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J. Indian Fish. Assoc., 32: 69-80, 2005 69
LARVAL REARING OF A FRESHWATER PRAWN MACRO BRACHIUM GANGETICUM
D. Roy*, V. K. Yadav and S. R. Singh
Department of Zoology, S.M.M. Town P G. College
Ballia- 277001, INDIA
ABSTRACT
Observations were made on the larval development of a freshwater prawn, Macrobracnium gangeticum, which revealed that hatching occurred in freshwater but the larvae failed to survive after 2nd molting. Salinity was necessary for survival of the larvae after 2nd moulting. The complete larval development involved nine larval stages and the lOth stage was considered to be the post-larva which measured between 4.5 and ~.0 mm in length. All the larval stages were completed within 26 days ofhatching. Specimens from each larval stage were taken out and examined under a microscope.
Keywords: Freshwater prawn, Macrobrachium gangeticum, Larval forms
INTRODUCTION
Out of 150 freshwater prawn species recorded worldwide, 40 species occur in the Indian subcontinent (Kumar and Pandey, 2003). Despite great significance of freshwater prawns as protein rich diet and a foreign exchange earner, the prawn culture programmes in India are restricted to the maritime states only and that too, is mainly through collection of juveniles from nature which is always a mixture of desirable and undesirable species (Parmeswaran,1999). Non availability of quality seeds is a major constraint to culture practice of freshwater prawns (Tripathi, 1999) in India, and as such, it is urgently needed to evolve indigenous
* - Corresponding author
technology for seed production of freshwater prawns. Sankoli et al., (1978) have discussed the potential of cultivable species of the Macrobrachium.
Macrobrachium gangeticum (earlier known as Macrobrachium choprai; Tiwari and Holthuis, 1996) is the only available economic variety of freshwater prawn in Uttar Pradesh, inhabiting middle and lower reaches of Ganga river. Its fishery has declined to a great extent, and immediate attention is needed to evolve methods for its propagation and culture.
The present work is, therefore, aimed at evolving techniques to breed it in captivity and rearing its larvae to
70 D. ROY ET AL.,
post larval stages under controlled conditions. Most freshwater prawns of the genus Macrobrachium are reported to require some salinity for their larval development. At places far distant from sea, larval development programmes are difficult because transportation of seawater involves huge cost and preparation of synthetic sea -water too is not economical. Therefore, an attempt has been made to develop optimum salinity desired for larval development with the help of rock salt and salt pan residue.
MATERIAL AND METHODS
Collection of animals, techniques adopted and precautions observed have been described along with the experiments conducted in different phases.
In the first experiment mature specimens were collected from the river Oanga near village Kotawan N arayanpur (about 35 km from the laboratory) and brought in earthen pots. They were kept in large plastic pools filled with well aerated tap water and allowed to
acclimatize to laboratory conditions.
They were fed daily on goat liver/ pellets made of dried and crushed Macrobrachium lamerrei niixed with wheat flour.
Large aquaria (3' x 1.5' x 1.5') and plastic pools fitted·with aerators, were filled with tap water. One mature male and two mature females were kept in each aquarium. Feeding was continued. The water was changed every alternate day. Once the females were berried, the males were taken out from the aquaria. Physico-chemical conditions of the water (Table-1) were regularly observed by using electronic water analysis kit and as per standard methods, APHA (1985).
On the 20th day hatching occurred early in the morning. Spent females were removed from the aquaria after hatching. The larvae were fed on plankton. But unfortunately, the larvae could not survive beyond 8th day of hatching during which they underwent three moults.
In th~ second phase of this
Table 1: Physico-chemical conditions of culture media (Experiment 1)
Parameters Range of Variation
Water temperature coc) 24 30.2 pH 8.4 9.3 Dissolved Oxygen (ppm) 7.2 13.0 Conductivity (mhos) 1.131 1.691 Total dissolved solids (ppm) 52.0 180.0 Total alkalinity 105 132.0
LARVAL REARING OF MACROBRACHIUM GANGETICUM 71
experiment three happas (2mx2mx1m) made of thinly meshed nylon cloth were fixed in marginal flood waters of the Ganga river. In each happa two berried females collected from the river were placed. Hatching took place in all cases and the spent females were removed from the happa. River water from outside the happa was poured gently into the happa twice daily to reduce the probability of any planktonic food shortage. However, here also the population of larvae dwindled completely after 4th/5th day.
