First report on captive breeding of threatened snakehead Channa gachua

(Hamilton, 1822) for species conservation

M.JAMES MILTON1 M.A.HANIFFA2,
College of Fishery, Key Lab of Agricultural Animal Genetics, Breeding and
Reproduction of Ministry of Education, Huazhong Agricultural University, Wuhan,
Hubei, 430070, PR China.
Centre for Aquaculture Research and Extension (CARE),St.Xaviers Collge
(Autonomous),Palayamkottai- 627002, Tamilnadu, India.

Abstract
Channa gachua (HamiltonBuchanan) has witnessed a drastic decline due
anthropogenic stress and is listed under vulnerable (VU) category. Captive breeding
was undertaken in C. gachua collected from wild by injecting three different
intramuscular doses (1000IU, 2000IU and 3000IU/kg body weight) of Human
Chorionic Gonadotropin (HCG). Distinct spawning behaviour was noticed in the
brooders injected with hormone. Total number of eggs spawned were estimated as
2740 and the fertilized eggs were spherical in shape (1.16mm 0.05 mm), floating
and pale yellow in colour. Fertilization rate varied from 62% - 70%. The eggs hatched
out between 24 36h after fertilization and about 1,420 hatchlings were produced
from one set. Newly hatched larvae measured 4.23mm 0.03 mm and 0.037g 0.008
g in weight and the mean diameter of yolk sac was 1.1 0.08 mm. Yolk sac was
absorbed after three days and the percentage survival of hatchlings varied from 40% to 56 %. Preflexion larvae showed a large head with a conspicuous occipital
melanophore and terminal mouth. The head, oil globule and the yolksac together
appeared as a bulb like structure. Melanophores were scattered on the yolk and a few
were present on the unpaired fin. The length of the young one at flexion stage was
6.2 7.1 mm from day 8 14 (n=29). Caudal fin began to separate;

faint

pigmentation of eyes was noticed, alimentary tract was distinct and pectoral fin bud
appeared. During Post flexion stage the length of the larvae was 7.1 14.0 mm from
day 14 30. The development of pectoral and pelvlilc fins was completed on day 26
(11.3 mm), whereas caudal fin completed its development on day 16 (7.2mm). The

number of rays pectoral , caudal and pelvic fins were 14, 13 15 and 6 7
respectively.
Introduction
Snakeheads (Perciformes: Channoidei: Channidae) are obligatory airbreathing freshwater fishes possessing accessory respiratory apparatus and are
distributed in African and Asian continents (Li et al., 2006). Snakeheads are
important food fishes in many countries within their distributional
range, and some colorful species are even common in the aquarium
trade (Ng and Lim, 1990; Musikasinthorn, 2003; Courtenay and
Williams, 2004). Snakeheads commonly called murrels consist two genera,
Channa and Parachanna. The Asian genus Channa, presently contains 26 valid
species and is widely distributed in Iran and southern Asia (Musikasinthorn, 2003;
Berra, 2001; Musikasinthorn and Taki, 2001; Zhang et al., 2002; Courtenay and
Williams, 2004); about 8 10 species have been reported in India (Haniffa, 2010).
Till date no standard reports including IUCN are not available regarding the
threatened status of snakeheads however the Workshop on Conservation Assessment
Management Plan (CAMP) evaluated the status of Channa species of India, in 1997
and categorized them as " Low risk near threatened, Low risk-least concern,
Vulnerable, and Critically endangered" due to restricted distribution, loss of habitat,
over exploitation, destructive fishing practices and trade (Anon., 1998).
Channa gachua is commonly called dwarf snakehead and even though widely
distributed in Asia, declined drastically (vulnerable) in India (CAMP, 1998; Anjan
Kumar Prusty et al., 2007) and endangered in Asia (Lim and Ng, 1990). C. gachua is
threatened due to habitat loss by introduction of alien species, high fishing pressure
and is prone to epizootic ulcerative syndrome (EUS). It inhabits rivers, lakes, ponds
and streams (Lee & Ng, 1994). C. gachua is important as food fish and is well known
in the aquarium trade as welldue to its beautiful colouration (Talwar and Jhingran
1992). Life history of threatened Channa species is still unknown and the knowledge
on Channa gachua is almost fragmentary. Captive breeding and the release of captive
bred individuals into the wild are one among the techniques used for the conservation
of rare and endangered fish species. In the present study C.gachua was successfully

