Revista de Biología Tropical, ISSN: 2215-2075, Vol. 71 (S1): e54926, abril 2023 (Publicado Abr. 30, 2023)

Ex situ culture of coral species Porites lobata (Scleractinia: Poritidae) and

Pocillopora damicornis (Scleractinia: Pocilloporidae), Costa Rica: first

assessment and implications

José A. Marín-Moraga1*; https://orcid.org/0000-0001-8499-5620

Jonathan Chacón-Guzmán2; https://orcid.org/0000-0002-8449-2334

Mauricio Méndez-Venegas3; https://orcid.org/0000-0003-4855-3731

Ronny A. Hernández-Mora4; https://orcid.org/0000-0001-6225-7096

Jorge Cortés5; https://orcid.org/0000-0001-7004-8649

1. Raising Coral Costa Rica, San José, Costa Rica; jamm@raisingcoral.org (*Correspondence)

2. Programa Parque Marino del Pacífico, Universidad Nacional, Puntarenas, Costa Rica;

jonathan.chacon.guzman@una.cr

3. Sistema Nacional de Áreas de Conservación (SINAC), Costa Rica; mauricio.mendez@sinac.go.cr

4. Centre for Earth Observation Sciences, Department of Earth and Atmospheric Sciences, University of Alberta,

Edmonton, Alberta, Canada; ronnyale@ualberta.ca

5. Centro de Investigación en Ciencias del Mar y Limnología (CIMAR), Ciudad de la Investigación, Universidad de

Costa Rica, San Pedro, 11501-2060 San José, Costa Rica; jorge.cortes@ucr.ac.cr

Received 30-IX-2022. Corrected 31-I-2023. Accepted 16-II-2023.

ABSTRACT

Introduction: Coral reefs worldwide decline has prompted coral restoration as a viable strategy to rewild vul-

nerable, foundational coral species. Stony corals are now propagated by thousands in both in-water and ex situ

(land-based) coral nurseries, the latter being unexplored in Costa Rica, despite their potential benefits as a reef

management tool.

Objective: To analyze the viability of ex situ culturing of the Pacific reef-building corals Porites lobata and

Pocillopora damicornis at Parque Marino del Pacífico (PMP), Puntarenas, Costa Rica, aquaculture facilities.

Methods: From May to October 2018 a total of 180 coral fragments were kept in an aquaculture recirculated

system. Survival, growth, and fragment yield in relation to culture medium (physicochemical parameters) were

recorded.

Results: Survival and growth rate varied between species and culture tanks. On average, surviving P. lobata

fragments (68.89 %) placed in Tank 1 (T1) grew 216 %, while fragments placed in Tank 2 (T2) had a survival

rate of 71.11 % and an increase of 277 % in live tissue area. P. damicornis fragments survival, basal and crown

area percentage increase were: 71.11 %, 980 % and 366 % in T1, and 100 %, 976 % and 287 % in T2. Although

fragments survival and growth were net positive, the yield in terms of culture was low, due to culture conditions

in the tanks not meeting coral culture optimal requirements.

Conclusions: Survival and growth of both species varied depending on the tank in which they were placed.

Survival was similar to that found in other ex situ studies and growth was similar to those reported in the wild,

however culture performance in terms of yield was low. Aquaculture systems at PMP constitute a good base

for the cultivation of corals, however for the culture effort to achieve maximum yield, current systems must be

optimized according to the requirements of the target coral species.

Key words: coral aquaculture; Parque Marino del Pacífico; Porites lobata; Pocillopora damicornis; Costa Rica;

survival; growth; culture yield.

https://doi.org/10.15517/rev.biol.trop..v71iS1.54926

SUPPLEMENT

2Revista de Biología Tropical, ISSN: 2215-2075 Vol. 71 (S1): e54926, abril 2023 (Publicado Abr. 30, 2023)

INTRODUCTION

Current reef degradation is mostly a con-

sequence of human-related activities. Rising

sea temperatures and land-produced pollu-

tion and sedimentation threaten reefs capac-

ity to provide invaluable ecosystem services

to coastal communities (Matthews & Wynes,

2022). Scleractinian corals, mainly reef-build-

ing species, and by extension reefs, are in a

ticking race against time, as recent models sug-

gest irreversible damage if serious committed

actions to achieve 1.5 °C goal are not taken

before the year 2050 (Kleypas et al., 2021).

While we as a society develop the strategies

and the technology to mitigate the root cause

(carbon emissions), it is paramount to buy time,

by fostering reef recovery and corals’ adaptive

capacity (Baums et al., 2019).

Coral restoration is a novel management

tool, that combines theories and concepts of

marine ecology with terrestrial reforestation

methods. In practice, restoration takes the form

of coral gardening, a science-based two-step

process consisting of (1) cultivating foun-

dational coral species and (2) transplanting

them in to denude parts of the reef (Rinkevich,

1995). The former can be done in site or in situ

(typically involving some form of underwater

nursery) or out of it —a.k.a. ex situ coral aqua-

culture—. Ex situ coral culture has proved to

be a valuable restoration tool as it enables the

mass production of coral fragments and larvae.

