70 J. Mar. Biol. Ass. India, 49 (1) : 70 - 75, JanuaryI.

-Rukmini
June 2007Sirisha and P. Yedukondala Rao

Reproductive biology of half – rough – back pufferfish, Lagocephalus spadiceus

(Richardson, 1844) off Visakhapatnam, east coast of India
I.Rukmini Sirisha and P. Yedukondala Rao
Department of Marine Living Resources, Andhra University,Visakhapatnam – 530003.
[email protected]
Abstract
The reproductive biology of the half- rough- back pufferfish Lagocephalus spadiceus off Visakhapatnam
(Latitude 17° 41’N, Longitutude 83°17’E) is reported. The observations showed that males are smaller than
females. Sex-ratio indicated that males were dominant in the catches. A scale of six stages of maturity of
gonads has been identified. Fecundity varied from 69,707 to 6, 16,956. Spawning takes place throughout the
year with a peak during January to March. The size at first maturity is found to be 170 mm total length.
Keywords: Reproductive biology, pufferfish, Lagocephalus spadiceus

Introduction
Lagocephalus spadiceus (Family:tetraodontidae) com- microscope with the help of an ocular micrometer with a
monly known as half-rough-back pufferfish is a bentho magnification of 0.02 mm to each division. In immature,
–pelagic species, found in tropical and temperate waters. mature and ripe ovaries (Stages I-V), about 400-500 ova
They are caught mainly by trawl nets and purse seines. were measured from each ovary. For fecundity estima-
These fishes are known to produce a virulent neurotoxic tions (Simpson, 1959), intact ovaries of stage V were
substance called ‘tetradotoxin’ that could yield valuable collected and weighed to the nearest milligram (0.01 mg)
and potential biomedical compounds. Even though these using Sartorius electronic balance. Small pieces from
fishes are highly toxic, they are consumed in countries anterior, middle and posterior portions of ovary were
like Japan, after some special processing. The present taken, weighed and counted on a binocular microscope.
study deals with the reproductive biology of L. spadiceus. The number of ova present in the ovary was calculated
Materials and methods using the formula,

The study was based on 649 specimens (377 males Fecundity = (wt of ovary/wt of sample) x No. of eggs
and 272 females) ranging in size from 80 mm to 287 mm in the samples
TL (10 to 314 g wt) collected from the commercial trawl Percentage occurrence of mature fishes and
catches off Visakhapatnam at regular intervals (thrice in gonosomatic index were used to determine the spawning
a month) from July 2004 to June 2005. During May, season. The GSI was calculated using the formula,
there were no samples because of fishing holidays from
15th April to 31st May. GSI = (wt of the gonad/wt of the fish) x 100

The collected samples were immediately brought to The morpho-histological study of gonads were
the laboratory for further analysis. After measuring the TL carried out adopting the methods given by Dixit and
(from tip of snout to tip of caudal fin) and weight (nearest Agarwal (1974b), Donato et al. (1980), De Vlaming
1g) of each specimen, the belly was cut open to note the (1972), Agarwal (1996).
sex, colour and general appearance of the gonads. The Results
gonads were then carefully removed and preserved in 5%
formalin for further analysis. Maturation
Measurements of ova diameter (Clark, 1934; Naik Testes: The testes are paired and elongated structures
and Jalihal, 1998) was recorded by taking sections from lying on both sides of the air bladder and remain attached
middle of the ovary to determine the spawning frequen- to the body wall. Six stages of maturity for testes have
cies. Their diameter was measured under a compound been identified.

Journal of the Marine Biological Association of India (2007)

