EurAsian Journal of BioSciences

Eurasia J Biosci 5, 110-118 (2011)

DOI:10.5053/ejobios.2011.5.0.13

Research Note

Embryonic and larval development of critically

endangered riverine catfish Rita rita
Muhammad Fazlul Aw al Mollah1 , Khanam Taslima1* , Harunur Rashid2 ,
Zakir Hossain1 , Muhammad Nasif Sarow ar1 , Muhammad Rakibul Kabir Khan3
1 Fisheries Biology and Genetics Department, Faculty of Fisheries, Bangladesh
Agricultural University, Mymensingh, Bangladesh
2 Fisheries Management Department, Faculty of Fisheries, Bangladesh Agricultural
University, Mymensingh, Bangladesh
3 Surgery and Obstetrics Department, Faculty of Veterinary Science, Bangladesh
Agricultural University, Mymensingh, Bangladesh

* Corresponding Author: taslima_bau@yahoo.com

Abstract
The present study w as carried out to investigate the embryonic and larval development of
freshw ater catfish Rita rita. The mature eggs and sperms w ere collected by using artificial
insemination technique and fertilized eggs w ere incubated in mini circular hatchery w ith
provision of continuous w ater supply. The fertilized eggs w ere transparent, demarsal,
spherical, non-adhesive and brow nish in colour w ith a diameter ranging betw een 1.3 to 1.6
mm. First cleavage occurred w ithin 25-30 min post-fertilization at temperature of 28 1 C.
Hatching started 22 h post-fertilization and completed w ithin 24 h at the same temperature
range. New ly hatched larvae w ere 2.0 mm in length devoid of mouth and pigmentation and
started feeding w ithin 48-60 h post-hatching. To date, this is the first time the early embryonic
and larval development of freshw ater catfish R. rita is described. Thus the findings of the
present study provide valuable information that may help establishing the large scale seed
production technique of Rita.
Keyw ords: Embryo development, larva, metamorphosis, Rita rita.
Mollah MFA, Taslima K, Rashid H, Hossain Z, Sarow ar MN, Khan MRK (2011) Embryonic and
larval development of critically endangered riverine catfish Rita rita. Eurasia J Biosci 5: 110118.
DOI: 10.5053/ejobios.2011.5.0.13

INTRODUCTION
Bangladesh is a land of high potential
w at er resources w here inhabit s 2 60
f reshw at er f ish species, 2 4 f reshw at er
praw ns, 475 marine f ish species, 36 marine
and brackish w at er shrimps and 16 exotic
species w hich supplying around 58% of
animal protein (Anonymous 2010). At least
55 species of freshw ater cat fishes belonging
t o 35 genera have been recorded in
Bangladesh (Rahman 2005). Rita rita is one of
them popularly know n as Rit a in Bangladesh
and also in India. The fish being greenish
brow n above, brow nish w hit e below w ith an
extremely slimy body w hich pref ers muddy to
clean w at er as their habitat.
5 4 f ish species of inland w at er of
Bangladesh are considered as eit her in
endangered or critically endangered situation.
Rita rit a (Hamilton, 1822) is categorized as
EurAsian Journal of BioSciences, 2011

critically endangered fish (Anonymous 2000).

