J. Env. Bio-Sci., 2016: Vol.

30 (1):33-37
(33) ISSN 0973-6913 (Print), ISSN 0976-3384 (On Line)

OPTIMIZING BREEDING, EMBRYONIC DEVELOPMENT AND REARING OF KOI CARP,

CYPRINUS CARPIO, UNDER COLDWATER CONDITIONS AT BHIMTAL, UTTARAKHAND
FOR MASS SCALE SEED PRODUCTION
Rabindar S. Patiyal*, Javaid I. Mir, Neha Saxena and Vijay K. Singh
Directorate of Coldwater Fisheries Research, Bhimtal 263136;
[Corresponding Author E-mail*: rspatiyal01@gmail.com]

Received: 04-01-2016 Accepted: 18-05-2016

The induced breeding of Koi Carp (Cyprinus carpio carpio) was successfully carried out at the Directorate of Coldwater Fisheries
Research, Bhimtal, India. Induced breeding and rearing conducted on the fish revealed that the fish can be breed at low temperature
using sGnRH analogue and dopamine antagonist (Ovaprim). Spawning was observed eighteen hrs after the injection, fertilization
and hatching and fry survival rate were observed in fishes spawned in April (70%, 75% and 42%, respectively) than fishes spawned
in August (80-85%, 90-95% and 80% respectively).The incubation period was 120 hours and 84 hours at temperature 15-18o C(April)
and 20-26oC (August), respectively. Present study revealed that favourable conditions on hill region for mass seed production of koi
carp in the month of April.

Ornamental fish is a generic term used to describe aquatic Information on the early life history of a fish is very important
animals which are kept in the hobby aquarium1. These fishes for optimization of its large scale seed production, culture and
act as important commercial component of aquaculture management practices in hill region therefore, present
providing for aesthetic requirements and upkeep of the experiment was carried out to study some aspects of the early
environment2. The USA is the largest importer of ornamental life history, the development biological clock of koi carp under
fishes followed by Europe and Japan. The emerging markets the coldwater condition of Uttarakhand at Bhimtal.
of ornamental fisheries are China and South Africa3. In global
MATERIAL AND METHODS
ornamental fish trade India's share is estimated to be less
than 1% with major share from wild collection.The overall The fishes were purchased from Lucknow local market during
domestic trade in this field cross Rs 1000 lakh and is 2012.Same day, the fishes were transported to the Fish Farm,
reportedly growing at the rate of 20 per cent annum3. Directorate of Coldwater Fisheries Research (DCFR), Bhimtal.
At the farm after disinfection, all fishes were reared in a
Common carp (Cyprinus carpio) is one of the most important
cemented pond. The fishes were fed with floating pellets
cultured fish in the world belonging to family Cyprinidae. More
containing crude protein 28%, crude fiber 11.1%, and
than 2.7 million tonnes of common carp were produced in
carbohydrate 33% (Table-1). After proper acclimatization and
20004. Koi carp is an ornamental variety of domesticated
maintenance, the healthy and mature breeders (90-550g) were
common carp (Cyprinus carpio) that are kept for the decorative
selected according to sexual dimorphism and transferred to
purpose in the aquarium as well as outdoor ponds or water
hatchery shed in FRP tank of size 200cm X 200cm X 30cm
gardens. It is one of the most popular and favorite ornamental
with flow through arrangement of water system.The females
fishes amongst all ornamental fish species and it has high
are usually easier to identify, as the belly of a mature female
market value for its excellent color. The color and scale pattern
is generally large and bulging, whereas male's remains
of the species is highly variable like white, black, red, yellow,
streamlined and more torpedo shaped8. The sex ratio of the
blue and cream. Like all cyprinids, koi carp is also an egg
spawners was kept at 2:1 for male and female. The breeding
layer. They produce adhesive eggs. This species exhibits
programme was carried out using salmon Gonadotropin
gonochorism, external fertilization with varied spawning
releasing hormone analogue and domperidone injection
frequencies5 and considered as batch spawner 6.They grow
(ovaprim, Syndel laboratories INDIA Pvt.ltd). Brooders were
up to a size of 100 cm with an elongate body. In their natural
administered hormone intra peritoneal in evening hours (0.6
habitat,koicarp can live up to 15-24 years7.
NAAS Rating (2016)-4.20
 OPTIMIZING BREEDING, EMBRYONIC DEVELOPMENT AND REARING OF KOI CARP (34)

