Pocillopora spp. growth analysis on restoration 1 structures in an Eastern Tropical Pacific upwelling 2 area 3 4 5 Lisa Combillet1, Sònia Fabregat-Malé2, Sebastián Mena3, José Andrés Marín-Moraga4, Mónica 6 Gutiérrez5 & Juan José Alvarado3,6,7 7 8 1 Programme de Master Sciences pour l'Environnement, parcours Gestion de l’Environnement et 9 Écologie Littorale, Université de La Rochelle, La Rochelle, France 10 2 Posgrado en Biología, Sistema de Estudios de Posgrado, Universidad de Costa Rica 11 3 Escuela de Biología, Universidad de Costa Rica. 12 4 Raising Coral Costa Rica; San José, Costa Rica 13 5 Sostenibilidad, Península Papagayo; Guanacaste, Costa Rica 14 6 Centro de Investigación en Ciencias del Mar y Limnología (CIMAR), Universidad de Costa 15 Rica; 2060-1000 San Pedro de Montes de Oca, Costa Rica 16 7 Centro de Investigación en Biodiversidad y Ecología Tropical (CIBET) (previously Museo de 17 Zoología), Universidad de Costa Rica. 18 19 Corresponding Author: 20 Lisa Combillet1 21 112 rue Mac Carthy, 33200, Bordeaux, France 22 Email address: lisa.combillet@gmail.com 23 24 Abstract 25 Coral reefs in Culebra Bay (North Pacific of Costa Rica) are threatened by multiple 26 anthropogenic disturbances including global warming, overfishing, eutrophication, and invasive 27 species outbreaks. It is possible to assist their recovery by implementing ecological restoration 28 techniques. This study used artificial hexagonal steel structures, called “spiders” to compare 29 growth of Pocillopora spp. coral fragments of different sizes. Three initial fragment class sizes 30 were used: 2, 5 and 8 cm, with each class size having 42 initial fragments. Changes in fragment 31 length, width and area were measured monthly from January to December 2020. Results showed 32 an overall survivorship of 70%, and no significant differences in survivorship and linear growth 33 rate were detected between class sizes. The linear growth rates are 4.49 ± 1.19 cm yr-1, 5.35 ± 34 1.48 cm yr-1 and 3.25 ± 2.22 cm yr-1 for the 2, 5 and 8 cm initial class sizes, respectively. Results 35 do not show significant differences in growth rates between the different initial fragment sizes. 36 However, since small fragments (2 cm) presented higher mortality during the first month, using 37 larger fragments is recommended. In addition, coral fragments grew 48% more during the non-38 upwelling season, which may suggest that it might be more effective and safer to start the 39 restoration efforts during this period. 40 41 Introduction 42 Coral reefs are highly diverse ecosystems that provide essential goods and services to hundreds 43 of millions of people (Knowlton et al., 2021), such as food, livelihoods through fisheries and 44 tourism, protection from coastal erosion and storms, and cultural practices (Woodhead et al., 45 2019). Nevertheless, in the last decades, many reefs around the world have collapsed, and live 46 coral cover has declined due to several factors, such as climate change, acidification and 47 unplanned coastal development (Hughes et al., 2017; El-Naggar, 2020; Knowlton et al., 2021). 48 The rapid deterioration of these ecosystems threatens the stability of marine environments and 49 human well-being (Eddy et al., 2021). 50 Due to this intense degradation of coral reefs worldwide and in the face of future climate change, 51 ecological restoration of coral reefs is becoming an increasingly important management 52 approach (McLeod et al., 2021). Restoration of degraded coral reefs can be achieved through 53 different means, using either sexual or asexual coral recruits in order to enhance coral 54 populations (Rinkevich, 1995; Rinkevich, 2019). During the last 20 years, several restoration 55 techniques have been developed, and coral gardening has been one of the most widely used. This 56 approach is based on the asexual propagation of corals by the fragmentation of wild donor 57 colonies. The collected fragments are later put into coral nurseries, where they grow until they 58 become larger colonies which are later outplanted onto a degraded reef (Rinkevich, 2006). A 59 wide variety of structures have been used as coral nurseries, from floating (suspended in the 60 water column) to fixed structures (on the seafloor) (Shafir & Rinkevich, 2010; Rinkevich 2019). 61 Most restoration projects have been developed in the Caribbean and Indo-Pacific (Boström-62 Einarsson et al., 2020). In the Eastern Tropical Pacific (ETP), however, coral reef restoration is 63 still in its infancy, and very few projects are based on coral gardening (Bayraktarov et al., 2020). 64 Conditions in the ETP are different from those in the Caribbean and Indo-Pacific. Coral reefs are 65 relatively small (a few hectares), discontinuous, and are built by few coral species, 66 predominantly of the genera Pocillopora, Porites and Pavona (Guzmán & Cortés, 1993; Glynn 67 et al., 2017). The region comprises three seasonal upwelling areas (Gulf of Tehuantepec, Gulf of 68 Papagayo and Gulf of Panama), with incursions of deep, cold and nutrient-rich waters (Cortés, 69 1997; Fiedler & Lavín, 2017). The ETP is also affected by the El Niño-Southern Oscillation 70 (ENSO), which causes an increase in sea surface temperatures that can lead to coral bleaching 71 and high mortality, with loss of live coral cover (Glynn, 1984; Guzmán et al., 1987; Jiménez et 72 al., 2001; Jiménez & Cortés, 2001; Brainard et al., 2018). 73 The North Pacific coast of Costa Rica was considered as one of the best regions for the 74 development of coral reefs in the country (Cortés & Jiménez, 2003; Alvarado et al., 2018). 75 Within it, the reefs in Culebra Bay (Fig. 1) were considered as the most diverse, but in the last 76 two decades various disturbances caused severe degradation that caused the collapse and loss of 77 many reefs around the bay. Red tides and macroalgal proliferation lead to coral bleaching and 78 mortality (Cortés et al., 2010). The following increase in sea urchin populations (Diadema 79 mexicanum) resulted in high bioerosion rates and caused the loss of the reefs structural 80 complexity and framework (Alvarado et al., 2012; Alvarado et al., 2016), which in turn had an 81 impact on diversity of reef-associated organisms and ecosystem functions (Arias-Godínez et al., 82 2019; Salas-Moya et al., 2021). 