For example, Shafir et al. (2001) carried out

fundamental work, evaluating the feasibility of

using 821 “nubbins” or buds (tiny fragments

of no more than 15 polyps), as an effective

method in the production of propagative coral

RESUMEN

Cultivo ex situ de las especies de coral Porites lobata (Scleractinia: Poritidae) y Pocillopora damicornis

(Scleractinia: Pocilloporidae), Costa Rica: primera evaluación e implicaciones.

Introducción: El declive mundial de los arrecifes de coral, ha impulsado a la restauración coralina como una

estrategia viable para recuperar especies de coral fundacionales, en estado vulnerable. Los corales pétreos se

propagan por miles, tanto en viveros subacuáticos como ex situ (en tierra). Siendo el segundo método poco explo-

rado en Costa Rica, a pesar de sus potenciales beneficios como medida como herramienta de manejo arrecifal.

Objetivo: Analizar la viabilidad del cultivo ex situ de las especies de coral constructoras de arrecifes Porites

lobata y Pocillopora damicornis en el módulo de acuicultura del Parque Marino del Pacífico (PMP), Puntarenas,

Costa Rica.

Métodos: Desde el 17 de mayo hasta el 17 de octubre de 2018, se mantuvieron un total de 180 fragmentos de

coral en un sistema de recirculación de acuicultura. Se registraron la supervivencia, el crecimiento y el rendi-

miento de los fragmentos en relación con el medio de cultivo (parámetros fisicoquímicos).

Resultados: La tasa de supervivencia y crecimiento varió entre especies y tanques de cultivo. En promedio, los

fragmentos de P. lobata supervivientes (68.89 %) colocados en el tanque 1 (T1) crecieron un 216 %. En con-

traste con los fragmentos colocados en el tanque 2 (T2) que mostraron una tasa de supervivencia del 71.11 % y

un aumento del 277 % en el área de tejido vivo. En el caso de P. damicornis, los porcentajes de supervivencia,

de aumento del área basal y del área de la corona fueron: 71.11 %, 980 %, y 366 %, y 100 %, 976 %, y 287 %

para los fragmentos colocados en T1 y T2, respectivamente. Aunque la supervivencia y el crecimiento de los

fragmentos fueron positivos, el rendimiento en términos de cultivo fue bajo, debido a que las condiciones en los

tanques no cumplían con las condiciones ideales para el cultivo de corales.

Conclusiones: La supervivencia y el crecimiento de ambas especies variaron en función del tanque en el que

se colocaron. La supervivencia fue similar a la observada en otros estudios ex situ y el crecimiento fue similar

al reportado en la naturaleza, pero el rendimiento del cultivo fue bajo. Los sistemas de acuicultura del PMP

constituyen una buena base para el cultivo de corales, sin embargo, para que el esfuerzo de cultivo alcance un

máximo de rendimiento, los sistemas actuales deben optimizarse en función de los requisitos de las especies de

coral objetivo.

Palabras clave: acuicultura de coral; Parque Marino del Pacífico; Porites lobata; Pocillopora damicornis; Costa

Rica; supervivencia; crecimiento; rendimiento de cultivo.

Revista de Biología Tropical, ISSN: 2215-2075, Vol. 71 (S1): e54926, abril 2023 (Publicado Abr. 30, 2023)

material. Pillay et al. (2012) compared the

growth rates and survival of Pocillopora dami-

cornis (Linnaeus, 1758) in an in situ and an ex

situ nursery for two years, resulting in a signifi-

cantly higher survival at the land base nursery.

Although most propagation studies focus on

fast-growing branching species, Forsman et

al. (2006) investigated the effect of initial cut-

ting size on the cultivation of massive coral

fragments (Porites compressa Dana, 1846 and

Porites lobata Dana, 1846), and found a sig-

nificant positive relation between fragment size

and growth, but no evidence of size-specific

mortality. In 2015, Forsman and colleagues,

expand this notion, characterizing not only

coral fission properties but also exploring coral

fusion restoration potential.

Parque Marino del Pacífico (PMP) is a

department of the Ministry of Environment

and Energy of Costa Rica (MINAE), design to

bolster marine social-focused endeavors for the

sustainable development of the Pacific coast

(https://parquemarino.org). These includes,

among other activities, the promotion of proj-

ects and research in marine aquaculture with

emphasis on the production of commercial fish

species through the Laboratory of Biotechnol-

ogy and Marine Aquaculture.

The objective of our study was to describe

ex situ coral culture viability (in terms of sur-

vival and growth) of the two-main reef-build-

ing species of the Pacific coast of Costa Rica,

a massive species, P. lobata, and a branching

coral, P. damicornis. We also described a meth-

od to account for production yield and present

coral culture recommendations.

MATERIALS AND METHODS

Collection site: Playa Blanca fringing

reef, with depths between 2-14 m, limits to

the east with Playa Matapalo and to the west

with Punta Matapalito (Méndez-Venegas et

al., 2021). Five healthy ~15 cm2 P. damicornis

coral colonies and one ~12 cm2 Porites lobata

colony were collected during April 2018 by

Scuba diving at Playa Blanca (10°32’04.5’’N,

85°45’47.6” W), Guanacaste. The colonies

were transported in a cooler with seawater and

aeration to the Marine Aquaculture Research

and Marine Biotechnology facilities at the

Parque Marino del Pacífico, Puntarenas. Once

in the wet lab, colonies were acclimatized

(28°C) for one week and then fragmented

(i.e., micro fragments between 1.5-2.0 cm2).