Reproductive biology of half-rough-back pufferfish 71

Stage – I (Immature): whitish, ribbon-shaped occupy- cavity, ova irregular and transparent. Eggs are not visible
ing ¼th of the body cavity; Stage-II (Early maturing): to the naked eye. The ova attain a modal size of 1-2 md
whitish, occupying 2/3rd of the body cavity; Stage-III (0.02-0.04 mm). Stage-II (Early Maturing): yellowish,
(Maturing): whitish, occupying 2/3rd to ¾th of the body occupying ¼ th to 2/3rd of the body cavity, ova round and
cavity; Stage-IV (Mature): creamy white, occupying the filled with yolk. Ova are small and not visible to the
entire body cavity; Stage-V (Ripe / Oozing): in the oozing naked eye and attain a modal size of 3-4 md (0.06-
stage; and Stage-VI (Spent): flaccid, occupying about ½ 0.08mm). Stage-III (Maturing): yellowish, occupying 2/
of the body cavity. 3 rd to 3/4 th of the body cavity. They are small and round
with a model size of 5-6 md (0.10-0.12mm) and com-
Ovaries: The ovaries lie close to the peritoneum. They
pletely filled with yolk. Blood capillaries are not distinct.
are paired and elongated structures. A scale of six stages
Stage-IV (Mature): yellowish, occupying nearly entire
of maturity of ovaries has been observed in the study.
body cavity, ova visible outside. They attain a model size
Stage-I (Immature): pinkish, occupying ¼th of the body
of 7-8 md (0.14-0.16mm). Yolk vacuolated, perivitelline
space present and blood capillaries are
distinct. Stage-V (Ripe/Oozing): pale yel-
low to creamish in colour occupying entire
body cavity. Blood capillaries distinct and
clearly visible. Ova visible to the naked eye
with a model size of 9-10 md (0.18-0.20mm)
and are in the oozing stage. Stage – VI
(Spent): flaccid, with blood vessels promi-
nent all over the surface. Occupy not more
than ½ of the body cavity.
Histology of the ovary: Histology
reveals that the ovary has a wall with
numerous developing follicles. The wall
consists of 3 layers namely peritoneum, tunica
albuginea and germinal epithelium. Each
oogonium passes through a series of stages
to form ripe egg involving complicated
changes occurring in the cytoplasm and
nucleus. On the basis of these changes, the
oocyte development can be divided into 5
stages (Fig.1). Stage I - Chromatin nucleolus
stage: major portion of an oocyte at this stage
is occupied by the centrally placed nucleus
which is surrounded by a thin layer of
basophilic cytoplasm (Fig. 1A).
Stage II - Peri nucleolus stage: A charac-
teristic feature of this stage is the presence of
large number of nucleoli of different sizes
lying inner to the nuclear membrane (Fig.1 B).
Stage III - Yolk vesicle stage : This
stage is designated mainly by the first ap-
pearance of minute vacuoles in the cortex of
cytoplasm which are termed as cortical al-
veoli or yolk vesicles (Fig. 1C).
Stage IV - Yolk stage : During this
stage, an extensive development of yolk
Fig.1. Different stages of ovaries (A- stage I Chromatin nucleolus stage; globules is observed, increasing in size and
B- Stage II, peri nucleolus stage; C – Stage III, yolk vesicle stage; occupying whole of the ooplasm around the
D- Stage IV, yolk stage; E-Stage V, ripe egg stage) nucleus (Fig. 1D).

Journal of the Marine Biological Association of India (2007)

72 I. Rukmini Sirisha and P. Yedukondala Rao

Stage V - Ripe egg stage : At this stage, the egg is fully Spawning frequency : The ova are distributed around
grown and completely packed with yolkmass (Fig. 1E). a model value of 1-2 md (0.02-0.04mm) in stage I. In
stage II and III the model values were at 3-4 md (0.06-
Size at first maturity: The size at first maturity for
0.08mm) and 5-6 md (0.10-0.12mm) respectively. In
females has been determined. The average size at which
stage IV, one mode is formed around 7-8 md (0.14-
50% of individuals attain maturity falls in the length
0.16mm) compared to one mode around 9-10 md (0.18-
groups of 160-180mm. The size at first maturity has been
0.20mm) in stage V. Thus it appeared that L. spadiceus
given as 170mm (Fig. 2).
spawns throughout the year releasing the eggs each time
in a single batch during the spawning season (Fig. 4).

Fig. 2. Size at first maturity of L.spadiceus

Spawning population: The percentage frequency

distribution of females in different stages of ovary and in
different length groups indicated that females in range of
160-180mm TL formed the spawning population (Table I).
Spawning season : The availability of more number
of mature and ripe ovaries (Stage IV & V) during January
to March showed peak spawning season. Monthly per-
centage occurrence of females in different stages of ma-
turity and GSI are taken into consideration (Table 2). The
GSI has been found to be high during July and December
2004; January, February and March 2005. A sudden
increase of GSI during January, February and March
indicated spawning activity during these periods (Fig. 3).