bred for the first time under captivity and captive bred populations are being
maintained at CARE Aquafarm.
Materials and methods
Channa gachua brooders (10.6 cm - 17.1 cm ; 35g 91g) were collected from
the river Tamirabarani (8.44N, 77.44 E) during April 2008February 2009 and were
reared in CARE stocking ponds. The brood fishes were fed with small live tadpoles
and semimoist feed (liver, rice bran, soy flour and fish oil) (2:1) at 4% of body weight
upto 4 months. After 4 months of rearing, they were found mature for captive
breeding. The female brooder showed bulged abdomen than male and in the case of
female the vent was reddish colour and fully mature female showed freely oozing
eggs when pressed gently (Fig .1). Eight sets each with 2 males (average weight 54g)
and one female (average weight of 65g) were selected from the available brood stock
for breeding experiments and a control set was maintained for each experiment. Three
different doses (1000IU, 2000IU and 3000IU) of HCG were injected intramuscularly
to both male and female. After HCG injection, each set was released into separate
breeding tanks. Hydrilla verticilata was introduced into the breeding tanks for hiding
purposes .
Diameter of eggs was measured using Magnus Pro Software with accuracy
level of 0.01 mm. Hatching rate was estimated on fourth day after spawning. After 14
days hatchlings were transferred to glass tanks (50l capacity) for further rearing. Paste
of boiled egg yolk and plankton (Rotifers and Daphnia) were supplied as
supplementary feed. Survival of hatchlings was calculated randomly by taking
samples from glass tank upto 15 days at 7 days interval. Spent fishes were removed
from the breeding pool, washed in KMnO4 solution and released back into stocking
pond. Data were analysed using statistical software package SPSS version 11.5. A
probability level of 0.05 was applied to account for the statistical difference between
the means.
Results
Chasing behaviour was observed for male after 14 18 h of injection and
female released eggs followed by male releasing milt. The fertilized eggs were clearly
distinguished from unfertilized ones; the former were larger, spherical and pale yellow
in colour.Three different doses of HCG administered in Channa gachua highlighted

that spawning occurred in the fishes injected with the doses of 2000IU and 3000IU/kg
body weight after 2024 h latency period as compared to 1000IU /kg where it took
2529 h. The former doses resulted in complete spawning whereas partial spawning
was noticed in the later. No spawning activity was observed in the control set. The
spawning pairs were seen moving together in search of suitable breeding place until
spawning. The total numbers of eggs spawned ranged from 1,160 - 2,740 / pair (Table
1).
Fertilization ranged from 55% - 70% and in the case of 1000IU dosage
fertilization rate was very poor (55%) The fertilized eggs were pale yellow in colour
and spherical in shape (1.16 0.05 mm) and floating. The eggs hatched out between
24h 32 h after fertilization. Changes in color of eggs and other characteristics were
noticed during embryonic development. The pale yellow eggs (015 h) became light
yellowish (15 - 20 h) and finally turned yellowish black (21h - 32h). Average hatching
ranged from 60% 73%.
Increase in dosage of hormone produced elevated values in hatching rate. For
instance maximum hatching rate of 73% was recorded as a function of 3000IU / kg
when compared to 60% in the case of 1000IU / kg. Analysis showed that relationship
between hormone dose versus latency period, number of eggs spawned, fertilization
rate and hatching rate was significant (p < 0.01) whereas the dosage of HCG versus
latency period was significant but negatively correlated.
Spawning took place approximately 20 - 26 h after hormone injection. Newly
hatched larvae showed large oval yolksac. The yolksac was ovoid with a single oil
globule located at its anterior margion bordering on ventral aspect of bent head. The
length of yolksac on day 1 was 1.1 mm and the yolk rapidly decreased from day 1 to 3
(0.72 0.12 mm). At this stage the head and tail ends of embryo became
distinguishable. The mouth was open at the end of the yolksac, when the eyes have
become pigmented. Yolk moved dorsally above abdominal cavity beside upper base
of the pectoral fin and was completely absorbed by day 3.
Preflexion larvae showed a large head with a conspicuous occipital
melanophore and terminal mouth. The head, oil globule and the yolksac together
appeared as a bulb like structure. Melanophores were scattered on the yolk and a few
are present on the unpaired fin. The length of the youngone at flexion stage was 6.2
7.1 mm from day 8 14 (n = 29). Caudal fin began to separate; faint pigmentation of
eyes, alimentary tract distinct and pectoral fin bud appeared. During Post flexion stage