83 The particular environmental conditions, combined with the relatively low experience on coral 84 reef restoration in the region, means that little is known about restoration techniques and specific 85 considerations about the species used. However, some studies (mostly in Mexico and Colombia) 86 have been carried out using the coral genus Pocillopora (Liñán-Cabello et al., 2011; Tortolero-87 Langarica et al., 2014; Nava & Figueroa-Camacho, 2017; Lizcano-Sandoval et al., 2018; Ishida-88 Castañeda et al., 2020; Vargas-Ugalde et al., 2020). The restoration project implemented in 89 Culebra Bay, which was initiated in 2019, could help determine the optimal initial coral fragment 90 size and the suitability of a new technique and thus, help respond to specific research questions 91 for the development of coral restoration projects in the ETP. Coral fragments of Pocillopora spp. 92 of three different initial sizes were attached to the structures, and their growth was monitored 93 monthly for one year. The aim of the present study is to determine whether coral fragment 94 growth and survival is affected by initial fragment size and presence of upwelling, in order to 95 establish the optimal fragment size and best period to start restoration efforts for Pocillopora 96 dominated reefs. 97 98 Materials & Methods 99 1.Study area 100 101 Culebra Bay, in the Gulf of Papagayo, is located in the Guanacaste province of Costa Rica, in the 102 Northwest Pacific of the country. This bay consists of a series of islets, beaches, cliffs and 103 estuaries with important economic marine resources, and it is subject to a seasonal upwelling 104 between December and April, which brings up colder and nutrient-rich waters (Jiménez, 2001; 105 Alfaro & Cortés, 2012). During this period, seawater temperatures can decrease by 8 to 9 °C 106 from the annual average (27.9 ºC) (Alfaro & Cortés, 2012; Alfaro et al., 2012). The bay is 107 naturally exposed to lower pH (pH = 7.8) than other regions, with high temporal variability 108 following the dry and rainy seasons (Sánchez-Noguera et al., 2018a). Even during the non-109 upwelling season (from May to November) there is a reduced pH that impacts photosynthesis, 110 respiration, and calcification processes (Rixen et al., 2012; Sánchez-Noguera et al., 111 2018a). Sedimentation in the area is low (3.0 ± 0.78 mg cm-2 day-; Fernández-García et al., 112 2012) and without any sign of human stress (Rogers, 1990). Coral reefs in the bay are dominated 113 by the genus Pocillopora, which forms monospecific patches that used to cover several hectares 114 along the bay in the 1990s. On some reefs, corals covered between 40 and 80% of the substrate 115 (Jiménez, 2001; Cortés & Jiménez, 2003). In 2010, however, live coral cover was only 1 to 4% 116 (Sánchez-Noguera et al., 2018b). The main Pocillopora species in the region are Pocillopora 117 damicornis (Linnaeus, 1758) and Pocillorpora elegans (Dana, 1846). In this study, these two 118 species were grouped under the name of Pocillopora spp. because the morphologies are similar, 119 with intermediate shapes, which make their precise identification in the field difficult. The 120 experiment took place in the coral reef patch in front of Playa Jícaro (10.619830°N, 121 85.675810°W) (Fig. 1). 122 123 2.Experimental design 124 125 Pocillopora spp. fragments (n = 126) were obtained from colonies on three sites around Culebra 126 Bay: Palmitas, Marina and Güiri-Güiri (Fig. 1). Healthy large donor colonies (>30 cm in 127 diameter and without observable injuries) were randomly selected at depths between 3 to 8 m, 128 and no more than three fragments were obtained from each donor. Three different initial 129 fragment sizes categories were considered: small (2 cm, 2.57 ± 0.38 cm), medium (5 cm, 5.35 ± 130 0.78 cm) and large (8 cm, 8.26 ± 1.63 cm). Forty-two fragments from each size class were 131 attached using plastic cable ties to three “spider” restoration structures, one for each fragment 132 size class. These hexagonal metallic structures are 90 cm high and have three levels, 30 cm apart 133 and 25, 35 and 45 cm long from top to bottom. Arrangement of the coral fragments within the 134 structure (for each of the six sides of the “spider”, 2 fragments on the top level, 2 in the middle 135 and 3 in the lowest) was based on fragments having enough space to grow and not competing 136 with each other (Fig. 2a). This design also allows coral fragments to grow on the external side of 137 the structure and if they break and fall from it, they can continue to grow surrounded by other 138 fragments on the seafloor, forming a three-dimensional structure. With this method, corals are 139 not necessarily destined to be outplanted to the reef afterwards, but to stay on the structures, 140 where they can keep growing. Thus, “spiders” have a double purpose, as they can act as both a 141 nursery and a substrate on which to permanently attach corals to contribute to the structural 142 complexity of the reef. The three “spiders” were placed at 6 m depth, on the front reef area. 143 144 3.Data collection 145 146 The experiment was conducted from January to December 2020. March 2020 is excluded from 147 the results because of the COVID-19 sanitary crisis, which prevented data collection in Costa 148 Rica. The study site was visited monthly and each Pocillopora fragment was photographed with 149 an underwater Nikon COOLPIX W300 camera, using a calliper as a scale (Fig. 2b). Photographs 150 were later analysed using ImageJ software, which allows for a 0.001 cm precision, and height 151 (cm), width (cm) and area (cm2) of each fragment was determined. This allowed us to estimate a 152 growth rate in terms of linear extension (cm year-1) and tissue area (cm2 yr-1). Linear extension 153 was calculated by measuring the vertical length between the two longest coral branches, while 154 the area of the coral was estimated by outlining the contour of the coral fragment, and 155 subsequently calculating the average. Mortality was visually determined; a fragment was 156 considered dead if it had no living tissue left and/or was covered by other organisms such as 157 algae, barnacles or ascidians. If the fragment was partially dead, only the part with living tissue 158 was measured. The number of dead fragments was established and used to calculate fragment 159 survival rates. Seawater temperature in the restoration area was recorded using HOBO® data 160 loggers, which were set to record data every 30 min. 161 162 4.Data analysis 163 164 Survival rates of each initial size class were calculated and compared with a Chi-squared 165 contingency test in order to determine the influence of the initial size of the fragment. Lost 166 fragments were excluded from this calculation since it is not possible to establish whether they 167 survived. Means of fragments length, width, and area at