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Revista de Biología Tropical, ISSN electrónico: 2215-2075 Vol. 69(S1): 464-473, March 2021 (Published Mar. 10, 2021)

Photoperiod in aquaculture of the sea urchin Arbacia dufresnii

(Echinodermata: Echinoidea): Effects on gamete production and maturity

Lucas R. Sepúlveda

1,2

Jimena Pía Fernandez

1,2

Mercedes Vera-Piombo

1,2

Florencia Belén Chaar

2,3

Tamara Rubilar

1,2

1. Biological Oceanography Laboratory, Centro para el Estudio de Sistemas Marinos, Centro Nacional Patagónico,

Consejo Nacional de Investigaciones Científicas y Técnicas, Bv. Alte. Brown 2915, Puerto Madryn, Argentina;

lsepulveda@cenpat-conicet.gob.ar, jpfernandez@cenpat-conicet.gob.ar, mercedes.verapiombo@gmail.com

2. Laboratory of Chemistry of Marine Organisms. Instituto Patagónico el Mar. Faculty of Natural Sciences and Health

Sciences. National University of Patagonia San Juan Bosco. Puerto Madryn. Argentina;

rubilar@cenpat-conicet.gob.ar (*Correspondence).

3. National University of Patagonia San Juan Bosco, Bv. Alte. Brown 3051, Puerto Madryn, Argentina;

chaar@gmail.com

Received 17-VII-2020. Corrected 25-VIII-2020. Accepted 05-X-2020.

ABSTRACT

Introduction: Photoperiod is, together with temperature and food availability, one of the main stimuli in the

regulation of gametogenesis in a wide variety of species. Objective: To evaluate the effect of photoperiod on

the production of mature gametes in cultured Arbacia dufresnii. Methods: An experiment was carried out with

three varying light-dark regimes/treatments: constant light (24 h light), neutral photoperiod (12 h light, 12 h

darkness), and constant darkness (24 h darkness). Twenty females were used in each treatment. All were induced

to spawn and, ten randomly selected females from each treatment were induced to spawn again after 30 days.

After 60 days, spawning was induced in the remaining females. The gametes were collected in filtered seawater,

fixed in Davidson solution, quantified and measured per individual in triplicate in a Sedgewick-Rafter chamber.

To determine maturation, fertilization success was evaluated 30 minutes after fertilization. Results: Our results

showed that in the aquaculture system, after only two months, mature gametes were obtained, and in the neutral

light regime there were 10 times more gametes than the number produced in wild sea urchins during the spawn-

ing period in question. We also found that with a greater exposure to light, a lower number of mature gametes

was produced. Conclusions: This study suggests the viability of the production of mature gametes in a short

period of time as regards Arbacia dufresnii.

Key words: gonad productivity; echinoderm; echinoid; aquaculture; mature gametes.

Sepúlveda, L.R.,

Fernandez, J.P., Vera-Piombo, M., Chaar,

F.B., & Rubilar, T. (2021). Photoperiod in aquaculture

of the sea urchin Arbacia dufresnii (Echinodermata:

Echinoidea): Effects on gamete production and

maturity. Revista de Biología Tropical, 69(S1), 464-

473. DOI 10.15517/rbt.v69iSuppl.1.46386

Sea urchin aquaculture is carried out for

a variety of purposes in different parts of the

world. The most common purpose of this activ-

ity is aimed at the gastronomic industry, that is,

the human consumption of sea urchin gonads.

However, aquaculture practices also take place

with the aim of enhancing the quality of the

gonads of wild individuals (Pearce, Daggett, &

Robinson, 2004; Walker et al., 2015; Rubilar et

al., 2016), as well as with the aim of producing

DOI 10.15517/rbt.v69iSuppl.1.46386

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juveniles that can repopulate populations (Cár-

camo, 2004) and, last but not least, with the

aim of being used as model organisms in

developmental studies (Harris & Eddy, 2015;

Unuma, Sakai, Agatsuma, & Kayaba, 2015).

In addition, the development of technology for

the production of marine non-food organisms

has taken place globally, and there is, today,

a broad range of applications, including, for

example, nutraceutical, cosmeceutical, phar-

maceutical, biofuel, and conservation products.

