Egg development and fecundity estimation in deep-sea red ... - redmic

Short communication
Fisheries Research 109 (2011) 373–378
Contents lists available at ScienceDirect
Fisheries Research
journal homepage: www.elsevier.com/locate/fishres
Egg development and fecundity estimation in deep-sea red crab, Chaceon affinis
(Geryonidae), off the Canary Islands (NE Atlantic)
Victor M. Tuset a,c,∗ , Domingo I. Espinosa b , Antonio García-Mederos a , José I. Santana a , José A. González a
a Departamento de Biología Pesquera, Instituto Canario de Ciencias Marinas (ICCM-ACIISI), Grupo de Biología Pesquera, P.O. Box. 56, E-35200 Telde (Las Palmas), Canary Islands,
Spain
b Departamento de Biología Animal, Universidad de La Laguna, c/Astrofísico F. Sánchez s/n, La Laguna, Tenerife, Spain
c Instituto de Ciéncias del Mar (ICM-CSIC), Passeig Marítim 37-49, 08003, Barcelona, Cataluña, Spain
article info
Article history:
Received 5 November 2010
Received in revised form 3 February 2011
Accepted 28 February 2011
Keywords:
Egg development
Fecundity
Chaceon affinis
NE Atlantic
1. Introduction
abstract
Geryonid deep-sea red crabs occur widespread throughout
the world and many of them have interest to fisheries such as,
e.g., Chaceon quinquedens Smith, 1879 and Chaceon fenneri (Manning
and Holthuis, 1984), Chaceon maritae (Manning and Holthuis,
1981), Chaceon bicolour (Manning and Holthuis, 1989) and Chaceon
notialis Manning and Holthuis, 1989, Chaceon chilensis Chirino-
Gálvez and Manning, 1989 (Elner et al., 1987; Erdman and Blake,
1988; Melville-Smith, 1988; Lindberg and Wenner, 1990; Arana,
2000; Steimle et al., 2001; Smith et al., 2004) and Chaceon affinis
(Manning and Holthuis, 1989). This last species inhabits the eastern
Atlantic Ocean (64 ◦ N–15 ◦ N) from Iceland to Senegal, including the
Azores, Madeira, and the Canary and Cape Verde Islands between
130 and 2047 m depth (Kjenerud, 1967; Samuelsen, 1975; Manning
and Holthuis, 1981; Sánchez and Olaso, 1985; Dawson and Webber,
1991), and also know from the Mid-Atlantic Ridges i.e., Menez
Gwen, Lost City (Biscoito and Saldanha, 2000; Biscoito, 2006). In
the Canary Islands, several studies demonstrated the relative abundance
of this species at depths between 600 and 1000 m (López
Abellán et al., 1994; González, 1995) and some biological aspects
∗ Corresponding author at: Instituto de Ciéncias del Mar (ICM-CSIC), Passeig Marítim
37-49, 08003, Barcelona, Cataluña, Spain. Tel.: +34 928 132 900; fax: +34 928
132 908.
E-mail addresses: victorta@iccm.rcanaria.es, vtuset@icm.csic.es (V.M. Tuset).
0165-7836/$ – see front matter © 2011 Elsevier B.V. All rights reserved.
doi:10.1016/j.fishres.2011.02.021
The goal of this work was to elaborate on reproductive knowledge of the deep-sea red crab, Chaceon affinis,
off the Canary Islands by providing information regarding its egg development and fecundity. Six stages
for eggs were observed, from fully filled with yolk to the embryo occupying almost all the space inside
the egg. A correlation was established between egg stage and the colour of the egg mass. Morphological
analyses indicated that the eggs are spherical in shape and that they increase the 10.7% in maximum
diameter from the initial to the final stage. Fecundity (AFE, annual fecundity estimation), defined as
the number of eggs borne per females, was calculated with two methods, manual by hand counter and
using an automated morphology system. No statistical differences for the relationships carapace width
(CW)–AFE and total weight (TW)–AFE were detected between the two methods. The number of eggs
ranged from 199,690 to 566,956 (105–160 mm CW). A positive correlation was obtained between AFE
and CW (r 2 = 0.672) and TW (r 2 = 0.785).
© 2011 Elsevier B.V. All rights reserved.
