REPRODUCTIVE BIOLOGY OF RIDGED SWIMMING CRAB,

Charybdis natator (HERBST, 1794), FROM ESTANCIA, ILOILO,

WESTERN PHILIPPINES

REY MALVIN A. PALLOMINA

An Undergraduate Special Problem Presented

to the Faculty of the Division of Biological Sciences
College of Arts and Sciences
University of the Philippines Visayas

In Partial Fulfillment of the Requirements

for the Degree of
Bachelor of Science in Biology

June 2018
 CHAPTER 1

INTRODUCTION

Fisheries is one of the major industries in the Philippines, an archipelagic

country, in the Pacific Ocean. The fisheries in the Philippines are mainly composed of
fishes, and shellfishes. Crabs are one of the major marine life products that are being
marketed. One commercially important crab is the Charybdis natator (Herbst, 1794)
commonly known as the “ridged swimming crab”, which belongs to Family
Portunidae, a group of marine crabs widely distributed across Asia, Australia, and
Africa (Wee and Ng, 1995). In Iloilo, Philippines, C. natator is locally known as
“kalintugas”. Charybdis natator is characterized by brown to orangish coloration on
the dorsal side of the carapace while the ventral side is found to have a bluish
coloration, mottled with white and pale red. Legs are dark and reddish brown in color.
The carapace is densely covered with short pubescence and is absent on the distinct
transverse ridges in the anterior side. The ridged swimming rock crab has a distinct
pattern from other crab species of having ridges located dorsally on its carapace
(Figure 1.1). The same pattern could be observed in both sexes of C. natator.
Although uncommon and is not a major commercially-fished species (Atlas of Living
Australia) in comparison to other more abundant and commercially important crabs,
such as the mud crabs (Scylla serrata) and sand crabs (Portunus pelagicus), the ridged
swimming rock crab also contribute to crab fisheries in Asia and Australia (Sumpton,
1990).

This study aims to determine establish the breeding cycle of C. natator

collected from Estancia, Iloilo, Western Philippines. The study also intends to identify
the monthly stages of gonad development of C. natator, through morphological and
histological analysis. Specifically, the study aims to determine: (1) the morphological
appearance of the gonads of C. natator at each stage of development, (2) size at
sexual maturity of the crab, (3) the percent occurrence of each gonadal maturation
stage, (4) the percent occurrence of male and female crabs, (5) determine the
gonadosomatic index (GSI) of both males and females, (6) compute for the gonad
index (GI) of both males and females, (7) establish the relationship of the different
gonadal maturation stages with gonadosomatic index (GSI) and gonad index (GI)
from September 2017 to February 2018.

1
 The information that will be gathered will be significant for the
implementation of regulatory measures for the management and conservation of the
remaining stock in the wild. There were a lot of studies on the reproductive biology of
distinct species of crabs but there is no or little information known about C. natator,
thus, the need for the present study. The results of this present investigation will also
be useful for the brood stock management in the future hatchery culture of this crab
species.

2
Figure 1.1 Ridged-pattern, located dorsally on its carapace, of Charybdis natator that
makes it distinct from other crab species.

Figure 1.2 Dorsal carapace of both male (A and B) and female (C and D) crabs
showing no difference on the carapace pattern present only in C. natator.

3
 CHAPTER 2

REVIEW OF RELATED LITERATURE

2.1. Taxonomy

Charybdis natator (Herbst, 1794) or ridged swimming rock crab or locally

known as “kalintugas” in the Western Visayas belongs to the family Portunidae which
belongs to the order Decapooda, class Malacostraca, and phylum Athropoda.
Synonyms of this species is Cancer natator which is also authored by Herbst (1794)
(WoRMS Editorial Board, 2018). Charybdis natator which is under family
Portunidae, along with families Cancridae or the rock crabs, Gecarcinidae or the land
crabs, Geryonidae or the golden crabs, Menippidae or the stone crabs, and
Ocypodidae or the ghost crabs, is a member of true crab species (Tavares, n.d.).

2.2. Description

Charybdis natator (Herbst, 1794) belongs to Family Portunidae, a typical

group of marine crabs which is widely distributed across Asia, Australia, and Africa
(Wee and Ng, 1995). The family Portunidae are groups of crabs which are mainly
comprised of swimming crabs (Bowling, 2012). Swimming crabs can be
differentiated from walking crabs because of their hind limbs, last pair of legs, which
are flattened, acts as propellers for these types of crabs to swim efficiently. They are
called ridged swimming crabs because of the ridges in their carapace (Samuel, 2014).

Figure 2.1 Global distribution of Charybdis natator (Source: http://www.marine

species.org/aphia.php? p=taxdetails&id=208817#distributions)
4
2.3. Morphology

According to Wee and Ng (1995) in “The Raffles Bulletin of Zoology”, the

crab species Charybdis natator can be describe through the following: Carapace,
uniformly pilose, sparse granules on anterolateral surface; anterior carapace ridges
present except frontals, epibranchials interrupted by unbroken metagastric ridge,
posterior with one pair of cardiac and three short pairs of mesobranchial ridges; six
frontal lobes, medians on lower plane, projecting beyond egually broad submedians,
laterals acute, separated from sub medians by deeper V-shaped notch; inner
supraorbitallobe broder than frontals, bluntly triangular; six anterolateral teeth, first
tooth truncate, second to fourth subequal with acute tips, last spiniform and least
prominent. Basal antennal segment bearing short granular ridge. Chelipeds unequal,
granular and pilose; anterior border of merus with three to four spines; carpus with
strong spine on inner angle and three spinules at outer angle; manus with four spines
on upper surface and a spinule at distal end of outer border, lower surface with
transverse squamiform ridges; fingers stout, deeply grooved. Propodus of natatory leg
serrated on posterior border. Second to fifth segment of male abdomen keeled,
penultimate segment with lateral borders parallel then converging distally. G1 distal
tip slender and elongate, abdominal surface bears two rows of short terminal bristles
ending proximal to lip region, outer suface with row of longer bristles starting near tip
and extending proximally as widely spaced bristles. Pubescence of dorsal surface
brownish, granules bright red. Ventral surface bluish, mottled with white and pale red.