(Costa-Leal, Rocha, Rosa, & Calado, 2016). In

Argentina, there is a startup with a focus on the

non-traditional species in aquaculture, Arbacia

dufresnii, aiming to produce mature gametes

with a high concentration of pigments and fatty

acids for application in nutraceutical, cosme-

ceutical, pharmaceutical and veterinary prod-

ucts (www.arbacia.com.ar). Arbacia dufresnii

is a temperate and abundant sea urchin species

with a wide distribution in South America,

from Río de la Plata in Argentina (35° S) on

the Atlantic coast to Puerto Montt in Chile

(42° S) on the Pacific coast, at a depth range of

0-315 m (Brogger et al., 2013). The species is

a summer spawner with two peaks of spawning

activity, one partial peak in the austral spring

(September-October) and another major peak

at the end of austral summer (March) (Brogger,

Martinez, & Penchaszadeh, 2010; Epherra et

al., 2014). The species was previously studied

to assess whether its gonadal quality could be

improved through aquaculture methods, dem-

onstrating the possibility to modify the produc-

tion of the gonadal mass and the composition

of cell content through the use of formulated

feed (Rubilar et al., 2016).

Eggs are the final product of gametogen-

esis in the female sea urchin. This process

involves the accumulation of nutrients, the

proliferation of oogonium, the differentiation

of gametes, maturation of gametes and, finally,

the spawning of gametes. Frequently, there is a

period of quiescent reabsorption of the residual

gametes and, then, the process starts all over

again (Mercier & Hamel, 2009; Walker, Lesser,

& Unuma, 2013; Epherra et al., 2014). There is

a control mechanism in the gametogenic cycle

triggering the end of the resting period and the

subsequent re-starting of gametogenesis. This

control is thought to be determined by both

endogenous and exogenous factors (Mercier

& Hamel, 2009). In sea urchins, gametogen-

esis is initiated in response to external factors,

such as temperature and photoperiod (Fuji,

1967; Pearse & Walker, 1986; Bay-Schmidt &

Pearse, 1987; Byrne, 1990; Unuma, Konishi,

Furuita, Yamamoto, & Akiyama, 1996; Meidel

& Scheibling, 1998; Walker & Lesser, 1998;

Spirlet, Grosjean, & Jangoux, 2000; Unuma,

2002; Kirchhoff, Eddy, & Brown, 2010;

Gianguzza, & Bonaviri, 2013; Wangensteen,

Turon, Casso, & Palacín, 2013; Díaz-Martínez,

Carpizo-Ituarte, & Benítez-Villalobos, 2019).

To be successful, any sea urchin aquaculture

must take into account the factors regulating

gametogenesis. In addition, the majority of sea

urchins have annual reproductive cycles (Mar-

zinelli, Bigatti, Giménez, & Penchaszadeh,

2006; Walker et al., 2013; Epherra et al., 2014)

which constraints the period of exploitation to

a narrow annual season which has to be well

controlled in order to harvest the gonads at the

appropriate time. Various strategies have been

proposed to extend the harvest season for high

quality gonads by focusing on the prolongation

of the growth period of the nutritive phagocytes

and/or by delaying gametogenesis (Unuma

& Walker, 2010). Due to the fact that the

photoperiod and temperature can be manipu-

lated in aquaculture, such manipulation can be

used to promote, or delay, gonad development

(Walker & Lesser, 1998; Spirlet et al., 2000;

Pearce, Daggett & Robinson, 2002; Kirchhoff

et al., 2010). All of the research undertaken

to manipulate gametogenesis in sea urchin

aquaculture is aimed at generating the gonadal

production of good quality for the gastronomic

market. One of the key factors in the quality of

gonads is the presence of fewer gametes rela-

tive to somatic cells (Unuma & Walker, 2010).

In this sense, sea urchin aquaculture could

produce market quality gonads for a longer

period during the year and could replace the

wild-harvested sea urchins. To do so, there are

two strategies to extend the period in which

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gonads are harvested in the aquaculture sys-

tem: to extend the period of somatic cells, or to

suppress gametogenesis (Walker et al., 2015).

However, the A. dufresnii aquaculture system

incorporates a completely different strategy.

Here, the goal is to obtain the largest number

of mature gametes in the shortest period pos-

sible in order to harvest mature gametes several

times during the year.

In this context the manner in which the

photoperiod affects this species gametogen-

esis is, still, unknown. The aim of the present

work is to evaluate the effect of photoperiod

on gamete production and maturation in the A.

dufresnii aquaculture system.

MATERIALS AND METHODS

Collection of sea urchins: Adults sea

urchins (x

= 30.69 ± 2.52 S.D. mm diameter)

were collected (N = 120) on July 1, 2019 from

Nuevo gulf (42º46’44’’ S & 64º59’52’’ W)

and transported to the Experimental Aquar-

ium at Centro Nacional Patagónico, Con-

sejo Nacional de Investigaciones Científicas

y Técnicas (CENPAT-CONICET) in Puerto

Madryn, Argentina.