In the second experiment large aquaria (3' X 1.5' X 1.5') fitted With aerators were filled with water containing-10 _ppt salinity developed with rock salt. Side by side, a little below the level of aquaria, plastic pools containing the water of the same salinity and fitted with water cooler pumps were installed to pump fresh saline water into
aquaria through plastic pipes. Another pipe was used to return back the water from culture media to plastic pool on simple siphon system. Thus, a system of recycljng of water was established. Continuous aeration was ensured in both the containers.
One female berried in the laboratory itself was placed in each aquarium and fed as in previous experiment. The water of the culture media was recycled daily. Hatching occurred after 18-20 days in batches. Spent females were removed after hatching.
No feed was supplied to the larvae on the first day. Planktonic food was supplied to them from 2nd day onward. The physico-chemical parameters of the culture media were estimated daily (Table-2).
The larvae were active swimmers, gregarious in nature and remained
Table 2: Physico-chemical conditions of culture media (Experiment -2)
Parameters Range of Variation
Water temperature (oc) 23.6 28.0 pH 8.0 8.4 Dissolved Oxygen (ppm) 7.2 7.8 Free carbon dioxide (ppm) 2.0 3.6 Total dissolved Solids (ppm) 68.0 70.0 Total alkalinity (ppm) 30.2 58.6 Salinity (ppt) 6.0 10.0 Nitrates (ppm) 0.08 1.35 Nitrite (ppm) 0.06 2.43 Ammonia (ppm) 0.05 1.30 Conductivity (mhos) 1.691 5.551
72 D. ROY ET AL.,
usually close to the surface of water. The larvae passed through morphologically different nine zoeal and a post-larval stages. However, the mortality rate was too high and only a few larvae survived to attain juvenile stage.
In the third experiment, the salinity of the culture media was developed by using rock salt in combination with liquid residue of salt pans (waste product) for larval development. The rock salt, a raw salt produced in salt farms, contains only macronutrients (Nacl, MgS0
2 and
CaCl2). The liquid residue on the other
hand contains various other components in traces viz., KCl (1.7%), NaHC0
3
(0.49% ), KBr (0.07% ), SrCl2(0.04% ),
Al/S02) (0.001 %), ZnS0/0.0002%),
properly cleaned plasitc pools and were transferred to the aquaria. Recycling of water was established as in the earlier experiment.
One berried female was transferred to each aquarium. Hatching occurred as in previous experiments. On the first day larvae were not supplied with food. From the second day onward they were supplied with artemia larvae.
Permanent mounts of all stages of larvae were prepared and their morphological details were examined with the help of a microscope. Physicochemical conditions were daily monitored as in previous experiments (Table 3). In this experiment, mortality of larvae was highest at 15ppt salinity,
Table 3: Physico-chemical conditions of culture media (Experiment -3)
Parameters
Water temperature (oc) pH Dissolved Oxygen (ppm) Free carbon dioxide (ppm) Total dissolved Solids (ppm) Total alkalinity (ppm) Conductivity (mhos)
Kl(0.0002%) etc. Mixing of freshwater, rock salt and liquid residue waste in the ratio of 200:2:1 gives a medium resembling ±15ppt sea water (Shree Prakash, 1988). Keeping the above in view, water of salinities 8.0, 10.0 and 15.0 ppt was prepared separately in
Range of Variation
20.6 28.9 8.2 8.9 9.2 12.2 0.0 3.2
61.0 106.7 85.0 118.0
1.342 1.674
while it was lowest in case of 8 ppt salinity.
RESULTS
The morphological details of the various stages of larvae are described in table 4. In all, nine larval and a postlarval stage were recorded.
Tab
le 4
: M
orph
olog
ical
Cha
ract
eris
tics
of L
arva
l For
ms
Sta
ges
Car
apac
e &
E
yes
Ant
enna
R
ostr
um
ST
AG
E I
C
arap
ace
smoo
th
Ses
sile
R
udim
enta
ry
(1-1
.1m
m)
wit
h a
New
ly h
atch
ed
pter
ygos
tom
al
spm
e;
Ros
trum
dir
ecte
d fo
rwar
d,0.
15-0
.2
mm
inle
ng
th
ST
AG
E I
I C
arap
ace
Sta
lked
A
nten
na!