The distinct reddish coloration of the anterolateral and frontal teeth, on the
granules and transverse carapace ridges are what make this species easily recognized.
It is similar to that of Charybdis granulata. Leene (1938), but it has since been
considered as a distinct species by Sakai (1976) and Miyake (1983). The under
surface of the manus of the chelipeds vary from having a transverse squamiform
arrangement to that of a surface with scattered granules (Wee and Ng, 1995).
Charybdis natator can grow its carapace ranging from 5 cm to 17 cm (Samuel, 2014).

5
2.4. Habitat and Ecology

Ridged swimming rock crabs, as the name implies, inhabits the bottom of the
ocean on rocky substrates or on coral reefs. They live ranging from 5 to 4 meters
below the sea levels (Samuel, 2014). They can also live on muddy or sandy substrate.
But according to Sakai (1976), this species is found on the bottom of rocks, pebbles or
sand at depths of 15-35 meters.

Although uncommon and is not a major commercially-fished species (Atlas of

Living Australia) in comparison to other more abundant and commercially important
crabs, such as the mud crabs (Scylla serrata) and sand crabs (Portunus pelagicus), the
C. natator also significantly contribute to crab fisheries in India and Australia
(Sumpton, 1990).

2.5. Reproductive Biology

In the study conducted by Sumpton (1990), he used trapping methods to

collect samples of Charybdis natator in Moreton Bay, Queensland. Based from his
results, he observed that during the month of June and July there are more females
than males, but in other months the ratio of males to females is two to one. The lesser
number of females maybe because of the spawning season of the C. natator during
June and July where females migrate to deeper waters. He also found two major
spawning peaks for C. natator. During winter the low proportion of gravid females
and high proportion of females with inactive gonads indicate that C. natator does not
spawn year- round in the subtropical waters of Moreton Bay. Pillai and Nair (1976)
found that the closely related Charybdis feriatus bred throughout the year in
southwestern Indian waters, although gravid females were more common during
January and February. It is thus likely that the cooler conditions during winter in
Moreton Bay limit the spawning activity of C. natator. The ridged swimming crabs
can ovulate up to three times annually and it was confirmed by ovary examination that
indeed the C. natator are inactive during winter season. Also, he indicated in his study
that the fecundity of a female crab ranges from 181,000 to 976,000. The reproductive
biology of C. natator has similarities to other crab species under the Family
Portunidae (Sumpton, 1990).

6
2.6. Gonad characterization

The macroscopic technique in identifying the stage of gonadal development of

both male and female crabs involves only the observation and examination of the
different shapes, sizes, and color of the gonads by means of the naked eye.

There are five macroscopic gonadal developmental stages of female crabs

(Minagawa et al., 1992; de Souza et al, 2009). The five stages of the female ovaries
are: (1) Stage 1 or Immature Stage, (2) Stage 2 or Early Maturing Stage, (3) Stage 3
or Late Maturing Stage, (4) Stage 4 or Fully Matured Stage, and lastly (5) Stage 5 or
Post-Spawning Stage. The Immature stage is described as a translucent ovary showing
a ribbon-like structure, slender and flaccid lobes in a tubular form, with white-yellow
coloration. The next stage, Early Maturing stage, the ovaries is compressed dorso-
ventrally and the anterior lobes covers almost all hepatopancreas and the stomach
region of the crabs, meaning there is increase in size of the crabs’ ovaries. The color
of this stage is light orange or peach in color. In the third stage, the Late Maturing
stage, the ovaries change its pigmentation form light orange or peach into orange, and
also the anterior lobes cover the totality of the hepatopancreas. In a Fully Matured
ovary, the color of the ovary becomes deep orange or scarlet, the maximum size of
ovary is attained, and it covers the whole hepatopancreas and the stomach area.
Lastly, the spent or post-spawning stage, the ovaries appears in a deep brown color
and sometimes it regresses back to its former color, white-yellow. This is due to the
release of mature oocytes and the previtellogenesis stage can be observed again, in
which it indicates a new beginning in the ovarian cycle. At this stage eggs can be
observed in the abdominal flaps of the female crabs because they have already
undergone fertilization.

Ikhwanuddin et al. (2012) described the morphological characteristics of four

stages of ovarian maturation in P. pelagicus. The four stages are Immature, Early
Maturing, Late Maturing and Mature. Immature ovaries are translucent in color,
showing ribbon or thread-like structure which is sometimes difficult to recognize
from other tissues. In the Early Maturing stage, ovary is yellow and shows increase in
size. For the Late Maturing stage, the color of the ovary significantly changes into
yellowish to light orange that increases massively in size. And for the mature stage,

7
individual eggs are visible, the color of the ovary become dark orange or red, and the
lobules are swollen with large ova

In the male crabs, there are only three stages of gonadal development for the
sperm (de Souza et al., 2012; Soundarapandian et al., 2013), (1) Immature, (2)
Maturing, and (3) Fully Matured. The gonads of the Immature crabs are relatively
small, with a cream color, located lateral to stomach. In a maturing stage, the gonads
possess a creamy white color. The testes appear as a coiled tube placed laterally and
posterior to the stomach, while the vas deferens extends laterally to the heart. The last
stage, Fully Matured stage, of the male gonads appear as a milky white color. The
male gonads have an enlarged testis, with the vas deferens becomes coiled, and it
completely covers the full body cavity.

8
 CHAPTER 3

MATERIALS AND METHODS

3.1. Collection site and sampling

A six-month survey of C. natator crabs was done from September 2017 to

February 2018. The samples were collected from the waters of Estancia Iloilo, stored
in Styrofoam box with ice, transported to the wet market of Miagao, Iloilo. Transport
travel was four hours Ten samples were randomly bought from the market every first
week of the month. The samples have representatives of different sexes with sizes
ranging from the smallest to the largest.

Crabs that were purchased from the market were stored in the ice box for
preservation, so that the tissues of the crabs will not deteriorate, and for the crabs to
be anesthetized. It is recommended that the samples should not be placed in the
freezer for the gonads will not be of use during the histological analysis because it
would be too stiff to be prepared as thin sections for histological analysis, and the
gonads will give off a different color once it has reacted to the different dyes:
haemotoxylin and eosin.

3.2. Determination of sexes

Male and female crabs were described and were differentiated

morphologically through the different shapes of their abdominal flaps shown in
Figure 3.1 and Figure 3.2.

