m o re o v e r s u c h h ig h fee d in g r a te s m ig h t
E sp e c ia lly in la te r la r v a l s ta g e s (Z 4-Z 5), th e r e w as,
b e e c o n o m ic a lly u n r e a lis tic . W e c a n th e re fo re c o n ­
h o w ev er, a te n d e n c y to w a r d s h ig h e r s u r v iv a l w ith
c lu d e t h a t fe e d in g 3 0 r o tife rs m L - 1 is e n o u g h fo r o p ­
in c r e a s in g ra tio n . I n t h i s re s p e c t, it m ig h t be b e n e fi­
ti m a l la r v a l p e rfo rm a n c e . I n p ra c tic e , h o w ev er, th e
c ia l to in c r e a s e t h e A r te m ia d e n s ity b y c ra b s ta g e fro m
in te r m e d ia te d e n s ity o f 4 5 r o tife rs m L _ 1 w a s fre ­
10 to 15 m L - 1 . H ig h live feed d e n s itie s w o u ld in ­
q u e n tly u s e d fo r fe e d in g e a rly la r v a l sta g es. O th e r
c r e a s e t h e c h a n c e fo r e a r ly la r v a e to e n c o u n te r a n d
s tu d ie s
d e n s itie s
c a p tu r e feed o r g a n is m s (Z eng & Li 1999) a n d t h e r e ­
( 3 0 - 8 0 m L - x ) a r e r e q u ire d fo r o p tim a l g r o w th a n d
fore w o u ld im p ro v e t h e la r v a l p e r fo r m a n c e (B rick
in d ic a te d
th a t
h ig h
ro tife r
s u r v iv a l o f S. p a ra m a m o sa in (D ju n a id a h , M ardjono,
1974; H e a s m a n & F ie ld e r 1983; Q u in itio et al. 2001).
L av en s & W ille 1998; Z e n g & Li 1999) a n d S. serrata
O n t h e o th e r h a n d , o ld e r la rv a l h a v e a h ig h e r in g e s ­
(S u p ra y u d i, T a k e u c h i, H a m a s a k i & H iro k a w a 2002).
tio n cap ac ity . O p tim a l r a tio n s s h o u ld th e re fo re b e d e ­
For
S.
p a ra m a m o sa in
and
te r m in e d fo r e a c h la r v a l s ta g e se p ara te ly . I n th is
6 0 ro tife rs m L ~ 1 re s u lte d in a sig n ific a n tly h ig h e r
re s p e c t, s tu d ie s o n in d iv id u a l la rv a e a r e v e ry u s e fu l
s u r v iv a l
c o m p a re d
la rv a e ,
w it h
15 r o tif e r s m L “ 1 ( D ju n a id a h
fee d in g
feed in g
30
o n ly
to d e te r m in e p re y c o n s u m p tio n . A c c o rd in g to o u r
et al. 2001). T h e se
p re v io u s e x p e r im e n ts (N g h ia 2 0 0 4 ), e a c h Z3, Z4, Z5
a u t h o r s f o u n d t h a t th e in d iv id u a l d r y w e ig h t o f Z 5-
a n d m e g a lo p a la r v a w a s c a p a b le o f c o n s u m in g o n
fe d 15 r o tife rs m L ~ 1 w a s s ig n ific a n tly lo w e r t h a n
a v e ra g e
t h o s e o f Z5 fed w ith h ig h e r r o tif e r d e n sitie s. P r a c ti­
A rtem ia d a y - 1 respectively. T h e re fo re , a t a s to c k in g
cally, fe e d in g 3 0 ro tife rs m L " 1 a t Z l a n d in c r e a s in g
o f 100 la r v a e L - \ th e d a ily A rte m ia fe ed in g d e n s itie s
g r a d u a lly to 4 5 m L " 1 a t Z2 p ro v e d to be su fficien t
th e o r e tic a lly s h o u ld be a t le a s t 1.5, 2.5, 3.7 a n d
15.
