Aquaculture Reports 2 (2015) 50–57

Contents lists available at ScienceDirect

Aquaculture Reports
journal homepage: www.elsevier.com/locate/aqrep

Mating behaviour of the orange mud crab, Scylla olivacea: The effect of
sex ratio and stocking density on mating success
Khor Waiho, Muhamad Mustaqim, Hanafiah Fazhan, Wan Ibrahim Wan Norfaizza,
Fadhlul Hazmi Megat, Mhd Ikhwanuddin ∗
Institute of Tropical Aquaculture, Universiti Malaysia Terengganu, 21030 Kuala Terengganu, Terengganu, Malaysia

a r t i c l e i n f o a b s t r a c t

Article history: Mud crabs from the genus Scylla have high commercial value and are considered as one of the highly
Received 11 January 2015 sought luxury seafood items. Thorough understanding about their biology and mating behaviour is vital
Received in revised form 17 July 2015 in providing important information for a sustainable exploitation and future incorporation into the aqua-
Accepted 19 August 2015
culture industry. The mating process of S. olivacea lasted 82.0 ± 10.8 h was divided into four phases:
Available online 29 August 2015
precopulation, molting, copulation, and postcopulation. Courtship displays and fighting were shown by
mature males while they were courting females. Precopulatory position lasted for 55.2 ± 10.8 h before the
Keywords:
pairs disengaged for the female to molt. The molting process was 4.6 ± 0.3 h. Copulation (mean duration
Orange mud crab
Mating behaviour
was 6.6 ± 0.5 h) occurred while the female’s exoskeleton was still soft. Postcopulatory guarding lasted
Sex ratio for 13.6 ± 0.6 h. Separation of the mating pairs indicates the end of postcopulation phase. Mating success
Stocking density percentage was unaffected by sex ratio, but inversely affected by stocking density. Cumulative mortality
Scylla increased with increasing stocking density and unequal sex ratios. Postcopulatory guarding duration was
significantly shorter in treatment with 1 male:2 females ratio and treatment with the lowest stocking
density (2 crabs m−2 ). We proposed rearing of mud crab broodstocks for mating purpose using sex ratio
of 1 male:2 females and stocking density of 6 crabs m−2 to maximize output (successful mating pairs)
while maintaining low mortality percentage and shorter postcopulatory guarding duration.
© 2015 The Authors. Published by Elsevier B.V. This is an open access article under the CC BY-NC-ND
license (http://creativecommons.org/licenses/by-nc-nd/4.0/).

1. Introduction Understanding of the mating behaviour is a paramount com-

ponent of the biology of any organism. Mating in mud crabs is
Mud crabs of the genus Scylla (Brachyura: Portunidae) are dis- known to take place in intertidal zones and estuaries before the
tributed throughout the Indo-Pacific region and are commonly females migrate as far as 50 km offshore to spawn their eggs
found in intertidal zones of mangrove forests and estuaries (Keenan (Koolkalya et al., 2006; Robertson and Kruger, 1994). The mating
et al., 1998). These crabs are of high demand both in the local strategies in eubrachyuran crabs can be divided into six groups
and international markets because of the sweet and delicate taste based on the difference in mating-molting link (soft-shell mat-
of its meat. Out of the four species of Scylla (S. serrata, S. oli- ing or hard-shell mating), growth pattern (indeterminate growth
vacea, S. tranquebarica and S. paramamosain) S. olivacea is popularly or determinate growth) and seminal receptacle structure (dorsal
used for aquaculture because of its hardy nature and ease of cap- or ventral seminal receptacle) (McLay and López Greco, 2011).
ture. Recently, mud crab farming has gained much attention in These 6 groups are classed into two main groups based on their
Malaysia. However, most of these farming activities involve only growth patterns, separating those with determinate growth (i.e.,
crab fattening and the production of soft-shelled crabs, with the Portunoids and Majoids) and indeterminate growth (i.e., Eriphioids,
use of captured juveniles from the wild (Ikhwanuddin et al., 2013). Xanthoids, Cancroids, and Grapsoid-Ocypodoids). Portunoids, hav-
This dependency on wild caught mud crab juveniles is due to the ing dorsal seminal receptacle and mate during soft-shell condition
unavailability of adequate knowledge on the biology and larval are grouped separately from Majoids (possess ventral seminal
rearing of mud crabs (Williams and Primavera, 2001). receptacle and mate during hard-shell condition). Among the mat-
ing strategies employed by members in Portunidae family are the
display of courtship behaviors, prolonged precopulatory and post-
∗ Corresponding author. Fax: +60 9 6683390. copulatory guarding, the presence of sperm plug, and the absence of
E-mail address: [email protected] (M. Ikhwanuddin). hinged opercula in vulvae (McLay and López Greco, 2011). Previous