A total of 180 coral fragments where gener-

ated and fixed to ceramic plugs. The surface

of the ceramic plugs surrounding coral tissue

was carefully scraped two times per week

with a nylon toothbrush in order to prevent

algal growth or biofouling accumulation. The

experimental trial lasted for 152 days.

Fragments and tanks set-up: Fragments

were randomly placed in one of six table-like

grid structures (60 x 60 cm2), each holding

15 fragments of P. lobata and 15 fragments of

P. damicornis. Coral tables were distributed

between two tanks, for a total of 90 fragments

(45 per species) in each tank. The 10 m3 circular

fiberglass tanks were placed on a concrete slab

under a mesh cover with 80 % sunlight filtra-

tion. Each tank was connected to a semi-closed

recirculation system with 5 % continuous flow

of fresh seawater from a common reservoir.

The recirculation flow rate was 60 L/min. The

systems were composed of a mechanical filter

model Astral pool Aster 99, a biological filter

(plastic canisters with 60 L of filtering material

from plastic biobarrels), a three-phase centrifu-

gal pump 230 v, 0.5 HP, a UV filter brand UV

STERILIZIER and a foam fractionator RK2,

model RK10AC-PF. In addition, two aeration

stones and one HOBO temperature sensors

per tank were placed. It was assumed that the

tanks had the same conditions and therefore

one would be the back-up of the other in

case of fragments collapsed. Photosynthetic

active radiation or PAR was measured using a

Sper Scientific luxmeter model L860152. To

determine the nutrient content (ammonium,

phosphates, nitrates, nitrites) per tank, water

samples were taken weekly and then analyzed

using a Continuous Flow Autoanalyzer (FIA),

at the Chemical Oceanography Laboratory,

CIMAR, University of Costa Rica. Alkalinity,

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calcium, and magnesium were measured using

commercial Aquaforest®. Salinity was mea-

sured at convenience using a refractometer.

Growth and survival: Photographs were

taken every 15 days from May 17th to October

17th 2018 with an Olympus (TG-5) waterproof

camera. Survival and fragment growth (as a

conservative estimate of change in live tissue

area) (Forsman et al., 2006) were estimated

from these photographs, using ImageJ 1.45 free

software (Collins, 2007) by calibrating pixel

value statistics using fixed ceramic plug area

(7.02 cm2), grid length (1.6 cm) and ceramic

plug (0.5 cm). In the case of P. lobata, the area

calculated from top view photographs cor-

responds to the total live tissue covering the

ceramic plug, while for P. damicornis the area

corresponds to the basal area (BA) of tissue

attached to the ceramic plugs. The length (the

maximum distance between the distal branch-

es) and width (the measure perpendicular to

the length) of P. damicornis branches were

also calculated from top-view photographs.

The height of each P. damicornis fragment

was calculated from lateral photographs. Out

of these three metrics, the crown area (CA)

(Baums et al., 2019) and ellipsoidal volume

(Vol) (Salinas-Akhmadeeva, 2018) for each P.

damicornis fragment were calculated.

Survival percentage was calculated by

subtracting the number of dead coral fragments

from the total sample. Growth rate (GR) was

calculated for the surviving fragments, as the

difference between current (afraC) and prior

measurement (afragP) divided by the days

passed between each other (tC-Tp). [GR (cm2

d-1) = (afragC - afragP) / (tC - tP)].

Porites lobata culture yield: culture yield

(Y) for this species was calculated as the yield

obtained proportional to the expected yield,

which equals 7.07 cm2 of live-coral tissue

covering the ceramic plug. In order to achieve

this, the following equations modified from

Schippers et al. (2012) were used:

Where Resp is the expected yield (the

desired harvest expressed in cm2), Nfrag is the

total initial number of fragments to be grown,

and Pfrag is the average harvest productivity

per fragment during the culture period; L is

suggested as the lag factor considering the sur-

vival (100 - % of surviving fragments that did

not reach the desired harvest size/100), and Ft

is the fragment size at the end of the harvesting

period of duration t.

Statistical analyses: Wilcoxon rank sum

test was used to establish significant differ-

ences between variables (except salinity, which

was not systematically measured) per tank.

Survival among species was compared using

the Kaplan-Meir method (Lee, 1992). Growth

data was transformed to log10 and fit into gen-

eral linear regression models. Spearman’s cor-

relation analysis was conducted to determine

the relationship between crown area (CA) and

volume (Vol). All statistical analyses were con-

ducted at 95 % confidence level using Rstudio

free software (Grömping, 2015).