ml/kg and 0.3 ml/kg body weight to female and male, reasons that caused the increase in nutrients in water body,
respectively) (Table-2). Injected brooders were released into leading to increase in abundance of plankton16. Therefore more
FRP tank of 3000 L capacity having provision for flow through food availability for fry will be during August month.
water system. Aquatic macrophyte (Hydrilla) was introduced
In present study, the embryonic development was divided into
into breeding tank for holding adhesive eggs9. The embryonic
zygote, cleavage, blastula, gastrula and hatching period (Fig.-
development was studied using light microscope with a digital
1) and each stage is characterized by typical feature. The
camera (Nikon ECLIPSE E100). The egg hatching and larval
comparative account of embryonic development of April and
rearing upto yolk sac absorption was carried out in the same
August were given in Table-4. Result of present study showed
tank. After yolk sac absorption, 1000 larvae of 2.5-3.5 mm
fast embryonic development in August month. The cleavage
average length were reared in three differentculture systems
was meroblastic and the first division (2 celled stage) occurred
i.e. polytank, cemented tank and earthen tank for 6 months.
45 minutes after fertilization, followed by second cleavage 1
The larvae stocked under different culture systems were fed
hour 20 minutes after fertilization. The 16 celled stage was
with egg yolk and live plankton for first 15 days followed by
reached 2 hours 10 minutes after fertilization.Further cleavage
gradually replaced with supplementary diet of 30% protein
increased cell number and reached at stage morula. At this
level, comprising ground nut oil cake, fish meal and rice bran
stage, a cap like structure was seen over the animal pole,
in equal proportion. The water quality was monitored
which gradually increases in size.The blastoderm further spread
throughout the year using HANNA HI 9828. All the statistical
over the yolk and the formation of germinal ring around yolk
analysis was done in SPSS 16.0 software. One way ANOVA
was clearly visible within 8 hours after fertilization. The yolk
and Duncan's multiple test (at 5%) were applied to evaluate
invasion completed after 12 hours and 45 minutes after
the growth of larvae under different culture systems.
fertilization. Further during yolk plug stage the head and tail
RESULTS AND DISCUSSION ends of the embryo became distinguishable and the blastopore
was almost closed. The notochord was clearly visible at 15
The water quality parameters are given in Table-2. In the present
hours and 15 minutes after fertilization. Further the embryo
study, spawning was noticed after 12-20 hours of hormone
was elongated and encircled the whole yolk material within 27
injection. The fertilized eggs were 0.8-1.1 mm in diameter,
hours 15 minutes after fertilization. At this stage, the anterior
yellowish in colour, adhesive and demersal in nature.
posterior axis was distinguishable in the broader cephalic region
Comparatively, lower fertilization rate and hatching rate were
with distinct forebrain and narrow end as tail region.After 38
observed in fishes spawned in April (70% and 75%,
hours and 15 minutes of fertilization cephalic region became
respectively) than fishes spawned in August (80-85% and 90-
prominent, optic lens starts differentiating and mesodermal
95% respectively) (Table-3). Similar more incubation period
somites (16-18) were highly visible. A heart beat (80-91) per
and less fry survival was reported in April month (120 hours
minutes were observed at this stage. The caudal region started
and 42%, respectively) than August month (84-90 hours and
detaching from yolk and head further elongated in size showing
80%, respectively) (Table-4).This may be attributed to
all parts of brain, heart, lens and 22-25 somites after 56 hours
difference in water temperature among both the months as
and 15 minutes after fertilization. The beating of heart intensified
temperature of water is higher in August (22 ±1.6oC)than April
130-140 beats per minutes and tail showed rhythmic movement
(16 ±1.3oC). Hence, the results of present study showed that
on both side one by one. At 72 hours and 10 minutesafter
water temperature play an important role on development and
fertilization lens fully formed and pectoral fin bud was clearly
hatching success of koi carp. Similarly, Yang and Chen10 stated
visible. In final stage of embryonic development, the growing
that temperature is as one of the most decisive environmental
embryo occupied the entire previtelline space. The lashing
variables affecting embryonic development in fish eggs.
movements, which gradually become vigorous and egg
Generally, lower temperature retards the rate of egg
capsules, were weakened and ruptured. The embryo ruptured
development and higher temperature accelerates it11-15. Other
the egg shell by the continuous movement and hatched out at
than water temperature, rainfall may be an important factor
84hours after fertilization at 22 ± 1.6oC.The hatchlings were
for higher fry survival in August month. As rainfall is one of the
(35) PATIYAL, MIR, SAXENA AND SINGH

