CHAPTER 1

INRTODUCTION

Fisheries is one of the major industries found here in the Philippines, given the
location of the country, since it is found in the Pacific Ocean. The fisheries in the
Philippines are mainly composed of fishes, and shellfishes. One of the major marine
life products that is being marketed are crabs. Charybdis natator (Herbst, 1794)
commonly known as the “ridged swimming crab”, belongs to Family Portunidae, a
typical group of marine crabs which is widely distributed across Asia, Australia, and
Africa (Wee and Ng, 1995). In the 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. 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 Asutralia (Sumpton, 1990).

Crabs live in burrows in sandy beaches, mud in the rivers, and below the rocks
at the bottom of the ocean. But during mating season, large population of mature
crabs migrate to the ocean to copulate and sometimes this can be intervened by the
fisherfolk, and this could be a huge problem because if crabs fail to copulate, there
will be lesser population of crabs in the next generation.

This study aims to determine and describe the reproductive stages of C.

natator 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) the
histological appearance of the gonads of C. natator at each stage of development, (3)
size at sexual maturity of the crab, (4) the percent occurrence of each gonadal
maturation stage, and (5) the percent occurrence of male and female crabs from
September 2017 to February 2018.

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.
 CHAPTER 2

REVIEW OF RELATED LITERATURE

Description

Charybdis natator (Herbst, 1794) commonly known as the “ridged swimming

crab”, 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, which are flattened, that acts as propellers for them to swim
efficiently. They are called ridged swimming crabs because of the ridges in their
carapace (Samuel, 2014).

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).

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.
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 contribute to crab fisheries in India and Asutralia (Sumpton, 1990).

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. This is
maybe because of the spawning season of the C. natator during June and July. 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 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).

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; Ikhwanuddin et al., 2012). 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.

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.

Histological examination

In histological examination there is a certain protocol that must be followed on

how to prepare the histological slides of the gonads of each crab. After the
macroscopic analysis of the gonads, portions of each stage of the gonads will be
placed in small vials half-filled with Bouin’s solution and will be stored for 24 hours
at room temperature. The gonads will be dehydrated through a series of increasing
alcohol concentration. Toluene will be added to remove the alcohol. The gonads will
then be placed in soft Paraffin wax for two hours. The gonads will be embedded in
hard Paraffin wax for several hours and the excess wax will be trimmed afterwards.
Thin sections will be prepared using a microtome (5µm), tissues will be stained using
haematoxylin and eosin, and will be left to dry overnight. Each section will be
mounted in slides, will be covered with cover slips and will be labeled with respective
date and crab number.

In female crabs, there are four histological stages that can be observed (de
Souza et al., 2009). The four stages of the female gonad development are: (1)
Previtellogenic stage, (2) Early-Vitellogenic Stage, (3) Late Vitellogenic Stage, and
lastly (4) Post-Spawning Stage. First, the previtellogenesis stage, oogonia may be
observed undergoing preliminary stages of meiotic division. The next stage, early-
stage vitellogenesis, the germinative zone can be seen that it is being compressed by
its surrounding previtellogenic oocytes. The oocytes may be observed as it is
undergoing early vitellogenesis. In the third stage, the mature vitellogenesis, the
ovaries are mostly filled with oocytes undergoing late-stage vitellogenesis. Mature
ovaries appear to be uniform and completely filled with mature oocytes, and lastly the
germinative zone is barely seen. Lastly, in the post-spawning stage, the ovaries can be
observed in a disarrayed arrangement due to the release of the mature oocytes outside
the system. Several previtellogenic oocytes and empty cavities can be observed in the
ovarian stroma along with a few counts of hamatocytesm fibers, and follicular cells.
The lining also assumes a distinct wavy appearance. There is also a restoration of the
ovarian arrangement along with the reduction of empty cavities and hematocytes.

In the male, the three stages of development that were identified histologically
are: (1) presence of spermatogonia in immature stage, (2) Presence of spermatogonia
and spermatocytes in maturing stage, and (3) formation of primary and secondary
spermatocytes, spermatids and spermatozoa in mature stage (Islam et al., 2013).

Fecundity

Fecundity is an index of reproductive capacity and is estimated by the number

of eggs produced by an organism (Baylon and Tito, 2012 “from” Reeby et al. 1990).
The eggs attached to the pleopods of ovigerous females will be scraped off to
determine the mean number of eggs per egg mass. Three replicates of 1 g sub-samples
will be taken and the number of eggs in each replicate will be counted under a
dissecting microscope (Baylon and Tito, 2012). The fecundity of each female crab
will be estimated by multiplying the mean number of eggs in the three replicates by
the total wet weight of the egg mass (Baylon and Tito, 2012 “from” Krajangdara and
Watanabe, 2005). To show the relationship of fecundity versus size of the female
crab, regression analysis will be used (Baylon and Tito, 2012).
 CHAPTER 3

MATERIALS AND METHODS

Collection site and sampling

A six-month survey of crab sampling was held during September 2017 to

February 2018. The sample was taken from Estancia Iloilo, delivered and was bought
in the wet market of Miagao, Iloilo. Ten samples were gathered in every first week of
the month. The samples shall have representatives of different sexes with sizes
ranging from the smallest to the largest for the determination of the fecundity and
gonad developmental stages.