(1.1
-1.3
mm
) de
velo
ps
flag
ellu
m
2 da
ys o
ld-
supr
aorb
ital
and
pr
esen
t br
anch
iost
egal
sp
ines
als
o;
Ros
trum
0.1
6-0
.2m
min
le
ngth
,ben
t do
wnw
ard
ST
AG
E I
II
Ros
trum
wit
h E
xten
ded
No
dist
inct
(2
.0-2
.5 m
m)
epig
astr
ic s
pine
sl
ight
ly
chan
ge
4 da
ys o
ld
Ant
ennu
le
Per
iopo
ds
mid
Ple
opod
s
Rud
imen
tary
F
irst
3 p
airs
of
umra
mou
s pe
riop
ods
pres
ent
Tw
o-1s
t and
2nd
segm
ente
d pe
riop
ods
ante
nnul
ar
beco
me
pedu
ncle
bi
ram
ous,
pr
esen
t ex
opod
4
segm
ente
d bu
t en
dopo
d un
segm
ente
d
No
dist
inct
3r
d an
d 4t
h
chan
ge
peri
opod
s be
com
e bi
ram
ous,
Sth
umra
mou
s
Tel
son
&U
rop
od
Not
dis
tinc
t
Rud
imen
ts o
f te
lson
and
ur
opod
pre
sent
R
udim
ents
of
tels
on a
nd
urop
od p
rese
nt
Tel
son
nano
ws,
ur
opod
pa
rtia
lly
deve
lope
d Con
td ...
. I
~ JJ ~ r JJ ~ JJ
z G) 0 T1 ~ ~ @
?2 ~ ~ ~ ~ :::l
() ~ '-
I c.v
Stag
es
Car
apac
e &
E
yes
Ant
enna
R
ostr
um
STA
GE
IV
R
ostr
um m
ore
Mor
e F
lage
llum
two
(2.4
-2.7
mm
) de
velo
ped
wit
h ex
tend
ed
segm
ente
d,
7 da
ys o
ld
rudi
men
t of o
ne
exop
od w
ith
dors
al t
ooth
se
tae
and
spin
e
ST
AG
EV
N
o di
stin
ct
No
dist
inct
F
lage
llum
thre
e (2
.7.-
2.95
) ch
ange
ch
ange
se
gmen
ted
10 d
ays
old
STA
GE
VI
Ros
trum
Sl
ight
ly
Exo
pod
wit
h (2
.95-
3.1m
m)
deve
lops
en
larg
ed
20 s
etae
and
1
13 d
ays
old
rudi
men
t of 2
nd
sp
me
dors
ol t
ooth
--------
--------
----
Ant
ennu
le
Per
iopo
ds
and
Ple
opod
s
Inne
r 5t
h pe
riop
od
ante
nnul
ar
beco
mes
fl
agel
lum
with
bi
ram
ous,
set
ae
swol
len
base
, de
velo
ped
at
dist
al s
egm
ent
ends
w
ith
seta
e
Ant
ennu
lar
Per
iopo
ds w
ell
pedu
ncle
de
velo
ped
deve
lope
d a
plum
ose
seta
e,
one
spin
e at
pr
oxin
al
segm
ent
Ant
ennu
lar
4th
& 5
th
pedu
ncle
wit
h pe
riop
ods
2 se
tae
on in
ner
beco
me
4 an
d 2
flag
ellu
m, o
uter
se
gmen
ted
flag
ellu
m w
ith
resp
ecti
vely
8
seta
e an
d 1
spm
e
Tel
son
&U
ropo
d I
Tel
son
!
elon
gate
d w
ith
I
term
inal
spi
nes;
: ur
opod
sho
ws
I
exop
od a
nd
I
endo
pod
wit
h j
seta
e
Tel
son
I
narr
ower
&
I
rect
angu
lar,
ur
opod
wit
h 1
spin
e &
12
I se
tae
I
Tel
son
as
I
abov
e,
urop
od m
uch
deve
lope
d.
wit
h se
tae
Con
td. .
..