initial time (January 2020) and every 168 month until December 2020 were estimated for each “spider'' and then, the relative growth 169 between initial and final time was calculated. These estimations considered only fragments that 170 survived until the last month of the experiment and excluded fragments that broke during the 171 course of the experiment. Means of fragment length, width and area of each month are compared 172 with a one-way ANOVA followed by Tukey HSD post-hoc tests. To compare the absolute 173 growth and growth rate between the three different initial class sizes, a two-way ANOVA test 174 was used followed by a Tukey HSD post-hoc test. Finally, a t-test was used to compare the 175 difference in growth between two periods: from January to April and from May to December, 176 according to the presence and absence of seasonal upwelling, respectively. Monthly average, 177 minimum and maximum seawater temperature was calculated from temperature data. Statistical 178 analyses were performed using R (R Development Core Team 2020), including the package 179 “stats” (R Core Team, 2018). 180 181 Results 182 1.Coral fragment survival 183 184 At the end of the experiment, 66 fragments survived (52.38%), 28 died (22.22%), and 32 were 185 lost (25.39%) due to fragmentation or cable tie break during the experimentation (Table 186 1). Excluding lost fragments, coral fragment survival is not affected by the initial fragment size 187 (Χ² = 3.993, df = 2, p > 0.05). The overall survival rate from January to December 2020 is 188 70.21%. The highest number of death fragments (9) appeared in February, whilst the number of 189 dead fragments during other months ranged from 0 to 5. In order to determine whether upwelling 190 had an effect on coral mortality, a Pearson’s chi-squared test was also performed between 191 upwelling season (January to April) and non-upwelling season (May to December). No 192 significant differences in mortality were observed between the two periods (Χ² = 1.5345, df = 1, 193 p > 0.05). The test also showed no statistical differences when considering initial fragment size ( 194 Χ² = 3.247, df = 2, p > 0.05 and Χ² = 0.812, df = 2, p > 0.05). 195 196 2. Coral fragment growth 197 198 During the period of observation, Pocillopora fragments grew significantly in terms of length 199 (F10,953 = 35.2, p > 0.001), width (F10,935 = 40.8, p > 0.001) and area (F10,953 = 46.5, p > 0.001), 200 independently of their initial class size (Fig. 3). On average, fragments grew 4.12 ± 2.77 cm yr-1 201 and quadrupled their surface over one year (438%). Pocillopora fragments grew more in terms 202 of length than width (Table 2). For some fragments, negative growth between months was 203 observed. Growth rate in length and width does not significantly differ by initial class size, but it 204 does for area measurements: A is significatively different from B and C, and B is significatively 205 different from C (Table 2). 206 207 3. Comparison between upwelling and non-upwelling periods 208 209 Coral growth is significantly impacted by seasonal upwelling for the 2 and 5 cm initial class size. 210 However, no significant difference between periods was observed for 8 cm coral fragments. 211 (Table 3). Regardless of the initial size, Pocillopora fragments grow 48% faster on average 212 during the non-upwelling season, coinciding with a higher mean temperature during this period 213 (Fig. 4). 214 215 Discussion 216 Coral reef management is a key issue in the current context of global change. Assessing the 217 resilience of coral species and identifying sites conducive to the survival of corals is thus crucial 218 in order to improve management actions (McLeod et al., 2021). While whether corals will have 219 the ability to acclimate rapidly enough to the new environmental conditions is still under debate 220 (Maynard et al., 2008; Eakin, 2014; Torda et al., 2017; Coles et al., 2018), active coral reef 221 restoration is emerging worldwide as a tool for assisting coral reef recovery and rehabilitation 222 (Rinkevich, 2019). Several restoration strategies have been developed, such as structural 223 complexity enhancement by artificial substrates, which increase coral recruitment and can be 224 used as an alternative or addition to coral transplantation for reef restoration purposes (Yanovski 225 & Abelson, 2019; Hein et al., 2020). This type of structures has mainly been used in the Indo-226 Pacific, specifically in the Maldives and Thailand, where metallic structures called “frames” 227 were set up (Hein et al., 2018, 2020). However, their use is recent, the coral species used are not 228 the same, and environmental conditions in those areas are different from those in the ETP 229 (Kench, 2009; Lizano & Alfaro, 2014). This makes comparisons difficult and obtained data are 230 not necessarily transferable to other reefs in other oceanic regions (Sherman, Gilliam & Spieler, 231 2001). Therefore, it appears necessary to generate data on the performance of this kind of 232 structures under the conditions in the ETP, in order to assess their viability in this oceanic region. 233 Evaluating this strategy involves monitoring fragment mortality and growth, and associating the 234 data with environmental information from the area. In this study, Pocillopora fragment mortality 235 was not significantly influenced by initial fragment size. The month with the highest mortality 236 was February, just one month after the fragmentation event and start of the experiment, with 9 237 dead fragments, 66% of which were 2 cm long. A positive relationship between coral fragment 238 survival and size has been established in several studies (Connell, 1973; Hughes, 1984; Lizcano-239 Sandoval et al., 2018; Ishida-Castañeda et al., 2020). Research on Pocillopora has found that 240 smaller coral fragments are more vulnerable to detrimental factors due to their greater 241 surface/volume ratio. This means that a lesion on the coral tissue can cause greater damage than 242 in larger fragments, and thus it makes them more sensitive to manipulation, competition with 243 other organisms and predation (Raymundo & Maypa, 2004; Lizcano-Sandoval et al., 2018; 244 Ishida-Castañeda et al., 2020). According to the micro fragmentation theory, this can be 245 compensated by small fragments growing more rapidly at first compared to larger fragments or 246 colonies, so that they can quickly reach a size which makes them less vulnerable to impacts 247 (Forsman, Rinkevich & Hunter, 2006; Page, Muller & Vaughan, 2018; Tortolero-Langarica et 248 al., 2020). However, our