http://dx.doi.org/10.1016/j.aqrep.2015.08.004
2352-5134/© 2015 The Authors. Published by Elsevier B.V. This is an open access article under the CC BY-NC-ND license (http://creativecommons.org/licenses/by-nc-nd/4.
0/).
 K. Waiho et al. / Aquaculture Reports 2 (2015) 50–57 51

studies (Davis, 2004; Koolkalya et al., 2006; Robertson and Kruger, the opposite sex. Molting phase involves crabs showing no move-
1994) have looked into the spawning biology and characteristics of ment, followed by shedding of their old carapaces. Copulation
wild-caught berried females from of the genus Scylla, but little has process is identified when the male started to flip over the newly-
been published on their mating behaviour prior to spawning. molted female and subsequently positioned his sternum within the
Sex ratio and stocking density are important elements affecting female’s opened abdomen, and the male’s gonopods were inserted
the mating success of mud crabs when reared in a confined area into the female’s gonopores. The postcopulatory guarding was
where space is a limiting factor (Naranjo-Paramo et al., 2004). This recorded when the male released the female and positioned her
is because mud crabs are well known for their territorial, aggressive underneath his abdomen, caging her with his legs.
and cannibalistic behaviors (Mirera and Moksnes, 2013). A thor-
ough understanding on the mating behaviour of mud crabs and the 2.3. Sex ratio and stocking density
effect of different sex ratio and stocking densities on the mating
process is essential to ensure selective breeding and for mating of To test the effect of sex ratio on the mating success of S. olivacea,
mud crabs to be successful in hatchery conditions. 27 pairs of mud crabs (n = 54) were randomly divided into three
This study describes the mating behaviour of orange mud crab, different male:female sex ratios, i.e., 1:1 (9 males:9 females), 2:1
S. olivacea under controlled laboratory conditions and the effect of (12 males:6 females) and 1:2 (6 males:12 females).
different sex ratios and stocking densities on mating success. The effect of stocking density on the mating success of S. olivacea
was separately investigated as well by stocking 2, 6 or 10 pairs of
2. Materials and methods crabs in each tank (n = 36). This is equivalent to approximately 2, 6,
or 10 crabs m−2 . A sex ratio of 1 male:1 female was applied.
2.1. Sampling Both the sex ratio and stocking density experiments were
conducted in triplicate, using a total of 270 crabs. Disinfection
S. olivacea were collected from Matang Mangrove Forest and quarantine period were the same as described in the previ-
Reserve, Perak, Malaysia (4◦ 45 58 N; 100◦ 37 31 E). Standard crab ous experiment. Environmental conditions, water level, feeding
pots with trash fish as bait were set up during low tides, left regime, tank dimensions, marking of individual crabs and water
overnight, and the crabs were harvested the next day during high exchange were similar to that used in mating behaviour exper-
tide. The identification of the S. olivacea species was based on iment. Crabs were monitored daily throughout the experiment.
the morphological description provided by Keenan et al. (1998). Dead or weak crabs were recorded, removed from the tanks and
The carapace width (CW) of selected mature males ranged from replaced with new crabs to compensate for mortality and maintain
110.23 mm to 124.49 while the CW of immature juvenile females stocking density. The experiment was carried out with a maximum
ranged from 65.33 mm to 78.25 mm (Ikhwanuddin et al., 2010, rearing period of 60 days.
2011). Immature juvenile females that were about to undergo Copulated females were transferred to individual tanks after
pubertal moult were used to ensure that they were unmated. The their male counterparts released them from post-copulatory guard-
specimens were transported back in dry condition on the same ing. They were maintained for at least a week before being brought
day to marine hatchery of Institute Tropical Aquaculture, Universiti into laboratory to check for the presence of sperm in the females’
Malaysia Terengganu, Malaysia. seminal receptacle (i.e., spermatheca). Successful sperm deposition
by males into females after copulation was considered as mating
success.
2.2. Mating behaviour