RESULTS

Survival: During the third week of Octo-

ber 2018 all fragments in tank 1 (T1) died

due to freshwater entering the culture system

as a consequence of extreme rainfall events

(Morera-Rodríguez, 2018). Prior to this event,

the overall survival for P. lobata fragments was

68.89 % in T1 compared to 71.11 % in tank

2 (T2). In the case of P. damicornis, 71.11 %

of fragments survived in T1, while in tank 2

Revista de Biología Tropical, ISSN: 2215-2075, Vol. 71 (S1): e54926, abril 2023 (Publicado Abr. 30, 2023)

survival was 100 %. Regardless of the species,

a significant difference was found in the total

survival of the fragments placed in T1 with

respect to those placed in T2.

Porites lobata growth: Average initial area

of P. lobata was 1.31 ± 0.37 cm2 and 1.36 ±

0.31 cm2 for fragments placed in T1 and T2,

respectively. After 152 days, fragments in T1

had doubled their area to a total of 2.83 ± 1.07

cm2 (216 % increase), at an average growth

rate of 0.010 ± 0.052 cm2 d-1. It is worth not-

ing that rapid coenosarc formation and tissue

expansion (~ 1 cm2) occurred during the first

month where fragments grew the fastest (0.069

± 0.026 cm2 d-1) until reaching a plateau at day

44, from which both growth and growth rate

decreased slightly but steadily until the end of

the experimental essay (Fig. 1).

The fragments of P. lobata in T2 expe-

rienced a rapid increase in area from 1.36 ±

0.31 cm2 to 2.06 ± 0.69 cm2 at 0.039 ± 0.053

cm2 d-1 rate during the first month; reaching

a maximum of 4.43 ± 1.55 cm2 at day 113,

from which shortly after, growth started to

decreased, resulting in a final area of 3.78 ±

1.55 cm2 (277 %) (Table 1). In addition, seven

T2 fragments were able to fully cover the

ceramic plug area in ~ 90 days.

Linear regression indicates that change in

area through time, for P. lobata fragments var-

ied significantly (p-value < 0.005) as a function

of the tank in which they were placed, although

the residence time in the tanks alone explained

only 16 % of the variation in growth (r2 = 0.16),

and 18 % of growth rate (r2 = 0.18) (Fig. 2).

Porites lobata culture yield: Yield (desired

harvest of live tissue coral area) was calculated

for T2 fragments that reached harvest size (7.07

cm2 or a fully cover ceramic plug). For an ini-

tial number of 45 Porites fragments (desired

harvest) would be 318.15 cm2. Lag value (Z)

was 0.78 (100-78 % of surviving fragments

Fig. 1. An example of the growth of a Porites lobata fragment placed in T1; A. Expansion of the coenosarc (13 days); B.

tissue with visible polyp formation (42 days); C. loss of tissue in the periphery of the fragment (55 days).

TABLE 1

Change in total number, percent survival, and growth values associated with P. lobata during the experimental period.

Tank Initial N Final N Survival* Initial fragment

area* (cm2)

Final fragment

area*(cm2)

Average

increased (%)

Growth rate*

(cm2 d-1)

T1 45 31 68.89 % 1.31 ± 0.37 2.83 ± 1.07 216 0.010 ± 0.052

T2 45 32 71.11 % 1.36 ± 0.31 3.78 ± 1.55 277 0.016 ± 0.0.43

Statistical significance

(p-value)

5 x10-08 0.002 0.0008

* Symbol denotes significant differences (P-value < 0.05) between tanks.

6Revista de Biología Tropical, ISSN: 2215-2075 Vol. 71 (S1): e54926, abril 2023 (Publicado Abr. 30, 2023)

that didn’t reach culture size during the experi-

mental period/100). Average productivity per

fragment was 1.55 (Pfrag = 1-0.78 x 7.07 cm2).

Actual yield obtained was 69.75 cm2 (45 x

1.55); 21 % of total biomass that formerly enter

the aquaculture system (Fig. 3).

Pocillopora damicornis growth: Growth

pattern varied between fragments. Some frag-

ments’ basal area managed to cover the major-

ity of the ceramic plug, but had poor branch

development; while the opposite was true for

other fragments. Hence, the high standard

Fig. 2. Porites lobata fragments growth (upper panel, P-value < 2.2 x10-16, r2 = 0.16) and growth rate (lower panel, P-value<

2.2 x10-16, r2 = 0.18) related to days at tank 1 and tank 2.

Revista de Biología Tropical, ISSN: 2215-2075, Vol. 71 (S1): e54926, abril 2023 (Publicado Abr. 30, 2023)

deviation for growth metrics values. Basal area

for T1 fragments increased from 0.57 ± 0.15

cm2 to 4.36 ± 2.01 cm2 at an initial growth rate

of 0.032 ± 0.030 cm2 d-1 that notably decreased

(0.001 ± 0.001 cm2d-1) towards the end of the

experiment (Table 2). T2 fragments increased

their BA from 0.59 ± 0.24 cm2 to 5.76 ± 1.29

cm2 at an initial growth rate of 0.026 ± 0.024

cm2 d-1 which then decreased to 0.005 ± 0.015

cm2d-1 (Fig. 4).