results show no significant differences between linear growth rates of 2 249 cm fragments and the other class sizes. The bay possesses a great productivity (Fernández-250 García, 2007; Stuhldreier et al., 2015a) making the structures a suitable substrate for the 251 settlement of benthic, fast-growing, opportunistic species, such as barnacles, ascidians, and 252 sponges. These benthic organisms compete with coral fragments and can affect their growth and 253 survival (Glynn et al., 2017). Although monthly maintenance of the structures limits this effect, 254 their great abundance and presence on the “spiders” could have had an effect on coral fragments, 255 especially on the smaller class size, because of their limited surface. These smaller coral 256 fragments might not have been able to compete for space against these other organisms. Due to 257 their small size, several of their energy reserves were probably not available, and therefore were 258 presumably being used to recover from fragmentation stress, and not for growth and defense 259 against competitors (Leuzinger et al., 2003; Henry & Hart, 2005). Hence, it is assumed that the 260 small fragments used in this study were the most fragile and affected by these detrimental 261 factors, and thus did not resist the stress of fragmentation and change in environment during the 262 first weeks of the experiment. 263 The loss of 32 coral fragments during the course of the experiment could be explained by several 264 reasons: (i) cable ties being either too tight and resulting in fragment break, or too loose cable 265 ties, causing them to fall, especially small (2 cm) fragments; or (ii) clumsiness during the manual 266 cleaning and maintenance of the restoration structures. These lost fragments are not included in 267 the survivorship results, since it is not possible to determine whether they survived in the reef or 268 not. The observed decrease in growth and fragment size between two consecutive months in 269 some coral fragments can possibly be a result of intrinsic variations of the colony, either by 270 partial mortality of coral tissue, or natural fragmentation processes. 271 The growth rates of P. damicornis and P. elegans in Culebra Bay were determined in 1995-1996, 272 with a mean of 5.3 ± 0.4 cm yr-1 and 4.1 ± 0.6 cm yr-1, respectively, using 13 cm long fragments 273 (Jiménez & Cortés, 2003). Under stressful conditions, in the presence of the competitor 274 macroalgae Caulerpa sertularioides, Pocillopora corals (no initial size reported) in the bay show 275 a lower growth rate (2.5 cm yr-1) than without it (4.2 cm yr-1) (Fernández-García, 2007). In the 276 present study, a length growth rate of 4.49 ± 1.19 cm yr-1, 5.35 ± 1.48 cm yr-1 and 3.25 ± 2.22 cm 277 yr-1 respectively for the 2, 5 and 8 cm initial size was established (Table 2), along with and an 278 overall growth rate of 4.12 ± 2.77 cm yr-1. These results seem to follow the rate calculated by 279 Jiménez & Cortés (2003) on the reef in the 1990s, when coral reef ecosystems in the bay were 280 considered healthier. This means that fragments on the “spiders” are growing at a similar rate to 281 corals growing naturally in the reef. These results are quite high compared to other reefs of the 282 ETP: for example, in Caño Island (South Pacific of Costa Rica), the rate for 15-25 cm long 283 fragments is 2.9 ± 0.3 cm yr-1 for P. damicornis and 3.17 ± 0.3 cm yr-1 for P. elegans (Guzmán 284 & Cortés, 1989). In the Central Mexican Pacific, the growth rate is 3.5 ± 0.6 cm yr-1, with no 285 initial size being mentioned (Tortolero-Langarica et al., 2017). The lowest Pocillopora growth 286 rates reported for the ETP are in the Gulf of Chiquiri (Panama) and Colombia, with 2.6 cm yr-1 287 (initial size = 6.3 ± 1.4 cm) (Randall et al., 2020) and 2.3 cm yr-1 (no initial size mentioned) 288 (Zapata & Vargas-Angel, 2003), respectively. It is hypothesized that the higher growth rate in 289 Culebra Bay is linked to the specific conditions of the bay, with the seasonal upwelling bringing 290 up more productive waters, which could lead to an increase of the corals heterotrophic feeding 291 (Jiménez & Cortés, 2003). These results also show that the corals of Culebra Bay are particularly 292 acclimated to the specific environmental conditions of the bay, which make them an example of 293 growth under suboptimal conditions, with incursions of colder and more acidic waters (Rixen et 294 al., 2012; Sánchez-Noguera et al., 2018b). 295 Understanding the best initial fragment size is vital for efficient restoration activities. Even 296 though this has been established for many species in other regions, information on Pocillopora 297 corals and under ETP conditions is limited (but see Lizcano-Sandoval et al., 2018; Ishida-298 Castañeda et al., 2020). Moreover, even though initial fragment size seems to be an important 299 factor for coral growth, most studies do not consider it in their analysis. Based on our results, it 300 can be assumed that 2 cm fragments are not of an optimal size when rearing as many corals as 301 possible onto the reef, since they experience high mortality during the first months after 302 fragmentation, and are more fragile and prone to breaking. On the other hand, it was found that 303 larger fragments (between 5 and 8 cm) grow at a similar rate while experiencing lower mortality. 304 However, extracting larger fragments and repeated fragmentation of coral colonies can 305 compromise the survival of these donor colonies, and it may lead to reduced sexual reproduction 306 (Zakai, Levy & Chadwick-Furman, 2000), which could in turn impact the development of the 307 whole coral reef ecosystem. The recovery of donor colonies after fragmentation events should 308 also be assessed in order to evaluate the impact of extracting large coral fragments. 