2.4. Sperm examination

Crabs were first disinfected with 150 ppm formaldehyde for
30 min and acclimatized for 3 days in tank with a salinity of 30 ppt
Copulated females were anesthetized by dipping into ice water
(Baylon, 2011). The crabs were marked using water-resistant mark-
for 20 min. They were then dissected to examine the presence
ers on their carapace and divided into 2 tanks (2 m length × 1 m
of sperm packets in their seminal receptacle which are located
width × 1 m height). Each tank consisted of 10 males and 5 females,
inside the gonopores. The deposited spermatophores appeared to
hence a total of 30 crabs were used. They were reared until spawn-
be small, round, solid and whitish in colour when viewed with the
ing occurred, with a maximum rearing period of 60 days. All males
naked eye. For confirmation, they were diluted with few drops of
were used only once and promptly removed from tank after mating
saline solution and examined under stereozoom microscope (Nikon
with a female had ended. The water level had a depth of 50 cm and
AZ100 M).
5 PVC pipes with open ends (diameter = 10 cm, length = 20 cm) per
tank were provided as shelter. Optimum environmental conditions
were maintained throughout the experiment (salinity 25 ppt; tem- 2.5. Data analysis
perature 26–30 ◦ C; natural photoperiod). Feeding with a mixture of
squid Loligo sp. and trash fish Decapterus sp. at approximately 10% of The effect of sex ratio and stocking density on mating
crab’s body weight was done twice a day while water exchange was success, cumulative mortality and duration of copulation and post-
conducted once every 3 days. Uneaten feed and excretory wastes copulation guarding stages in S. olivacea were compared. All data
were siphoned out 1 h after each feeding period. Observation for were checked for normality of distribution and homogeneity of
any mating behaviour was carried out twice daily (12 h interval). variance using Wilks-Shapiro and Bartlett’s test, respectively. Mat-
Once mating behaviour was observed, continuous observation of ing success percentage data were arcsine transformed and analysed
the mating pairs was conducted. During observations, PVC pipes using One-Way Analysis of Variance (ANOVA) to find out if there
were gently pulled away for a brief period to reveal the hiding pairs. was significant differences among the different treatments. This
Mating behaviour and duration of mating phases (hours ± standard was followed by Tukey’s test to find out which treatments were
error) for each mating pair were described and recorded via direct significantly different from each other (Sokal and Röhlf, 1981; Zar,
observation with the aid of another 4 researchers in shifts. Total 1984). Cumulative mortality of crabs in relation to time for each
mating duration was inclusive of precopulation, molting, copula- treatment (sex ratios and stocking densities) were fitted via poly-
tion and postcopulation process. The observation for precopulation nomial regression and compared graphically. Statistical analyses
process started when any crabs showed mating behaviors towards were performed using Microsoft Excel 2010.
52 K. Waiho et al. / Aquaculture Reports 2 (2015) 50–57

Fig. 1. The mating behaviour of S. olivacea. (A) Precopulatory caging. (B) Male guarding over newly molted female prior to copulation. (C) Male flipping female over, with
female’s abdomen readily opened. (D) Copulation between male and female. (E) Insertion of male’s gonopods into female’s gonopores. (F) Postcopulatory guarding.

3. Results towards the female. At the same time, the male walked slowly
towards her and tried to grab her using his walking legs. The female
3.1. Mating behaviour and duration either fought back or walked away by extending her chelipeds and
acting aggressively as a sign of rejection if she was not receptive.
The whole mating process of mud crab can be divided into four Receptive females became submissive, with chelipeds and all walk-
phases, namely (i) precopulation, (ii) molting, (iii) copulation, and ing legs remaining close to her abdomen. In some instances, other
(iv) postcopulation. Total mating sequence of S. olivacea (from pre- mature males tried to interrupt the courting process by joining
copulation to the end of postcopulation) lasted for 82.0 ± 10.8 h. in and fighting with the courting male. The male that success-
Out of all the mating phases, only the duration of the first phase of fully warded off other males was generally larger in size and more
mating (precopulation) varies largely among individuals. Seven out aggressive. The male crab then moved closer to the female and
of 10 pairs of mud crabs (from both tanks) successfully mated and grabbed her using his second walking leg and positioned her neatly
observations of mating behaviour were based on these successful under his abdomen. This position is known as the precopulatory
copulations. caging (Fig. 1A). Pairs stayed in precopulatory position even during
feeding. A male in this position always extended his front chelipeds
outwards and was very aggressive in warding off other competing
3.1.1. Precopulation
males. There were occasions when other males managed to sep-
Mature males were attracted to immature females as early as
arate the pair from their precopulatory position and replaced the
24 h after they were placed together in the tanks. The male began
initial courting male. The submissive female normally remained
to court the female by extending and waving its front chelipeds
 K. Waiho et al. / Aquaculture Reports 2 (2015) 50–57 53

100 the triangular and light coloured abdomens when they were imma-
ture. There were instances where the male aided the molting female
80 to extricate herself from her old exoskeleton by pushing her old
exoskeleton gently using his chelipeds. This male aiding behaviour
Mang success (%)