-...J ~
~
:0
0 -< ~ )::,. s-
Stag
es
Car
apac
e &
E
yes
Ant
enna
A
nten
nule
R
ostr
um
ST
AG
E V
II
Ros
trum
wit
h A
s ab
ove
Inne
r fla
gell
um
Out
er fl
egel
lum
(3
.2-3
.72m
m)
two
dors
al t
eeth
al
so d
evel
ops
wit
h te
n se
tae
15 d
ays
old
wit
h fo
ur s
etae
ST
AG
E V
III
No
dist
inct
Sl
ight
ly
As
abov
e A
s ab
ove
(3.6
-3.9
mm
) ch
ange
en
larg
ed
19 d
ays
old
ST
AG
E I
X
Fur
ther
elo
n-N
o di
stin
ct
Inne
r an
d ou
ter
No
dist
inct
(4
-4.5
mm
) ga
ted
wit
h tw
o ch
ange
fl
agel
la s
plit
ch
ange
23
day
s ol
d te
eth
ST
AG
E X
R
ostr
um w
ith
Slig
htly
S
egm
ente
d S
egm
ente
d (P
OS
T
thre
e te
eth
enla
rged
fl
agel
la
flag
ella
LA
RV
A)
( 4.5
-5.0
mm
) 26
day
s ol
d
Per
iopo
ds
and
Ple
opod
s
1st &
2nd
peri
opod
s be
com
e ch
elat
e, 5
th
peri
opod
lo
nges
t p l
eo p
od b
uds
appe
ar
Per
iopo
ds
mor
e de
velo
ped,
pl
eopo
d bu
ds
bira
mou
s
Per
iopo
ds a
s ab
ove,
ple
opod
bu
ds b
iram
ous
Fiv
e pa
irs
of
bira
mou
s pl
eopo
ds
Tel
son
& U
ropo
d
Tel
son
mor
e na
rrow
ed,
urop
od m
uch
deve
lope
d
Uro
pod
mor
e de
velo
ped
Tel
son
wit
h th
ree
pair
s o
f se
tae,
uro
pod
bilo
bed
Tel
son
wit
h se
tae
and
spin
es,
urop
od
fully
dev
elop
ed I
s;: :0
~ r :0
m
)>
:n z (j
) 0 "Tl ~ g @
~ ~ ~ ~ <:
G
) 111
:::l
C
') ~ -....!
()1
76 D. ROY ET AL.,
The newly hatch~"d larva was the first stage zoea which Jf1uerwent moulting after 2 days and formed the stage II zoea. Second moulting occurred after 4 days resulting into stage III zoea larva. Thereafter, seven moultings occurred one each on 7th, 1Qth,I3t\15th, 19t\20th
Fig. 1: Larval Stage: Zoea I
3: Larval Staf!e: Zoea JJJ
Fig. 5: Larval Stage: Zoea V
and 26th days of hatching resulting into IVth, Vth, Vlth, VIIth, VIIIth,IXth, and Xth (post larva) zoea larval stages respectively Fig.l-10. The growth pattern of larvae is depicted in Fig.ll.
The initial larval stages were transparent with few ch~'1matoohores
Fig. 2: Larval Stage: Zoea II
Fig. 4: Larval Stage: Zoea IV
Fig. 6: Larval Stage: Zoea VI
A-J a-t a
b e
LARVAL REARING OF MACROBRACHIUM GANGETICUM
Fig. 8: Larval Stage: Zoea VIII
Fig. 7: Larval Stage: Zoea VII
Fig. 9: Larval Stage: Zoea IX Fig. I 0: Post Larvae
Abbriviations used in the figures 1-10.
Larval stages X 30 (reduced by 25%) Appendages of larval stages x 60 (reduced by 25%) antennule antenna eye
p-I to PI-V r
Pleopods I-V rostrum
p-I to p-V Periopods I-V t tel son
77
78 D. ROY ET AL.,
5
4
E' E ~E 3 £E g'~ (]) II _.E ~(.) roN
2 Gi ~
1
04-~--~--~------~--.--.--.--.--.--.--.--. 0 2 4 6 ,_8 10 12 14 16 18 20 22 24 26 28
Age (in days) 1 em= 2 days
Fig: 11: Growth pattern o/{arvae of Macro brachium gangeticurn.
spread on their bodies, particularly on abdomen, telson, periopods and eyes talk. Gradually the chromatophores increased in number and branched.
The post larvae metamorphosed into juveniles which ceased to have pelagic life, settled to the bottom ,assumed benthic behavior and were noted to cling on submerged vegetation and other objects.
DISCUSSION
The newly hatched larva of M. gangeticum is a typical protozoea and exhibits close relation to that of other species of the genus which shows a long larval history . The process of hatching
shares a number of common features observed by Ling (1962) a1 Rajyalakshmi (1960) witn M. rosenbergii and some other species of palaemonid prawns respectively.
It was observed in the present study that M.gangeticum passes through ten larval stages during its embryonic history.The larvae of almost all the stages exhibited strong attraction for light and were seen for most of the time at the surface. Reeve (1970) has also reported positive phototaxis in palaemonid larvae.