309 Corals from Culebra Bay have already been confronted by stressing episodes which have had an 310 effect on the coral reef ecosystem (Jiménez, 2001; Alvarado et al., 2012; Fernández-García et 311 al., 2012). Nonetheless, ETP reefs have shown a high resilience to stressing events (Romero-312 Torres et al. 2020), which would allow large-scale rehabilitation even after severe disturbances, 313 such as El Niño events (Williams et al., 2018). Culebra Bay is located in one of the three 314 seasonal upwelling areas of the ETP, which from December to April brings colder and more 315 acidic waters to the surface, with a higher concentration of nutrients (Rixen et al., 2012; 316 Stuhldreier et al., 2015a, Sánchez-Noguera et al., 2018b). The decrease in seawater temperature 317 and increase in productivity can have an effect on coral growth and survival (Clausen & Roth, 318 1975; Coles & Jokiel, 1978). Our results show a difference in growth between the upwelling and 319 non-upwelling periods: coral growth increased 48% on average during the non-upwelling period 320 (May to December) compared to the upwelling period. Similar results were obtained in the bay 321 when comparing the growth rate of Pocillopora spp. during seasons, with higher rates occurring 322 during the non-upwelling season (Fernández-García, 2007). However, 8 cm fragments were not 323 found to be significantly impacted by the presence of the seasonal upwelling. These fragments 324 also correspond to the size class with the lowest growth rate. It is thus hypothesised that since 325 these coral fragments are already large, they allocate less energy to their growth rather than in 326 other physiological processes. The lower temperatures during upwelling, with incursions of 18.5 327 ºC waters, could also be responsible for the higher mortality of coral fragments during the first 328 months of the experiment, coinciding with the possible stress caused by fragmentation and 329 smaller size of the coral fragments. Studies on the effect of cold water on branching corals have 330 found that in the short term, low temperatures can be more damaging than warm temperatures, 331 but acclimatation is possible after a few weeks, and corals can recover quickly when 332 temperatures rise back (Jokiel & Coles, 1977; Roth, Goericke & Deheyn, 2012; Rodríguez-333 Troncoso et al., 2014). Even though it has been suggested that in case of stress, corals 334 preferentially use heterotrophic feeding and use lipids stored in their tissues (Grottoli et al., 335 2004, 2006; Rodríguez-Troncoso, Carpizo-Ituarte & Cupul-Magaña et al., 2010), it seems that 336 this type of feeding is less efficient in terms of nutrition than autotrophy. Considering this, we 337 suggest that restoration activities in Culebra Bay, such as fragmentation of new corals, should 338 take place after the upwelling season; thus, smaller newly generated fragments would have 339 higher chances of surviving the initial months and could reach a larger size before the upwelling 340 begun and temperatures decreased again. Restoration efforts in these areas of the ETP where 341 seasonal upwelling is present should thus take into account these considerations for the optimal 342 growth of Pocillopora spp. fragments. 343 Considering our results, the use of “spiders” is a viable option for coral reef rehabilitation and 344 restoration, and their effect could be scaled up by increasing the number of structures in order to 345 cover a greater extension and add more structural complexity to the reef. Even though 47.61% of 346 initial coral fragments were either lost or died during the experiment, it allowed us to determine 347 the most resistant fragment sizes, and thus those that should be used on future restoration efforts. 348 Other factors must also be considered, such as the location of the structures and the donor sites 349 for coral fragments. Beyond the technical aspects of a restoration project, two main limiting 350 factors exist: the economic aspect - which includes the costs of setting up and maintaining the 351 structures (Dunning, 2015; Hesley et al., 2017) - and the communication about conservation 352 strategy (Dunning, 2015). These structures have a relatively lower cost to other underwater coral 353 nurseries, only costing around US$25 per structure (US$0.66 per coral fragment, excluding 354 indirect costs). Moreover, they require less time to clean and maintain: one “spider” can be 355 cleaned by one diver in around 15 minutes, which is considerably less time than what is needed 356 for other structures in the same restoration project, such as rope nurseries or PVC and glass fiber 357 coral trees (1.6 m long x 1.2 m wide) (S. Fabregat-Malé, pers. comm.). These limitations can be 358 bypassed with the involvement of local communities and tourists by creating a participative 359 program (Hein et al., 2019) in which the cost of the project will be reduced and there will be an 360 increase in public awareness and workforce, allowing for larger-scale restoration efforts. The 361 restoration project in Culebra Bay, which started on August 2019 with coral gardening 362 techniques (Fabregat-Malé et al., in prep.), is now complemented by the use of artificial 363 structures in this project, leading towards its expansion through greater restoration efforts and the 364 implementation of a participatory program. This study complements those already carried out 365 and in progress, allowing an improvement of the techniques used to optimise the restoration 366 efforts of reefs in Culebra Bay. 367 368 Conclusions 369 Active restoration has become a key management tool to rehabilitate anthropogenically 370 deteriorated coral reefs. In Culebra Bay, North Pacific of Costa Rica, coral reefs have suffered 371 several degrading episodes in the last decades but are currently subject to ecological restoration 372 actions. Various transplantation techniques are used with the genus Pocillopora, including the 373 coral gardening approach. Here, a new technique in the ETP was tested, consisting in rearing 374 coral fragments on artificial structures (“spiders”), which not only work as a nursery and 375 substrate for coral fragments to grow on, but also add structural complexity to the reef. Our 376 findings show that small Pocillopora fragments are especially vulnerable and sensitive to 377 environmental stresses during the first months after fragmentation, which results in higher 378 mortality rates. Even though we found no significant differences in linear growth between size 379 classes, the smallest class size appears to be less optimal than larger ones if restoration efforts are 380 to be scaled. The presence of a seasonal upwelling in the bay has an effect on coral growth, most 381 likely due to cold temperatures. The upwelling brings up deeper and nutrient-rich waters, 382 resulting in the proliferation of opportunistic and highly competitive benthic organisms 383 (Fernández-García et al., 2012; Stuhldreier et al. 2015b), which could potentially have an effect 384 on coral growth and survival. This information is key in order to plan restoration activities in 385 areas affected by seasonal upwelling, since fragments will grow more optimally if transplanted at 386 the end of the upwelling season, and will be robust enough to cope with the next upwelling 387 period as they will have reached a larger size. Our data also show how corals can survive under 388 suboptimal conditions when acclimated to such an environment. Studying the particular 389 characteristics of these areas is essential to understanding, optimising and innovating reef 390 restoration strategies at local scales, especially in the ETP region, where information is still 391 scarce. 