Male crabs possess a narrow and triangular abdominal flap, while the female
crabs have broader and rounder abdominal flaps which covers the whole sternum of
the female crabs (Nemenzo, 1976). In males, they possess a pair of gonopods which is
located in the anterior part, inside the abdominal flap. Gonopods are long tubules that
are used to facilitate the sperm into the gonopore of the female crabs (Zinski, 2010).
Gonopores are small opening in the female crabs that is located ventrally in the first

9
segment, in between the first pair of pereiopods (Wilkin, 2002). Another distinct
characteristic of a female crab is that they have hairy appendages that functions as an
attachment of the developing embryos (Nemenzo, 1976). Size of the crabs can also be
used to determine the gender of the crab; female crabs are smaller than male crabs.

Figure 3.2 Ventral side of male Charybdis natator. Male crabs can be identified by its
narrow and triangular abdominal flaps (encircled in the figure).

10
Figure 3.2 Ventral side of female Charybdis natator. Female crabs can be identified
by its broad and round abdominal flaps (encircled in the figure) that almost
cover its sternum.

3.3. Body Measurement

For each crab, the body weight (BW), and carapace width (CW) was
measured. Before the body weight was measured, the crab should be patted dry using
clean cloth or tissue after removing it from the refrigerator. This is to avoid errors in
the results. The crab was then placed on an analytical balance and the weight was
recorded to the nearest tenth of a gram. Carapace width was measured using a Vernier
caliper and the measured value was recorded to the nearest millimeter. Carapace
width (CW) is the distance between anterior lateral spine and the most posterior
lateral spine (Brown, 2009).

CW

Figure 3.3 Measurement of carapace width (CW) on the widest part of the carapace
laterally from the left margin to the right

3.4. Dissection

Using a pair of sharp scissors, the dissection of crab proceeded by cutting the
dorsal carapace from the posterior end of the carapace moving in counterclockwise
direction, making a square shaped incision. Then using scalpel and forceps, the thin

11
black membrane covering the cephalothoracic cavity was removed to expose the
gonads. A Y-shaped structure of gonads, called the anterior horn, was observed. It lies
dorsal to the hepatopancreas. The hepatopancreas has a fingerlike appearance with
light yellow to red orange in coloration. It is located on both sides of the gastric mill
on the anterior part of the cephalothoracic cavity. The hepatopancreas is extended
posteriorly, connected to the gastric mill.

Figure 3.6 Crab dissection proceeding with a square-shaped incision at the dorsal
carapace. Figure 3.6.A shows the part of the carapace where the square-shaped
incision was made, and Figure 3.6.B shows the dissected portion of the carapace.

3.5. Gonad analysis

The gonad developmental stages for all sexes of Charybdis natator were
investigated using macroscopic technique which involves the description of the
external morphology of the gonads.

The macroscopic technique involved the examination of the shape, size and
color of the gonads by using the naked eye.

The five gonadal development stages of female C. natator based on

macroscopic observation according to Ikhwanuddin (2010) were the following: (1)
Immature, (2) Early Maturing, (3) Late Maturing, (4) Fully Mature, and (5) Spent.
The shape, size and color of the gonads was analyzed based on this classification. The
male C. natator was analyzed based on the three stages of gonadal development
according to Soundarapandian (2013): (1) Immature, (2) maturing, and (3) mature.

3.6. Gonad analysis

12
 The percent occurrence of each gonad maturation stages was calculated by
using the formula:

no . of individuals within that stage

% occurrence of each gonad maturation stage= x 100 %
total no . of individuals∈that sex

The percent occurrence of male and female was calculated by using the formula:

total no . of individuals of a sex

% occurrence of each sex= x 100 %
total no . of samples

Gonad analysis included computations of the gonadosomatic indices and

gonad indices. The quantitative gonadosomatic index (GSI) was calculated using the
equation by Krajangdara and Watanabe (2005):

Gonad Weight (g)

Gonadosomatic index ( GSI ) = x 100 %
Body Weight (g)

The mean value of gonad indices (GI) or the index of sexual maturity was then
calculated using the equation:

Gonad index ( GI )=
∑ (n x WV )
N

Where n is the number of individuals in each developmental stage; WV is the

pre-weighted value for the developmental stage; and N is the total number of crabs
per monthly sample. Pre-weighted values (WV) will be assigned to the female gonad
stages as: undeveloped and recovering - 1, developing - 2, ripe and spawning – 3. For
the male testes, the values to assign will be: Immature - 1, and mature - 2. Values of
mean gonad indices per month were plotted against time (months) to determine the
spawning pattern or reproductive cycle (Baylon and Tito, 2012)

13
 CHAPTER 4

RESULTS

From September 2017 to February 2018, a total of 60 crabs were examined for
body weight (BW) and carapace width (CW) with a total of 26 males and 34 females
from Estancia, Iloilo. All 60 samples were dissected and examined for gonad
maturation stages.

4.1. Morphological appearance of male and female gonads

A total of 26 males and 34 females from September 2017 to February 2018

were dissected to examine the macroscopic appearance of the different stages of their
gonads. The determination of stages was based on color, shape and volume of the
gonads. The progress of the ovarian maturation was classified in four stages.

1) Stage 1 (Immature). The ovary showed translucent to dirty-white color which can
be difficult to distinguish from its hepatopancreas since the ovaries are minute, thin,
elongated and its appearance were slightly convoluted (Figure 4.1).

2) Stage II (Early Maturing). The color of the ovary ranged from pale orange to
peach. The ovaries were more ostensible and it can be easily determined from the
hepatopancreas but do yet extend to the hepatic region (Figure 4.2)

3) Stage III (Late Maturing). The ovary showed a significant increase in size and
changer in appearance. The ovary changed also to a bright orange coloration. The
anterior horns of the ovary form a distinct M shape, and the ovary started occupying
the lateral parts of the branchial region, and the lobules become more visible (Figure
4.3).

14
4) Stage IV (Fully Mature). The ovary showed a deep orange to scarlet coloration.
There was massive enlargement of the size of ovary which now occupied the gastric,
posterior and intestinal cavities. The middle lobes of the ovary were observed to cover
the whole stomach area. Ovarian lobules appeared swollen (Figure 4.4).