(reproduction, sizes at maturity, population structure) have been
studied (Fernández-Vergaz et al., 2000; López Abellán et al., 2002).
Models of population dynamic used in the management of
fishery resources require, among other factors, an estimation of
fecundity. Fecundity is a general term used to describe the number
of eggs produced by individual females (Arshad et al., 2006). In the
red crabs, eggs are laid and held attached to the female pleopods
under the abdomen for up to nine months until the eggs hatch
and the larvae are released into the water column (Haefner, 1978;
Erdman et al., 1991). Studies suggest that these species may not
spawn annually, due to long intermoult, period for adult females
(5–7 years), although it is possible that sperm could be stored
for intermoult spawning (Hines, 1982; Lux et al., 1982: Erdman
et al., 1991; Biesiot and Perry, 1995). The goal of this paper is to
increase our knowledge on the reproduction of C. affinis in the
Canary Islands previous to its possible economic exploitation by
describing egg development and by estimating its fecundity. In
addition, the application of an automated image analysis system
for estimation fecundity and for egg diameter measurements in
crustaceans was tested.
2. Materials and methods
2.1. Sampling
A total of 41 berried females were captured from two experimental
research cruises, made in February–March 2005 (n = 20) and

374 V.M. Tuset et al. / Fisheries Research 109 (2011) 373–378
November–December 2006 (n = 21), coinciding with the spawning
period October–May (López Abellán et al., 2002). Individuals were
recollected using bottom traps at depths of 600–1000 m (Carvalho
et al., 2007) to the south of Gran Canaria (Canary Islands, northeastern
Atlantic) (Fig. 1). Only fresh samples, the last day of collecting,
were taken to the laboratory for the posterior study: 14 females
we used for the morphological study of eggs and 30 females for the
fecundity estimation. Carapace width (CW in mm) and total weight
(TW in g) were taken from each crab.
2.2. Egg development
The fresh egg mass was separated from the pleopods, placed on
a mesh of 100 m, and washed with seawater under pressure to
detach all the eggs. Then some eggs were placed in two Petri dishes
with seawater under a compound microscope with reflected light.
Four images for each Petri dish (one by quadrant) were taken in
each female with a digital video camera (Basler A310, Vision Technologies,
Basler AG, Ahrensburg) for the automated morphology
analysis (Fig. 2). Approximately, 40 eggs were measured in each
quadrant. Image-Pro Plus version 4.1.0 software (Media Cybernetics,
Inc.) was used to calculate the following shape parameters and
indices: projected area (A in mm 2 ), maximum diameter (D in mm),
minimum diameter (d in mm), roundness (4A/( D 2 )), and aspect
ratio (D/d)(Russ, 1990). Egg morphology was described and classified
according to Arshad et al. (2006) and Walker et al. (2006) for
other brachyuran crabs, and colour changes of the egg mass considering
previous studies in Chaceon spp. (Wigley et al., 1975; Haefner,
1978).
2.3. Estimation of fecundity
The gravimetric method was applied to determine fecundity
(AFE, annual fecundity estimation), which was defined as annual
egg production, considering that spawning might occur only once
a year, using a subsample of 5% by wet weight of the egg mass
(Lowerre-Barbieri and Barbieri, 1993). Two methodological procedures
were applied: (a) in method 1 (February–March sample), the
fresh egg mass was separated from the pleopods, placed on a mesh
of 100 m, and washed with seawater under pressure to detach all
the eggs. Then the eggs were placed in Petri dishes with seawater
Fig. 1. Location of the study area.
under a compound microscope with reflected light, to count the
eggs using a hand counter. (b) In method 2 (November–December
sample), fresh egg masses were stored in 4% buffered formalin,
detached from the pleopod filaments using tweezers, placed on
Fig. 2. Digitalized image of eggs for morphological analysis: (A) original image; (B)
modified binary image.

V.M. Tuset et al. / Fisheries Research 109 (2011) 373–378 375
Fig. 3. Chromatic variation of eggs attached to pleopods until their hatching: (A) red-orange or light purple; (B) burgundy; (C) dark purple; (D) brownish; (E) brown; (F)
grey-black. (For interpretation of the references to color in this figure legend, the reader is referred to the web version of the article.)