Fig. 4. Ova-diameter frequency distribution in ovaries of

different stages of maturation in L.spadiceus

Sex Ratio: Sex ratio in relation to season indicated

that males dominated the catches during July, August,
September, November and December 2004; January and
March 2005 (Table 3). Females dominated the catches
during the remaining months. The sex ratio of male to
female during the period of study was 1:0.72.
Chi-square analysis indicated that there is a significant
difference between the sexes in occurrence. Sex-ratio in
Fig. 3. Gonosomatic Index for female L.spadiceus relation to length indicated that males dominated the catches

Journal of the Marine Biological Association of India (2007)

Reproductive biology of half-rough-back pufferfish 73

Table 1 . Length – wise percentage frequency distribution of females of L. spadiceus in different stages of maturation
Length No. of
groups specimens Stage Stage Stage Stage Stage Stage
(TL in mm) examined. I II III IV V VI
101- 110 2 3.57 8.34 - - - -
111- 120 5 17.85 - - - - -
121- 130 16 25 16.65 9.75 - 3.48 -
131- 140 14 10.72 8.34 14.63 6.66 1.16 1.44
141- 150 17 10.72 16.65 2.43 6.66 8.13 2.89
151- 160 28 7.14 25 14.63 10.0 10.47 10.14
161- 170 30 10.72 - 17.04 13.35 15.11 4.34
171- 180 25 3.57 8.34 4.88 13.35 13.95 7.25
181- 190 32 10.71 8.34 14.63 16.66 10.47 28.98
191- 200 39 - 8.34 9.75 16.66 10.47 28.98
201- 210 19 - - 4.88 6.66 10.47 8.69
211- 220 11 - - 2.43 - 4.65 8.69
221- 230 11 - - - 3.34 3.5 10.14
231- 240 6 - - 2.43 - 3.5 2.89
241- 250 7 - - 2.43 6.66 2.32 2.89
251- 260 - - - - - - -
261- 270 1 - - - - - 1.44
271- 280 - - - - - - -
281- 290 1 - - - - 1.44

Table 2. Monthly percentage distribution of females in different stages of maturation (2004 - 2005)

Month No. Stage Stage Stage Stage Stage Stage

examined I II III IV V VI
Jul. 17 - - 11.7 5.8 76.4 5.8
Aug. 11 - 9.1 18.1 36.3 36.3 -
Sep. 26 19.2 30.7 7.6 19.2 23.0
Oct. 43 20.9 9.3 2.3 23.2 44.1
Nov. 33 18.1 15.1 15.1 9.0 9.0 33.3
Dec. 23 - 4.3 8.6 34.7 8.6 43.4
Jan. 15 - 6.6 13.3 - 66.6 13.3
Feb. 36 - 2.7 11.1 19.4 38.8 27.7
Mar. 30 3.3 3.3 16.6 3.3 46.6 26.6
Apr. 18 38.8 5.5 11.1 5.5 27.7 11.1
May. - - - - - - -
Jun. 20 15.0 5.0 25.0 15.0 30.0 10.0

at 80-190 mm TL(Table 4). Females dominated the catches Fecundity: Sixty mature specimens (Stage V) were
from 191-250 mm during the study period. The sex ratio collected during the study period from July 2004 to June
showed a marked trend from smaller size groups to larger 2005. The fecundity ranged from 69,707 to 6,16,956 in
size groups. fish of size 147-250 mm. The fecundity of L. spadiceus

Journal of the Marine Biological Association of India (2007)

74 I. Rukmini Sirisha and P. Yedukondala Rao

Table 3. Sex ratio in L. spadiceus during July 2004 to was studied in relation to fish-length, fish weight and
June 2005 estimated by using the regression equation,
Month Total Sex ratio F = a Lb or in logarithmic form,
Nos. Male Female Male : Female Log F = Log a + b Log L
Jul. 62 45 17 1: 0.37 The number of ova increased with increase of fish
Aug. 50 39 11 1: 0.28 size, the relationship between the fecundity and length
Sep. 68 42 26 1: 0.61 showed linearity (Fig. 5). The linear form of the relation-
Oct. 64 21 43 1: 2.04 ship between fecundity and total length is
Nov. 85 52 33 1: 0.63 log F = 3.4684 + 0.8062 log TL
Dec. 60 37 23 1: 0.62 The relationship between fecundity and weight of the
Jan. 60 45 15 1: 0.33 fish (Fig. 6) is also observed to be linear indicating that
Feb. 53 17 36 1: 2.11
Mar. 84 54 30 1: 0.55
Apr. 25 7 18 1: 2.57
May - - - -
Jun. 38 18 20 1: 11
Total 649 377 272 1: 0.72