Experimental design: A week after col-

lection (on July 7) and without being fed to

ensure depletion of the gut and to standardize

the nutritional status of all individuals (Watts,

Lawrence, & Lawrence, 2013), the sea urchins

were placed and distributed into three aquaria

(N = 20 per aquarium) and three different light-

dark regimes (treatments) were established:

Constant Darkness (CD = 24 hours of daily

darkness), Neutral Photoperiod (NP = 12 hours

of light and 12 hours of darkness), and Con-

stant Light (CL = 24 hours of daily light). Each

aquarium was 90 l in volume, with a semi-

closed aquaculture recirculating system. The

seawater was pumped through a decanter, a

physical filter, a biofilter and then recirculated

into the aquaria. Extra air pumps were placed

in each aquarium to ensure good oxygenation

of the water. Twenty-five percent of the water

volume was changed twice a week, and in

conjunction with this, any feces and uneaten

feed was siphoned off, and the animals were

also fed at this time, that is, twice a week. The

water quality was maintained within the opti-

mal parameters and checked every week on the

basis of an aquarium test (Tetra) at a salinity of

35 ppt and a temperature between 14 °C and 16

°C (similar values as those found in field con-

ditions). The sea urchins were fed every three

days with a weighed amount of formulated feed

(500 mg per individual) (Table 1) produced

by the Group for Research and Technological

Development in Aquaculture and Fisheries

(GIDTAP) at the National Technological Uni-

versity (UTN).

TABLE 1

Composition of the formulated feed

Ingredients

Percentage weight

(as is or as fed basis)

Wheat Starch 4 %

Cornstarch 19 %

Soy protein 11 %

Lecithin 0.6 %

Marine Ingredients 36 %

Non-Marine Ingredients 28.2 %

Vitamin premix 0.6 %

Minerals premix 0.6 %

To determine the sex of the individuals,

each sea urchin was induced to spawn their

gametes at the beginning of the experiment.

For the induction of spawning, sea urchins

were injected with 0.3 ml of 0.55 M KCl solu-

tion (Strathmann, 1987). Only females were

selected for the experiment as the production

of mature eggs was the focus of the study.

Males sea urchins were introduced into the

broodstock of the experimental aquarium at

the Centro Nacional Patagónico. After 30 days,

ten random females were selected and once

again, spawning was induced. After 60 days,

the experiment was finalized and spawning

was induced in the remaining females. The

gametes were collected on filtered seawater,

fixed in a Davidson solution, photographed

and quantified per individual in triplicate in

a Sedgewick-Rafter chamber using a Leica

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DM 2500 microscope, Leica ICC50W digital

camera and LAS EZ B4.5.0.418 software.

Using the Image-J program, the female gam-

etes were measured.

The maturity of the gametes was assessed

on the basis of fertilization success, for this

purpose gametes from five females from each

treatment were fertilized. Sperm was obtained

from males of the broodstock. Fertilization was

carried out by using a 1:100 000 v/v dilution

of sperm (Fernández, Epherra, Sepúlveda, &

Rubilar, 2019). After 30 minutes, the fertiliza-

tion success was verified under microscope by

observing the fertilization membrane in the

eggs (Fig. 1). The percentage of fertilized eggs

was calculated.

permutations using unrestricted permutation

of the raw data. For pairwise tests where there

were < 20 unique permutations, the Monte

Carlo P-value was used, as recommended in

Anderson et al. (2008). Fertilization success

was performed applying a one-way analysis of

variance (ANOVA). The ANOVA assumptions

were tested and all of the statistical analyses

were carried out using the Statistica software

(version 7).

RESULTS

The number of gametes changed signifi-

cantly during time and treatments. There was

no interaction between time and treatments

(Table 2). The SIMPER analysis showed that

total gametes had > 65% dissimilarity over

time. At the beginning of the experiment, the

number of gametes per female was between

250 000 and 450 000. After 30 days of treatment,

the number of gametes per female increased in

every treatment from 400 000 to 700 000 gam-

etes. At this point, the sea urchins in the neutral

photoperiod treatment (NP) showed the highest

total number of gametes per female. At the end

of the experiment, after 60 days of the differ-

ent light regimes, the total number of gametes

per female increased notably, up to 3 500 000

gametes. The treatment with constant darkness

(CD) resulted in the lower value, with a median

of 1 395 000 gametes. The SIMPER analysis

showed that total gametes in the CD treatment

had 38 % of gametes that constant light (CL)

treatment, and NP treatment had 62 % of gam-

etes that CL (Fig. 2, Table 2).