60 was observed in 2 mating pairs. However, complications may arise

during molting, causing unsuccessful molt which results in the
40 death of the female. Two out of 10 females experienced such con-
ditions and died before being able to completely molt out of their
old exoskeleton. It was observed that the guarding male would
20
promptly abandon the carcass of the dead female and other mud
crabs would then start to devour her.
0
If molting of the female crab was successful, some males would
1:1 2:1 1:2
cradle the newly molted female in the previous precopulatory
Sex rao (male : female)
caging position while others would guard their females by always
Fig. 2. The mating success percentage of S. olivacea in different sex ratios being close by without cradling them.
(male:female). Error bars represent standard deviation. Sample size for sex ratio
1:1 = 9 males:9 females, 2:1 = 12 males:6 females and 1:2 = 6 males:12 females.
3.1.3. Copulation
inactive while other males fought to get hold of her. However, there Right after molting, the male flipped the body of the female crab
were 2 occasions where the females rejected the winning male and over using his chelipeds and walking legs while her exoskeleton
moved away when he tried to cradle her. The rejected males in was still soft. Both of the crabs faced each other, with the male crab
both occasions were smaller in size compared to the other available on top and holding the female below him using his second and
males. On all occasions, males brought the females into a secluded third walking legs (Fig. 1C and D). During this time, female opened
shelter (PVC pipes in this experiment) for a more efficient guard- up her abdomen, exposing her gonopores to allow the insertion of
ing. The time required for males to successfully pair with a female the male’s gonopods and the transfer of sperm (Fig. 1E). The male
ranged from 1 to 15 days in captivity. The average time when the eventually turned the female back over which implies the end of
pairs were in precopulatory positions until right before molting was copulation. The whole copulation process lasted about 6.6 ± 0.5 h.
55.2 ± 10.8 h.

3.1.2. Molting 3.1.4. Postcopulation

The pairs disengaged from the precopulatory embrace position Males guard their female partners even after copulation to
when the female was ready to molt. Molting normally took place ensure that the females were able to harden their exoskeleton
within the shelter though some occurred in the open, with the male safely by cradling the females underneath their body (Fig. 1F). This
aggressively guarding over the female while she was in the pro- is also known as postcopulatory guarding. In some instances, males
cess of shedding her old exoskeleton (Fig. 1B). The female crabs do allowed females to harden in the shelter while they stood guard
not feed 1–2 days prior to molting. The molting process lasted for nearby. The hardening of new exoskeleton in newly molted females
4.6 ± 0.3 h. This pubertal molt represents the transition of female took 1–2 days. During this time, only the male exhibited locomo-
from immature to mature female. Sexual maturity was confirmed tion using his available walking legs while the female’s walking
by the change in the shape and coloration of the female’s abdomen. legs were tucked in towards her abdomen. The guarding males
Mature females have darkened and wide abdomens compared to were aggressive during this period. Postcopulation which lasted for

35

30
Cumulave mortality (individuals)

25

20
1:1
15 2:1
1:2
10

0
0 10 20 30 40 50 60
Time (days)

Fig. 3. Cumulative mortality of S. olivacea in different sex ratios (male:female) throughout the experiment (60 days). Curves were fitted with second degree polynomial
regressions for each sex ratio. Sample size for sex ratio 1:1 = 9 males:9 females, 2:1 = 12 males:6 females and 1:2 = 6 males:12 females.
54 K. Waiho et al. / Aquaculture Reports 2 (2015) 50–57

20 a
100
18
a
a
16 80
a

Mang success (%)

14
Duraon (hours)

12 60
b
10 copulaon b

8 40
6 postcopulatory guarding

4 20
2
0 0
1:1 2:1 1:2 2 6 10
Sex rao (Male:Female) Stocking density (crabs/m2)

Fig. 4. The copulation and postcopulatory guarding duration of S. olivacea in dif- Fig. 5. The mating success percentage of S. olivacea in different stocking densities.
ferent sex ratios (male:female). Sample size for sex ratio 1:1 = 9 males:9 females, Error bars represent standard deviation. Sample size for stocking density of 2, 6, and
2:1 = 12 males:6 females and 1:2 = 6 males:12 females. 10 crabs m−2 is 2, 6, and 10 pairs of crabs, respectively.

3.3. Stocking density

13.6 ± 0.6 h was considered to be completed when the exoskeleton
of the female had hardened and the male finally abandoned her. Stocking density significantly affected mating success rate
(F = 10.10, P = 0.01) (Fig. 5). Post hoc Tukey test showed that treat-
ment with stocking density of 10 crabs m−2 had significantly lower
mating success rate compared to treatments with lower stock-
3.2. Sex ratio ing densities. Mate displacement, fighting and cannibalism were
frequently observed as the stocking density was increased. Cumu-
Sex ratio did not have a significant effect over mating success lative mortality increased with time and stocking density (Fig. 6).
rate (F = 2.47, P = 0.16) (Fig. 2). Cumulative mortality increased with Mortality percentages at the end of the experiment were 16.7%,
time (Fig. 3). After 60 days, mortality was highest in treatment with 30.6% and 50% for treatments with stocking density of 2 crabs m−2 ,
ratio of 2 males:1 female (59.3%) and lowest in treatment with ratio 6 crabs m−2 and 10 crabs m−2 , respectively.
of 1 male:1 female (35.2%). Cumulative mortality was intermediate An increasing trend of male postcopulatory guarding dura-
in treatment with ratio of 1 male:2 females (40.7%). tion was observed as the stocking density increases, although the
The mean duration of copulation process which ranged from 6.1 duration of copulation showed no significant difference between
to 6.8 h showed no significant variation among different sex ratios. different stocking density treatments (Fig. 7). Male crabs spent less
However, the duration of postcopulatory guarding significantly dif- time in postcopulatory guarding in the treatment with the low-
fered between sex ratios (Fig. 4). A post hoc Tukey test showed est stocking density (2 crabs m−2 ) compared to the other two
that the treatment using male:female ratio of 1:2 was significantly treatments with higher stocking densities (6 or 10 crabs m−2 ,
shorter compared to the other two sex ratio treatments although respectively) while there is no significant difference in postcopu-
no significant difference was found between the two treatments latory guarding duration between the two treatments with higher
using male:female ratios of 1:1 and 2:1. stocking densities.