Table 4. Sex ratio of males and females in relation length

groups during July 2004 to June 2005
Lt.groups Male Female Sex ratio
(Tl mm) M: F
80-90 3 - 1:0
91-100 7 - 1:0
101-110 3 2 1:0.7
Fig. 5. Relationship between fecundity and fish length in
111-120 11 5 1:0.5 L.spadiceus
121-130 22 15 1:0.7
131-140 37 14 1:0.4 fecundity increased with increase in weight of the fish.
141-150 44 18 1:0.4 The equation obtained is
151-160 59 26 1:0.4
log F = 4.1945 + 0.5620 log W
161-170 60 31 1:0.5
171-180 60 29 1:0.5
181-190 42 33 1:0.8
191-200 19 38 1:2
201-210 1 18 1:18
211-220 4 11 1:2.8
221-230 2 11 1:5.5
231-240 - 10 1:10
241-250 - 6 1:6
251-260 - - 1:1
261-270 1 1 1:1
271-280 1 - 1:1
281-290 - 1 1:0.7
Fig. 6. Relationship between fecundity and fish weight in
Total 377 272 L.spadiceus

Journal of the Marine Biological Association of India (2007)

Reproductive biology of half-rough-back pufferfish 75

Discussion cated a significant difference between the sexes in their

occurrence. In the present study, males dominated the
In the present investigation on L. spadiceus, a
catches throughout the year except in October, February,
six stages of maturity are described more or less adhering
April and June. According to Naik and Jalihal (1998), X2
to the scale given by International Council for Exploration
analysis revealed a significant deviation from the 1:1 ratio
of Seas (ICES) corresponding to the scale given by
in October besides September and December. Fecundity
Lovern and Wood (1937) and later followed by Naik and
ranged from 69,707 to 6,16,956 ova. Naik and Jalihal
Jalihal (1998). The size at first maturity has been given
(1998) estimated fecundity in L. spadiceus and ranged
as 170 mm for species off Visakhapatnam. According to
from 0.75 x 105 to 3.5 x 105 ova.
Naik and Jalihal (1998), the size at first maturity for both
males and females is 95 mm TL for this species on the References
West Coast. The difference may be due to geographical
Agarwal, N.K. 1996. Fish reproduction. APH-Publishing
variations and selectivity of gear. Based on ova-develop- Corporation, New Delhi. 147pp.
ment, it appeared that this fish spawned throughout the
year with peak in January to March releasing the eggs in Clark, F.N.1934. Maturity of the California Sardine
(Sardina caerulea), determined by ova diameter mea-
a single batch. Naik and Jalihal (1998) also reported that
surements. Fish.Bull.Calif.Div.Fish.Game, 42: 1-49.
this species spawns throughout the year. Histology of the
ovary revealed that the oocyte undergoes different devel- De vlaming, V.L.1972. The functional morphology of
opmental stages to form ripe egg. Present investigation is teleost gonads. In: Fish Physiology. Bull.Jpn.Soc.
more or less similar to that of work done by Agarwal Sci.Fish., 31: 123-132.
(1996). Dixit, R.K.and N. Agarwal. 1974 b. Studies on the devel-
opmental rhythm in the oocyte of Puntius sophore.
Tetraodontids have a relatively extended spawning Acta anat., 90: 133-144.
season. The availability of more number of mature and
ripe ovaries (Stages IV & V) during January to March Donato, A., A.Contini, A. Maugeri and S. Fasulo. 1980.
showed peak spawning seasons in the coastal waters off Structural and ultra structural aspects of the growing
oocyte of Chromis chromis (Teleostei, Labridae). Riv.
Visakhapatnam, even though the fish spawns throughout
Biol. Norm. Patol., 6:31-66.
the year. Naik and Jalihal (1998) reported the spawning
season in L. spadiceus to be from February to March and Lovern, J.A. and H.Wood. 1937. Variations in the chemi-
September to November off the West Coast of India. cal composition of herrings. J. Mar. Biol. Ass. U.K.,
22:281-293.
GSI has been found to be high during January to
Naik, S.K. and D.R.Jalihal. 1998. Biological observations
March indicating that spawning activity takes place during
on the pufferfishes of South Konkan Coast with spe-
this period. According to Naik and Jalihal (1998), high cial reference to the net damaging species L. spadiceus
GSI values are indicated once during September-Novem- (osteichthyes, Tetraodontidae). Indian J. Mar. Sci.,
ber and again in February-March, suggesting the spawn- 27:426-432.
ing of L. spadiceus during those months on the West
Simpson, A.C. 1959. The spawning of the plaice
Coast of India. The sex ratio of male to female during (Pleuronectides platessa) in the North Sea. Fish.Invest.
July 2004 to June 2005 is 1:0.72. According to Naik and Ser., 222:111pp.
Jalihal (1998), the sex-ratio of L. spadiceus is 1:1.02 on
the West Coast of India. Chi-square (X2) analysis indi- Received: 21 November 2006
Accepted: 6 August 2007

Journal of the Marine Biological Association of India (2007)