25,
37
and
114
n e w ly
h a tc h e d
fo r a s to c k in g d e n s ity o f 1 0 0 Z l L ~ 1in o u r tr ia ls i n la r ­
11.4 m L ~ 1 for Z3, Z4, Z5 a n d m e g a lo p a s ta g e s r e s p e c ­
g e r r e a r in g ta n k s ( 5 0 0 - 1 0 0 0 L). In c re a s in g th e ra ­
tively. F or Z l, Z2 a n d Z3 s ta g e s o f S. serrata, t h e
© 2007 The Authors
Journal Compilation © 2007 Blackwell Publishing Ltd, Aquaculture Research, 38, 1539-1553
1549
Improved rearing techniques for m ud crab larvae T T N ghia et al.
n u m b e r o f A rte m ia n a u p lii in g e s te d b y t h e la rv a e a t a
A quaculture Research. 2007, 38, 1539-1553
c u l t u r e s y s te m s u s e d i n o t h e r e x p e r im e n ts . T h e s m a ll
lo w e r food d e n s ity o f 2.5 m l - 1 w a s c o m p a ra b le to
n a u p lii siz e o f th e A r te m ia s t r a i n (V in h C h a u s tr a in )
t h a t a t 5 m L - 1 , a n d fo r Z 4 -Z 5 , a t 5 m L - 1 , it w as
u s e d i n o u r s tu d y c o u ld b e a n o t h e r r e a s o n t h a t led
c o m p a ra b le to 10 m L - 1 (B ay lo n et al. 2004). I n th a t
to in c r e a s e d in g e s tio n . I n p r a c t ic e , ( la rg e r sized)
study, A rte m ia w as, h o w e v e r, co -fe d w ith ro tife rs a t a
h ig h ly u n s a t u r a t e d f a tty a c id - e n r ic h e d A rte m ia w e re
d e n s ity o f 1 5 - 2 0 m L ~ \ I f A rte m ia w a s th e o n ly food,
n o r m a lly u s e d i n o r d e r to r e d u c e t h e p re y a m o u n t to
th e o p tim a l A rte m ia r a tio n w o u ld th e re fo re p ro b ab ly
5 - 1 0 m L - \ M e g a lo p a p ro b a b ly s h o u ld b e fed m o re
b e h ig h e r t h a n 2 .5 - 5 m L - 1 . I n a n o t h e r s tu d y o n S.
f re q u e n tly a n d , to w a r d s t h e e n d o f t h a t d e v e lo p m e n ­
serrata, a d a ily o p tim u m food c o n c e n tr a tio n o f 10 Ar­
t a l sta g e , a n o n - m o v in g food m a y b e better.
tem ia n a u p lii m L " 1 w a s e s ta b lis h e d fo r zo e a su rv iv a l
(B rick 1974). I n t h e m a s s s e e d p ro d u c tio n o f S. serrata,
n e w ly h a tc h e d A r te m ia a r e g iv e n s ta r tin g la te Z2 a t
P ro p h yla ctic chem icals
0 .5 - 3 m L - 1 a n d 5 - 7 - d a y - o ld A rte m ia a r e r o u tin e fed
L a b o ra to ry c u l tu r e s o f c ra b la r v a e o f te n su ffe r sev ere
fro m la te Z5 to e a r ly m e g a lo p a (Q u in itio & P a ra d o -
m o r ta lity f ro m d ise a se , p a r t i c u l a r l y fro m epib io tic
E sta p a 200 3 ). O ld er A r te m ia p ro v id e d a la rg e r-siz e d
b a c te r ia a n d la r v a l m y co sis ( A rm s tro n g , B u c h a n a n
p re y fo r z o e a e to m e g a lo p a e a n d h e n c e , th e d e n s ity
& C a ld w ell 1976; H a m a s a k i & H a ta i 1 9 9 3 a, b). A s tu d y
w a s re d u c e d .
o n S. serrata in d ic a te d a s ig n if ic a n tly h ig h e r s u rv iv a l
F or m e g a lo p a e o f S. p a ra m a m o sa in , w e f o u n d a
u p to D A H 7 (o v er 9 0 % ) w h e n u s i n g O x y tetracy clin e,
th re e fo ld h i g h e r n u m b e r o f in g e s te d n e w ly h a tc h e d
w h e r e a s a lm o s t c o m p le te m o r ta lit y o c c u r r e d in t h e
A rtem ia n a u p lii c o m p a r e d w it h Z5 (114 a n d 37
c o n tr o l t r e a t m e n t ( M a n n 2001). T h e a u th o r c o n s id ­
A rtem ia resp ec tiv ely )
e re d t h a t p o te n tia lly u p to 8 0 % o f t h e la rv a l m o r ta lity
fo r a s im ila r p re y d e n s ity
(N g h ia 2 0 0 4 ). T h is m e a n s t h a t m e g a lo p a e a r e vora­
c o u ld b e a ttr ib u te d to b a c te r io lo g ic a l c a u s e s. T h e r e ­
c io u s p re d a to rs , c a p a b le o f c h a s in g th e ir p re y actively
s u lts o f o u r s tu d y a ls o in d ic a te d t h a t b a c te r ia are,
a n d c o n s u m e la r g e a m o u n t s o f feed in a s h o r t tim e.