The maturity of gametes was affected

by the light regime applied. After 30 days of

experiment, sea urchins in the neutral photo-

period treatment showed the lowest fecundity

success rate, less than 15 % (F

= 10.03, P =

0.0003). After 60 days under light regimes, the

fecundity success rate increased in all of the

treatments and showed significant differences

= 3.28, P = 0.044). NP evidenced the high-

est maturity of gametes, with 94 % fecundity

success rate, and the CL treatment resulted in

Fig. 1. Fertilized egg with the fertilization membrane (left)

and unfertilized egg (right).

Data analysis: Comparisons between

the quantity of gametes produced for each

treatment was made using Primer v7.0.13

(Primer-E, Quest Research Ltd). Data were

untransformed, converted into similarity matri-

ces using Bray-Curtis distances. Multivariate

two-way permutational analysis of variance

(PERMANOVA) was performed in Primer

v7.0.13 with the PERMANOVA + 1 add-on

(Anderson, Gorley, & Clarke, 2008). Significant

differences of PERMANOVA routine compar-

ing distances between centroids in treatments

and time, resulted in further analysis using a

similarity percentages (SIMPER) routine. The

PERMANOVAs were conducted using Type

III (partial) sums of squares, treatment as fixed

effects, and time as random effects with 9 999

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the lowest value, with 64 % fecundity success

rate (Fig. 3).

The egg diameter showed that by the end

of the experiment, all of the light regimes pre-

sented mature gametes (> 50 µm). However,

the NP treatment also showed a few immature

gametes, more so than with the other treatments

(Fig. 4).

DISCUSSION

The manipulation of gametogenesis in

aquaculture is not a new concept, in fact it is

widely used to control reproductive timing in

a number of different aquacultured fish and

invertebrate species (Devauchelle & Mingant,

1991; Bromage, Porter, & Randall, 2001). The

generation of an “out of season” or “perma-

nent” harvest season in sea urchin aquaculture

is a main goal (Walker & Lesser, 1998). Using

photoperiod to modify sea urchin gameto-

genesis is not a novel idea; however, it can

produce a variety of effects. For example, in

Strongylocentrotus droebachiensis in the U.S.,

it was possible to manipulate the production of

mature gametes by maintaining the spring light

regime (12L:12D). Here it should be stated

that the reproductive cycle was advanced by

six months. Viable gametes could not be pro-

duced in a large number prior to nine months

(Kirchhoff et al., 2010). In the same species in

Norway, in an aquaculture system, sea urchins

were able to produce mature gametes over a six

months period by simulating a summer pho-

toperiod (James, Siikavuopio, & Mortensen,

Fig. 2. Total gametes spawned in order of thousands through time period and light-darkness regimes in Arbacia dufresnii.

CD = Constant darkness. NP = Neutral photoperiod. CL = Constant light.

TABLE 2

PERMANOVA of total number of gametes in relation to the light regime (treatment) and time (day)

Factor S.S. D.F. M.S. Pseudo-F P (perm)

Time 66892 2 33446 29.143 0.0001

Treatment 5180.4 2 2590.2 4.5599 0.00328

Day*Treatment 2170.4 4 542.6 0.47279 0.911

Residuals 1.2624E+05 110 1147.7

Fig. 3. Fecundity success after 30 and 60 days of light

treatment as regards Arbacia dufresnii. CD = Constant

darkness. CL = Constant light. NP = Neutral photoperiod.

The * refers to the treatments showing significant

differences.

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Fig. 4. Egg diameter of day 60 of light/darkness treatment in Arbacia dufresnii. CD = Constant darkness. NP = Neutral

photoperiod. CL = Constant light. Each figure represents an individual.