GM
HP HP
OO OO
H
G
G
C C

Figure 4.1 Immature (Stage I) ovary of female C. natator is, thin and has creamy
white coloration, slightly convoluted which extends dorsolateral to its
hepatopancreas. (HP = hepatopancreas; OO = ovary; GM = gastric mill; G =
gills; C = cartilage; H = heart)

15
 HP HP
GM
OO
OO
H
G G

C C

Figure 4.2 Early Maturing (Stage II) ovary of female C. natator is enlarged compared
to its previous stage, color range from pale orange to peach, and runs
dorsolateral to the hepatopancreas. (HP = hepatopancreas; OO = ovary; GM =
gastric mill; G = gills; C = cartilage; H = heart)

GM HP
HP
OO OO

H
G
G
C C

Figure 4.3 Late Maturing (Stage III) ovary of female C. natator has increased its size
compared to its previous stage, has a distinct M-shaped, bright orange in color,
now extends up to the branchial region. (HP = hepatopancreas; OO = ovary;
GM = gastric mill; G = gills; C = cartilage; H = heart)

16
Figure 4.4 Fully Mature (Stage IV) ovary of female C. natator has significantly
increased its size from its previous stage, has also a distinct M-shaped, deep
orange in color, covers most of the hepatopancreas and extends up to the
branchial region. (HP = hepatopancreas; OO = ovary; GM = gastric mill; G =
gills; C = cartilage; H = heart)
On the other hand, the male gonad maturation stages were classified into
three: Immature, Maturing, and Mature.

1) Stage I (Immature) The testes were thin, translucent to opaque white in color,
which lie on either side of the stomach. Testes and the vas deferens were slightly
convoluted and were difficult to distinguish from each other (Figure 4.5).

GM HP
HP

T T
VD VD

H G
G
C C

Figure 4.5 Immature (Stage I) gonads of male C. natator is difficult to distinguish

from the hepatopancreas because of its thin and convoluted structure, testes and vas

17
 deferens are indistinct, creamy white in color, and runs dorsal to the
hepatopancreas. (HP = hepatopancreas; T = testis; VD = vas deferens; GM =
gastric mill; H = heart; G = gills; C = cartilage)

2) Stage II (Maturing). The testes were slightly enlarged compared to the previous
stage and remained to be creamy white in color. The testes and vas deferens were
more well developed and were distinguishable from each other. The testes, posterior
to the gastric mill, extends laterally to the hepatopancreas. Vas deferens was further
enlarged, the structure became highly-coiled, and the vas deferens further extends
towards the heart (Figure 4.6).

3) Stage III (Mature). There was further enlargement of testes and vas deferens,
acquiring a more prominent white coloration. They were coiled and swollen which
occupied the body cavity (Figure 4.7).

GM HP
HP
T
T VD
VD

G G
H

C C

Figure 4.6 Maturing (Stage II) gonad of male C. natator is more recognizable. The
testes and vas deferens are distinct from one another. Testes runs dorsolateral
to the hepatopancreas, and vas deferens, anterior to the heart, shows a coiled
structure. (HP = hepatopancreas; T = testis; VD = vas deferens; GM = gastric
mill; H = heart; G = gills; C = cartilage)

18
 GM HP
HP
T T
VD VD

G G
H

C C

Figure 4.6 Mature (Stage III) gonads of male C. natator with testis which has further
enlarged and extended, and an enlarged highly coiled vas deferens. (HP =
hepatopancreas; T = testis; VD = vas deferens; GM = gastric mill; H = heart;
G = gills; C = cartilage)

4.2. Minimum size at sexual maturity

The size composition and size frequency distribution of each gonad maturation
stages are shown in Figure 4.8 for female C. natator and Figure 4.9 for male C.
natator.

The carapace width (CW) of the 34 C. natator female crabs ranged from 71 to
93 mm with a mean of 82.5 mm, while CW of the 26 males ranges from 72 to 111
mm with a mean of 85 mm.

All the 34 samples of C. natator females that were dissected were used for the
data on the size composition and size frequency distribution. The greatest percentage
of the females with Immature ovary (Stage I) was in the range of 71 to 75 mm CW,
while the highest percentage of female crabs under Early Maturing ovary (Stage II)
was in the range of 81 to 85 mm CW. At Late Maturing ovarian stage (Stage III), it
was determined that the highest percentage where this stage is at was at the range of
86 to 90 mm CW, and lastly females with Fully Mature ovaries or at its spawning
stage (Stage IV) was found at the range of 86 to 95 mm CW, but only attained its
peak at 86 to 90 mm CW and further increase in the carapace width (CW) showed

19
also an increase in the number of individual female crabs attaining a higher gonadal
maturation stage, meaning there is also an increase of occurrence of female crabs
attaining Fully Mature ovary. The smallest female C. natator with a Fully Mature
gonadal stage has 86 mm CW (Figure 4.8).

A sample size of 26 male C. natator were used for the data on the size
composition and size frequency distribution of the different gonadal maturation
stages. The highest percentage of males having Immature gonads (Stage I) was in the
range of 72 to 75 mm CW, while male C. natator possessing maturing gonads (Stage
II) where found highest at the range of 82 to 85 mm CW. The occurrence of mature
testes (Stage III) was divided in 4 carapace widths (CW) having the smallest size
range at 105 to 111 mm and the highest size range at 90 mm, meaning that the
carapace width (CW) of the smallest male C. natator with mature gonads is 90 mm
and the largest mature crab has a CW of 111 mm (Figure 4.9).

There was variation in the population size along the increase in the carapace
width of female crabs. Starting off with a large population size in the smallest size
range (70 to 75 mm CW) with a gradual decrease of population as the carapace width
also increases, until at the size range of 86 to 90 mm CW a sudden increase of
population was determined. It was different for the male C. natator where the trend
was a gradual increase of population, was shown, as the carapace width increase only
up to its peak in the size range of 86 to 90 mm and continually decreased onward as
the carapace width increased.

20
 10
9
8
NO. of OCCURANCE 7
6
5
4
3
2
1
0
70 - 75 76 - 80 81 - 85 86 - 90 91 - 95

CARAPACE WIDTH (MM)

Immature Early Maturing Late Maturing Fully Mature

Figure 4.8 Size composition and size frequency distribution of different stages of
ovarian development in female C. natator were collected from Estancia, Iloilo
(n=34). Females with Fully Mature ovary (Stage IV) were only observed from
86 to 90 mm carapace width (CW).
8

6
NO. of OCCURANCE

0
70 - 75 76 - 80 81 - 85 86 - 90 91 - 95 96 - 100 101 - 105 106 - 110 111 - 115

CARAPACE WIDTH (MM)

Immature Maturing Mature

Figure 4.8 Size composition and size frequency distribution of different stages of
testes development in male C. natator were collected from Estancia, Iloilo (n=26).
Males with Fully Mature testes (Stage III) were observed from 86 to 111 mm
carapace width (CW) with the peak at the size range of 86 to 90 mm.