the length of the larvae was 7.1 14.0 mm from day 14 30. The development of
pectoral and pelvic fins was completed on day 26 (11.3mm), whereas caudal fin
completed its development on day 16 (7.2mm). The number of rays in pectoral,
caudal and pelvic were 14 rays, 13 15 rays and 6 7 days.
Discussion
In the present study, a single intramuscular injection of natural hormone, HCG
resulted in successful spawning of Channa gachua. The results showed that complete
spawning of Channa gachua occurred when injected with 2000IU and 3000 IU/kg
and the dose of the hormone significantly affected the percentage of fertilization, egg
output, hatching rate and survival of hatchling (Tables 1 and 2). Higher latency period
due to 1000IU /kg indicated the difference in the mode of action of the hormone.
Similar observation was reported by Sahoo et al. (2007) in Clarius batrachus. The
latency period was more (24h) for the fish injected with high dosage of HCG
(3000IU). Haniffa and Sridhar (2002) observed a latency period of 28 34hr for
C.punctatus and fertilization of eggs varied from 75 80%. Marimuthu et al (2007)
reported a latency period of 23 27hr of C.striatus and fertilization rate of 60 -70%
was achieved in fish injected with ovatide. Francis (1996) observed a higher latency
period in H.fossilis (20 21h) treated with HCG. Human chorionic gonadotropin
(HCG) has been suceessfully used in catfishes such as C. macrocephalus, C.
batrachus and Pangasius suitchi and H.fossilis also showed a high latency period
(Manob Tangtrongpiros et al., 1988; Zarin et al., 1999). Spawning was complete in
the case of medium and high doses of HCG injected brooders whereas partial
spawning was observed at low dose. In terms of fertilization (70%) and hatching
(73%) rates, high dose (3000IU) showed better results, which is comparable to earlier
reports in C. punctatus (75.5%) using HCG (Haniffa et.al.2006) and C. striatus
(58.83%) using PG hormones (Hossain et al 2008). Hatching took place about 24 - 32
h after fertilization. Marimuthu and Haniffa, (2007) reported incubation period of the
eggs in C.striatus varied from 23 - 24 h. Young et al., (1989) used HCG to C.fuscus at
the rate of 2-4IU/g body weight. Kim et al., (1990) bred Ictalurus Punctatus using
HCG at the rate of 1100 IU / kg body weight.
Just 1-2 h before hatching, the embryo of C.gachua

showed twisting

movements inside the egg envelops. The similar hatching behaviour is reported in the
H.fossilis by Puvaneswari et al., (2010), and commonly observed in other catfish
species, M.montanus (A.J.A.Raj et al., 2003), M.cavasius (Rahman et al., 2004) C.

batrachus (Thakur 1980) and P. sutchi (Islam 2005).The fertilized egg diameter of
C.gachua ranged from 1.16 mm, which is more or less similar to C.striatus (1.4-1.6
mm) observed by Parameshwaran and Murugesan (1976b) . High variation in egg
sizes are also recorded in African catfish, C. gariepinus by different authors (Bruton
1979, Zaki and Abdula 1983, Herath 1988, Verreth et al., 1993).According to Ogunji
and Rahe (1999) the length of newly hatched larvae in H.longifilis recoreded the
length of 4.09-4.9 mm. Parameswaran and Kamal (1988) the length of newly hatched
murrel hatchling was as follows; 3.88 4.47 mm (Channa marulius) 2.81 3.22 mm
(C.striatus) and 2.49 2.7 mm (C.punctatus). C.gachua newly hatched larvae was
found to be 2.5 2.8mm and yolksac was fully resorbed by the third day, when the
larvae measured 5.0-5.5 mm in length.
In the present study pectoral fin buds and mouth cleft appeared in 48 h after
hatching. But in H. longifils Ogunji and Rahe (1999) reported the mouth opening 3-4
hr after hatching. Larvae commenced feeding at 5.4 mm length (48 hr), and its first
feeding took place 12 hr after the mouth opening. Ogunji and Rahe (1999) also
reported first feeding in H. longifilis larvae at 48 hr after hatching. In the present
study alimentary canal was observed 48 hr after hatching. However it may not have
been fully developed physiologically. The yolksac of H. longifilis is fully resorbed on
55 hr after hatching, (Ogunji and Rahe, 1999).
In the present study of captive breeding of C.gachua using three different
doses of HCG, the high dose (3000IU / Kg BW) showed better results in terms of
spawning, rate of fertilization and rate of hatching. Hence it is obvious to recommend
a dose of 3000IU / kg BW of HCG for higher fecundity. This success of breeding
experiments will be of immense help in conservation programmes of this threatened
murrel species.
Acknowledgement:
This research work was funded by CSIR Emeritus Scientist grant (No.21
(0670)/07/EMR-II). We are grateful to Dr. Alphone Manickam, Principal, St.Xaviers
College, Tamilnadu, India for providing necessary facilities.

Table 2.
Hormone

Fish Wt

Dosage
(Kg/BW)

Latency

Fertilization

Hatching rate

Ref.

550 625

0.2

period (h)
(%)
Channa striatus
26.0 0.5a
65.3 7.0a

560 620

0.4

23.0 0.8b

79.5 3.0b

PG

620 - 750
480 560

0.6
45

23.0 50.7b
12

79.0 5.0b
35

45

Hossain et al.,

Hormone

500 650

40

10

55

60

2008

600 - 700

60

60

65

60 70

1000IU

9
C.punctatus
-

Haniffa and

70 80

2000IU

28 34

75.5

65.5

Sridhar, 2002

65 - 85

3000IU

28 - 34

78.0

70.5

HCG

HCG

(%)
Haniffa et al.,
2000