35

30
Cumulave mortality (individuals)

25

20
2

15 6
10

10

0
0 10 20 30 40 50 60
Time (days)

Fig. 6. Cumulative mortality of S. olivacea in different stocking densities throughout the experiment (60 days). Curves were fitted with second degree polynomial regressions
for each sex ratio. Sample size for stocking density of 2, 6, and 10 crabs m−2 is 2, 6, and 10 pairs of crabs, respectively.
 K. Waiho et al. / Aquaculture Reports 2 (2015) 50–57 55

16 has molted and become receptive. Females benefit from this mat-
14 b
b
ing behaviour by having a chance to select a more appropriate male
in terms of size and aggressiveness to safeguard her during molt-
12
ing, copulation and postcopulation phases. It is however, not an
Duraon (hours)

10
a
assurance that the male that guards a female during precopulation
8 phase will end up copulating with the secured female. As seen in
6
copulaon this study, even males in precopulatory embrace position can still
be challenged and substituted by other rival males, especially those
4
postcopulatory guarding larger in size. A strategy adopted by the male partners in reducing
2 their chances of encountering other challenging males is to bring
0 the embraced female to a secluded shelter with limited opening.
2 6 10 The positioning of female right underneath the male’s body dur-
Stocking density (crabs/m2)
ing precopulatory guarding is common in other brachyurans, such
Fig. 7. The copulation and postcopulatory guarding duration of S. olivacea in dif-
as the swimming crab Arenaeus cribrarius (Pinheiro and Fransozo,
ferent stocking densities. Error bars represent standard deviation. Sample size for 1999), sand crab Portunus pelagicus (Soundarapandian et al., 2013a),
stocking density of 2, 6, and 10 crabs m−2 is 2, 6, and 10 pairs of crabs, respectively. flower crab Charybdis feriata (Soundarapandian et al., 2013b), shore
crab Carcinus maenas (Krång and Ekerholm, 2006), burrowing crab
Neohelice granulata (Sal Moyano et al., 2014) and pill-box crab Hali-
4. Discussion carcinus cookii (Van Den Brink and McLay, 2010).
Copulation of S. olivacea occurs right after the female has under-
Seminal receptacle in S. olivacea female acts as both sperm stor- gone her puberty molt. This is consistent with most brachyurans
age and the place where fertilization occurs (Diesel, 1991; McLay (Hartnoll, 1969; McLay and López Greco, 2011), although some
and López-Greco, 2011). As shown in this study, the process of crabs such as H. cookii (Van Den Brink and McLay, 2010), Amari-
sperm transfer into S. olivacea female’s seminal receptacle can only nus spp. (Lucas, 1980) and Halicarcinus planatus (Richer de Forges,
occur when the female has just molted and is in its soft-shelled con- 1977) are able to copulate before their pubertal molt and the
dition. Due to this small window of opportunity to ensure paternity, stored eggs are deposited under the newly formed abdominal brood
S. olivacea males often paired up with females in precopulatory chamber right after female’s pubertal molt. Van Den Brink and
position after sensing that the females will reach their pubertal McLay (2010) postulated that this precocious mating and contin-
molt soon, either based on external cues, chemical cues or the inter- uous reproduction may be due to the species’ small body size and
action of both (Herborg et al., 2006). Mating males are all mature short life span.
males in their intermolt phase with hard exoskeletons, as observed The postcopulation duration of S. olivacea (13.6 ± 0.6 h) is sim-
in S. olivacea and other brachyuran crabs (Van Den Brink and McLay, ilar to that of other portunids such as P. pelagicus (13.16 ± 0.60 h)
2010; Brockerhoff and McLay, 2005a; Countreras-Garduño et al., (Soundarapandian et al., 2013a) and the hymenosomatid H. cookii
2007; Herborg et al., 2006; Krång and Ekerholm, 2006; Paul and (mean 18.3 h) (Van Den Brink and McLay, 2010). Postcopulatory
Paul, 1996; Wada et al., 2000). guarding right after copulation is seen as an effective way to protect