Once t he abundance of this fish w as very rich
in A f ghanist an, Pakist an, India, Nepal,
Bangladesh and Mayanmar (Tripat hi 1996).
But the availabilit y of this f ish is decreasing
day by day due to the ecological changes of
the breeding ground. As a result breeding
grounds are losing t heir suit ability t o be used
by t he species posing a great t hreat of
extinction and till now it is not possible to
collect the fry of this f ish f rom the river
system w here t hey breed nat urally.
Although Rit a is a tasty, commercially
important, crit ically endangered catfish of
Indian subcontinent, published report s on its
induced breeding and rearing are quite scanty
and no report s are readily available on t he
Received: July 2011
Accepted: October 2011
Printed: November 2011
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development al biology and larvae rearing
technique of this species. Earlier att empts
have been made to study it s length-w eight
(Lal and Dw evedi 1969), age and grow th
(Devi et al. 1990), environmental impact
assessment
(Mukhopadhyay
1 99 4 ),
morphometric charact ers (Devi et al. 1991),
f ood (Devi et al. 19 9 2), mat urit y and
fecundit y (Saxena 1972), structure of t he skin
(Mital 1968) and reproductive biology of R.
rita (Amin et al. 2008). Recent success in
induction of breeding of R. rit a w as performed
by using PG (Mollah et al. 2008).
Considering the enormous importance of
Rit a fish, early lif e hist ory informat ion is an
essential requirement f or optimization of large
scale
seed
product ion,
cult ure
and
management . The present w ork is t he f irst
ever preliminary att empt and is expect ed to
serve as a basis f or furt her and more int ensive
future studies. An att empt w as made to
conduct t his st udy t o investigat e and to
provide t he det ailed inf ormat ion of t he
embryonic and larval development of crit ically
endangered fish (R. rita).
MATERIAL AND METHODS
This experiment al w orks w ere carried out
in t he Mini Hatchery cum Breeding Complex
and t he laboratory of t he Department of
Fisheries Biology and Genet ics under the
Facult y of Fisheries, Bangladesh Agricultural
Universit y (BAU) Mymensingh. The collected
mature broods of Rita rita (Hamilt on, 1822)
w ere
induced
to
spaw n
t hrough
hypophysat ion previously described by Mollah
et al. (2008). Aft er that, the eggs w ere
fertilized by the normal milt . For collection of
milt , the t est es from male w ere dissect ed out
from it s body cavit y and w ere crushed by
scissor. Fertilized eggs w ere w ashed several
times w it h st erile dist illed w at er and f inally
transf erred t o and spread as homogeneously
as possible in mini circular hatchery (50 L
capacit y) w it h cont inuous w at er supply
throughout the incubation phase. A set of
fertilized eggs w ere collect ed very caref ully
from t he mini plastic circular hat chery w here
they w ere in const ant mot ion by using a
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Mollah et al.

dropper and siphoning process. Aft er t hat , it

w as observed under the microscope (Olympus
CX41) fit ted w it h a soft w are (Magnus MIPSMicrosoft Image Processing Syst em). Each
sample w as observed f our times to ident if y
t he development al st ages (Hanif f a et al.
2003).
Fertilized eggs w ere collected up to morula
st age every 15 min and t hen every one hour
int erval until hatching and every f our hours for
t he next. The hat ching of f ertilized eggs w as
completed w ithin 24 h. Formalin (5%, v/v)
w as used for t he immobilization of the larvae
during st udy periods. The diamet ers of the
eggs w ere measured by using an eye piece
micromet er. The size of t he larvae w as
measured by placing on the petri dish placed
on a 1 mm graph paper (Miah et al. 2009).
The w hole experimental w orks w ere done at a
t emperat ure of 28 1 C.
RESULTS
Embryonic development
The embryonic period starts w hen t he egg
is fertilized by a sperm and ends w hen the
embryo has att ained the generalized organ
syst ems as they appear in common in all the
f ish (Table 1).
Unfert ilized and f ertilized eggs
The unf ert ilized eggs of R. rit a w ere
opaque, demarsal, spherical and w hitish in
colour measuring 1.0 to 1.3 mm in diameter
(Fig. 1a). While t he fert ilized eggs w ere
t ransparent , demarsal, spherical, nonadhesive and brow nish colour (Fig. 1b).
Immediately af ter fert ilizat ion the diamet er of
t he egg increased ow ing to slight sw elling of
t he egg w hich ranged bet w een 1.3 t o 1.6
mm. Fert ilized eggs had a reddish spot
(blastodisc) and readily recognizable w it h the
naked eye (Fig. 1c).
Formation of embryo
The f irst cleavage t hat divided t he
blastodisc into tw o blast omeres w as occurred
w ithin 25 t o 30 min of post -fertilization (Fig.
1d). The cleavage w as typically meroblastic.
While t he second and t hird cleavage (4 and 8
cell st age) w as appeared f or 60 min and 90
min af ter fertilization respect ively (Fig. 1e and
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Mollah et al.

Table 1. Brief descriptions of the embryonic developmental stages of R. rita.