m o re t h a n a n y o t h e r fa c to r te s te d , a m a in c a u s e o f
F ro m th is , it c o u ld be b e n e fic ia l if m e g a lo p a e a r e fed
la r v a l m o rta lity . A n tib io tic s m o re t h a n d o u b le d s u r ­
fre q u e n tly s m a lle r r a tio n s in o r d e r to o p tim iz e feed
v iv a l u p to t h e c r a b sta g e.
q u a lity a n d r e d u c e c a n n ib a lis m . G e n o d e p a et al
H ow ever, a n tib io tic s h a v e n o t a lw a y s b e e n u s e d in
(2 004) s im ila rly in d ic a te d t h a t in c o n tr a s t to e a rlie r
a r e s p o n s ib le m a n n e r in a q u a c u ltu r e . A m a jo r c o n s e ­
la rv a l sta g e s, w h i c h c a n b e fe d o n c e p e r day, S. serrata
q u e n c e o f u s in g a n tib io tic s h a s b e e n t h e p ro life ra tio n
m e g a lo p a e m a y n e e d to b e fed m o re o fte n to m ax i­
o f r e s is ta n t b a c te r i a a n d t h e tr a n s m is s io n o f r e s is ­
m iz e in g e s tio n . T h e s e a u t h o r s fo u n d n o sig n ific a n t
t a n c e to o t h e r b a c te r ia l sp e c ie s (B e n s o n 1998). T h e
d ifferen ces in t h e i n g e s tio n r a te o f m e g a lo p a e fed m i­
d e v e lo p m e n t o f a n tib io tic r e s is ta n c e b y p a th o g e n ic
c r o b o u n d d ie ts a t r a tio n s r a n g in g fro m 12.5% to
b a c te r ia is c o n s id e re d to b e o n e o f t h e m o s t se rio u s
100% o f t h e s ta n d a r d r a t io n (e q u iv a le n t to 5 A rtem ia
risk s to h u m a n h e a lt h a t t h e g lo b a l lev el (FAO 2002).
n a u p lii m L “ 1 in 1 h). B a y lo n et al (2 0 0 4 ) a lso fo u n d a
F o rm a lin is m o re a c c e p ta b le t h a n a n tib io tic s a s it
h ig h in c r e a s e in A r te m ia in g e s tio n i n t h e first few
s h o w s n o a c c u m u la tio n in a n im a l tis s u e s (Jung,
d ay s o f t h e p la n k to n ic p h a s e o f t h e m e g a lo p a stage.
K im , J e o n & L ee 2001). R ecently, h o w ev er, J a p a n h a s
L a te r o n , m e g a lo p a e b e c o m e m o r e b e n th ic a s th e y
s tr ic tly b a n n e d t h e u s e o f fo rm a lin i n a q u a c u lt u r e a s
p re p a re fo r th e s e c o n d m e ta m o r p h o s is to first crab.
it m a y c a u s e c a n c e r in h u m a n s , r e d u c e s o x y g en
S w im m in g A rte m ia a r e n o lo n g e r accessib le, a n d
levels i n t h e w a te r a n d c a u s e s a lg a e to d ie off (VASEP
m in c e d s h r im p o r m u s s e l m e a t a r e a m o re su ita b le
2003). M oreover, in o u r e x p e rim e n ts , f o r m a lin d id n o t
feed.
s ig n ific a n tly im p ro v e la r v a l s u r v iv a l c o m p a r e d w ith
I n c o n c lu s io n , a r a tio n o f 10 A rte m ia n a u p lii m L “ 1
t h e n e g a tiv e c o n tro l. P a th o g e n ic b a c te r ia a r e c o n s id ­
a p p e a rs to b e su ffic ie n t fo r t h e o p tim a l p e rfo rm a n c e
e re d to be o n e o f t h e m o s t s e rio u s c a u s e s fo r th e h ig h
o f Z3 la rv a e . A n in c r e a s e in p r e y d e n s ity i n th e Z 4 -Z 5
m o r ta lity o f e a rly c ra b la rv a e . I t c a n b e sa fely a s­
sta g e
s u m e d t h a t a ll i n p u ts (se a w a te r, b ro o d sto c k , live feed
m ay,
h o w e v e r,
be
b e n e fic ia l.