Crown area increased from 1.50 ± 0.76

cm2 to 5.49 ± 4.01 cm2 at a rate of 0.023 ±

0.053 cm2 d-1 in the case of T1 fragments;

change in CA for T2 fragments was from 0.59

± 0.24 cm2 to 4.20 ± 3.12 cm2 at 0.019 ± 0.048

cm2 d-1. Growth rate for both sets of fragments

decreased notably towards the end of the exper-

iment. Final growth rate was 0.001 ± 0.010 cm2

d-1 and 0.015 ± 0.022 cm2 d-1 for T1 and T2,

respectively (Fig. 5). Obtain values for volume

behaved analogously to the CA values (Fig. 6).

Contrary to CA and Vol, no significance

difference was found in BA values between

tanks. Initial CA, Vol and growth rates values in

T1 fragments were slightly higher than T2 frag-

ments. However, growth rate in T1 decreased

over time, as in T2 remained constant. As with

Porites fragments linear regression indicates

that tank factor alone is not a sufficiently

robust predictive variable to explain differ-

ence between growth (T1/T2: r2 = 0.23) and

growth rate (T1/T2: r2 = 0.006) among tanks.

As expected, the same trend is observed for Vol

(Fig. 4, Fig. 5, Fig. 6).

The majority of the physicochemical vari-

ables showed significant differences between

tanks (P-value = 0.005), except for alkalinity.

A significant difference was obtained between

temperature and PAR, both conservative prop-

erties. The majority of physicochemical param-

eters oscillated around the recommended lower

limit, with the exception of temperature (Table

3, Table 4).

DISCUSSION

Porites lobata survival rates for both tanks

were consistent with those reported in Forsman

et al. (2006) which range between 78–92 %. In

the case of P. damicornis, Pillay et al. (2012)

reported survival rates of 100 %; comparable

to the survival of fragments in T2 in this study.

Fragment survival varied between coral

species and culture tank. Fragment death

occurred during the first two weeks of culture,

regardless of the species. This is not surprising

since the period subsequent to subclonation

of donor colonies is one of most sensitive for

the survival of small fragments, susceptible to

predation, sedimentation and/or disease (For-

rester et al., 2013; Lizcano-Sandoval et al.,

2018; Rinkevich, 2005; Shafir et al., 2001).

The latter was especially true for P. damicornis

in T1, which presumably suffered from rapid

tissue necrosis, a common disease in the realm

of coral husbandry, and for which there is no

known trigger; although it is typically associat-

ed with stress (Calfo, 2001; Luna et al., 2007).

Fig. 3. An example of the growth of a P. lobata fragment placed in T2. From left to right, growth from May 17 to August

9 (82 days).

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Fig. 4. Pocillopora damicornis basal area growth (upper panel; P-value = < 2.2 x10-16, r2 = 0.56) and growth rate (lower

panel; P-value = < 2.2 x10-16, r2 = 0.14) related to days at tank 1 and tank 2.

Revista de Biología Tropical, ISSN: 2215-2075, Vol. 71 (S1): e54926, abril 2023 (Publicado Abr. 30, 2023)

The intraspecific variability in P. damicor-

nis survival could be due to intrinsic factors,

such as genetics differences, physiological

state, previous health status, and even the area

of the colony from which the fragments were

extracted (Kenkel & Matz, 2016; Tagliafico

et al., 2018; Yap, 2004). In his coral hus-

bandry compilation manual, Bartlett (2003)

warns that there can be significant differences

between species within the same genus, and

even between colonies of the same species.

For ex situ reared corals, mortality is

usually the result of incidental herbivory by

unwanted commensals, mainly nudibranch

gastropods and Drupella snails (Gochfeld &

Aeby, 1997; Forsman et al., 2006). No com-

mensal organisms were observed during the

152 days of the experimental trial, corrobo-

rating the effectiveness of the filter system.

Another reason related to potential mortality

is the direct effect of nutrients on the algal

growth; Yap & Molina (2003) report survival

rates for Porites as low as > 20 %, attributed to

competition derived from excessive growth of

macroalgae in culture tanks with high nutrient

levels. Although proliferation of macroalgae

inside the tanks was notorious, the frequent

cleaning of the ceramic plugs avoided possible

fragment demise derived from excessive algal

overgrowth.

Coral bleaching and death of Stylophora

pistillata (Esper, 1792) fragments has been

recorded after exposure to salinity levels of

15-18 PSU for 24h (Kerswell & Jones, 2003).

Corals in T1 bleached and die over the course

of 48 hours, presumably as the result of osmot-

ic shock (Hoegh-Guldberg, 1999), due to heavy

rainfall exposure which caused salinity to drop

to 17 PSU.

Regarding growth, P. lobata fragments

increased their area 216–277 % in 152 days;

concurring with previous studies that report

increases in area up to 228 % in 119 days

and 357 % in 205 days (Forsman et al., 2006;

Forsman et al., 2015). While it is true that the

increase in area in this study and that reported

by Forsman et al. (2006) are similar, the

path that led to them was highly contrasting.

TABLE 2

Change in total number, percent survival, basal area (BA) and crown area (CA) values associated with P. damicornis during the experimental period.