1f ), the 16 and 32 cell-stages w ere observed

bet w een 13 0 min and 16 0 min post fert ilizat ion, respectively (Fig. 1g and 1h).
Gradually t he successive cleavages w ere
found, but the blast omeres w ere decreased in
size. The morula stage w as completed w it hin
t he durat ion of 2.7 5 t o 3 .30 h af t er
fert ilizat ion (Fig. 1i and 1j). During 4.0 t o 4.3
h of post -f ert ilizat ion, t he t op of t he
blast oderm st art ed spreading over the yolk
sphere f ormed a thin layer and the embryo
reached t he blastula stage (Fig. 1k and 1l).
Aft er 6.0 h of fert ilizat ion, the blast oderm
flat tened dow n ont o t he yolk-sphere. The
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out er surf ace f ollow ed the curvature of t he

yolk sphere resulted a dome shape st ruct ure
(Fig. 1m). The blast oderm began to expand
(30% epiboly, about 1/4 of the yolk sphere)
over the upper surface of t he yolk sphere
during 7.30 h of post fertilizat ion (Fig.1n).
Within 9.30 h of post-fertilizat ion, more t han
half a portion of t he yolk (50% epiboly,
Fig.1o) w as invaded by t he embryonic shield
reached t he lat e gastrula stage (Fig.1p). Then
af t er 1 1.3 0 h of f ert ilizat ion, t he head
(rudimentary brain) w as made anteriorly and
the rudimentary t ail w as created post eriorly in
the distinct embryonic body (Fig.1q). During
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13 .3 0 h post -f ert ilizat ion, some of t he
mesodermal cells divided t he body of the
embryo in a number of somit es and also
visibly show ed optic cups. Antero-posterior
end w as dist inguished, the cephalic portion
became broader and ant erior end formed t he
head fold, and posterior end f ormed t he t ail
fold (Fig.1r). Aft er t hat , 18-20 h old embryo
occupied more t han t hree f ourths of the egg
peripherical space and mesodermal somit es
increased in number and became more dist inct
and the pigment ation w as developed in t he
somites area. The tubular heart w as appeared
underneath t he head and the heart act ively
st arted t he blood circulat ion (Fig.1s). Bef ore
hat ching (Fig.1t), t he embryo start ed t w isting
movement inside the egg and cont inuously
beat the egg shell by the caudal region
especially around t he middle part of t he body.
This movement gradually became vigorous
and the egg capsules w ere w eakened. By
rupt uring the egg capsule, the middle part of
the embryo gradually disconnect ed f rom t he
egg capsule. The egg membrane w as broken
dow n from t he caudal region and t he larva
first ly emerged w ith its t ail port ion. At least 12 h w as required f or complet e emerging of t he
hat chling because t he entire embryo did not
hat ch out at a time.
Larval development
A larva (Latin; plural larvae) is a young
form of animal w ith indirect development
going t hrough or undergoing metamorphosis
(f or example, insect s, amphibians or
cnidarians). The larva looks complet ely
diff erent from t he adult form (Table 2).
Hatchling
New ly hat ched larvae w ere st raight ,
slender, t ransparent , lat erally compressed
body and gradually tapering tow ards t he tail.
The larvae w ere brow nish-colour and 2.0 mm
in length. The hat chlings had unpigment ed
eyes and devoid of distinct mout h and f ins.
The head w as very small and t he yolk sac w as
initially oval-shape w it h pale-brow nish colour,
w hile numerous small chromatophores at t he
dorsal region gave it a dark appearance. The
larvae could not sw im and floated passively
on t he w ater and irregularly appeared upside
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Mollah et al.

dow n movement . The numbers of body

somites ranged betw een 40 and 46. The
w hole hat ching experiment al w orks w ere
done at t emperature of 28 1 C (Fig. 2a).
8 h old larva
Average length of the larvae w as found
about 2.1 mm and brow nish in colour. Mouth
w as not yet developed but only a conspicuous
depression identified the position of mout h. A
depression at the post erior end of the yolk sac
w as visible w hich w as ident if ied as anal pore.
Heart became more distinct . Circulat ion of
body f luid w as continued and barbels had not
yet developed (Fig. 2b).
16 h old larva
Sixteen hours aft er hat ching, the larvae
w ere found of 2.2 mm in lengt h. At this
st age, yolk sac w as partially reduced and
gradually became elongated. Heart and brain
w ere clearly distinct. A tube like structure
w it hin t he body w as represent ed t he
aliment ary canal (Fig. 2c).
24 h old larva
One day old larva w it h reduced yolk sac
w as approximately 2.5 mm length. Eye spot
w ith a dark pigmented area w as appeared on
t he anterior part of t he head. Barbels w ere
f ound in t he f orms of t iny knobs. Pect oral fin
buds w ere seen. Mout h w as not opened and
t he anal opening w as st ill closed.
Pigment ation w as gradually extended all over
t he body and t he blood circulat ion syst em
w as fully developed (Fig. 2d).
36 h old larva
The average length of 36 h old larva w as
2.8 mm in length. Dark pigmented eyes w ith
spherical shape w ere visible. Distinct heart
w as visible and f unct ioned act ively. Reddish
blood w as seen around t he heart region. Fully
f ormed upper and low er jaw s w ere visible and
gills w ere covered by the operculum. Mouth
w as formed as an opening and t he anal pore
w as also opened and caudal f in bud w as
f ormed. Vigorous movement s of t he larvae
w ere seen (Fig. 2e).
48 h old larva
Tw o days old larva w as 3.0 mm in lengt h
and part ial yolk sac w as present ed. Dorsal
region became dark due to t he format ion of
numerous
chromat ophores
and
t he
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Mollah et al.