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2015). In Paracentrotus lividus, an advanced

photoperiod, in combination with a constant

temperature, was used successfully to promote

“out of season” gonadal maturation. However,

this required at least three months and there

is no information as to whether the gametes

were mature (Kelly, Carboni, Cook, & Hughes,

2015). In addition, constant darkness reduced

the gonad production (McCarron, Burnell, &

Mouzakitis, 2010), as we also found in our

study. In populations of the same species in

Israel, long day treatments were able to delay

gametogenesis in comparison with darkness

and shorter days (Shpigel, McBride, Marciano,

& Lupatsch, 2004). In Strongylocentrotus pur-

puratus, oogenesis can be inhibited by long

days (16L:8D) and activated by short days

(8L:16D) and a neutral photoperiod (12L:12D)

(Bay-Schimith & Pearse, 1987). In Eucidaris

tribuloides, a tropical species, the change in

the photoperiod was the key to producing out

of season gametogenesis and to increasing

the number of gametes. Shorter days were

more effective (9L:15D) (McClintock & Watts,

1990). The females of Psammechinus miliaris

are incapable of completing gametogenesis in

short photoperiods and the photoperiod appears

to be the primary stimulus for gametogenesis

(Kelly, 2001). As regards Tripneustes gratilla,

a private Australian company, a spin-off from

research activity at Macquarie University, was

able to maintain sea urchins at a specific

reproductive stage through manipulation of the

photoperiod (Vaïtilingon & Williamson, 2008).

In spite of this, not every species seems to be

influenced by the photoperiod in this manner.

For example, Loxechinus albus in the Beagle

Channel showed a negative correlation with

photoperiod (Pérez, Boy, Morriconi, & Calvo,

2010). Pseudechinus magellanicus, a spe-

cies coexisting with A. dufresnii, also showed

a negative correlation with the photoperiod

(Marzinelli et al., 2006). In the aquaculture of

Strongylocentrotus intermedius in Japan, pho-

toperiod manipulation is not used to promote

gametogenesis, as temperature seems to be

the most important factor (Musgrove, 2005;

Unuma et al., 2015), and, in China, photoperiod

control is not widely used to condition sea

urchin aquaculture (Liu & Chang, 2015).

Wild A. dufresnii start the gametogenesis

during autumn (April-May) when the days are

shortening, followed by spawning during the

spring and summer when there are lengthen-

ing days. A positive correlation between gonad

weight and photoperiod was previously sug-

gested (Brogger et al., 2010; Epherra et al.,

2014). Surprisingly, in aquaculture systems of

A. dufresnii, only two months are needed to

develop mature gametes regardless of the light

regime. In this study, the highest quantities of

gametes were found at constant light; however,

only the sea urchins with a neutral photope-

riod evidenced mature gametes in excess of

90 %. The constant light treatment diminished

the fecundity success rate by 30 % and also

generated a problem with the seawater qual-

ity as water exchange and cleaning had to be

undertaken more often due to the growth of

microalgae. Constant darkness, on the other

hand, even though it produced mature gametes,

resulted in an amount of gametes equivalent

to 50 % of the amount produced by the other

light regimes. This indicates that periods of

darkness are needed to successfully generate

gametes in good numbers in A. dufresnii. We

present the first study on the effect of photope-

riod on fecundity in the sea urchin A. dufresnii.

Our results suggest that the reproductive cycle

of this species can be altered by modifying

the photoperiod. We found that by using a

12L:12D photoperiod in the aquaculture sys-

tem, mature gametes were obtained in only two

months, and the number of gametes was almost

10 times in excess of the amount produced by

wild sea urchins. This study suggests the fea-

sibility of the production of mature gametes in

a short period of time in the novel aquaculture

species A. dufresnii.

Ethical statement: authors declare that

they all agree with this publication and made

significant contributions; that there is no con-

flict of interest of any kind; and that we fol-

lowed all pertinent ethical and legal procedures

and requirements. All financial sources are

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fully and clearly stated in the acknowledge-

ments section. A signed document has been

filed in the journal archives.

ACKNOWLEDGMENTS

We are grateful to the diver Ricardo Bebo

Vera for the collection of the sea urchins, to

Mariano Moris, for helping with the experi-

ments, and to Kathleen C. Anderson for the

revision of the English. This work was done

with funds from PIP 0352/14, PICT 2018-1729.

The sea urchins were collected on the basis

of the Provincial Permit N°586/18. The first

author has a PhD Scholarship from CONICET

and Chubut Province.

RESUMEN

Fotoperiodo en la acuicultura del erizo de mar

Arbacia dufresnii (Echinodermata: Echinoidea):

El efecto en la producción y madurez de gametas

Introducción: El fotoperiodo es, junto con la tempe-

ratura y la disponibilidad de alimentos, uno de los principa-

les estímulos para el desarrollo de la gametogénesis en una

amplia variedad de especies. Objetivo: Evaluar el efecto

del fotoperiodo en la producción de gametas maduras de

Arbacia dufresnii en un sistema de recirculación cerrado

para determinar el mejor fotoperiodo para una acuicultura

novedosa, enfocada en la producción de gametas con alta

concentración de pigmentos para usos biotecnológicos.

