Journal of Marine Science and Engineering Article Prospecting the Effects on Abalone (H. discus) Growth under Low-Salinity Stress after Feeding Citrus Peel (CP) and Ecklonia cava disuse (ECD) as Feed Additives Min-Seok Jwa 1 and Chang-Yu Hong 2,* 1 Department of Future Strategy Research, Jeju Research Institute, Busan 63147, Korea; [email protected] 2 Division of Global & Interdisciplinary Studies, Pukyong National University, Busan 48513, Korea * Correspondence: [email protected]; Tel.: +82-51-629-5623 Abstract: This study addressed the effects of adding citrus peel (CP) and Ecklonia cava disuse extract (ECD) in the diet of abalones, which are a highly utilized marine species in Korea. As interest in abalone farming is increasing around the fishing community, the installation of land tanks and fish farms on the southern coast and Jeju Island in South Korea is spreading remarkably. In order to create the best opportunities for survival and reproduction, immunity to disease, and growth potential of abalones, we must consider the diet implemented in abalone farming. The survival rate of abalone has shown positive results when their diet is supplemented with CP and ECD. Our research also concluded that the addition of ECD may have a significant effect on the abalone growth and physiology. Keywords: Abalone (H. discus); Ecklonia cava; citrus peel; low salinity; survival rate; diets Citation: Jwa, M.-S.; Hong, C.-Y. Prospecting the Effects on Abalone (H. discus) Growth under Low-Salinity 1. Introduction Stress after Feeding Citrus Peel (CP) and Ecklonia cava disuse (ECD) as As interest in abalone marine life has increased, means of catching and storing them Feed Additives. J. Mar. Sci. Eng. 2021, have also become significantly more common along the southern coast of South Korea [1]. 9, 707. https://doi.org/10.3390/jmse Abalone (H. discus) is rich in protein and taurine, so it has benefits when used in skin care, 9070707 postpartum cooking, fatigue recovery, and in the prevention of myocardial infarction. Making a suitable environment for abalone is the most important consideration. An Academic Editor: Gualtiero Basilone ideal environment can potentially aid in growth and their ability to fight off disease. For these benefits, the feeding supply should also be considered. Unlike the environment and Received: 8 May 2021 the sources of diseases, which cannot be controlled totally at the farm, diets are under Accepted: 24 June 2021 full control. Published: 26 June 2021 In particular, diet is an important factor that accounts for more than half of the operating costs of aquaculture [2]. It is one of the more important factors that can be Publisher’s Note: MDPI stays neutral modified artificially. with regard to jurisdictional claims in Raw seaweed and raw kelp are supplied as food in abalone aquaculture. The supply published maps and institutional affil- of these natural foods is not only unstable, depending on the crop conditions, but the price iations. fluctuations are also severe and the effect on growth is reported to be lower than that of nurtured feeds [3–6]. Accordingly, some studies on the development of abalone feed have already been carried out in some countries such as Australia, China, Japan, Chile [7–11]. These researchers aim to develop economically blended feeds suitable for the domestic Copyright: © 2021 by the authors. aquacultural situation. Citrus peel, which is produced in the citrus agriculture industry—a Licensee MDPI, Basel, Switzerland. major primary industry in Jeju Island, is a byproduct generated after processing citrus This article is an open access article fruits. It is a great financial burden for the agricultural households of Jeju because it costs a distributed under the terms and lot to dispose of citrus peel. conditions of the Creative Commons Citrus fruits are known to contain various flavonoid compounds, which are polyphe- Attribution (CC BY) license (https:// nols, such as naringin, hesperidin, naringenin, hesperetin, nobiletin, and tangeretin; thus, creativecommons.org/licenses/by/ they have good physiological activity [12,13]. It has also been reported that citrus peel, a 4.0/). J. Mar. Sci. Eng. 2021, 9, 707. https://doi.org/10.3390/jmse9070707 https://www.mdpi.com/journal/jmse J. Mar. Sci. Eng. 2021, 9, 707 2 of 10 byproduct of citrus fruits, contain a large amount of flavonoids and thus have antioxidant activity. Research to extract these functional substances is being actively conducted [14]. In addition, Ecklonia cava is a perennial brown algae belonging to the taxonomic rank Phaeophyta, Phaeophyceae, Laminariales, Laminariaceae. It is one of the largest seaweed species in