4.3. Monthly percent occurrence of each gonad maturation stages

21
 The monthly percent occurrence of each gonadal stages is illustrated in Figure
4.10 for the female C. natator and Figure 4.11 for the male C. natator.

From September 2017 to February 2018, a total of 34 female crabs were

analyzed for their different ovarian maturation stages. In September, 80% of the
samples were Immature (Stage I) and the remaining 20% that were examined
belonged in the Late Maturing stage (Stage III). In October, 60% of the ovaries
observed was Immature (Stage I), 20% were at Early Maturing stage (Stage II) and
the last 20% observed were Late Maturing stage (Stage III). In November, Immature
(Stage I) female crabs were only at 33.33% whereas the remaining 66.67% were at
Late Maturing stage (Stage III). In December, 50 % of the female crab samples were
Immature (Stage I) and 50% were at Early Maturing stage (Stage II). Three stages
were observed during January, 40% of which were Immature (Stage I), 40% were
Early Maturing (Stage II), and the last 20% were Late Maturing (Stage III). And lastly
for the month of February, 28.57% of the ovaries of female C. natator were Early
Maturing (Stage II), Late Maturing stages (Stage III) were at 28.57% occurrence, and
42.86% were with Fully Mature gonads (Stage IV). Fully Mature (Stage IV) ovaries
were observed only during the month of February (Figure 4.10)

On the other hand, a total of 26 male C. natator were examined for the
different gonad maturation stages. In September, 20% were Immature (Stage I), 40%
were Maturing (Stage II), and the remaining 40% of the male crabs were Matured
(Stage III). During October, 40% of the male crabs observed were still Immature
(Stage I) and 60% of the male C. natator were Maturing (Stage II). November showed
no signs of Immature (Stage I) crabs with 75% of the population were Maturing
(Stage II), and the remaining 25% were already Mature (Stage III). Majority of the
crabs during December were Immature (Stage I) with 75% occurrence and 25% were
Mature (Stage III) male C. natator. In January, 20% of the male crabs were Immature
(Stage I), 60% were still Maturing (Stage II), and the other 20% of the male crabs
were Mature (Stage III). February 2018 showed an equally portioned stages of the
male gonadal maturation, with all the gonad stages, from Immature (Stage I) to
Mature (Stage III) stages, showed 33% occurrence. Fully Matured (Stage III) male
crabs were observed most of the months except during October, with the highest
number of occurrence, among all the sampling month period, on September (Figure
4.11).

22
 100%
90%
80%
% OCCURANCE 70%
60%
50%
40%
30%
20%
10%

0%
September October November December January February

Month

Immature Early Maturing Late Maturing Fully Mature

Figure 4.10 Percent occurrence of the different stages of ovarian maturation in female
C. natator from September 2017 to February 2018 on crabs collected from
Estancia, Iloilo (n=34).
100%
90%

80%
70%
% OCCURANCE

60%
50%

40%
30%

20%
10%

0%
September October November December January February

Month

Immature Maturing Mature

Figure 4.10 Percent occurrence of the different stages of gonad maturation in male C.
natator from September 2017 to February 2018 on crabs collected from
Estancia, Iloilo (n=34).

4.4. Monthly occurrence of male and female C. natator

23
 During the six-month sampling from September 2017 to February 2018, a total
of 60 crabs were examined for the identification of sexes. For the duration of the
sampling period a total of 34 female crabs and 26 male crabs were determined. The
monthly occurrence of the different sexes is shown in Figure 4.12.

The female population has a slow but steady increase during the first few
months on September 2017 up to December 2017 from 50% to 60%, then it dropped
back again to 50% during the next month, January 2018. The lowest point of
occurrence was observed during the first two months, September and October 2017,
having 50% population of the female crabs. The female C. natator achieved its peak
during the last month, February 2018, with 70% occurrence.

Meanwhile the male C. natator observed a slow decrease in its population in

comparison to its opposite sex. The occurrence of male crabs slowly decreased from
50% to 40% during September 2017 up to December 2017. Then the population
increased from December 2017 (40%) to January 2018 (50%) by 10% and finally
reaching its lowest point in February 2018 where a 30% occurrence of male crabs
were observed.

100%
90%
80%
70%
% OCCURANCE

60%
50%
Female
40%
Male
30%
20%
10%
0%
September October November December January February

MONTH

Figure 4.12 Percent occurrence of male and female C. natator from September 2017
to February 2018 on crabs collected from Estancia, Iloilo (n=60).

4.5. Gonadosomatic Index (GSI)

24
 The Gonadosomatic Indices (GSI) were only obtained for the months of
September 2017 to February 2018. The values of the monthly GSI of both female and
male C. natator were plotted and compared in Figure 4.13.

The mean gonadosomatic index of the females for the month of October 2017
was 1.82. The GSI showed a sharp increase during the month of November 2017 with
a value of 3.53, then a sudden drop during the month of December attaining its lowest
peak having a mean GSI value of 0.97 and then a continuous increase was observed
throughout the following months. It slightly increased in January 2018 with a mean
GSI value of 1.59 then a drastic increase was observed in the final month, February
2018, attaining its highest peak of 3.67 mean GSI value.

A slight increase of GSI value were observed between the months of October
2017 (1.10) and November 2017 (1.54) then a continuous decrease of mean GSI
values throughout the following months were observed. The month of December 2017
attained the lowest peak for the male C. natator having 0.35 GSI value and an
increase was also shown in the next month, January 2018 (0.99). In February 2018, a
decrease was observed with 0.79 mean GSI for this month. The highest peak was
attained during November 2017 (1.54). No mean GSI value for the month of
September because there were no data for gonad weight were recorded.

3.5

2.5
mean gsi %

2
MALE
1.5 FEMALE
1

0.5

0
Octo ber Nov ember December J a nua ry F ebr ua ry

Month

Figure 4.13 Comparison of the monthly mean gonadosomatic indices (GSI) of male
and female C. natator for October 2017 to February 2018 from Estancia, Iloilo
(n=60).