Collected crabs is stored in the ice box for preservation, so that the tissues of
the crabs will no deteriorate, and for the crabs to be anesthetized since crabs tend to
have cannibalistic instincts. 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.

Determination of sexes

Male and female crabs can be described, and can be differentiated

morphologically through the different shapes of their abdominal flaps. 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 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.
Body Measurement

For each crab, the body weight (BW), carapace width (CW), and carapace
length (CL) will be measured. The body weight will be measured by patting the crab
dry using clean cloth or tissue after removing it from the refrigerator. This is to avoid
errors in the results. The crab will be placed on a digital electronic balance and the
weight will be recorded to the nearest tenth of a gram. Carapace width and length will
be measured using a Vernier caliper and will be recorded to the nearest millimeter.
Carapace width (CW) is the distance between anterior lateral spine and the most
posterior lateral spine (Brown, 2009). Carapace length (CL) is the distance between
the centers of the frontal interorbital carapace margin and the posterior margin.

Dissection

Using a pair of sharp dissecting scissors, the dissection of crab will proceed by
cutting the dorsal carapace from the posterior end of the carapace moving in
counterclockwise direction, making a square shaped incision. Then using a scalpel,
forceps, and dissecting needle, the thin membrane covering the cephalothoracic cavity
will be removed to expose the gonads. A Y-shaped structure of gonads, called the
anterior horn, will be 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 arterior part of the
cephalothoracic cavity. The hepatopancreas is extended posteriorly, connected to the
gastric mill.

Gonad analysis

The gonad developmental stages for all sexes of Charybdis natator will be
investigated using two methods: (1) macroscopic technique which will involve the
description of the external morphology of the gonads and (2) microscopic technique
which will involve histological examination of the gonads.

Morphological characterization
 The macroscopic technique will involve 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 Quinitio et al (2007) 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 will be analyzed based on this classification. The
male C. natator will be analyzed based on the three stages of gonadal development
according to Silva et al (2012): (1) immature, (2) maturing, and (3) mature. (more
detailed…)

Gonad analysis

Gonad analysis shall include computations of the gonadosomatic indices and

gonad indices. The quantitative gonadosomatic index (GSI) will be calculated using
the equation: GSI = gonad weight (g)/body weight (g) x 1000 (Krajangdara and
Watanabe, 2005). 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. The mean
value of gonad indices (GI) or the index of sexual maturity will then be calculated
using the equation: GI = Σ(n x WV)/N, where n is the number of individuals in a
given developmental stage; WV is the pre-weighted value for the developmental
stage; and N is the total number of crabs per monthly sample. Values of mean gonad
indices per month will then be plotted against time (months) to determine the
spawning pattern or reproductive cycle (Baylon and Tito, 2012). (separate the
equations from text)
 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 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.

The immature (Stage I) 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).

Female crabs of the early maturing stage (Stage II) showed change in the color
of the ovary which range from pale orange to peach. The ovaries were more
ostensible and it can be easily determined from the hepatopancreas but is still not yet
extending to the hepatic region (Figure 4.2)

Late maturing (Stage III) ovary showed severe increase in size and
appearance. The ovary appeared to have a color change as well, having a bright
orange coloration. The anterior horns of the ovary had 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).

The last observed stage of the ovarian maturation development showed a fully
mature (Stage IV) ovary showing a deep orange to scarlet coloration. A massive
enlargement of the ovary was visible, which the ovary 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; O = ovary; GM = gastric mill; G = gills;
C = cartilage; H = heart)

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; O = 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)
Figure 4.4 3 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. Immature (Stage I) gonads of male crabs are
thin, translucent to opaque white in color which lies on either side of the stomach.
Testes and the vas deferens are slightly convoluted and are difficult to distinguish
from each other (Figure 4.5).

Gonads of male crabs with maturing (Stage II) stage are slightly enlarged and
are creamy white in color. The testes and vas deferens are more well developed and
are distinguishable from the other. The testes, posterior to the stomach, extends
laterally to the hepatopancreas. Vas deferens is further differentiated into three
separations based on its location: anterior (AVD), middle (MVD), and posterior vas
deferens (PVD). AVD is enlarged and dorsal to MVD and PVD. MVD and PVD
further extends towards the heart (Figure 4.6). Male crabs of mature (Stage III)
gonadal stage showed further enlargement of testes and vas deferens, with a more
prominent white color, and are coiled and swollen which occupies the body cavity
(Figure 4.7).

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
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.
 12

10

OCCURANCE 8 2
1

6
2
5 1
4 4
7
3
2 4
1
2
1 1
0
70 - 75 76 - 80 81 - 85 86 - 90 91 - 95

CARAPACE WIDTH (MM)

immature early maturing late maturing spawning

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).