Korea, along with other members of the genus Ecklonia. As phlorotannin is known to have excellent antioxidant, antihypertensive, antithrombotic, and anticancer properties, it is emerging as an important ingredient in the “health food” and pharmaceuti- cal industries [15,16]. However, because the Ecklonia stalk, which accounts for approximately 25% of Ecklonia cava weight, is discarded without being used as a processing byproduct, a method of utilization is required. Because polyphenolic substances have been reported to impart high antioxidant activity in the stems of Ecklonia cava [17], we expect the use of Ecklonia byproducts in the feed of abalones, which contain seaweed, to create new added value as well as aid environmental preservation. Therefore, this study was conducted to evaluate the effects of citrus fruit and Ecklonia byproducts, on growth and salinity stress, as additives for abalone blended feed, which is a major aquaculture target species in Korea. 2. Materials and Methods 2.1. Experimental Materials and Breeding Management The abalone (H. discus) used in the experiment was purchased and used at a private marine life farm near Jeju City, Jeju Province. Individual abalones with an average length of 3.8 ± 0.2 cm, a width of 2.5 ± 0.2 cm, and a weight of 6.4 ± 0.1 g were used. The experimental abalones were bred at 18 ± 0.3 ◦C for one week beforehand, and after they were acclimatized, bred in a tank with a circulatory filtration system. At this time, the salinity was maintained at approximately 33.5 psu and air was supplied by putting a shelter through a PVC pipe that was cut vertically (Æ 20 cm, L 20 cm). During the preliminary breeding period, raw seaweed, weighing 2% of the abalone weight, was put in the tank as a food source and 80 abalones were placed in each tank. 2.2. Production of Feed for Abalones In this breeding experiment, a total of four types of feed were prepared and each experimental group had three replicates. Casein was used as the main protein source, and fish oil and soybean were used as the dextrin lipid sources. The amount of carbohydrate sources was controlled using oil, and wheat flour was added to control the change in carbohydrates that occurred because of the citrus peel (CP) and the fermented Ecklonia cava disuse extract (ECD). For the test feed, the test feed was added so that the concentration of CP and ECD was 6% and mixed at a ratio of 1:1. The composition of the experimental feed is shown in Table1. 2.3. Survival Rate and Growth Rate Measurement Experiment According to Feed To measure the survival rates, abalones that naturally died at weeks 0, 1, 2, 3, 4, 8, and 12 were recorded in a timely manner. The dead abalones that were not attached to the shelter or tank wall were counted. Subjects that did not respond even when their feet and tentacles were stimulated with a glass rod were considered dead. In addition, to measure the growth of the abalones, we measured their change in size immediately after the start of the experiment, and on the 4th, 8th, and 12th weeks. A digit caliper (0.01 mm) was used for the shell length and shell width (Figure1), and a mobile scale (0.01 g) was used for the weight. Following the addition of ECD and CP to the abalone diet, the differences in growth were then compared and analyzed. J. Mar. Sci. Eng. 2021, 9, 707 3 of 11 J. Mar. Sci. Eng. 2021, 9, 707 3 of 10 Casein 20 20 20 20 Sodium alginate 23 23 23 23 Mineral 4 4 4 4 Table 1. Ingredients (%) of the experimental diets. Vitamin 2 2 2 2 Choline 0.5 0.5 Experimental Diets0.5 (%) 0.5 Soybean oil Ingredients1.5 1.5 Control CP1.5 ECD1.5 ECD + CP Fish oil CP2 (Citrus Peel)2 62 2 3 Total ECD (100.0Ecklonia cava disuse) 100.0 100.0 6100.0 3 Proximate analysis (%) Fish meal 15.5 15.5 15.5 15.5 Crude protein 28.8Dextrin 28.8 25 2528.5 2528.6 25 Crude lipid 1.47Wheat flour2.22 6.5 0.51.77 0.51.99 0.5 Casein 20 20 20 20 Crude carbohydrate 4.97 5.72 5.27 5.49 Sodium alginate 23 23 23 23 Mineral 4 4 4 4 2.3. SurvivalVitamin Rate and Growth Rate Measurement 2 Experiment 2 According 2 to Feed 2 To measureCholine the survival rates, 0.5 abalones that naturally 0.5 died at 0.5 weeks 0, 1, 2, 0.5 3, 4, 8, and 12 wereSoybean recorded oil in a timely manner. 1.5 The dead 1.5 abalones that 1.5 were not attached 1.5 to the shelter orFish tank oil wall were counted. 2 Subjects that 2did not respond 2 even when their 2 feet and tentaclesTotal were stimulated with 100.0 a glass rod 100.0were considered 100.0 dead. In addition, 100.0 to measure the growth of the abalones, we measured their change in size immediately after Proximate analysis (%) the start of the experiment, and on the 4th, 8th, and 12th weeks.