T h e se
food
a m o u n ts a p p e a r to b e h i g h e r t h a n w h a t m o s t o th e r
a n d d a ily m a n a g e m e n t i n h a tc h e rie s ) in to th e c u l­
s tu d ie s r e c o m m e n d ( 2 .5 -1 0 A r te m ia n a u p lii m L -
t u r e t a n k a r e p o te n tia l s o u r c e s o f in f e c tio n (Black-
B rick 1974; B a y lo n e t al. 2 0 0 4 ). P e r h a p s t h e r e c irc u la t­
s h a w 2001). S tric t h y g ie n e a t a ll s te p s is alw ay s
in g s y s te m u s e d i n th i s s tu d y r e s u lte d in a g r e a te r loss
a d v is e d fo r h a t c h e r y a c tiv itie s. H ow ever, th is ad v ice
o f p re y o r g a n is m s (e.g. m o r e A r te m ia w e re e n tra p p e d
is
o n th e o v e rflo w sc re e n ) t h a n i n th e s m a ll b a tc h
h a tc h e rie s . T h e re fo re , o t h e r te c h n iq u e s s h o u ld be
1550
not
a lw a y s
fo llo w ed ,
esp e c ia lly
in
b a c k y a rd
© 2007 The Authors
Journal Compilation Ç) 2007 Blackwell Publishing Ltd, Aquaculture Research, 3 8 , 1539-1553
Aquaculture Research, 2007, 38, 1539-1553
Improved rearing te c h n iq u e s for m ud crab larvae T T N g h ia et al.
in v e s tig a te d a s a lte r n a tiv e s fo r t h e u s e o f c h e m ic a ls .
p ro d u c ts, m o re o v e r, a r e n o t e n c o u r a g e d fo r c o m m e r­
O z o n a tio n a n d p ro b io tic s c o u ld b e in te r e s tin g in th is
c ia l m u d c ra b l a r v i c u l tu r e a s th e y a r e u n s a fe . D ire c t
re s p e c t (Davis 2003; N g h ia 2 0 0 4 ). O z o n e is a p o w e r­
o z o n a tio n a s a n a lte r n a t iv e to p ro p h y la c tic c h e m ic a ls
fu l o x id a n t a n d is b e c o m in g m o r e a n d m o re p o p u la r
is w o r th in v e s tig a tin g .
in v a rio u s a q u a c u ltu r e s y s te m s fo r d is in f e c tio n a n d
im p ro v in g w a te r q u a lity b y o x id a tio n o f in o r g a n ic
a n d /o r o rg a n ic c o m p o u n d s (T an g o & G a g n o n 2003).
I n o u r study, d ir e c t a p p lic a tio n d id n o t s ig n ific a n tly
A c k n o w led g m en ts
im p ro v e s u rv iv a l c o m p a r e d w it h a n e g a tiv e c o n tro l.
T h is s tu d y w a s s u p p o r te d by th e E u r o p e a n C o m m is­
H ow ever, th e r e w a s a te n d e n c y o f a r e s id u a l 0 3 c o n ­
s io n (INCO-DC) t h r o u g h t h e p ro je c t g r a n t No IC A 4-
c e n tr a tio n o f 0.0 6 m g L ~ 1 to im p ro v e la r v a l p e rfo r­
C T -2001-10022 ‘C u lt u r e a n d m a n a g e m e n t o f Scylla
m a n c e . V ariab ility
w as,
sp e c ie s’, t h e F le m is h In te r-U n iv e rsity C o u n c il (VLIR-
h o w ev er, v e ry h ig h , w h ic h c o u ld be in d ic a tiv e o f th e
IUC) a n d th e I n t e r n a ti o n a l F o u n d a tio n fo r S cien ce
fa c t t h a t d o sin g w a s n o t c a r e f u l e n o u g h . L o n g e r O3
(IFS) th r o u g h th e r e s e a r c h g r a n t a g r e e m e n t N o A /
e x p o s u re tim e s ( 4 - 1 0 - m in e x p o s u re , e q u iv a le n t to
2505-1 ‘I m p r o v e m e n t o f la r v ic u ltu r e o f t h e m u d crab
0 .1 2 -0 .1 9 m g L _1 re s id u a l 0 3) a ll d e c r e a s e d o v erall
(Scylla p a ra m a m o sa in ) in th e M ekong D elta, V ie tn a m ’.
w it h in
th is
tre a tm e n t
su rv iv a l. T h e s e h ig h 0 3 c o n c e n tr a tio n s p ro b a b ly
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