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.