Fig. 1. Embryonic developmental stages of R. rita. (a) unfertilized egg, (b) fertilized egg, (c) blastodisc and
(d) 2 cell stage, (e) 4 cell stage, (f) 8 cell stage, (g) 16 cell stage (h) 32 cell stage (i) early morula
stage and (j) late morula stage, (k) early blastula stage, (l) late blastula stage, (m) pre- early gastrula
stage (n) 30% epiboly (o) mid gastrula stage and (p) late gastrula stage, (q) head and tail bud
formation, (r) somite stage, (s) 18-20 h old embryo and (t) just before hatching.
(EM- Egg membrane, VM- Vitelline membrane, PS- Perivitelline space, Y- Yolk, BD- Blastoderm, BM- Blastomere, APAnimal pole, EVL- Enveloping layer, VP- Vegetal pole, YSL- Yolk syncytial layer, DBL- Dorsal blastoporal lip, VBL- Ventral
blastoporal lip, H- Head portion, T- Tail portion, YS- Yolk sac)

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Table 2. Summary of the larval developmental stages of R. rita.

t ransparent vent ral region w as visible.

Pect oral fin f olds became dist inct and t he
rudiment ary rays developed in the caudal f in.
Barbels became elongat ed and mouth w as
w ell formed. The alimentary tract became
short, st raight and distinct and a small pouch
like st omach w as f ormed. Blood circulation
w as w ell observed in t he head, heart and t ail
region. At the end of t his st age, yolk sac w as
absorbed to half size and the larvae w ere
f ound smoot hly sw imming and f eeding
exogenously (Fig. 2f).
DISCUSSION
R. rit a is a crit ically endangered f ish
species and t heir cult ure t echnique in
controlled environment is not pract iced. To
prot ect t his species and develop their cultural
t echnique, it is essent ial t o assemble
inf ormat ion about embryonic and larval
development of this crit ically endangered f ish.
But there are no report s about the embryonic
and larval development of t his species.
Therefore, t he aim of t he present st udy
potent ially invest igat ed and provided det ailed
information about t he embryonic and larval
development of R. rit a species.
The eggs of R. rit a reached rapidly in their
embryonic and larval stages. The fertilized
eggs of
Rit a are non-adhesive but
adhesiveness of eggs is the special character
found in other catf ishes such as Claries
gariepinus (Khan and Mollah 1998, Osman et
al. 20 08 ), C. bat rachus (Brut on 1 97 9),
115

Mystus montanus (Arockiaraj et al. 2003) and

Pangasius sut chi (Islam 2005). The diamet er
of the f ertilized eggs ranged betw een 1.0 and
1.3 mm. In similar study, some researchers
show ed dif ferent egg size of C. gariepinus
(Osman et al. 2008). These diff erences might
be att ribut ed to the species variation and
brood size (Puvanesw ari et al. 2009). First
cleavage of eggs of Rit a w as observed w it hin
25-30 min post -f ert ilization at t he w at er
t emperat ure of 27-28 C, but a discrepancy of
t ime w as observed in case of C. gariepinus,
Mystus cavasius w hich w as w it hin 40-50 min
post -fert ilizat ion report ed f rom Khan and
Mollah (1998) and Rahman et al. (2004) at
28.5 and 26 C respect ively. This variation
might be due to species diff erence and other
environmental fact ors. Just 1-2 h before
hatching, t he embryo of R. rit a show ed
t w ist ing movement s inside t he egg capsule
w hich w as also f ound in ot her fishes reported
by Puvanesw ari et al. (2009), Khan and
Mollah (1998) and Osman et al. (2008). In the
present study, hatching began aft er 22 h
post-fert ilizat ion and completed w ithin 24 h at
27-29 C w hich is similar t o other f indings
reported f rom Puvanesw ari et al. (2009).
Another report from Mollah and Tan (1982)
show ed t hat in Clarias macrocephalus, incubation period w as 22 h at 30 C and 34 h at
25 C but no hat ching w as observed at 20 C.
Time requirement of egg hat ching w as inversely related to their incubation temperature
(Rana 1990). Lengt h of the new ly hat ched
larvae of this species w as around 2.0 mm
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Mollah et al.