Petri dishes, and counted using an automated imaging system
(Leica QWin Standard, Leica Microsystems GmbH). Berried females
in embryo stage (see results, stage VI) were eliminated from the
analysis, because some embryos could have hatched already. In
both methods, power regressions were fitted to fecundity versus
carapace width and total weight (Haddon, 1994). Analysis of covariance
(ANCOVA) was used to determine the effect of the method on
the relationships CW–AFE and TW–AFE, in which the dependent
variable was the natural logarithm of the fecundity, the fixed factor
the method, and the covariate the natural logarithm of CW or TW.
3. Results
Six development stages were observed and correlated with the
colour of the egg mass (Figs. 3 and 4): stage I, egg still undivided,
fully filled with yolk, red-orange or light purple colour; stage II,
the free region of yolk is just visible, burgundy colour; stage III, a
slight pigmentation of the eyes appears, dark purple colour; stage
IV, pigmented eyes enlarging in moon shape, brownish colour;
stage V, visible pigmented structures enlarging eyes, segmented
appendages and abdomen appearing, brown colour; stage VI, eyes
acquiring oval shape, embryo occupies almost all the space inside
the egg, grey-black colour. After the last stage, larvae are present.
Egg development seems to be not completely synchronous and
sometimes two colour patterns can be observed simultaneously,
one in the inner part, and another in the outer part of the egg mass
(Fig. 3A). The eggs present a spherical shape, the mean sizes (maximum
diameter, minimum diameter, and area) increasing from
newly spawned ones (e.g., 0.585 mm D) to hatching (0.655 mm D)
(Table 1).
Fecundity ranged between 264,492 (118 mm CW) and 566,956
eggs (143 mm CW) using method 1 (n = 10) and from 199,690
(105 mm CW) to 559,308 eggs (160 mm CW) with method 2 (n = 20).
The ANCOVA test did not show significant differences in the
relationships CW–AFE (F = 1.474, P = 0.235) and TW–AFE (F = 0.188,
P = 0.668) between methods, therefore relationships were calculated
for the whole set of data. Regression analysis showed a
significant, positive correlation between fecundity and carapace
width (AFE = 3.180 × CW 2.407 , r 2 = 0.681, P < 0.001) and fecundity
and total weight (AFE = 684.486 × TW 0.977 , r 2 = 0.764, P < 0.001)
(Fig. 5).
4. Discussion
The egg development of C. affinis studied herein was very similar
to the general embryonic pattern observed in many brachyuran
crabs, including an increase in egg size from newly spawned ones
to just prior to hatching (Arshad et al., 2006; Walker et al., 2006).
However, changes in colour patterns differ greatly among these
deep-sea red crabs, making it necessary to develop a macroscopic
scale for each one separately. The eggs of C. fenneri are initially
light purple or burgundy, becoming dark purple and purple-brown,

376 V.M. Tuset et al. / Fisheries Research 109 (2011) 373–378
Fig. 4. Phases of egg development. (For interpretation of the references to color in this figure legend, the reader is referred to the web version of the article.)
while in C. quinquedens they are red-orange, then brown, purple,
and finally black (Wigley et al., 1975; Haefner, 1978; Erdman and
Blake, 1988).
Crabs of genus Chaceon inhabit deep waters and seem to have
continuous reproduction along year (Elner et al., 1987; Erdman and
Blake, 1988; Steimle et al., 2001; Pinho et al., 2001; Smith et al.,
2004). Berried females migrate to shallower and warmer waters
enhancing eggs that are swept up surface by the current (Haefner,
Table 1
Morphological values (mean ± standard deviation) of stages of egg development.
1978). The larvae phase is pelagic and consists in four zoeal stages
and a final megalopa. In the first postmegalop instar stage the carapace
width is relatively large, which may be an adaptation to slow
post-settlement growth (see Steimle et al., 2001). The eggs are the
largest known for crabs with planktonic development: 600 m in
maximum diameter in C. fenneri (Erdman and Blake, 1988; Hines,
1988; Erdman et al., 1991) and C. bicolour (Smith et al., 2004),
680 m D in C. maritae (Melville-Smith, 1987), 789 m D in C.