392 393 Acknowledgements 394 The present investigation would not have been possible without the support of Centro de 395 Investigación en Ciencias del Mar y Limnología (CIMAR) of the University of Costa Rica and 396 students who helped during fieldwork and data collection for this project. We would also like to 397 thank José Francisco Cascante, who built the structures used in this experiment. We especially 398 thank Península Papagayo for their economic and logistical support. Fieldwork would not have 399 been possible without the help from Carlos Marenco and the staff of Marina Papagayo, to whom 400 we are really thankful. 401 402 403 References 404 Abelson A. (2006) “Artificial reefs VS coral transplantation as restoration tools for mitigating 405 coral reef deterioration: benefits, concerns, and proposed guidelines”. Bulletin of Marine 406 Science, 78, 151-159. 407 408 Alfaro E.J., Cortés J. (2012) “Atmospheric forcing of cool subsurface water events in Bahia 409 Culebra, Gulf of Papagayo, Costa Rica”. Revista de Biologia Tropical, 60, 173-186. 410 411 Alfaro E.J., Cortés J., Alvarado J.J., Jiménez C., León A., Sánchez-Noguera C., Nivia-Ruiz J., 412 Ruiz E. (2012). “Clima y temperatura sub-superficial del mar en Bahía Culebra, Golfo de 413 Papagayo, Costa Rica”. Revista de Biología Tropical, 60, 159-171. 414 415 Alvarado J.J., Beita-Jiménez A., Mena S., Fernandez-Garcia C. (2018) “Cuando la conservación 416 no puede seguir el ritmo del desarrollo: Estado de salud de los ecosistemas coralinos del Pacifico 417 Norte de Costa Rica”. Revista de Biología Tropical, 66, 280-S308. 418 419 Alvarado J.J., Cortés J., Guzman H.M., Reyes-Bonilla H. (2016) “Bioerosion by the sea urchin 420 Diadema mexicanum along Eastern Tropical Pacific coral reef”. Marine Ecology, 37, 1088-1102. 421 422 Alvarado J.J., Cortés J., Reyes-Bonilla H. (2012). “Reconstruction of Diadema mexicanum 423 bioerosion impact on three Costa Rican Pacific coral reefs”. Revista de Biología Tropical, 60, 424 121-132. 425 426 Arias-Godinez G., Jiménez C., Gamboa C., Cortés J., Espinoza M., Alvarado J.J. (2019) “Spatial 427 and temporal changes in reef fish assemblages on disturbed coral reefs, north Pacific coast of 428 Costa Rica”. Marine Ecology, e12532. 429 430 Bayraktarov E., Banaszak A.T., Montoya Maya P., Kleypas J., Arias-González J.E., Blanco M., 431 Calle-Trivino J., Charuvi N., Cortés-Useche C., Galvan V., Garcia Salgado M.A., Gnecco M., 432 Guendulain-Garcia S.D. et al. (2020) “Coral reef restoration efforts in Latin American countries 433 and territories”. PLoSONE, 15(8): e0228477. 434 435 Boström-Einarsson L., Babcock R.C., Bayraktarov E., Ceccarelli D., Cook N., Ferse S.C.A. 436 Hancock B., Hein M., Shaver E., Smith A., Suggett D., Stewart-Sinclair P.J., Vardi T., McLeod 437 I.M. (2020) “Coral restoration - A systematic review of current methods, successes, failures and 438 future directions”. PLoS ONE, 15: e0226631. 439 440 Brainard R.E., Oliver T., McPhaden M.J., Cohen A., Venegas R., Heenan A. Vargas-Angel B., 441 Rotjan R., Mangubhai S., Flint E., Hunter S.A. (2018) “Ecological impacts of the 2015/16 El 442 Nino in the central equatorial Pacific”. In: Explaining extreme events of 2016 – from a climate 443 perspective, Special supplement to the Bulletin of the American Meteorological Society, Chap.5, 444 21-26. 445 446 Clausen C.D., Roth A.A. (1975) “Effect of temperature and temperature adaptation on 447 calcification rate in the hermatypic coral Pocillopora damicornis”. Marine Biology, 33, 93-100. 448 449 Coles S.L., Jokiel P.L. (1978) “Synergistic effects of temperature, salinity and light on the 450 hermatypic coral Montipora verrucosa”. Marine Biology, 49, 187-195. 451 452 Coles S.L., Bahr K.D., Rodgers K.S., May S.L., McGowan A.E., Tsang A., Bumgarner J., Han 453 J.H. (2018) “Evidence of acclimatization or adaptation in Hawaiian corals to higher ocean 454 temperatures”. PeerJ, 6: e5347. 455 456 Connell J.H. (1973) “Population ecology of reef building corals”. In: Jones OA, Endean R, 457 editors. Biology and geology of coral reefs. Vol. 2. New York (NY): Academic Press, p. 205–458 245. 459 460 Cortés J. (1997) “Biology and geology of Eastern Pacific coral reefs”. Coral Reefs, 16, 39-S46. 461 462 Cortés J., Jiménez C. (2003) “Corals and coral reefs of the Pacific of Costa Rica: history, 463 research and status”. Latin America Coral Reefs, 361-385. 464 465 Cortés J., Jiménez C., Fonseca A.C., Alvarado J.J. (2010) “Status and conservation of coral reefs 466 in Costa Rica”. Revista de Biologia Tropical, 58, 33-50. 467 468 Dunning K.H. (2015) “Ecosystem services and community-based coral reef management 469 institutions in post blast-fishing Indonesia”. Ecosystem Services, 16, 319-332. 470 471 Eakin C.M. (2014) “Lamarck was partially right - and that is good for corals”. Science, 472 344(6186), 798–799. doi:10.1126/science.1254136. 473 474 Eddy T.D., Lam V.W.Y., Reygondeau G., Cisneros-Montemayor A.M., Greer K., Palomares 475 M.L.D., Bruno J.F., Ota Y., Cheung W.W.L. (2021) “Global decline in capacity of coral reefs to 476 provide ecosystem services”. One Earth, 4, 1278-1285. 477 478 El-Naggar H.A (2020) “Human impacts on coral reef ecosystems”. In Natural Resources 479 Management and Biological Sciences, Intech Open. 480 481 Fernández-García C. (2007) “Propagación del alga Caulerpa sertularioides (Chlorophyta) en 482 Bahía Culebra, Golfo de Papagayo, Pacífico norte de Costa Rica”. Master thesis, Universidad de 483 Costa Rica. San Pedro, Costa Rica. 484 485 Fernández-García C., Cortés J., Alvarado J.J., Nivia-Ruiz J. (2012) “Physical factors 486 contributing to the benthic dominance of the alga Caulerpa sertularioides (Caulerpaceae, 487 Chlorophyta) in the upwelling Bahía Culebra, north Pacific of Costa Rica”. Revista de Biología 488 Tropical, 60 (Suppl. 2), 93-107. 489 490 Fiedler P.C., Lavín M.F. (2017) “Oceanographic Conditions of the Eastern Tropical Pacific”. In: 491 Glynn, PW., Manzello, DP., & Enochs, IC. (Eds). Coral Reefs of the Eastern Tropical Pacific: 492 Persistence and Loss in a Dynamic Environment. Springer, Berlin, p.59-83. 493 494 Forsman Z.H., Rinkenvich B., Hunter C.L. (2006) “Investigating fragment size for culturing 495 reef-building corals (Porites lobata and P. compressa) in ex situ nurseries”. Aquaculture, 261, 496 89-97. 