25
4.6. Gonad Index (GI)

The mean gonad indices (GI) are plotted with the percent occurrences of the
different gonad maturation stages from September 2017 to February 2018 to show the
relationship of the GI to the presence of gonad stages in each month. The monthly
mean gonad indices (GI) are shown in Figure 4.14 for the females and Figure 4.15
for the males.

In September, the mean GI value of the females was 1.4 and increased to 1.6
in October this is due to the decrease of occurrence of samples with Immature ovary
stage. The following month, November showed an increase in the mean GI with a
value of 2.33 due to the absence of Immature ovary stage and an increase in the
occurrence of Late Maturing stage which significantly increased the mean GI value. A
decease was observed in December with a mean GI value of 1.5 because of the
absence of Late Maturing and Fully Mature ovarian stages and more occurrence of
Immature and Early Maturing stages. There was a slight increase in the GI in the
month of January, this was due to the higher occurrence of both early and Late
Maturing stage, with a GI value of 1.8. The last month, February, showed a sudden
increase of mean GI value attaining the highest peak with a GI value of 2.71 where
occurrence of Fully Mature female crabs and absence of Immature females were
observed in the particular month (Figure 4.14).

For the males, the mean GI in September was 1.8 which slightly decreased in
the next month, October, with a mean GI of 1.6 due to increase in the occurrence of
Immature testes. A slight increase of mean GI was seen in November due to the
absence of Immature testes, however, a drastic decrease of gonad index was observed
in the next month, December, with a mean GI value of 1.25 because of an increase of
Immature testes. In January, and increase was observed with a mean GI of 1.8.
Finally, in February, a decrease was observed with a mean GI of 1.67 due to the
decrease of mature testes in comparison to the previous month (Figure 4.15).

A high GI value means there were more females in later stages of

development, while low GI indicates a higher occurrence of females in earlier stages
of development.

26
 100% 1
90% 0.9
80% 0.8

GONAD INDEX (GI)

70% 0.7
% OCCURANCE

60% 0.6
50% 0.5
40% 0.4
30% 0.3
20% 0.2
10% 0.1
0% 0
September October November December January February

Month

Immature Early Maturing Late Maturing

Fully Mature GI

Figure 4.14 Percent occurrence and gonad indices (GI) of the different stages of
ovarian maturation in female C. natator retrieved from Estancia, Iloilo (n=34).
High GI values for females coincided with the months having high occurrence
of Late Maturing (stage III) and Fully Mature gonad stages (stage IV).

100% 1
90% 0.9
80% 0.8
GONAD INDEX (GI)

70% 0.7
% OCCURANCE

60% 0.6
50% 0.5
40% 0.4
30% 0.3
20% 0.2
10% 0.1
0% 0
September October November December January February

MONTH

Immature Maturing Mature GI

27
Figure 4.15 Percent occurrence and gonad indices (GI) of the different stages of
gonad maturation in male C. natator retrieved from Estancia, Iloilo (n=26). High GI
values for males coincided with the months having high occurrence of
maturing (stage II) and mature (stage III) gonads.

CHAPTER 5

DISCUSSION

In the present study, the breeding cycle of the Charybdis natator was
monitored for six months, the stages of gonad development of both male and females
were identified, and macroscopic appearance of the gonads were described. For the
female crabs, four stages of gonad development were identified namely: a) Immature
(Stage I), b) Early Maturing (Stage II), c) Late Maturing (Stage III), and d) Fully
Mature (Stage IV). For male C. natator, three stages of gonad development were
identified, Immature (Stage I), Maturing (Stage II), and Mature (Stage III). The
classification for the maturation of the female gonad stages of C. natator were based
on from the paper by Ikhwanuddin et al. (2010) which classified the ovarian
development of blue swimming crab, Portunus pelagicus, in Malaysia, while for the
male gonad maturation stages were based on the paper of Soundarapandian et al.
(2013) which was used to classify the gonad maturation stages of both male and
female flower crab, Charybdis feriata, in India.

The classification of the ovarian maturation stages of female crabs reported by

different authors on various crabs, was more or less identical with each other. The
classification of gonad maturation stages conducted by Liu et al (2014) on Portunus
pelagicus crabs, coincides with the present study. The stages IV and V identified by
Liu et al (2014) are merged as Fully Mature ovary (Stage IV) in the present study as
described by Ikhwanuddin et al (2010) on P. pelagicus. The Stage I or the resting
stage described by Batoy et al. (1983) on P. pelagicus, agrees with the Immature
Stage of Ikhwanuddin et al (2010) where gonads are degenerate, shrunken, and have
no to minimal coloration which makes the ovary difficult to distinguish from the
hepatopancreas. Stage II or developmental stage, which includes Early Maturing and
Late Maturing stages, described by Batoy et al. (1987), is similar to the description by
Ikhwanuddin et al. (2010) as Early Maturing (Stage II) and Late Maturing (Stage III)
stages and in the present study. The identified stage 3 or ripe stage by Batoy et al.

28
(1983) is similar to the identified Fully Mature stage (Stage IV) of the present study
which also includes Stage 4 (spawning and redeveloping stage) by Batoy et al (1983).
However, the last stage, spent stage, identified by Batoy et al (1987) was not
identified in this present study on Charybdis natator, which according to the
description by de Souza and Silva (2009) on Goniopsis cruentata, in the post
Spawning stage or the Spent stage, the ovaries revert back to being white which
makes it harder to distinguish from Previtellogenic stage or Immature stage of the
ovaries. Also, after mating, females migrate to high-salinity waters in lower estuaries,
and nearshore spawning areas (FWC-FWRI, 2006), which means that there is low
probability of female crabs with spent stage to be caught by the fishers since their
target area of catching crabs are in the pelagic zones where sea water is at the depth of
ten to fifteen meters since C. natator crabs are caught underneath the rocks.

The development of gonad maturation stages of male C. natator in the present

study are similar to those other crabs belonging to the family Portunidae such as the
Scylla serrata (Quinitio et al., 2007), Charybdis feriata (Soundarapandian et al.,
2013), and Portunus sanguinolentus (Sukumaran et al., 1998) where in three male
gonad stages were identified as a) Immature, b) Maturing, and c) Mature stages.