Stage n Maximum diameter (D, mm) Minimum diameter (d, mm) Area (A, mm 2 ) Roundness Ratio aspect
I 302 0.585 ± 0.031 0.551 ± 0.029 0.254 ± 0.026 1.075 ± 0.007 1.097 ± 0.031
II 2042 0.582 ± 0.039 0.541 ± 0.043 0.251 ± 0.034 1.124 ± 0.081 1.157 ± 0.062
III 616 0.580 ± 0.043 0.551 ± 0.045 0.257 ± 0.038 1.078 ± 0.009 1.115 ± 0.039
IV 326 0.590 ± 0.047 0.558 ± 0.048 0.262 ± 0.042 1.077 ± 0.008 1.126 ± 0.043
V 533 0.601 ± 0.033 0.566 ± 0.035 0.272 ± 0.030 1.081 ± 0.009 1.131 ± 0.050
VI 334 0.655 ± 0.038 0.627 ± 0.036 0.328 ± 0.036 1.083 ± 0.008 1.109 ± 0.040

Fig. 5. Relationships between fecundity and carapace width (A) and total
weight (B).
affinis (present study), and 846 m D in C. quinquedens (Haefner,
1978; Elner et al., 1987; Hines, 1988; Erdman et al., 1991). However,
large egg size has reproductive implications by decreasing
the number of eggs (Hines, 1982, 1988). Comparison of fecundity
estimates in deep-sea red crabs does not reveal great differences
among species, although it can be concluded that there is a positive
correlation female size and the number of eggs: 36,000–226,000
eggs (90–118 mm CW) inC. quinquedens (Hines, 1988; Steimle
et al., 2001), 15,592–288,312 eggs (98–133 mm CW) inC. bicolor
(Smith et al., 2004), 107,000–350,000 in C. maritae (Melville-
Smith, 1987), 160,000–375,000 eggs (110–143 mm CW)inC. fenneri
(Erdman and Blake, 1988; Hines, 1988), and 199,690–566,956 eggs
(105–160 mm CW)inC. affinis (present study). In all these species it
has been found that fecundity increased with the increase of carapace
width or wet weight of the individuals (Melville-Smith, 1987;
Erdman and Blake, 1988; Hines, 1988; Steimle et al., 2001; Smith
et al., 2004; present study), as also occurs in other brachyurans
(Haddon, 1994; Mantelatto and Fransozo, 1997; Litulo, 2004, 2005;
Arshad et al., 2006). However, in deep-sea red crabs it has been
noted that these variables are not good predictors of fecundity,
reaching variability values of 45–65% (C. quinquedens, C. fenneri,
and C. affinis) or at least 13–26% (C. bicolor). Also, it is known crabs
V.M. Tuset et al. / Fisheries Research 109 (2011) 373–378 377
can lose eggs during incubation through disease, egg predation, or
other kinds of natural failure of egg development (Perkins, 1971).
The high variability may be due to the large time that females
carry the eggs as the incubation period may last up to nine months
(Haefner, 1978; Erdman et al., 1991). It can be hypothesized that the
use of females with eggs in different stages of development could
influence the fecundity estimates. Nevertheless, Erdman and Blake
(1988) and Hines (1988) found a similar trend for Chaceon spp.
selecting only embryos in early stage. Moreover, our results prove
that the comparative study between automated system versus
manual counting, and also between fresh versus fixed samples,
were not causing this high variability. Consequently, this study
provides relevant information for stock assessment of this species,
moreover the analysis of the colour of the eggs and their development
may be used for a sustainable fishing due to the quick
observation.
Acknowledgements
The authors are indebted to José A. Pérez-Peñalvo, Rosa
Domínguez-Seoane and Eliseba García for skilful technical assistance.
Financial support was received from the EU ERDF in
the framework of the PIC INTERREG III B project PESCPROF-2
(03/MAC/4.2/M8) and from the Spanish Government in the framework
of project REDECA (CTM2005-07712-C03/MAR).
References
Arana, P.M., 2000. Estimación de abundancia y biomasa del cangrejo dorado (Chaceon
chilensis), en el archipiélago de Juan Fernández, Chile. Inv. Mar. 28, 53–68.
Arshad, A., Kamarudin, M.S., Saad, C.R., 2006. Study on fecundity and larval development
of blue swimming crab Portunus pelagicus (Linnaeus, 1758) under
laboratory conditions. Res. J. Fish. Hydrobiol. 1, 35–44.