7
3
6
OCCURANCE

3 6 6
1 5
2

1 2
1 1 1
0
70 - 75 76 - 80 81 - 85 86 - 90 91 - 95 96 - 100 101 - 105 106 - 110 111 - 115

CARAPACE WIDTH (MM)

type I type II type III

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

 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 crab species

were analyzed for their different ovarian maturation stages. In September 2017, 80%
of the samples were immature and the remaining 20% that were examined belonged
in the late maturing stage. In October 2017, 60% of the female crab samples observed
was immature, 20% were at early maturing stage and the last 20% observed were
under late maturing stage. In November 2017, immature female crabs were only at
33.33% whereas the remaining 66.67% were female crabs at late maturing stage. In
December 2017, 50 % of the female crab samples were immature and 50% were at
early maturing stage. Three stages were observed during January 2018, 40% of which
were immature, 40% were early maturing, and the last 20% were late maturing female
crabs. And lastly for the month of February 2018, 28.57% of the female C. natator
were early maturing, also female with late maturing stages were also observed at
28.57% occurrence, and 42.86% were fully mature female C. natator crabs with no
occurrence of immature crabs. Fully matured crabs 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 2017, 20% were immature, 40%
were maturing, and the remaining 40% of the male crabs were matured. During
October 2017, 40% of the male crabs observed were still immature and 60% of the
male C. natator were maturing with no occurrence of matured male crabs. November
2017 showed no signs of immature crabs with 75% of the population were maturing
and the remaining 25% were already mature. Majority of the crabs during December
2017 were immature with 75% occurrence and 25% were mature male C. natator. In
January 2018, 20% of the male crabs were immature, 60% were still maturing, and
the other 20% of the male crabs were mature. February 2018 showed an equally
portioned stages of the male gonadal maturation, with all the gonad stages, from
immature to mature stages, showed 33% occurrence. Fully matured male crabs were
observed most of the months except during October 2017, with the highest number of
occurrence, among all the sampling month period, on September 2017 (Figure 4.11).
 100%
90% 20% 20% 20%
80% 43%
50%
70% 20%
67%
% OCCURANCE

60% 40%
50%
29%
40% 80%
30% 60%
50%
20% 40%
33% 29%
10%

0%
Sep-17 Oct-17 Nov-17 Dec-17 Jan-18 Feb-18

Month

immature early maturing late maturing spawning

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%
25% 25% 20%
33%
80% 40%
70% 60%
% OCCURANCE

60%

50% 60% 33%

40% 40% 75% 75%
30%

20% 40%
33%
10% 20% 20%
0%
Sep-17 Oct-17 Nov-17 Dec-17 Jan-18 Feb-18

Month

type I type II type III

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
 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%
50% 50% 50%
70% 60% 60%
70%
% OCCURANCE

60%
50%
Female
40%
Male
30%
50% 50% 50%
20% 40% 40%
30%
10%
0%
Sep-17 Oct-17 Nov-17 Dec-17 Jan-18 Feb-18
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)
 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 drastic 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).

4 3.67
3.53
3.5

2.5
mean gsi %

1.83
2
1.54 1.59 MALE
1.5 FEMALE
1.10 0.99
0.97
1 0.79

0.5 0.35

0
Oc t -1 7 No v -1 7 D e c -1 7 J an -1 8 F e b -1 8

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).
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 2017, the mean GI value of the females is 1.4 and increased to
1.6 in October 2017 this is due to the decrease of occurrence of samples with
immature ovary stage. The following month, November 2017 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 following decease was observed in December 2017 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 were observed. There
was a slight increase in the GI in the month of January 2018, this is due to the higher
occurrence of both early and late maturing stage, with a GI value of 1.8. The last
month, February 2018, 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 2017 was 1.8 and then slightly
decreased in the next month, October 2017, 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 2017 due to the absence of immature testes, however, a drastic decrease of
gonad index was observed in the next month, December 2017, with a mean GI value
of 1.25 because of an increase of immature testes. In January 2018, and increase was
observed with a mean GI of 1.8. Finally, in February 2018, 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.
 1 1 1 1
100% 1 1 1
90% 20% 20% 20% 0.9
80% 43% 0.8
50%
70% 20% 0.7

GONAD INDEX (GI)

% OCCURANCE 67%
60% 40% 0.6
50% 0.5
40% 80% 29% 0.4
30% 60% 0.3
50%
20%
33% 40% 0.2
29%
10% 0.1
0% 0
Sep-17 Oct-17 Nov-17 Dec-17 Jan-18 Feb-18
MONTH

immature early maturing late maturing

spawning 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).

1 1 1
100% 1 1 1 1
90% 25% 25% 20% 0.9
33%
80% 40% 0.8
70% 60% 0.7

GONAD INDEX (GI)

% OCCURANCE

60% 0.6
50% 60% 33% 0.5
40% 40% 75% 75% 0.4
30% 0.3
20% 40% 0.2
33%
10% 20% 20% 0.1
0% 0
Sep-17 Oct-17 Nov-17 Dec-17 Jan-18 Feb-18
MONTH

type I type II type III GI

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
DISCUSSIONS