For the sex ratio, results showed that throughout the sampling period, there
were lesser number of males (n=26) than the occurrence of females (n=34).
Generally, the trend, as reported by Sumpton (1990), is population of male crabs
outnumbers the population of female crabs by ratio of 2 to 1. Findings of Germano et
al. (2006) coincides with the results identified by Sumpton (1990). Germano et al.
(2006) reported that males generally outnumbered female blue crabs which indicates
that males are generally active and more vulnerable to fishery. In present study
females outnumbered all the male crabs, especially on the month of February 2018,
wherein females (70%) dominates the population of males (30%) by thirty percent
which slightly coincides with that of Germano et al. (2006) where females were more
abundant in February, March, August, October and December. This inconsistency in
the monthly occurrence of male versus females, is due to bias of the fishermen as they
catch more female in this particular month because of the abundance of ovigerous
females. Fishermen tend to catch more gravid females because of their higher value in
the market because of the presence of crab eggs. In addition, the smallest size at
sexual maturity for both male and female C. natator in Estancia, Iloilo may be even

29
smaller because the samples that were collected were dependent on the crabs available
in the market which means that crabs of smaller size were set aside and were not sold
in the market since fishermen and market vendors would only benefit from crabs of a
larger marketable specimens which demand higher price than the small ones.

The minimum size at sexual maturity recorded in this present study coincides
with the previous study on Charybdis natator done by Sumpton (1990). According to
Liu et al. (2014), females with vitellogenic ovaries were considered to be mature
individuals (developmental stages III to V). Thus, for the present study, individuals
with Late Maturing stage of development were also considered to be mature. Thus,
for the present study, individuals with Late Maturing stage of development were also
considered to be mature. The carapace size of females having Late Maturing to Fully
Mature ovary ranged from 72 mm to 90 mm CW with the mean carapace size of 88
mm CW, while for the male C. natator, sizes of crabs having mature testes ranged
from 90 mm to 111 mm CW with the mean size of 99 mm CW. The minimum size of
both female and male C. natator coincides with that of Sumpton (1990) where in the
minimum size were both 71 mm carapace width (CW). The minimum size at sexual
maturity determined in the present study, involving C. natator, appears to be smaller
than that of the minimum size at sexual maturity for C. feriatus identified by
Soundarapandian et al. (2013) where they reported that the minimum carapace size of
a mature male C. feriatus measured 95 mm CW and for the females, they attained
sexual maturity at the size at 91 mm CW. In comparison with Soundarapandian et al.
(2013), the present study has the smallest size at sexual maturity for both male (90
mm CW) and female C. natator (72 mm CW). The differences in the results of the
present study from the results from previous studies could be due to sampling bias. In
the present study, the samples were not collected directly from the ocean, instead, the
samples obtained were the crabs available and were bought in the market, which was
pre-selected in the buying station where crabs were distributed.

In the present study, Late Maturing (Stage III) females and Fully Matured
(Stage IV) females were not present in all months of the sampling period, however,
high occurrence rate of Immature (Stage I) females were prominent. Fully Mature
(Stage IV) females were only present in the last month of sampling in February 2018,
in which is at its peak at forty-three percent. This is similar to that of Batoy et al.
(1987) which reported that the highest percentage of ripe individuals occurred in

30
February, while Germano et al. (2006) reported that the highest percentages of ripe
stages were found in February to May. Although, the results in the present study for
the highest peak of occurrence of Fully Mature (Stage IV) females coincides with
both of Batoy et al. (1987) and Germano et al. (2006), both studies reported that ripe
individuals and spawning female crabs, which are both identified in the present study
as Fully Mature (Stage IV), were observed throughout the year. However, the present
study did not coincide with the results reported by Sumpton (1990) where Fully
Mature female C. natator are at its peak during the months of October and March.
Due to the low occurrence of Fully Mature (Stage IV) female C. natator and the high
incidence of Immature (Stage I) females means that the present study indicates that C.
natator does not spawn year-round. The low occurrence of mature female is because
during the onset of rainy season, females with mature gonads would migrate to
higher-salinity waters in order to release their eggs (spawn) because this is a favorable
season for the larvae to feed and grow. Moreover, Fisher (1999) reported that crabs
mature faster as temperature and salinity increase. Heavy rainfalls during May could
have caused the water’s temperature to drop as well as dilute the water causing the
salinity to fall.

The male samples, however, did not coincide with the result of Baylon & Tito
(2012). The differences in the results of the present study maybe due to the small
monthly sample size gathered and examined for gonad maturation stages which is
only ten samples per month. This gives a low probability of having a high variation of
male to female ratio which will lessen the variation of the different gonad maturation
stages in male crabs.

Percentage distribution of gonadal stages serve to prove homogeneity or

heterogeneity of breeding habits of each species of crab (Batoy et al, 1987). In the
case of the present study, it has shown that C. natator display heterogeneity of the
breeding population because different gonad stages were found in every month in
both males and females, which is the same as the results of Batoy et al. (1987) and
Germano et al. (2006).

The gonadosomatic index (GSI) is a measure of the reproductive condition of

a population (Baylon & Tito, 2012). This means that the higher the GSI, the greater
the reproductive activity is for a given time. Both female and male GSIs showed
monthly fluctuations. The GSI trend for female C. natator agrees with previous

31
studies wherein high fluctuations can be observed due to the higher occurrence of
female C. natator with Immature (Stage I) ovary in almost every month which is also
present in the GSI trend of male C. natators. However, even if high fluctuations were
both reported in the present study, female GSI values is higher than males since male
gonad weight has minimal contribution in the total body weight of the crab. Females
generally have higher mean GSI compared to the males, which matched the results by
Germano et al. (2006) explaining that females have higher gonad weight relative to
body weight. Male GSI values reported in previous studies shows a constant trend
wherein there is zero to minimal fluctuations observed which differs from the present
study because of the low occurrence of mature male gonads and the low occurrence of
male crab population compared to females in monthly sample especially in the last
month wherein only three male crabs which was equally distributed among the three
gonad maturation stages. In addition, changes in the GSI can also be caused by certain
factors such as the salinity. Germano et al. (2006) reported that low GSIs of the
female blue crabs coincided with high salinity suggesting that salinity could be a
possible reproductive controlling factor. Environmental factors such as temperature
and salinity can also affect size at sexual maturity in crabs. Fisher (1999), who
investigated the effect of temperature and salinity on size at sexual maturity of female
crab (Soundarapandian, 2013). Physicochemical analysis, which includes salinity
determination, was not included in the scope of the present study, and therefore, we
cannot deduce that it be the factor that could have contributed to the aberration of the
result from previous related studies.