Biesiot, P.M., Perry, H.M., 1995. Biochemical composition of the deep-sea red crab
Chaceon quinquedens (Geryonidae): organic reserves of developing embryos and
adults. Mar. Biol. 124, 407–416.
Biscoito, M., 2006. Chaceon affinis. In: Desbruyères, D., Segonzac, M., Bright, M. (Eds.),
Handbook of Deep-sea Hydrothermal Vent Fauna, 2nd ed. Denisia, Linz, p. 458.
Biscoito, M.J., Saldanha, L., 2000. Occurrence of Chaceon affinis (Decapoda: Geryonidae)
in the vicinity of a hydrothermal vent site on the mid-Atlantic ridge. J.
Crust. Biol. 20, 128–131.
Carvalho, D., Delgado, J., Biscoito, M., Frietas, M., González, J.A., Santana, J.I., Tuset,
V.M., Isidro, E., Pinho, M.R., Consorcio Pescprof. 2007. Recursos Pesqueros de
Aguas Profundas del Atlántico Centro-Oriental: alternativas a la pesca en la
Macaronesia. Memoria científico-técnica final del Proyecto PESCPROF-2 (PIC
Interreg III B, 03MAC/4.2/M8). European Union, Regional Policy, FEDER. Telde
(Las Palmas), p. 154.
Dawson, E.W., Webber, W.R., 1991. The deep-sea red crab Chaceon (“Geryon”): a
guide to information and a reference list of the family Geryonidae. Nat. Mus.
New Zealand Miscell. Ser. 24, 1–83.
Elner, R.W., Koshio, S., Hurley, G.V., 1987. Mating behavior of the deep-sea red crab,
Geryon quinquedens Smith (Decapod, Brachyura, Geryonidae). Crustaceana 52,
194–201.
Erdman, R.B., Blake, N.J., 1988. Reproductive biology of female golden crabs Geryon
fenneri Manning and Holthuis, from southeastern Florida. J. Crust. Biol. 8,
392–400.
Erdman, R.B., Blake, N.J., Lockhart, F.D., Lindberg, W.J., Perry, H.M., Waller, R.S., 1991.
Comparative reproduction of the deep-sea crabs Chaceon fenneri and C. quinquedens
(Brachyura: Geryonidae) from the northeast Gulf of Mexico. Invertebr.
Reprod. Dev. 19, 175–184.
Fernández-Vergaz, V., López Abellán, L.J., Balguerías, E., 2000. Morphometric, functional
and sexual maturity of the deep-sea red crab Chaceon affinis inhabiting
Canary Islands waters: chronology of maturation. Mar. Ecol. Prog. Ser. 204,
169–178.
González, J.A., 1995. Catálogo de los Crustáceos Decápodos de las islas Canarias.
Publicaciones Turquesa, Santa Cruz de Tenerife.
Haddon, M., 1994. Size-fecundity relationships, mating behaviour, and larval release
in the New Zealand paddle crab, Ovalipes catharus (White, 1843) (Brachyura:
Portunidae). N. Z. J. Mar. Freshw. Res. 28, 329–334.
Haefner Jr., P.A., 1978. Seasonal aspects of the biology, distribution and relative abundance
of the deep-sea red crab Geryon quinquedens Smith, in the vicinity of the
Norfolk Canyon, Western North Atlantic. Proc. Natl. Shellfish Assoc. 68, 49–62.
Hines, A.H., 1982. Allometric constraints and variables of reproductive effort in
brachyuran crabs. Mar. Biol. 69, 309–320.
Hines, A.H., 1988. Fecundity and reproductive output of two species of deep-sea
crabs, Geryon fenneri and G. quinquedens (Decapoda, Brachyura). J. Crust. Biol. 8,
557–562.

378 V.M. Tuset et al. / Fisheries Research 109 (2011) 373–378
Kjenerud, J., 1967. A find of Geryon affinis Milne-Edwards and Bouvier, 1984 (Crustacea
Decapoda) off the coast of Norway. Sarsia 29, 193–198.
Lindberg, W.J., Wenner, E.L., 1990. Geryonid crabs and associated continental slope
fauna: a research workshop report. Florida Sea Grant College Tech. Pap. 58: 1–61.