Métodos: Se realizó un experimento con tres regímenes/

tratamientos diferentes de luz y oscuridad: luz constante

(luz durante 24 h), fotoperiodo neutro (12 h de luz, 12 h de

oscuridad) y oscuridad constante (oscuridad durante 24 h).

Se utilizaron veinte hembras en cada tratamiento. Se indujo

a todas las hembras a desovar al comienzo del experimento.

Después de 30 días, diez hembras seleccionadas al azar de

cada tratamiento fueron inducidas a desovar nuevamente.

Al final del experimento, después de 60 días, se indujo

el desove a las hembras restantes en cada tratamiento.

Las gametas se recolectaron en agua de mar filtrada,

se fijaron en solución de Davidson, se cuantificaron y

midieron por triplicado en una cámara Sedgewick-Rafter.

Para determinar la maduración, se evaluó el éxito de la

fecundación después de 30 minutos de fertilización, cal-

culando el porcentaje de huevos fertilizados. Resultados:

Nuestros resultados muestran que, en el sistema acuícola,

en solo dos meses se obtuvieron gametas maduras y casi

10 veces más la cantidad producida por los erizos de mar

en su ambiente natural usando el fotoperiodo neutro (12 h

luz:12 h oscuridad). También encontramos que la mayor

exposición a la luz produce la menor cantidad de gametas

maduras. Conclusión: Este estudio sugiere la viabilidad de

la producción de gametos maduros en un corto período de

tiempo en Arbacia dufresnii.

Palabras clave: productividad gonadal; equinodermo;

equinoideo; acuacultura; gametas maduras.

REFERENCES

Anderson, M.J., Gorley, R.N., & Clarke, K.R. (2008).

PERMANOVA+ for PRIMER: Guide to software

and statistical methods. Plymouth, United Kingdom:

Primer_E Ltd.

Bay-Schmith, E., & Pearse, J.S. (1987). Effect of fixed

daylengths on the photoperiodic regulation of

gametogenesis in the sea urchin Strongylocentrotus

purpuratus. International Journal of Invertebrate

Reproduction and Development, 11(3), 287-294.

Brogger, M.I., Martinez, M.I., & Penchaszadeh, P.E. (2010).

Reproduction of the sea urchin Arbacia dufresnii

(Echinoidea: Arbaciidae) from Golfo Nuevo, Argen-

tina. Journal of the Marine Biological Association of

the United Kingdom, 90(7), 1405-1409.

Brogger, M., Gil, D.G., Rubilar, T., Martinez, M., Díaz

de Vivar, M.E., Escolar, M., Epherra, L., Pérez, A.,

& Tablado, A. (2013). Echinoderms from Argen-

tina: Biodiversity, distribution and current state of

knowledge. In J.J. Alvarado & F. Solís-Marín (Eds.),

Echinoderm Research and Diversity in Latin America

(pp. 359-402). Berlín, Heidelberg: Springer-Verlag.

Bromage, N., Porter, M., & Randall, C. (2001). The envi-

ronmental regulation of maturation in farmed finfish

with special reference to the role of photoperiod and

melatonin. Aquaculture, 197, 63-98.

Byrne, M. (1990). Annual reproductive cycle of the com-

mercial sea urchin Paracentrotus lividus from an

exposed intertidal and a sheltered subtidal habitat

on the west coast of Ireland. Marine Biology, 104,

275-289.

Cárcamo, P. (2004). Massive production of larvae and

seeds of the sea urchin Loxechinus albus. In J.M.

Lawrence & O. Guzmán (Eds.), Sea urchins: Fishe-

ries and Ecology (pp. 299-306). Lancaster: DEStech

Publications Inc.

Devauchelle, N., & Mingant, C. (1991). Review of the

reproductive physiology of the scallop, Pecten maxi-

mus, applicable to intensive aquaculture. Aquatic

Living Resources, 4(1), 41-51.

Díaz-Martínez J.P., Carpizo-Ituarte, E.J., & Benítez-Villa-

lobos, F. (2019). Reproductive patterns of the black

starry sea urchin Arbacia stellata in Punta Banda,

Baja California, Mexico. Journal of the Marine

472

Revista de Biología Tropical, ISSN electrónico: 2215-2075 Vol. 69(S1): 464-473, March 2021 (Published Mar. 10, 2021)

Biological Association of the United Kingdom, 99(6),

1379-1391.

