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Echinodermata: Echinoidea) Belén Chaar, F., Fernández, J.P., Sepúlveda, L.R., & Rubilar, T. (2021). The influence of density on survival and larval development in the sea urchin Arbacia dufresnii (Echinodermata: Echinoidea). Revista de Biología Tropical, 69(S1), 334-345. DOI 10.15517/rbt.v69iSuppl.1.46365 DOI 10.15517/rbt.v69iSuppl.1.46365 The influence of density on survival and larval development in the sea urchin Arbacia dufresnii (Echinodermata: Echinoidea) Florencia Belén Chaar1,2,3 Jimena Pía Fernández2,3 Lucas R. Sepúlveda2,3 Tamara Rubilar 2,3* 1. National University of Patagonia San Juan Bosco, Bv. Almte. Brown 3051, Puerto Madryn, 9120, Chubut, Argentina; [email protected] 2. Laboratory of Chemistry of Marine Organisms. Instituto Patagónico del Mar. Faculty of Natural Sciences and Health Sciences. National University of Patagonia San Juan Bosco. Puerto Madryn headquarters. Argentina; [email protected]; [email protected], [email protected] (*Correspondence). 3. Biological Oceanography Laboratory- Centro para el Estudio de Sistemas Marinos- Centro Nacional Patagónico, Centro Científico Tecnológico del Consejo Nacional de Investigaciones Científicas y Técnicas- Consejo Nacional de Investigaciones Científicas y Técnicas de Argentina, Bv. Almte. Brown 2915, Puerto Madryn, 9120, Chubut, Argentina. Received 13-VII-2020. Corrected 03-VIII-2020. Accepted 18-IX-2020. ABSTRACT Introduction: Density is one of the critical factors in echinoderm larvae for aquaculture purposes. Echinoplutei larvae are very sensitive to overcrowding. High culture density can lead to problems with bacteria or protozoa, decreasing survival and generating abnormal morphotypes. Objective: To evaluate the effect of culture density on survival and larval growth in the sea urchin Arbacia dufresnii. Methods: Two days after fertilization of A. dufresnii we we kept treatments at 1, 3, 5 and 10 larvae.ml-1, with three replicates each. We recorded survival and abnormal morphotypes periodically, as well as growth:somatic rod length, total length, and length of the post oral arms,. we applied generalized linear models. Results: Survival is dependent on density, time and replicates, and their interactions. Larval growth depended on density and time, also with interaction between the variables. The treatment of 5 larvae.ml-1 had the highest survival and larval condition. Conclusions: Larval culture of A. dufresnii had the best results at 5 larvae.ml-1. Key words: Echinoplutei, growth, morphometry, aquaculture, treatments. Echinoderm embryos, particularly sea for ecotoxicology analysis (Rahman, Tsuchi- urchins and starfish, have been studied for ya, & Uechara, 2009) and molecular studies more than 170 years (Rubilar & Crespi-Abril, (McClay, 2011; Hinman & Burke, 2018). Addi- 2017). The culture is simple and has been used tionally, the development of simple protocols as a classical animal model for embryological facilitates the aquaculture of the entire life cycle studies (Williams & Anderson, 1975). More of sea urchins. This progress has been crucial in recently, echinoderm embryos have been used the further development of aquaculture and in 334 Revista de Biología Tropical, ISSN electrónico: 2215-2075 Vol. 69(S1): 334-345, March 2021 (Published Mar. 30, 2021) the implementation of repopulation projects in obtain a production of juveniles justifying the areas affected by overexploitation, or by mass required investment and efforts (Stotz, 2014). mortality, as found in Japan, Chile and China, For aquaculture purposes, the maximum viable among other areas (Unuma, Sakai, Agatsuma, density of a species is of particular importance. & Kayaba, 2015; Brady & Scheibling, 2006). In addition, when analyzing a culture, it is As a result, advances in this area are of major important to consider the morphometrics of interest for research, conservation and aquacul- the larvae, both in terms of size and as well as ture purposes. in terms of the proportion of its structures as The echinoderm larvae culture is crucial to these can provide information on the culture any conservation or aquaculture project. There conditions and the effects on development. is a need of having certain, optimal growing To estimate larval growth, both PO (post oral conditions in order to obtain good results in arms) and TL (total length) of sea urchin larvae the successful production of post settled juve- have been two of the most frequently used niles. Variables, such as temperature, salin- indicators in recent years. The PO is one of the ity (Astudillo, Rosas, Velásquez, Cabrera, & most frequently used measures as it is, gener- Maneiro, 2005; Domínguez, Rosas, Velásquez, ally, very sensitive to changes in environmental Cabrera, & Mata, 2007), feeding, and larval variables (Fenaux, Strathmann, & Strathmann, culture density (Astudillo et al., 2005; Buitrago 1994; Hart & Strathmann, 1994; Eckert, 1998; & Lodeiros-Seijo, 2005) are among the most Jimmy, Kelly, & Beaumont, 2003; Cárcamo, important factors. Candia, & Chaparro, 2005; Domínguez et al., There exists a wide variety of values 2007; Clark et al., 2009; Rosas, Velásquez, in the optimal culture density depending on Fernández, Mata, & Cabrera, 2009; Adams, the species, with values ranging from 0.025 Sewell, Angerer, & Angerer, 2011; Fernández, to 3 larvae.ml-1 (Buitrago & Lodeiros-Seijo, 2019). At the same time, measuring the length 2005; Salas-Garza, Carpizo-Ituarte, Parés-Sier- of the somatic rod (SO), can be an impor- ra, Martínez-López, & Quintana-Rodríguez, tant indicator of growth parameters related 2005; Kalam-Azad, Mckinley, & Pearce, 2010; to ambient factors (Clark et al., 2009; Feu- Díaz-Pérez & Carpizo-Ituarte, 2011). High nax et al., 1994). culture densities can affect the quality of the The sea urchin Arbacia dufresnii is abun- water, increasing the risk of contamination dant along the Argentinian and Chilean coasts, due to the increase in organic matter, as well inhabiting coastal areas from Río de la Plata as the increase in waste metabolic substances (35° S) in the Southwest Atlantic to Puerto (Lamare & Barker, 1999; Salas-Garza et al., Montt (42° S) in the South Pacific, including 2005; Domínguez et al., 2007; Clark, Lamare, the Malvinas Islands (Bernasconi, 1966). This & Baker, 2009; Kalam-Azad et al., 2010). High species has been the subject of several scientific densities can also generate stress or competition studies. Specific ecological aspects have been for space or food, affecting the normal develop- described, such as their abundance, distribu- ment of the organisms (Pechenik, 1999; García tion, role in the ecosystem and diet in different et al., 2005) Inadequate density cultures usually environments (Vásquez, Castilla, & Santelices, translate into decreased survival, changes in the 1984; Larraín, Mutschke, Riveros, & Solar, growth of organisms, and/or a higher incidence 1999; Penchaszadeh & Lawrence, 1999; Zaixso of abnormal phenotypes, such as the absence & Lizarralde, 2000; Teso, Bigatti, Casas, Piriz, of arms or very small bodies with very long & Penchaszadeh, 2009; Newcombe, Cárdenas, arms (Clark et al., 2009; Kalam-Azad et al., & Geange, 2012; Brogger et al., 2013; Castro, 2010). However, considering the natural mor- 2014; Epherra, 2016; Epherra, Martelli, Mor- tality during the larval stage and the specific san, & Rubilar, 2017). Descriptions of certain culture conditions, it is necessary that the initial physiological aspects of adult individuals have quantity of organisms is sufficient to be able to been described, such as the reproductive cycle Revista de Biología Tropical, ISSN electrónico: 2215-2075, Vol. 69(S1): 334-345, March 2021 (Published Mar. 30, 2021) 335 and gonadal composition (Brogger & Iva- 2004; Strathmann, 2014; Ettensohn, 2017). A nov, 2010; Epherra et al., 2015a; Epherra et total of 800 000 eggs were placed in a liter of al.,2015b; Parra et al., 2015; Zárate, Díaz de sea water filtered up to 1 µm and sterilized with Vivar, Ávaro, Epherra, & Sewell, 2016). Addi- UV light. Fertilization was carried out by using tionally, specific aspects of the early stages of a 1:100 000 v/v dilution of sperm according the life cycle of this species have also been to Fernández et al. (2019). After 30 min, the described, such as fertilization under controlled percentage of fertilization success was verified conditions (Fernández, Epherra, Sepúlveda, & and calculated under a light microscope (Leica Rubilar, 2019), complete embryonic develop- DM 2500) on the basis of observing the fertil- ment and specific stages of larval development ization membrane in the eggs. (Bernasconi, 1942; Brogger, 2005; Fernández, 2019). Furthermore, there have been descrip- Embryo culture: The fertilized eggs were tions of the response of the larvae to ocean kept for 48 h with constant and gentle aeration, acidification (Catarino, De Ridder, Gonzalez, at a temperature of approximately 17 °C and at Gallardo, & Dubois, 2012) and the monitoring a salinity of 34-35 ppm. At 24 h after fertiliza- of development in natural conditions (Kino, tion, the culture volume was tripled in order to 2010). These studies have served as a basis decrease the density of developing organisms. for the exploration of the aquaculture potential After 72 h, the larvae in the prism stage were of the species (Rubilar et al., 2016). However, transferred to a new container, quantified, and there are, still, few studies which have succeed- the corresponding dilutions were undertaken ed in determining the optimal larval culture for each treatment. density
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