Litulo, C., 2004. Fecundity of the pantropical fiddler crab Uca annulipes (H. Milne
Edwards, 1837) (Brachyura: Ocypodidae) at Costa do Sol Mangrove, Maputo
Bay, southern Mozambique. Western Indian Ocean J. Mar. Sci. 3, 87–91.
Litulo, C., 2005. Fecundity and size at sexual maturity of the fiddler crab Uca vocans
(Linnaeus, 1758) (Brachyura: Ocypodidae). Thalassa 21, 59–65.
López Abellán, L.J., Santamaría, M.T.G., Balguerías, E., 1994. Resultados de la campaña
experimental de pesca, realizada en aguas del suroeste de la isla de Tenerife,
Canarias 9206. Inf. Téc. Inst. Esp. Oceanogr. 147, 1–62.
López Abellán, L.J., Balguerías, E., Fernández-Vergaz, V., 2002. Life history characteristics
of the deep-sea crab Chaceon affinis population off Tenerife (Canary
Islands). Fish. Res. 58, 231–239.
Lowerre-Barbieri, S.K., Barbieri, L.R., 1993. A new method of oocytes separation
and preservation for fish reproduction studies. Fish. Bull. 91, 165–
170.
Lux, F.E., Ganz, A.R., Rathjen, W.F., 1982. Marking studies on the red crab Geryon
quinquedens Smith off southern New England. J. Shellfish Res. 2, 71–80.
Manning, R.B., Holthuis, L.B., 1981. West African brachyuran crabs. Smithsonian
Contrib. Zool. 306, 1–379.
Melville-Smith, R., 1987. The reproductive biology of Geryon maritae (Decapoda
Brachyura) off southwest Africa/Namibia. Crustaceana 53, 11–27.
Melville-Smith, R., 1988. Comparative population size estimates for a portion of the
red crab Geryon maritae stock off southwest African coast. South African J. Mar.
Sci. 6, 23–31.
Mantelatto, F.L.M., Fransozo, A., 1997. Fecundity of the crab Callinectes ornatus Ordway,
1863 (Decapoda, Brachyura, Portunidae) from the Ubatuba region, São
Paulo, Brazil. Crustaceana 70, 214–224.
Perkins, H.C., 1971. Egg loss during incubation from off-shore northern lobster
(Decapoda: Homaridae). Fish. Bull. 69, 451–453.
Pinho, M.R., Gonçalves, J.M., Martins, H.R., Menezes, G.M., 2001. Some aspects of the
biology of the deep-water crab, Chaceon affinis (Milne-Edwards and Bouvier,
1894) off the Azores. Fish. Res. 51, 283–295.
Russ, J.C., 1990. Computer-assisted Microscopy: the measurement and analysis of
images. New York, Plenum Press.
Samuelsen, T.J., 1975. The third record of Geryon affinis, A. Milne-Edwards
and Bouvier (Crustacea, Decapoda) from western Norway. Sarsia 59, 47–
48.
Sánchez, F., Olaso, I., 1985. Presencia de Geryon affinis Milne-Edwards y Bouvier,
1894, en el golfo de Vizcaya (Decapoda, Brachyura). Bol. Inst. Esp. Oceanogr. 2,
155–157.
Smith, K.D., Potter, I.C., Hall, N.G., 2004. Biological and fisheries data for managing the
deep-sea crabs Hypothalassia acerba and Chaceon bicolor in Western Australia.
Projects No. 1999/154 and 2001/055. FRDC Australia.
Steimle, F.W., Zetlin, C.A., Chang, S., 2001. Essential Fish Habitat Source Document:
Red Deepsea Crab, Chaceon (Geryon) quinquedens, Life History and Habitat
Characteristics. NOAA Tech. Memo. NMFS NE 163, 1–27.
Walker, A., Ando, S., Smith, G.D., Lee, R.F., 2006. The utilization of lipovitellin during
blue crab (Callinectes sapidus) embryogenesis. Comp. Bioch. Physiol. Part B 143,
201–208.
Wigley, R.L., Theroux, R.B., Murray, H.E., 1975. Deep-sea red crab, Geryon quinquedens,
survey off northeastern United States. Mar. Fish. Rev. 37, 1–27.