Most of the species of the genus Macrobrachium have been reported to complete their larval life within 25-30 days of rearing (Choudhuri, 1971; Ling
LARVAL REARING OF MACROBRACHIUM GANGETICUM 79
1962; Pillai and Mohammed, 1957).In the present study also, all the larval stages were completed within 26 days of hatching. This may be correlated with the smaller egg size of this species. J alihal et al., (1979, 1999) have also reported that Macrobrachium species equipped with smaller eggs have longer life histories. In contrast to it, Alvarez et al., (2002) and Mantel et al., (2005) have reported that species having larger eggs (M. tuxtlaense and M. hainanense respectively) have shorter larval histories.
It was observed that hatching is possible in freshwater but the species needs salinity to pass through all the larval stages. In freshwater, the larvae could survive only upto 2nct larval stage. The hatching phenomenon in this species occurred in about 18-20 days after the female became berried. This is in contrast to M. rosenbergii in which hatching is reported to occur within a day or two (Reddy, 1999). Moreover, it was observed in the present study that the larvae of M. gangeticum require relatively lower salinity (8ppt) as compared to those of M. rosenbergii (14-16ppt).
ACKNOWLEDGEMENT
The financial assistance received from Council of Science & Technology, Lucknow and Indian Council of Agricultural Research, New Delhi, is gratefully acknowledged.
REFERENCES Alvarez, F., Villalobos J. L. and
Robles R., 2002. Abbreviated larval development of Macrobrachium tuxtlaense reared In laboratory. Crustaceana, 75 (5) : 717-730
APHA., 1985. Standard Methods for the examination of water and waste water. American Public Health Association. American Water Works Association.
Choudhuri, P. C., 1971, Complete larval development of palaemonid shrimp Macrobrachium carcinus reared in laboratory. Crustaceana, 20 (1) :51 - 61.
Jalihal, D.R., Shenoy, S. and Sankoli, K.N., 1979. Laboratory culture studies in the freshwater prawns M.kistensis (Tewari) from Wai (Maharashtra). Bull. Fish Fac. Konkan Agr. Univ. India, 1 (1): 73-82.
J alihal D.R., Shenoy S. and Sankoli K.N., 1999. Larval development of Indian atyid shrimp Caridian kempi reared in laboratory, College of Fisheries, Ratn~giri.
Kumar, A. and Pandey, A. K., 2003, Neuroendocrine regulation of ovarian maturation in the giant freshwater prawn, Macrobrachium rosenbergii, Fishing Chimes, 23 (7) : 10-17.
80 D. ROY ET AL.,
Ling, S. W., 1962, Studies on the rearing of larvae and juveniles and culturing of adults of Macrobrachium rosenbergii. FA. 0. Current Affairs Bull., 35: 1-11.
Mantel, Kaur S., Dudgeon, David., 2005. Reproduction and Sexual dimorphism of the Palaemonid shrimp, M. hainanense in Hong Kong Streams. Journal of Crustacean Biology., 25 (3): 450-459.
Parmeswaran, S., 1999. Aquaculture of freshwater prawn-global scenario. Manual of summer school on Hatchery and Grow out Technology of Giant freshwater prawn, 1 - 9.
Pillai, N .N. and Mohammed K.H., 1957, Larval History ·of Macrobrachium idella reared in laboratory. 1. Mar. Biol. Assoc. India, 15 (1) : 150.
Rajyalakshmi, T., 1960. Observation on the embryonic and larval
. I
development of some palaemonid prawns. Proc. Nat. Inst. Sci. India., 26 B (6) : 395
Reddy, A. K., 1999. Larval rearing of giant freshwater prawn. Manual of Summer School on Hatchery
and Grow out Technology of Giant Freshwater prawn. 86-92.
Reeve, M. R., 1970. The laboratory culture of prawn Palaemon serratus. Fish Invest., London, 26 (1}: 1
Sankoli, K.N., Shenoy S., Jalihal D.R., 1978. Possible potentially cultivable new species of freshwaterprawns in the genus Macrobrachium. National Symposium on Shrimp Farming organized by MP EDA at Bombay.
Shree Prakash, 1988. On the successful reanng of Macrobrachium rosenbergii in medium prepared of common . salt and salt pan residue. Indian 1. Fish., 35 (1) : 41-45.
Tiwari, K. K~ and Holthuis, L.B., 1996. The identity of Macrobrachium gangeticum Bate, 1868. Crustaceana, 69 (7): 922-925.
Tripathi, S. D., 1999. Present status and prospects ofprawn farming in India. Manual of Summer School on Hatchery and Growout Technology of Giant Freshwater Prawn. 13-15.
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