Male courting behaviors such as extending and waving che- the soft-shelled female from predators and ensuring the propa-
lipeds towards a targeted female and walking slowly towards her as gation of the male’s progeny, although the guarding male has to
shown by S. olivacea males are common cues portrayed by brachyu- sacrifice his feeding time and potential mating opportunities (Van
ran crabs to signal their potential partners that they are attractive Den Brink and McLay, 2010). Other brachyurans exhibit a vast
for copulation (Donaldson and Adams, 1989; Goshima et al., 1996; range of postcopulation durations, ranging from 47.5 h in Menippe
Soundarapandian et al., 2013a). Cheliped waving as a directional spp. (Wilber, 1989) to no postcopulatory guarding in M. squinado
signal to attract females to male’s burrow for mating has been (González-Gurriarán et al., 1998).
reported in fiddler crab Uca perplexa (Murai and Backwell, 2005) Sex ratio did not have a major impact on the mating success of S.
but was not observed in S. olivacea. Crab species from the family olivacea, although unequal sex ratio was shown to increase mortal-
Grapsidae and Majidae on the other hand typically skip the court- ity (Fig. 3). The postcopulatory guarding duration was significantly
ing process (Brockerhoff and McLay, 2005b; González-Gurriarán shorter in sex ratio of 1 male:2 females but increases as the number
et al., 1998). This lack of courtship is rarely found in portunid crabs, of males increases, although the difference between the other two
including S. olivacea (Kamio et al., 2008). In S. olivacea, the court- treatments with sex ratio of 1 male:1 female and 2 males:1 female
ing was carried out by males, although in some species, females are was not significant. Van Den Brink and McLay (2010) reported an
the ones that display courtship behaviors, This was observed in Call- increase of postcopulatory guarding duration from 2.5 h to 20 h and
inectes sapidus where females would intentionally collide with the subsequently 29.6 h when pill-box crab H. cookii were subjected
targeted male, signalling him to cradle her (Glesson, 1982). Females to sex ratio of 1 male:2 females, 1 male:1 female and 2 males:1
of Neohelice granulata visited several male burrows and made swift female respectively, with the guarding duration in ratio of 1 male:2
contact with the chosen male partner using the dactyls of her last females significantly lower from the other two ratios. Compared to
three legs (Sal Moyano et al., 2014). Interruption of other males females, males invest less into each offspring and will be readily
during the brief courting period was frequently observed in present available for mating faster than females. The lower postcopulatory
study. It was postulated that the size and aggressiveness of compet- guarding duration by males when the ratio of females increases
ing males are the keys in securing access to females. However, the is presumably due to less competition and lower chances of the
final determining factor of appropriate mate and subsequently the females being cannibalized by rival males. Also, the guarding males
outcome of mating success are still based on the female’s behaviour are more likely to go after other females as potential mates, hence
and mate-selection (Countreras-Garduño et al., 2007). This is in the shorter duration of postcopulatory guarding as compared to
conformation with current study in which there were occasions when the numbers of male competitors is high. In addition, the ter-
when females refused to be cradled by a courting male, even though ritorial and cannibalism behaviors of mud crabs, especially between
he fought and won over other males. males make the usage of lower male: female sex ratio preferable.
Guarding of females prior to copulation is important as it guar- Lower number of males is also more economical as mature males
antees the male the best chance to mate with the female once she are comparatively more expensive than females.
56 K. Waiho et al. / Aquaculture Reports 2 (2015) 50–57