Fig. 2. Larval developmental stages of R. rita. (a) new ly hatched larva, (b) 8 h old larva, (c) 16 h old larva
and (d) 24 h old larva, (e) 36 h old larva and (f) 48 h old larva.
(M- Myomere, PFB- Pectoral fin bud, E- Eye, YS- Yolk sac, UJ- Upper jaw , LJ- Low er jaw , CFB- Caudal fin bud, B- Barbel,
MO- Mouth opening, V- Vertebra)

w hich is in t he ranges of the findings of Khan

and Mollah (1998). In the present st udy,
larvae w ere st arted t o feed at 48 h post hat ching w hich is similar to other previously
report ed st udies st at ed by Puvanesw ari et al.
(2009).
R. rit a w as selected f or t his study because
it is already declared as critically endangered
fish, so an att empt should be t aken to protect
this species from being extinct . Unfortunately
there is no remarkable inf ormation on t he

embryonic and larval development of t his f ish,

but t his unique research w ork w ill act as base
line for f urther study.
ACKNOWLEDGEMENTS
The aut hor t akes t he privilege t o
acknow ledge BARC (Bangladesh Agricultural
Research Council) authorit ies f or providing
financial assistance t o conduct t his research
w ork.

REFERENCES
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of Rita rita (Hamilton). Journal of Bangladesh Agricultural University 6(1): 159-163.

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Anonymous (2000) International Union for Conservation of Nature, Bangladesh. Bangladesher

Bipanno Prani, IUCN Bangladesh, The World Conservation Union, Dakka.
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Dakka.
Arockiaraj AJ, Haniffa MA, Seetharaman S, Singh SP (2003) Early development of a threatened
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Devi NT, Khumar F, Siddiqui MS (1991) Observations on the morphometric characters of the
catfish Rita rita (Ham.) of the river Yamuna. Journal of Inland Fisheries Society of India 23(1):
52-58.
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relation to its age and regional variations. Indian Journal of Zoology 9(2): 61-75.
Mollah MFA, Amin MR, Sarow ar MN, Muhammadullah (2008) Induceed breeding of the riverine
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Kritik Olarak Tehlike Altinda Olan Dere Kedi Baligi Rita rita' nin Embriyonik ve Larval Gelisimi
zet
Bu alisma, tatli su kedi baligi Rita rita' nin embriyonik ve larval gelisimini incelemek amaciyla
gereklestirildi. Olgun yumurta ve spermler yapay dllenme teknigi kullanilarak toplandi ve
dllenmis yumurtalar srekli su kaynagina sahip mini dairesel kulukahanede inkbe edildi.
Dllenmis yumurtalar; seffaf, demarsal, dairesel, yapiskan olmayan, renkleri kahverengimsi idi ve
aplari 1.3-1.6 mm arasinda degismekteydi. Ilk blnme, dllenmeyi takibeden 25-30 dak.
ierisinde 28 1 C' de gereklesti. Yumurtadan ikma islemi, ayni sicakliklarda, dllenmeden 22 s
sonra basladi ve 24 s ierisinde tamamlandi. Yumurtadan yeni ikan larvalar, 2.0 mm
uzunlugunda, agizdan ve pigmentten yoksundu ve beslenme, yumurtadan ikistan 48-60 s sonra
basladi. Tatli su kedi baligi R. rita' nin erken dnem embriyonik ve larval gelisimi ilk kez
tanimlanmaktadir. Bu yzden bu alismanin bulgulari, byk lekli Rita yavru balik retimi
gereklestirilmesi iin kiymetli bilgiler saglamaktadir.
Anahtar Kelimeler: Embriyo gelisimi, larva, metamorfoz, Rita rita.
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