Epherra, L., Gil, D.G., Rubilar, T., Perez-Gallo, S., Reartes,

M.B., & Tolosano, J.A. (2014). Temporal and spatial

differences in the reproductive biology of the sea

urchin Arbacia dufresnii. Marine and Freshwater

Research, 66, 329-342.

Fernández, J.P., Epherra, L., Sepúlveda, L., & Rubilar, T.

(2019). Desarrollo embrionario y larval del erizo de

mar verde Arbacia dufresnii. Naturalia Patagónica,

14, 44-58.

Fuji, A. (1967). Ecological studies on the growth and

food consumption of Japanese common littoral sea

urchin, Strongylocentrotus intermedius (A. Agassiz).

Memoirs of the Faculty of Fisheries Hokkaido Uni-

versity, 15(2), 83-160.

Gianguzza, P., & Bonaviri, C. (2013). Arbacia. In J.M.

Lawrence (Ed.), Developments in Aquaculture and

Fisheries Science (Vol. 38, pp. 275-283). Amsterdam:

Elsevier.

Harris, L.G., & Eddy, S.D. (2015). Sea urchin ecology and

biology. Echinoderm Aquaculture, 1-24.

James, P., Siikavuopio, S.I., & Mortensen, A. (2015). Sea

urchin aquaculture in Norway. Echinoderm Aquacul-

ture, 147-173.

Kelly, M.S. (2001) Environmental parameters controlling

gametogenesis in the echinoid Psammechinus milia-

ris. Journal of Experimental Marine Biology and

Ecology, 266(1), 67-80.

Kelly, M., Carboni, S., Cook, E., & Hughes, A. (2015). Sea

urchin aquaculture in Scotland. Echinoderm Aqua-

culture, 211-224.

Kirchhoff, N., Eddy, S., & Brown, N. (2010) Out-of-season

gamete production in Strongylocentrotus droeba-

chiensis: Photoperiod and temperature manipulation.

Aquaculture, 303(1-4), 77-85.

Costa-Leal, M., Rocha, R.J.M., Rosa, R., & Calado, R.

(2016). Aquaculture of marine non-food organisms:

What, why and how?. Reviews in Aquaculture, 10(2),

400-423.

Liu, H., & Chang, Y.Q. (2015). Sea urchin aquaculture in

China. Echinoderm Aquaculture, 127-146.

Marzinelli, E.M., Bigatti, G., Giménez, J., & Penchas-

zadeh, P.E. (2006). Reproduction of the sea urchin

Pseudechinus magellanicus (Echinoidea: Temno-

pleuridae) from Golfo Nuevo, Argentina. Bulletin of

Marine Science, 79(1), 127-136.

McCarron, E., Burnell, G., & Mouzakitis, G. (2010). An

experimental assessment on the effects of photope-

riod treatments on the somatic and gonadal growth

of the juvenile European purple sea urchin Paracen-

trotus lividus. Aquaculture Research, 41, 1072-1081.

McClintock, J.B., & Watts, S.A. (1990). The effects of

photoperiod on gametogenesis in the tropical sea

urchin Eucidaris tribuloides (Lamarck) (Echinoder-

mata: Echinoidea). Journal of Experimental Marine

Biology and Ecology, 139(3), 175-184.

Meidel, S.K., & Scheibling, R.E. (1998) Annual reproduc-

tive cycle of the green sea urchin, Strongylocentrotus

droebachiensis, in differing habitats in Nova Scotia,

Canada. Marine Biology, 131, 461-478.

Mercier, A., & Hamel, J.F. (2009). Endogenous and exoge-

nous control of gametogenesis and spawning in echi-

noderms. Advances in Marine Biology, 55, 1-302.

Musgrove, R.J.B. (2005). Aquaculture and Diet Develo-

pment Subprogram: Postharvest Enhancement of

Sea Urchin Roe for the Japanese Market. Australia:

FRDC Project 99/319, SARDI Aquatic Sciences

Publication No. RD03/0102.

Pearce, C.M., Daggett, T.L., & Robinson, S.M.C. (2002).

Optimizing prepared feed ration for gonad production

of the green sea urchin Strongylocentrotus droeba-

chiensis. Journal of the World Aquaculture Society,

33(3), 268-277.

Pearce, C.M., Daggett, T.L., & Robinson, S.M. (2004).

Effect of urchin size and diet on gonad yield and

quality in the green sea urchin. Aquaculture, 223(1-

4), 337-367.