497 498 Glynn P.W. (1984) “Widespread coral mortality and the 1982-83 El Nino warming event”. 499 Environmental Conservation, 11, 133-146. 500 501 Glynn P.W., Reyes-Bonilla H., Cortés J., Jiménez C., Maté J., Vargas A., Zapata F., Wieters 502 E.A., Navarrete S., Hubbard D.K., & Alvarado JJ. (2017) “Eastern Pacific coral reef regions, 503 coral community composition and reef structure: an overview”. In: Glynn, PW., Manzello, DP., 504 & Enochs, IC. (Eds). Coral Reefs of the Eastern Tropical Pacific: Persistence and Loss in a 505 Dynamic Environment. Springer, Berlin, p.107-176. 506 507 Grottoli, A.G., Rodrigues, L.J., Juarez, C. (2004) “Lipids and stable carbon isotopes in two 508 species of Hawaiian corals Porites compressa and Montipora verrucosa, following a bleaching 509 event”. Marine Biology, 145, 621-631. 510 511 Guzmán H.M., Cortés J. (1989) “Growth rates of eight species of Scleratinian corals in the 512 Eastern Pacific (Costa Rica)”. Bulletin of Marine Science, 44, 1186-1194. 513 514 Guzmán H.M., Cortés J. (1993) “Arrecifes coralinos del Pacifico Oriental Tropical: Revisión y 515 perspectivas”. Revista de Biologia Tropical, 41, 535-557. 516 517 Guzmán H.M., Cortés J., Richmond R.H., Glynn P.W. (1987) “Efectos del fenómeno de "El 518 Niño Oscilación Sureña" 1982/83 en los arrecifes coralinos de la Isla del Caño, Costa Rica”. 519 Revista de Biologia Tropical. 35, 325-332. 520 521 Hein M.Y., Beeden R., Birtles A., Gardiner N.M., Le Berre T., Levy J., Marshall N., Scott C.M., 522 Terry L., Willis B.L. (2020) “Coral restoration effectiveness: Multiregional snapshots of the 523 long-term responses of coral assemblages to restoration”. Diversity, 12, 153-175. 524 525 Hein M.Y., Birtles A., Willis B.L., Gardiner N., Beeden R., Marshall N.A. (2019) “Coral 526 restoration: Socio-ecological perspectives of benefits and limitations”. Biological Conservation, 527 229, 14-25. 528 529 Hein M.Y., Couture F., and Scott C.M. (2018) “Ecotourism and coral reef restoration: case 530 studies from Thailand and the Maldives''. In: Coral Reefs: tourism, conservation, and 531 management. Prideaux B. and Pabel A. (Ed.) Routledge, Abingdon, UK, 137-150. 532 533 Henry L.A., Hart M. (2005) "Regeneration from injury and resource allocation in sponges and 534 corals - a review". International Review of Hydrobiology, 90, 125-158. 535 536 Hesley D., Burdeno D., Drury C., Schopmeyer S., Lirman D. (2017) “Citizen science benefits 537 coral reef restoration activities”. Journal for Nature Conservation, 40, 94-99. 538 539 Hughes T.P. (1984) “Population dynamics based on individual size rather than age: a general 540 model with a reef coral example”. The American Naturalist, 123(6):778–795. 541 542 Hughes T.P., Kerry J.T.A., lvarez-Noriega M.A., lvarez-Romero J.G., Anderson K.D., Baird 543 A.H., et al. (2017) “Global warming and recurrent mass bleaching of corals”. Nature, 543: 373–544 377. 545 546 Ishida-Catañeda J., Pizarro V., Lopez-Victoria M., Zapata F.A. (2020) “Coral reef restoration in 547 the Eastern Tropical Pacific: feasibility of the coral nursery approach”. Restoration Ecology, 28, 548 22-28. 549 550 Jiménez C. (2001) “Arrecifes y ambientes coralinos de Bahía Culebra, Pacífico de Costa Rica: 551 aspectos biológicos, económico-recreativos y de manejo”. Revista de Biología Tropical, 49, 552 S215-S231. 553 554 Jiménez C., Cortés J. (2001) “Changes in reef community structure after fifteen years of natural 555 disturbances in the eastern Pacific (Costa Rica)”. Bulletin of Marine Science, 69(1), 133-149. 556 557 Jiménez C., Cortés J. (2003) “Growth of seven species of Scleratinian corals in an upwelling 558 environment of the Eastern Pacific (Golfo de Papagayo, Costa Rica)”. Bulletin of Marine 559 Science, 72, 187-198. 560 561 Jiménez C., Cortés J., Leon A., Ruiz E. (2001) “Coral Bleaching and mortality associated with 562 the 1997-98 El Nino in an upwelling environment in the Eastern Pacific (Gulf of Papagayo, 563 Costa Rica)”. Bulletin of Marine Science, 69, 151-169. 564 565 Jokiel P.L., Coles S.L. (1977) “Effects of temperature on the mortality and growth of Hawaiian 566 reef corals”. Marine Biology, 43, 201-208. 567 568 Kench P. (2009) "Maldives". Encyclopedia of Islands, edited by Rosemary Gillespie and David 569 Clague, Berkeley: University of California Press, pp. 586-587. 570 571 Knowlton N., Grottoli A.G., Kleypas J., Obura D., Corcoran E., de Goeij J., Felis T., Harding S., 572 Mayfield A., Miller M., Osuka K., Peixoto R., Randall C.J., Voolstra C.R., Wells S., Wild C., 573 Ferse S. (2021) “Rebuilding Coral Reefs: A Decadal Grand Challenge.” International Coral Reef 574 Society and Future Earth Coasts, 56 pp. 575 576 Leuzinger S., Anthony K.R.N, Williw B.L. (2003) "Reproductive energy investment in corals: 577 scaling with module size". Oecologia, 136, 524-531. 578 579 Liñan-Cabello M.A., Flores-Ramirez L.A., Laurel-Sandoval M.A., Garcia Mendoza E., Soriano 580 Santiago O., Delgadillo-Nuño M.A. (2011) “Acclimatation in Pocillopora spp. during a coral 581 restoration program in Carrizales Bay, Colima, Mexico”. Marine and Freshwater Behaviour and 582 Physiology, 44, 61-72. 583 584 Lizano O.G., Alfaro E.J. (2014) “Dinámica atmosférica y oceánica en algunos sitios del Área de 585 Conservación Guanacaste (ACG), Costa Rica”. Revista de Biología Tropical, 62(4), 17-31. 586 587 Lizcano-Sandoval L.D., Londoño-Cruz E., Zapata F.A. (2018) “Growth and survival of 588 Pocillopora damicornis (Scleractinia: Pocilloporidae) coral fragments and their potential for reef 589 restoration in the Tropical Eastern Pacific”. Marine Biology Research, 14, 887-897. 590 591 Maynard J.A., Anthony K.R.N., Marshall P.A., Masiri I. (2008) “Major bleaching events can 592 lead to increased thermal tolerance in corals”. Marine Biology, 155, 173–182. 593 594 McLeod E., Shaver E.C., Beger M., Koss J., Grmsditch G. (2021) “Using resilience assessments 595 to inform the management and conservation of coral reef ecosystems”. Journal of Environmental 596 Management, 277, 111384. 597 598 Nava H., Figueroa-Camacho A.G. (2017) “Rehabilitation of damaged reefs: Outcome of the use 599 of recently broken coral fragments and healed coral fragments of pocilloporids corals on rocky 600 boulders”. Marine Ecology, 38, 1-10. 601 602 Page C.P., Muller E.M., Vaughan, D.E. (2018) “Microfragmenting for the successful restoration 603 of slow growing massive corals”. Ecological Engineering, 123, 86-94. 