Table-1. Feed composition of diet fed to Koi Carp, Table-2. Physico-chemical characteristics of Koi carp,
Cyprinus carpio, Brood fish. Cyprinus carpio, breeding tank.
In g red ien ts P ercen tag e Param eter April August
H um idity 10% o
Tem perature ( C) 16.2 ±1.3 22.1 ±1.6
C rude protein 28% pH 7.9±0.40 7.7 ±0.39
C rude fibre 11% Dissolved oxygen (m g L -1) 10.9±0.42 8.6 ±0.49
A cidic insoluble m aterial 6% Alkalinity (m g L -1) 112±13.43 115±8.04
-1
C arbohydrate 33% Am m onia (m g L ) 0.026±0.002 0.027±0.004
Nitrite-Nitrogen (m g L -1) 0.027±0.005 0.061±0.005
N itrogen free extract 12%
Nitrate-Nitrogen (m g L -1) 0.133±0.01 0.145±0.01
V itam in A 5.0 I.U
V itam in B 2 0.025 I.U
V itam in D 3 1.00 I.U culture system, larvae reared in polytank showed comparatively
V itam in E 0.750 I.U better total length gain (78.56mm) than earthern (63.71 mm)
V itam in K 0.01gm
and cemented condition (37.41). Therefore, it can concluded
C alcium pantothenate 0.02gm
that the poly tanks are more suitable ponds for rearing koi
N icotinam ide 0.150gm
V itam in B-12 0.100gm carp fry under controlled conditions. Similar results were
C oline chloride 0.158gm obtained by Kumar et al. 21 in common carp in mid hill region.
C alcium 8.00gm In conclusion,our study indicate that koi carp can be easily
M anganese 0.275gm
matured and breed successfully under coldwater conditions
Iodine 0.010gm
in captivity similar to carps. Koi carp showed better breeding
Iron 0.075gm
Z inc 0.150gm performance in August month and the poly tanks are more
C opper 0.020gm suitable ponds for raising koi carp fry under controlled
C obalt 0.004gm conditions.The descriptive investigation into the embryonic
development and temperature tolerance should provide valuable
information about the ability of the species to handle low
transparent, oval head, short tail and measured 3.45-4.75 mm temperature condition. As there are no commercial approaches
in total length . The yolk got fully absorbed within 2-3 days of induced breeding and seed production of koi carp in the hill
and by this time mouth formation was complete and the larvae region of Kumaon Uttrakhand but there is high demand of this
started exogenous feeding. ornamental fish locally for its colorful and attractive appearance.
Hence, In spite of the long incubation period, the captive
Observation on overall embryonic and larval development
breeding, embryonic development and rearing protocol
stages of koi carp in present study are in aggrement with
described herein should provide a base for future studies on
pattern described for koi carp by other authors9,17-18. However,
koi carp and help in achieving conservation and commercial
variation in timing of ontogenic events, structure formation and
goals.
incubation period exist among species In present study,
embryo hatched out in 120 hrs after fertilization at 16 ± 1.3oC ACKNOWLEDGEMENTS
and 84-90 hours after fertilization at 22 ± 1.6 oC which was
The Department of Biotechnology, New Delhi is acknowledged
similar to the earlier findings 17-18. However, the results of
present study vary from Hanniffa et al. 19, who concluded that for funding this study. Thanks are due to Director, Directorate
72-73 hours are needed for hatching of Koi carp at 26-28°C. of Coldwater Fisheries Research, Bhimtal, Uttarakhand for
This can be attributed to different physical condition of brood providing necessary facilities for the study.
fish and lower temperature of water at the time of breeding20. REFERENCES
The comparative account of total length of Koi carp under 1. Livengood EJ, Chapman FA. (2009). In: The ornamental fish
different culture methodsis given in Table-5. Among all three trade: An introduction with perspective for responsible
 OPTIMIZING BREEDING, EMBRYONIC DEVELOPMENT AND REARING OF KOI CARP (36)