Tank Initial

Final

Survival*

(%)

Initial

BA (cm2)

Final BA

(cm2)

Average BA

increased (%)

Initial CA*

(cm2)

Final CA*

(cm2)

Average CA

increased (%)

Initial Vol*

(cm3)

Final Vol*

(cm3)

Average Vol

increased (%)

T1 45 32 71.1 0.57 ± 0.15 5.59 ± 1.86 980 1.50 ± 0.76 5.49 ± 4.01 366 11.12 ± 6.8 58.61 ± 53.75 527

T2 45 45 100 0.59 ± 0.24 5.76 ± 1.30 976 1.46 ± 0.93 4.20 ± 3.12 287 8.69 ± 6.24 37.0 ± 37.86 426

Statistical

significance (p-value) - - < 2.2 x10-16 0.8176 - 3.08e-10 - < 2.2 x10-16 -

* Symbol denotes significant differences (P-value < 0.05) between tanks.

10 Revista de Biología Tropical, ISSN: 2215-2075 Vol. 71 (S1): e54926, abril 2023 (Publicado Abr. 30, 2023)

Fig. 5. Pocillopora damicornis crown area growth (upper panel; P-value = < 2.2 x10-16, r2 = 0.23) and growth rate (lower

panel; P-value = 0.05582, r2 = 0.006) related to days at tank 1 and tank 2.

Revista de Biología Tropical, ISSN: 2215-2075, Vol. 71 (S1): e54926, abril 2023 (Publicado Abr. 30, 2023)

Fig. 6. Pocillopora damicornis volume growth (upper panel; P-value = < 2.2 x10-16, r2 = 0.26) and growth rate (lower panel;

P-value = 6.001e-06, r2 = 0.03) related to days at tank 1 and tank 2.

12 Revista de Biología Tropical, ISSN: 2215-2075 Vol. 71 (S1): e54926, abril 2023 (Publicado Abr. 30, 2023)

TABLE 3

Monthly value of physical-chemical parameters per tank.

Average values per tank (Tank 1/Tank

2) in a monthly basis

Temperature*

(°C)

Alkalinity-KH

(µmol/L)

Calcium

(mg/L)

Magnesium*

(mg/L)

PAR* (µ

E.m-2.s-1)

Reference values (Borneman, 2008;

Kleypas et al., 1999) 26-30 8 425 1250 250-1000

May T1 29.42 ± 0.44 7.9 ± 0.14 392.5 ± 45.96 1175 ± 49.49 445 ± 132.23

T2 29.06 ± 0.72 7.2 ± 0.14 392.5 ± 60.10 1175 ± 49.50 476 ± 129.40

June T1 29.18 ± 0.48 7.6 ± 0.5 400 ± 0.5 1260 ± 0.5 1212 ± 34.41

T2 28.80 ± 0.47 6.2 ± 0.5 410 ± 0.5 1240 ± 0.5 1361 ± 138.35

July T1 29.26 ± 0.04 7.15 ± 0.21 392.5 ± 10.60 1255 ± 7.07 688 ± 514.77

T2 28.69 ± 0.44 7.4 ± 0.55 390 ± 14.14 1230 ± 0.5 699 ± 617.53

August T1 29.23 ± 0.13 7.0 ± 0.5 355 ± 7.07 1185 ± 21.21 1060 ± 251.49

T2 28.97 ± 0.003 6.63 ± 0.40 373.3 ± 23.09 1210 ± 14.14 610 ± 16.26

September T1 29.10 ± 0.30 6.2 ± 0.5 340 ± 0.5 1230 ± 0.5 1440 ± 468.29

T2 28.86 ± 0.26 6.7 ± 0.5 360 ± 0.5 1220 ± 0.5 671 ± 147.07

October T1 27.60 ± 0.26 4.56 ± 0.87 280 ± 0.5 860 ± 180.83 1171 ± 537

T2 27.41 ± 0.41 6.2 ± 0.5 340 ± 0.5 1110 ± 0.5 1267 ± 486.96

Total

(average values

during experimental trial)

T1 29.09 ± 0.56 6.50 ±1.39 350.9 ± 51.80 1118 ± 188 895 ± 414.55

T2 28.74 ± 0.55 6.6 ± 0.87 368 ± 44.2 1156 ± 124.6 900 ± 410.31

Statistical significance (p-value) <2.2 x10-16 0.2133 <2.2 x10-16 0.0192 0.001693

* Symbol denotes significant differences (P-value < 0.05) between tanks.

TABLE 4

Monthly value of nutrients per tank.