The inverse relationship between mating success and cumula- the semi-terrestrial hermit crab Coenobita compressus (H. Milne Edwards). J.
tive mortality with stocking density (Figs. 5 and 6) in S. olivacea Crust. Biol. 27 (3), 411–416.
Davis, J.A., 2004. Spawning characteristics of the South African Mudcrab Scylla
might be due to the higher chances of encountering rival suitors serrata (Forskål) in captivity. J. World Aquacult. Soc. 35 (2), 121–133.
as the available space becomes increasingly limited. The duration Diesel, R., 1991. Sperm competition and the evolution of mating behaviour in
of postcopulatory guarding at a lower stocking density of 2 crabs Brachyura, with special reference to spider crabs (Decapoda, Majidae). In:
Bauer, R.T., Martin, J.W. (Eds.), Crustacean Sexual Biology. Columbia University
per m2 was shortest (8.6 ± 0.7 h) because the available space was Press, New York, pp. 145–164.
sufficient and clearly there was less possibility for other crabs to Donaldson, W.E., Adams, A.E., 1989. Ethogram of behavior with emphasis on
reach the newly hardened copulated female. Thus the guarding mating for the Tanner Crab Chionoecetes bairdi Rathbun. J. Crust. Biol. 9 (1),
37–53.
male readily releases the female when her carapace was almost
Glesson, R.A., 1982. Morphological and behavioural identification of the sensory
hardened. However, males in treatments with higher stocking den- structures mediating pheromone reception in the blue crab, Callinectes sapidus.
sities of 6 or 10 crabs m−2 were found to guard their females for Biol. Bull. 163, 162–171.
Goshima, S., Koga, T., Murai, M., 1996. Mate acceptance and guarding by male
a longer period of time (10.5–16.9 h) ensuring that the carapace
fiddler crabs Uca tetragonon (Herbst). J. Exp. Mar. Biol. Ecol. 196, 131–143.
of the females was fully hardened and the females were able to González-Gurriarán, E., Fernández, L., Muiño, F.R., 1998. Mating and role of seminal
fend for themselves from being cannibalized by other crabs before receptacles in the reproductive biology of the spider crab Maja squinado
the guarding males leave. This decrease in mating success and (Decapoda, Majidae). J. Exp. Mar. Biol. Ecol. 220, 269–285.
Hartnoll, R.G., 1969. Mating in Brachyura. Crustaceana 16 (2), 161–181.
lengthened postcopulatory guarding duration by males as stocking Herborg, L.-M., Bentley, M.G., Clare, A.S., Last, K.S., 2006. Mating behaviour and
density increases indicate that rival encounter rates affects both chemical communication in the invasive Chinese mitten crab Eriocheir sinensis.
the mating success and the guarding duration in S. olivacea. Stone J. Exp. Mar. Biol. Ecol. 329 (1), 1–10.
Ikhwanuddin, M., Bachok, Z., Mohd Faizal, W.W.Y., Azmie, G., Abol-Munafi, A.B.,
crabs Menippe spp. were found to increase their postcopulatory 2010. Size of maturity of mud crab Scylla olivacea (Herbst, 1796) from
guarding time as well (from 47.5 h to 80.1 h) when they were in the mangrove areas of Terengganu coastal waters. J. Sustain. Sci. Manage. 5,
presence of other males, suggesting the need to safeguard newly- 134–147.
Ikhwanuddin, M., Azmie, G., Juariah, H.M., Zakaria, M.Z., Ambak, M.A., 2011.
copulated females for a longer period to eliminate the possibility Biological information and population features of mud crab, genus Scylla from
of the females being cannibalized or re-copulated by other males mangrove areas of Sarawak, Malaysia. Fish. Res. 108 (2–3), 299–306.
(Wilber, 1989). Ikhwanuddin, M., Azmie, G., Zakariah, M.I., Abol-Munafi, A.B., 2013. Mud Crab:
Culture System and Practice in Malaysia. Penerbit Universiti Malaysia
Our findings described in detail the mating process of S. oli-
Terengganu, Terengganu. Malaysia.
vacea and suggest that mud crabs can copulate readily in captivity, Kamio, M., Reidenbach, M.A., Derby, C.D., 2008. To paddle or not: context
although sex ratio and stocking density do affect the mating suc- dependent courtship display by male blue crabs, Callinectes sapidus. J. Exp. Biol.
211 (8), 1243–1248.
cess, duration and mortality. These can be applied in aquaculture
Keenan, C.P., Davie, P.J.F., Mann, D.L., 1998. A revision of the genus Scylla de Haan,
sectors, especially in hatchery breeding operations. Even though 1833 (Crustacea: Decapoda: Brachyura: Portunidae). Raffles B Zool. 46 (1),
sex ratio did not affect mating success rate, sex ratio with higher 217–245.
number of females resulted in fewer mortalities and shorter post- Koolkalya, S., Thapanand, T., Tunkijjanujij, S., Havanont, V., Jutagate, T., 2006.
Aspects in spawning biology and migration of the mud crab Scylla olivacea in
copulatory guarding duration. Thus sex ratio of 1 male:2 females the Andaman Sea, Thailand. Fish. Manage. Ecol. 13 (6), 391–397.
should be favoured. Stocking density of 10 crabs m−2 and above Krång, A.-S., Ekerholm, M., 2006. Copper reduced mating behaviour in male shore
should be avoided due to low percentage of mating success rate crabs (Carcinus maenas (L.)). Aquat. Toxicol. 80, 60–69.
Lucas, J.S., 1980. Spider crabs of the family Hymenosomatidae (Crustacea:
and high mortality. Although, stocking density of 2 crabs m−2 Brachyura) with particular reference to Australian species: systematics and
showed highest survival, mating success and shorter postcopula- biology. Rec. Aust. Mus. 33, 148–247.
tory duration, it is not economical due to the large rearing space McLay, C.L., López Greco, L.S., 2011. A hypothesis about the origin of sperm storage
in the Eubrachyura, the effects of seminal receptacle structure on mating
required. Stocking density of 6 crabs m−2 would be more practical strategies and the evolution of crab diversity: how did a race to be first become
and cost-effective because it gives higher yield per m2 with moder- a race to be last? Zool. Anz. 250, 378–406.
ate mortality rate. Future research on the effect of available hiding Mirera, O.D., Moksnes, P.-O., 2013. Cannibalistic interactions of juvenile mud crabs
Scylla serrata: the effect of shelter and crab size. Afr. J. Mar. Sci. 35 (4), 545–553.
spaces in a confined environment on the mating success and mor-
Murai, M., Backwell, P.R.Y., 2005. More signalling for earlier mating: conspicuous
tality of mud crabs is recommended to address the need for large male claw waving in the fiddler crab, Uca perplexa. Anim. Behav. 70 (5),
rearing space. 1093–1097.
Naranjo-Paramo, J., Hernandez-Llamas, A., Villarreal, H., 2004. Effect of stocking
density on growth, survival and yield of juvenile redclaw crayfish Cherax
Acknowledgements quadricarinatus (Decapoda: Parastacidae) in gravel-line commercial nursery
ponds. Aquaculture 242, 197–206.
Paul, A.J., Paul, J.M., 1996. Observations on mating of multiparous Chionoecetes
The present study was financially supported by the Malaysia’s
bairdi Rathbun (Decapoda: Majidae) held with different sizes of males and
Ministry of Higher Education under Niche Research Grant Scheme one-clawed males. J. Crust. Biol. 16 (2), 295–299.
(NRGS) (Vot. No. 53131) and Prototype Research Grant Scheme Pinheiro, M.A.A., Fransozo, A., 1999. Reproductive behaviour of the swimming crab
(PRGS) (Vot. No. 54243). Special thanks are due to Institute of Trop- Arenaeus cribrarius (Lamarck, 1818) (Crustacea, Brachyura, Portunidae) in
captivity. Bull. Mar. Sci. 64, 243–253.
ical Aquaculture, Universiti Malaysia Terengganu for the space and Richer de Forges, B., 1977. Étude du crabe des îles Kerguelen Halicarcinus planatus
infrastructure provided. (Fabricius). CNFRA 42, 71–133.
Robertson, W.D., Kruger, A., 1994. Size at maturity, mating and spawning in the
portunid crab Scylla serrata (Forskål) in Natal. S. Afr. Estuar. Coastal Shelf Sci.
References 39, 185–200.
Sal Moyano, M.P., Gavio, M.A., McLay, C.L., Luppi, T., 2014. Habitat-related
Baylon, J.C., 2011. Survival and development of larvae and juveniles of the mud differences in the pre-copulatory guarding and copulation behaviour of
crab [Scylla olivacea Forskal (Crustacea: Decapoda: Portunidae)] at various Neohelice granulata (Brachyura, Grapsoidea, Varunidae). J. Sea Res. 87, 8–16.
temperature and salinities. Philipp. Agric. Sci. 94, 195–204. Sokal, R.R., Röhlf, F.J., 1981. Biometry. Freeman, W.H., San Francisco.
Brockerhoff, A.M., McLay, C.L., 2005a. Mating behaviour, female receptivity and Soundarapandian, P., Varadharajan, D., Anand, T., 2013a. Mating behaviour of Sand
male–male competition in the intertidal crab Hemigrapsus sexdentatus crab. Portunus pelagicus (Linnaeus) 2, http://dx.doi.org/10.4172/
(Brachyura: Grapsidae). Mar. Ecol. Prog. Ser. 290, 179–191. scientificreports.685.
Brockerhoff, A.M., McLay, C.L., 2005b. Comparative analysis of the mating Soundarapandian, P.P., Varadharajan, D., Ilavarasan, N., Kumar, J., Kumar, A., 2013b.
strategies in Grapsid crabs with special references to the intertidal crabs Mating behaviour of flower crab, Charybdis feriata (Linnaeus). J. Mar. Sci. Res.
Cyclograpsus lavauxi and Helice crassa (Decapoda: Grapsidae) from New Dev. 3, http://dx.doi.org/10.4172/2155-9910.1000127.
Zealand. J. Crust. Biol. 25 (3), 507–520. Van Den Brink, A.M., McLay, C.L., 2010. Competing for the last place: mating
Countreras-Garduño, J., Osorno, J.L., Códoba-Aguilar, A., 2007. Male–male behaviour in a pill-box crab, Halicarcinus cookii (Brachyura:
competition and female behaviour as determinants of male mating success in Hymenosomatidae). Zool. Anz. 249, 21–32.
 K. Waiho et al. / Aquaculture Reports 2 (2015) 50–57 57

Wada, S., Ashidate, M., Yoshino, K., Sato, T., Goshima, S., 2000. Effects of sex ratio Williams, M.J., Primavera, J.H., 2001. Choosing tropical portunid species for culture,
on egg extrusion frequency and mating behaviour of the spiny king crab domestication and stock enhancement in the Indo-Pacific. Asian Fish. Sci. 14,
Paralithodes brevipes (Decapoda: Lithodidae). J. Crust. Biol. 20 (3), 479–482. 121–142.
Wilber, D.H., 1989. The influence of sexual selection and predation on the mating Zar, J.H., 1984. Biostatistical Analysis. Prentice Hall, Englewood Cliffs.
and postcopulatory guarding behaviour of stone crabs (Xanthidae, Menippe).
Behav. Ecol. Sociobiol. 24 (6), 445–451.