Table-3. Detail of Induced Breeding trials of Koi carp, Cyprinus carpio.

Dose:
Name Latency Incubation Average Average Fry
Breeding Sex ovaprim Temp .
of period o period Fertilization hatching survival
trial (male/female) (ml/kg body ( C)
species (Hours) (Hours) Rate % Rate % %
weight)
1 (April Male 0.3
18-20 15-19 120 70 75 42
2013 Female 0.6
Koi Carp
2 (August Male 0.3
12 23-26 84 80 95 80
2013) Female 0.6
3 (August, Male 0.3
14-15 20-25 90 85 90 80
2015) Female 0.6

Table-4 . Comparative account of embryonic development of Koi carp, Cyprinus carpio, under different temperature.
Hours after fertilization
S tages /E vents April August
Tem perature (16 ± 2 oC) Tem perature (22 ± 1.6 o
C)
Fertilization 0 0
2-celled stage 1.00 45
4-celled stage 1:35 1:20
8-celled stage 2:00 1:50
16-celled stage 2:30 2:10
G erminal ring formed 15:03 8:00
Half, Yolk invasion completed 32:13 12:45
Notochord clearly seen 46:16 15:15
Cephalic region broadened with distinct forebrain 48:00 27:15
14 somites, cephalic region broaded 54.00 32:30
16-18 somites, optic lens starts diff erentiating 76:23 38:15
22-25 somites, lens formed in eye 101.00 56:15
Lens fully formed, pectoral fin bud seen 109.00 72:10
Hatching started 112.00 76:12
Hatching completed 120.00 84:00

Table-5 . Comparative account of total length of Koi carp, Cyprinus carpio, under different culture methods
T o t a l le n g t h la r v a e ( m m )
Age
P o ly t a n k E a r th e n ta n k C e m e n te d ta n k
3 .0 2 ± 0 .2 2 a 3 .0 2 ± 0 .2 2 a 3 .0 2 ± 0 .2 2 a
0 d a y o ld
( 2 .4 0 - 3 .7 5 ) ( 2 .4 0 - 3 .7 5 ) ( 2 .4 0 - 3 .7 5 )
a b c
6 .7 7 ± 0 .2 1 4 .7 5 ± 0 .0 8 3 .4 6 ± 0 .4 2
1 5 d a y s o ld
( 6 .0 0 - 7 .5 0 ) ( 4 .5 0 - 5 .0 0 ) ( 2 .4 6 - 5 .3 0 )
1 0 .4 ± 0 .7 0 a 5 .0 8 ± 0 .2 6 b 4 .5 5 ± 0 .1 7 b
1 m o n t h o ld
( 8 .0 0 - 1 2 .4 0 ) ( 4 .6 0 - 5 .9 0 ) ( 4 .1 - 5 .1 0 )
a b c
3 8 .8 6 ± 0 .3 4 2 7 .3 6 ± 1 .7 8 1 8 .2 7 ± 0 .8 9
3 m o n t h s o ld
( 3 8 .0 0 - 4 0 .1 0 ) ( 2 0 .2 0 - 3 1 .2 0 ) ( 1 5 .2 0 - 2 0 .5 0 )
a b c
8 1 .5 8 ± 1 .1 5 6 6 .7 3 ± 1 .1 9 4 0 .4 3 ± 1 .0 5
6 m o n t h s o ld
( 7 8 .0 0 - 8 5 .1 0 ) ( 6 3 .5 0 - 7 0 .1 0 ) ( 3 8 .1 0 - 4 5 .0 0 )
TLG 7 8 .5 6 6 3 .7 1 3 7 .4 1