Average values per tank

(Tank 1/Tank 2) in a monthly basis

Phosphates

(µmol/L)

Nitrates

(µmol/L)

Nitrites

(µmol/L)

Ammonium

(µmol/L)

May T1 0.37 ± 0.22 1.90 ± 0.31 1.53 ± 0.44 7.70 ± 0.04

T2 0.36 ± 0.21 1.49 ± 0.38 1.25 ± 0.28 6.10 ± 6.61

June T1 0.40 ± 0.12 1.97 ± 0.15 1.59 ± 0.12 8.67 ± 4.5

T2 0.40 ± 0.09 1.81 ± 0.41 1.48 ± 0.33 11.20 ± 5.1

July T1 0.33 ± 0.01 2.19 ± 0.39 3.68 ± 2.71 5.80 ± 1.78

T2 0.32 ± 0.06 1.65 ± 5.85 ± 6.38 6.99 ± 2.57

August T1 0.31 ± 0.01 nd 9.73 ± 1.59 6.36 ± 0.73

T2 0.37 ± 0.07 nd 8.17 ± 2.56 5.12 ± 0.94

September T1 0.37 ± 0.05 nd 8.96 ± 1.24 5.61 ± 2.19

T2 0.36 ± 0.05 nd 9.23 ± 1.46 5.20 ± 1.89

October T1 nd nd 7.85 ± 3.142 ±

T2 nd nd 4.98 ± 6.77 ±

Total T1 0.35 ± 0.05 2.0 ± 0.15 5.19 ± 3.6 6.34 ± 2.87

T2 0.39 ± 0.08 1.61 ± 0.38 5.34 ± 4.15 7.08 ± 4.23

Statistical significance (p-value) 0.8439 < 2.2 x10-16 3.378 x10-11 0.0001413

nd denotes not detectable.

Revista de Biología Tropical, ISSN: 2215-2075, Vol. 71 (S1): e54926, abril 2023 (Publicado Abr. 30, 2023)

In Forsman’s results growth remains steady,

whereas in this case, fragments grew exceed-

ingly fast after stimulation by cutting during

the fragmentation process (Page et al., 2018),

yet after 44 days in the tanks the newly-form

tissue began to slough off in most of the

fragments, reflecting and overall decrease in

growth and negative growth rate values. Thus,

it is conceivable that the larger area recorded

for fragments in T2 did not necessarily mean

that they grew faster than those in T1, but

that they deteriorated less as they were able to

(1) maintain tissue integrity longer and/or (2)

repair damage at the rate at which it accumulat-

ed (Kirkwood, 1981). It could be hypothesized

that, while the biological filters and protein

skimmer kept the water free of undesirable

biological hazards, they may have simultane-

ously limited the supply of organic nutrients

(i.e. zooplankton) and dissolved organic carbon

in the tanks, which could have affected coral

performance. Therefore, the inability of some

fragments to sustain their newly-produced tis-

sue could have been due to the lack of supple-

mentary nutrition, forcing corals to catabolize

their energy reserves, first lipids and in an

extreme case, tissue (Gates & Edmunds, 1999;

Kirkwood, 1981). This may have been the case

for P. lobata in this study, since under stress

conditions this species does not reduce meta-

bolic demand, but rather relies on its abundant

lipid reserve (Levas et al., 2013); a reserve that

could not be replenished in a culture medium

lacking a heterotrophic energy source.

When compared with wild colonies growth

rate, ex situ reared fragments grew 12.2 mm

year-1 (√ (0.013 cm2 per day x 365 days a year

/ π) x 10 mm; according to Forsman et al.,

(2006)) in contrast with 15.3–19.3 mm year-

1 (Guzmán & Cortés, 1989). Growth in the

culture tanks was comparable to growth in the

field, but was far from profitable in terms of

aquaculture, which is the premise upon invest-

ment in ex situ culture is based. Only seven

Porites fragments fully covered the ceramic

plug, reaching culture size at a rate of 25.05

mm year-1. This means a culture yield of just

21 % (69–75 cm2) of the desired coral harvest

(318.15 cm2). In a hypothetical restoration/pro-

duction scenario, this result would imply mayor

investment losses, as one would expect a mini-

mum yield of 200 % in order to sustain produc-

tion, as part of the harvest is used to replenish

the next production cycle (Leal et al., 2016;

Osinga et al., 2011; Schippers et al., 2012).

The speed at which corals can attach to

the substrate is often the first bottleneck in

in situ coral restoration projects (Forrester et

al., 2011; Guest et al., 2011; Tagliafico et al.,

2018). Attachment rate varies according to

region, environmental conditions, methodol-

ogy and species (Clark & Edwards, 1995). For

example, after one year of monitoring on the

island of Bali, Endo et al. (2013) reports an

attachment of 53 % for colonies of P. damicor-

nis, in contrast, Guzman (1991) observed the

formation of attachment points in less than 5

months for Pocillopora, placed on iron rods on

Caño Island. No significant differences were

found between tanks for attachment rate of

Basal area. On average, it took the fragments

70 days to cover 63 % of the ceramic plug,

which compares to Guest et al. (2011) findings,

whom reported tissue extension over 66 % of

the substrate in 82.5 days.

The gradual decrease in the growth rate of

Basal area responds to the fact that once fixed

and stable on the substrate, Pocillopora coral

colonies reorganize its shape until it reaches the

compact bush configuration that characterizes

it (Guest et al., 2011; Rinkevich, 2000).