Pearse, J.S., & Eernisse, D.J. (1982). Photoperiodic regu-

lation of gametogenesis and gonadal growth in the

sea star Pisaster ochraceus. Marine Biology, 67,

121-125.

Pearse, J.S., & Walker, C.W. (1986). Photoperiodic regu-

lation of gametogenesis in a North Atlantic sea star,

Asterias vulgaris. International Journal of Inverte-

brate Reproduction and Development, 9, 71-77.

Pérez, A.F., Boy, C., Morriconi, E., & Calvo, J. (2010).

Reproductive cycle and reproductive output of the

sea urchin Loxechinus albus (Echinodermata: Echi-

noidea) from Beagle Channel, Tierra del Fuego,

Argentina. Polar Biology, 33(3), 271-280.

Rubilar, T., Epherra, L., Deias-Spreng, J., Díaz De Vivar,

M.E., Avaro, M., Lawrence, A.L., & Lawrence, J.M.

(2016). Ingestion, absorption and assimilation effi-

ciencies, and production in the sea urchin Arbacia

dufresnii fed a formulated feed. Journal of Shellfish

Research, 35(4), 1083-1093.

Spirlet, C., Grosjean, P., & Jangoux, M. (2000). Optimiza-

tion of gonad growth by manipulation of temperature

and photoperiod in cultivated sea urchins, Paracen-

trotus lividus (Lamarck) (Echinodermata). Aquacul-

ture, 185(1-2), 85-99.

473

Revista de Biología Tropical, ISSN electrónico: 2215-2075, Vol. 69(S1): 464-473, March 2021 (Published Mar. 10, 2021)

Strathmann, R. (1987). Echinoderm larval ecology viewed

from the egg. Echinoderm Studies, 2, 55-136.

Unuma, T., Konishi, K., Furuita, H., Yamamoto, T., &

Akiyama, T. (1996). Seasonal changes in gonad of

cultured and wild red sea urchin Pseudocentrotus

depressus. Suisanzoshoku, 44(2), 169-175.

Unuma, T. (2002). Gonadal growth and its relationship to

aquaculture in sea urchins. In Y. Yokota, V. Matranga,

& Z. Smolenicka (Eds.), The Sea Urchin: From Basic

Biology to Aquaculture (pp. 115-127). Rotterdam:

A.A. Balkema Publishers.

Unuma, T. & Walker, C.W. (2010). The role of the major

yolk protein in sea urchin reproduction and its rele-

vance to aquaculture. In L. Harris, S.A. Böttger, C.W.

Walker, & M.P. Lesser (Eds.), Echinoderms: Durham

(pp. 437-444). London: Taylor and Francis Group.

Unuma, T., Sakai, Y., Agatsuma, Y., & Kayaba, T. (2015).

Sea urchin aquaculture in Japan. Echinoderm Aqua-

culture, 77-126.

Vaïtilingon, D., & Williamson, J.E. (2008). WIPO Patent

No. WO/2008/074084. Geneva, Switzerland: World

Intellectual Property Organization.

Wangensteen, O.S., Turon, X., Casso, M., & Palacín, C.

(2013). The reproductive cycle of the sea urchin

Arbacia lixula in northwest Mediterranean: potential

influence of temperature and photoperiod. Marine

Biology, 160(12), 3157-3168.

Walker, C.W., & Lesser, M.P. (1998). Manipulation of

food and photoperiod promotes out-of-season game-

togenesis in the green sea urchin, Strongylocentrotus

droebachiensis: implications for aquaculture. Marine

Biology, 132, 663-676.

Walker, C.W., Lesser, M.P., & Unuma, T. (2013). Sea

urchin gametogenesis-structural, functional and

molecular/genomic biology. In: C.A. Lawrence (Ed.),

Sea urchins: Biology and ecology (Vol. 3, pp. 25-44).

San Diego: Elsevier Academic Press.

Walker, C.W., Böttger, S.A., Unuma, T., Watts, S.A.,

Harris, L.G., Lawrence, A.L., & Eddy, S.D. (2015).

Enhancing the commercial quality of edible sea

urchin gonads-technologies emphasizing nutritive

phagocytes. Echinoderm Aquaculture, 263-286.

Watts, S.A., Lawrence, A.L., & Lawrence, J.M. (2013).

Nutrition. In: J.M. Lawrence (Ed.), Sea urchins: Bio-

logy and ecology (Vol. 3, pp. 155-169). San Diego:

Elsevier Academic Press.