604 605 R Core Team (2018) “R: A language and environment for statistical computing”. R Foundation 606 for Statistical Computing, Vienna, Austria. https://www.R-project.org/ 607 608 Randall C.J., Toth L.T., Leichter J.J., Maté J.L., Aronson R.B. (2020) “Upwelling buffers 609 climate change impacts on coral reefs of the eastern tropical Pacific”. Ecology, 101(2):e02918. 610 611 Raymundo L.R., Maypa A.P. (2004) “Getting bigger faster: mediation of size-specific mortality 612 via fusion in juvenile coral transplants”. Ecological Applications, 14, 281-295. 613 614 Rinkevich B. (1995) “Restoration strategies for coral reefs damaged by recreational activities: 615 the use of sexual and asexual recuits”. Restoration Ecology, 3, 241-251. 616 617 Rinkevich B. (2006) “The coral gardening concept and the use of underwater nurseries: lessons 618 learned from silvics and silviculture.” In: Coral reef restoration handbook, Precht W.F. (Ed.), 619 291-301. 620 621 Rinkevich B. (2019) “The Active Reef Restoration Toolbox is a Vehicle for Coral Resilience and 622 Adaptation in a Changing World”. Journal of Marine Science and Engineering, 7(7), 201. 623 624 Rixen T., Jiménez C., Cortés J. (2012) “Impact of upwelling events on the seawater carbonate 625 chemistry and dissolved oxygen concentration in the Gulf of Papagayo (Culebra Bay), Costa 626 Rica: Implications for coral reefs”. Revista de Biologia Tropical, 60(Suppl. 2), 187-195. 627 628 Rodríguez-Troncoso A.P., Carpizo-Ituarte E., Cupul-Magaña A.L. (2010) “Differential response 629 to cold and warm water conditions in Pocillopora colonies from the Central Mexican Pacific”. 630 Journal of Experimental Marine Biology and Ecology, 391, 57-64. 631 632 Rodríguez-Troncoso A.P., Carpizo-Ituarte E., Pettay D.T., Warner M.E., Cupul-Magaña, A.L. 633 (2014) “The effects of an abnormal decrease in temperature on the Eastern Pacific reef-building 634 coral Pocillopora verrucosa”. Marine Biology, 161, 131-139. 635 636 https://www.r-project.org/ Rogers C.S. (1990) “Response of coral reefs and reef organisms to sedimentation”. Marine 637 Ecology Progress Series, 62, 185–202. 638 639 Romero-Torres M., Acosta A., Palacio-Castro A.M., Treml E.A., Zapata F.A., Paz-Garcia D.A., 640 Porter J.W. (2020) “Coral reef resilience to thermal stress in the Eastern Tropical Pacific”. 641 Global Change Biology, 26, 3880-3890. 642 643 Roth M.S., Goericke R., Deheyn D.D. (2012) “Cold induces acute stress but heat is ultimately 644 more deleterious for the reef-building coral Acropora yongei”. Scientific Reports, 2, 240, 645 DOI:10.1038/srep00240. 646 647 Salas-Moya C., Fabregat-Malé S., Vargas-Castillo R., Valverde J.M, Vasquez-Fallas F., Sibaja-648 Cordero J., Alvarado J.J. (2021) “Pocillopora cryptofauna and their response to host coral 649 mortality”. Symbiosis, 84, 91–103. 650 651 Sánchez-Noguera C., Jiménez C., Cortés J. (2018a) “Desarrollo costero y ambientes marino-652 costeros en Bahía Culebra, Guanacaste, Costa Rica”. Revista de Biología Tropical, 66, S309-653 S327. 654 655 Sánchez-Noguera C., Stuhldreier I., Cortés C., Morales A., Wild C. Rixen T. (2018b) “Natural 656 ocean acidification at Papagayo upwelling system (north Pacific Costa Rica): implications for 657 reef development”. Biogeosciences, 15, 2349-2360. 658 659 Shafir S., Rinkevich B. (2010) “Integrated long-term mid-water coral nurseries: a management 660 instrument evolving into a floating ecosystem”. Mauritius Research Journal, 16, 365-379. 661 Sherman R.L., Gilliam D.S., Spieler R.E. (2001) “Site dependent differences in artificial reef 662 function: implications for coral reef restoration”. Bulletin of Marine Science, 69, 1053-1056. 663 664 Stuhldreier I., Sánchez-Noguera C., Roth F., Cortés J., Rixen T., Wild C. (2015a) “Upwelling 665 increases net primary production of corals and reef-wide gross primary production along the 666 pacific coast of Costa Rica”. Frontiers in Marine Science, 2:113. doi: 10.3389/fmars.2015.00113 667 668 Stuhldreier I., Sánchez-Noguera C., Roth F., Jiménez C., Rixen T., Cortés J., Wild C. (2015b) 669 “Dynamics in benthic community composition and influencing factors in an upwelling-exposed 670 coral reef on the Pacific coast of Costa Rica”. PeerJ, 3:e1434. 671 672 Torda G., Donelson J.M., Aranda M., Barshis D.J., Bay, L., Berumen M.L., Bourne D.G., Cantin 673 N., Foret S., Matz M., Miller D.J., Moya A., Putnam H.M., Ravasi T., van Oppen M.J.H., Vega-674 Thurber R., Vidal-Dupiol J., Voolstra C.R., Watson S.A., Whitelaw E., Willis B.L., Munday P.L. 675 (2017) “Rapid adaptive responses to climate change in corals”. Nature Climate Change, 7, 627-676 636. 677 678 Tortolero-Langarica J.J.A., Cupul-Magaña A.L., Rodriguez-Troncoso A.P. (2014) “Restoration 679 of a degraded coral reef using a natural remediation process: A case study from a Central 680 Mexican Pacific National Park”. Ocean & Coastal Management, 96, 12-19. 681 682 Tortolero-Langarica J.J.A., Rodriguez-Troncoso A.P., Cupul-Magaña A.L., Carricart-Ganivet 683 J.P. (2017) “Calcification and growth rate recovery of the reef-building Pocillopora species in 684 the northeast tropical Pacific following an ENSO disturbance”. PeerJ, 5e3191. 685 686 Tortolero-Langarica J.J.A., Rodríguez-Troncoso A.P., Cupul-Magaña A.L., Rinkevich B. (2020) 687 “Micro-Fragmentation as an Effective and Applied Tool to Restore Remote Reefs in the Eastern 688 Tropical Pacific”. International Journal of Environmental Research and Public Health, 17(18), 689 6574. 690 691 Vargas-Ugalde R., Gómez-Salas C., Pérez-Reyes C., Umaña-Vargas E., Acosta-Nassar M. 692 (2020) ““Jardinería” para la restauración coralina en el Golfo Dulce, Costa Rica: Una prueba 693 práctica.” UNED Research Journal, 12(1):e2809. DOI: 10.22458/urj.v12i1.2809 694 695 Woodhead A.J., Hicks C.C., Norström A.V., Williams G.J., Graham N.A. (2019) “Coral reef 696 ecosystem services in the Anthropocene”. Functional Ecology, 33, 1023-1034. 697 698 Williams S.L., Sur C., Janetski N., Hollarsmith J.A., Rapi S., Barron L., Heatwole S.J., Yusuf 699 A.M., Yusuf S., Jompa J., Mars F. (2018) “Large-scale coral reef rehabilitation after blast-fishing 700 in Indonesia”. Restoration Ecology, 27, 447-456. 701 702 Yanovski R., Abelson A. (2019) “Structural complexity enhancement as a potential coral-reef 703 restoration tool”. Ecological Engineering, 132, 87-93. 704 705 Zakai D., Levy O., Chadwick-Furman N.E. (2000) “Experimental fragmentation reduces sexual 706 reproductive output by the reef-building coral Pocillopora damicornis”. Coral Reefs, 19, 185-707 188. 708 709 Zapata F.A., Vargas-Angel B. (2003) “Corals and coral reefs of the Pacific coast of Colombia”. 710 In: Latin American Coral Reefs, J.Cortés (Ed.), 419-447. 711 712