Gonad index is a measure of sexual maturity of a population (Baylon & Tito,

2012). A high GI value means there were more individuals with gonads in later stages
of development, while low GI indicates a higher occurrence of individuals with
gonads in earlier stages of development. The Gonad Indices (GI) of females is lowest
in the September 2017 (1.4) and reached its peak in the last month of sampling,
February 2018 (2.71). The result of the present study did not coincide with the
previous study conducted by Sumpton (1990). GI values are calculated to determine
on which months have more individuals that attained a higher gonad maturation stage.
In the present study the September 2017 had the lowest GI value which did not
coincide with the results determined by Sumpton (1990), in which September and
February counted the most population of Fully Mature female crabs because of high

32
occurrence of gravid females in this particular months. For the male GI values, the
present study was not parallel to any previous studies which showed a minimal
variation in the monthly mean GI values. High variation of present studies means that
there were some inconsistencies in the number of mature male catch during the
monthly sample which was different to previous studies because there were always
mature male crabs available for every month. The high variation in the GI values and
the low occurrence of mature male crab may be because of the low sampling size
every month which lessened the probability on the discovery mature male crabs.
According to Batoy et al. (1987), the factors that contribute to the breeding, sexual
maturity and fecundity of P. pelagicus included in their study were temperature, water
transparency and dissolved oxygen content, which were reported to have slight
correlations with the monthly gonad index fluctuations but were not statistically
significant.

In the six-month sampling duration from September 2017 to February 2018,

the first week of the month sampling was not followed due to certain conditions that
were encountered, such as high tides and heavy rains accompanied by strong wind, in
the monthly catch of Charybdis natator. High tides affected the monthly harvest
because the sea water has increased which made it difficult for the fishermen to obtain
the trapping device to catch crabs beneath the substrate. According to the local crab
fishers in the community, there is usually more harvest during the late summer up to
the rainy months of May to July. Aside from the ridged swimming rock crab, there are
also other species that are included in the daily which are highly marketable than the
crab species used in the study such as Charybdis feriatus and Portunus pelagicus.
However, during April 2018 until May 2018, there zero occurrence of C. natator in
the markets according to local vendors.

Even though there is a constant amount of harvest every month, aside for
periods where there is interference, there is indeed a need for implementing a
regulation that should limit the catch of these crabs because even though there is still
no alert in the decrease of the population of Charybdis natator, management and
control over wild stock could protect the remaining stock from extermination. This
inconsistency in the monthly occurrence of male versus females, is due to bias of the
fishermen as they catch more female in this particular month because of the

33
abundance of ovigerous females. Fishermen tend to catch more gravid females
because of their higher value in the market.

CHAPTER SIX

CONCLUSION AND RECOMMENDATION

In this study, four stages of ovarian maturation for females, and three stages of
gonad maturation for the males, were determined. The female gonad maturation
stages are Immature, Early Maturing, Late Maturing and Mature. The male gonad
maturation stages were Immature, Maturing and Mature. The said stages were
classified macroscopically based on the color, shape, and size of the gonads.

Charybdis natator exhibited heterogeneity in its breeding habit since the

present study showed occurrence of different gonad stages per month which was
shown in the monthly percentage occurrence of each gonadal stage. Almost every
month there were Immature (Stage I) ovary present with a low percent occurrence of
Fully Mature females which was only present in the last month, February 2018, which
was also its peak with 43%. For males, mature gonad stages were present in all
months, except in October 2017, showing its peak at September 2017 at 40%.

The minimum size of females having Late Maturing to Fully Mature ovary
ranges from 72 mm to 93 mm carapace width (CW) with a mean carapace size of 83
mm CW. For males, minimum size at maturity ranges from 90 mm to 111 mm CW
with a mean size of 85 mm CW. The results coincide with the previous study on
Charybdis natator reported by Sumpton (1990) having the minimum size of maturity
at 71 mm up to its maximum of 105 mm CW for males and 95 mm CW for females.

There was lower occurrence of male crabs (n=26) than female crabs (n=34)
obtained in the samples which was different from the previous samples of other

34
researchers. Sumpton (1990) and Germano et al. (2006) reported that the population
of male crabs outnumbers the population of female crabs. Female crabs were
particularly higher in the month of February 2018 (70%) than the males (30%).

Female and male GSIs obtained from the present study showed monthly
fluctuations. Females generally have higher mean GSI compared to the males. This
means that females have higher gonad weight relative to body weight. The anomaly in
the monthly fluctuation of male GSI values was due to the low population of mature
male crabs than the Immature male crabs.

The Gonad Indices (GI) of females is lowest in September (1.40) and reached
its peak in the last month (2.71), while males attained the highest peak in September
and January (1.8), however, in the GI value in the month of January only tied with
that of September because of the higher occurrence of maturing males, and the
fluctuations in the plotted Gonad Indices were also because of the low occurrence of
mature male stages. Months with high GSI and GI values coincided with the months
having high occurrence of later stages of development and low values of GSI and GI
coincided with months with more individuals in their earlier stages of development.

It is recommended that this study on the breeding cycle of Charybdis natator

in Estancia, Iloilo be continued for six more months to complete the one-year data
which also includes the histological analysis of the gonads to further differentiate the
gonad development in microscopic level and to further differentiate post-spawning
stages from Immature stages since macroscopically both stages have the same
morphological characteristics. Another study should include analysis of growth and
breeding patterns as influenced by various environmental factors such as temperature,
abundance of food and salinity. Further studies should also be done on the fecundity
and as well as the data for the monthly population of ovigerous females in the catch.

The information obtained from the present study and could be obtained by
completing this study for six more months to complete the anual data of the breeding
cycle, would significantly help in implementing regulatory measures and municipal
protocols imposing off season and minimum legal-size limit on catching these crabs,
and also prohibiting the catch of gravid females. This will be beneficial for the
preservation and management of wild stock in the coastal areas of Estancia, Iloilo,
Western Visayas, Philippines

35
36