Correlation results suggest both crown

area and volume share the same degree of asso-

ciation with the independent variable “accu-

mulated days”. Since the majority of scientific

literature uses length and area, over volume as

dimensional metrics for coral growth, Pocil-

lopora growth results are discussed in terms

of crown area (cm2). Needless to say, the use

of one metric over the other will depend on

the research’s question. For example, studies

focusing on biota-assemblage associated with

coral colonies (Doszpot et al., 2019; Salinas-

Akhmadeeva, 2018) will benefit from using

volumetric measurements over area. The dif-

ference between T1 and T2 fragments initial

14 Revista de Biología Tropical, ISSN: 2215-2075 Vol. 71 (S1): e54926, abril 2023 (Publicado Abr. 30, 2023)

and final growth rate, indicates growth rate was

affected by the culture medium. T2 fragments

growth rate remained relatively constant, in

accordance with Kinzie & Sarmiento (1986)

hypothesis which states: P. damicornis growth

rate is independent of colony size, remaining

constant over time.

In contrast to these argument, Lizcano-

Sandoval et al. (2018) report that growth rate

is determined by fragment size, the larger the

size, the greater the growth, however, these

authors compare growth rate between different

size groups, rather than the change in growth

rate of fragments of the same group size as

they grew. Either way, studies focused on

P. damicornis growth kinetics will hopefully

settle this question.

Pocillopora genus is characterized by

encompassing fast-growing species (Muko &

Iwasa, 2011; Schlöder & D’Croz, 2004). On

Caño Island an annual growth of 29.8 mm

yr-1 was reported, similar to that one in Pan-

ama (27.8 mm yr-1) and Mexico (21.9 mm

yr-1) (Gómez, 2014; Manzello, 2010). Whilst

growth rates of 50 mm yr-1 were reported

for Bahía Culebra, the highest in the Eastern

Tropical Pacific (Jiménez & Cortés, 2003). In

the Mexican Pacific, Tortolero-Langarica et al.

(2019) and Tortolero-Langarica et al. (2020)

report growth rates for P. damicornis fragments

transplanted directly on the reef of 23.1 and

44.7 mm yr-1. When convert to length, estimate

growth rate (~ 15.5 mm yr-1; as a result of Fors-

man formula) in this study is low compared

with colonies growing in the wild, but similar

to the 13.1 ± 15 mm yr-1 reported for P. dami-

cornis growing in an open water flow culture

system (Pillay et al., 2012).

Given the multiplicity of factors having

a potential effect on coral fragments perfor-

mance, isolating and determining the indi-

vidual or combined effect(s) of any of these

variables on both species is beyond the scope

and objectives of this study. Nevertheless,

the differences observed between fragments

performance among tanks, can and should be

cautiously explored.

Both tanks were adjacent to each other and

received water from the same marine reservoir.

Therefore, the contrasting results between one

tank and the other may have been due to sig-

nificant differences in conservative properties,

such as temperature, PAR and precipitation,

all influenced by shading and tank exposure.

Another hint towards this hypothesis is that P.

damicornis in T1 (more exposed), developed

on average a bigger crown area compared

with T2 fragments (less exposed). In the long

run, small “overlooked” differences in these

physical variables would have had an effect

in non-conservative factors (e.g., ammonium

values change with biological activity) (Li et

al., 2017; Silva et al., 2009), causing notice-

able differences between sea water properties

in each tank, which would have had an effect

on coral fragments.

Nutrient and alkalinity (Kh) values were

found to be on the low end of recommended

range (Bartlett, 2013), and may have had

negatively influenced coral growth by dis-

turbing coral-zooxanthellae dynamics (Grover

et al., 2003) or/and via macro algae prolif-

eration enhancement.

This study describes base-line findings for

scleractinian coral culture at Parque Marino

del Pacífico aquaculture module. Both survival

and fragment growth varied between species

(P. lobata and P. damicornis) and culture tank,

only few fragments show significant stable

growth. Hence, culture tanks current set up

and water conditions provided a suboptimal

medium for effective coral rearing, there is a

need for more studies regarding the relation-

ship between species-specific growth kinetics,

optimal culture conditions and cost-effective

production. These results do not discredit coral

aquaculture efforts. On the contrary, we hope

they demonstrate the potential for refining

ex situ coral culture practice in Costa Rica,

and to be a starting point, from which to

optimize culture conditions and methods into

a viable tool for the country´s reef systems

effective management.

Revista de Biología Tropical, ISSN: 2215-2075, Vol. 71 (S1): e54926, abril 2023 (Publicado Abr. 30, 2023)

Ethical statement: the authors declare

that they all agree with this publication and

made significant contributions; that there is no

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

lowed all pertinent ethical and legal procedures

and requirements. All financial sources are

fully and clearly stated in the acknowledge-

ments section. A signed document has been

filed in the journal archives.

ACKNOWLEDGMENTS

We would like to thank Parque Marino del

Pacífico authorities and personal for allow-

ing us to conduct this study as part of the

first author’s Licenciatura thesis. María Fer-

nanda Valverde and Marino are deeply thanked

for helping in the maintenance of the corals

in the tanks. Thanks to CIMAR researchers

Eddie Gómez and Juan Guillermo Sagot for

the help with the chemical analyses, and to

Juan José Alvarado and Cindy Fernández for

facilitating the collection of specimens. We

thank association Raising Coral Costa Rica for

providing water temperature loggers. Special

thanks to Joanie Kleypas, Dave Vaughan and

Hernán Azofeifa for providing valuable guid-

ance and ideas.

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