Data presented as Mean ± standard error. Figures with same superscript letters in rows are not significantly different
from each other at p>0.05

aquariumcooperative extension service, institute of food and
agricultural science, university of Florida, Gainesville.
2. Swain SK, Singh SK, Routray P, Barik NK. (2008). e- planet
6(2): 2, 20.
3. Anonymous. (2006). In: Hand Book of Fisheries and Aquaculture.
Indian Council of Agricultural Research. New Delhi.
4. Food and Agricultural Organization (FAO) (2000). In: World status
of ornamental fish, 3: 5.
5. Balon, E.K. (1990). Guelph Ichthyology Reviews, 1:1.
6. Kailola, P.J., Williams, M.J., Stewart, P.C., Reichelt, R.E., McNee,
A. and Grieve, C. (1993). In: Australian fisheries resources.
Bureau of Resource Sciences, Canberra, Australia.
(37) PATIYAL, MIR, SAXENA AND SINGH

Figure-1. Different Embryonic development stages of Koi carp, Cyprinus carpio. A, Fertillized egg; B, Cleavage
stage; C -F, Blastula stage; G- N, gastrula stage; O, newly hatched larves. YK Yolk Sac , Bl blastoderm
, GD germinal disc,AR anterior Region , NC notochord, CR cephalic region, TR Tail region , OLF
optic lens formation, SM Somites
7. Kuroki, T. (1981). In: The latest manual to nishikigoi.Shin-Nippon 15. Das T, Pal A K, Chakraborty S K, Manush S M, Dalvi R S, Sarma
Kyoiku-Tosho Co. Ltd. Japan. K, Mukherjee S G (2006). Aquaculture 255: 536.
8. Mihalache A, Oprea L, Grecu I, Cristea V,Badalan C, Enache I, 16. Negi RK, Rajput V (2013). International Journal of Advanced
Ion S (2011). Journal of Environmental Protection and Ecology Research. 1(5): 171.
12(4): 1835. 17. Watson, C.A., Hill, J.E.,Pouder, D.B. (2004). In: Species Profile:
9. Haniffa MA, Benziger PSA, Arockiaraj AJ, Nagarajan M, Siby P. Koi and Goldfish; SRAC Publication No. 7201.
(2006). Journal of Bio-Science 14:121. 18. Ghosh A K, Biswas S, SarderLSabbir W, Rahaman S M B (2012).
10. Yang Z, Chen Y (2005). Aquaculture 246 : 173. Mesopot. J. Mar. Sci., 27 (1): 1.
11. Marangos, C., Yagi, H., Ceccaldi, H.J., (1986). Aquaculture 54:287. 19. Haniffa M A, Allen Benziger P S, Jesu Arockiaraj A, Nagarajan M,
12. Pepin, P., (1991). Canadian Journal of Fisheries and Aquatic Siby P (2007). Taiwania 52(1): 93.
Sciences 48: 503. 20. Valentin FN, do Nascimento NF, da Silva RC, Fernandes JB,
13. Mihelakakis, A., Kitajima, C., (1994). Aquaculture 126:361. Giannecchini LG, Nakaghi LS. (2013). Zygote. 2013 Nov 12:1.
14. Hart, P.R., Purser, G.J., (1995). Aquaculture 136:221.