Low‐density culture of red abalone juveniles,Haliotis rufescens Swainson 1822, recirculating aquaculture system and flow‐through system

Commercial abalone culture is carried out using flow‐through systems with a high water volume exchange in Baja California, Mexico. The objective of this work was to compare the growth rate and survival of red abalone cultured in two systems. Flow through (daily water exchange rate of 800%) and recirculating systems consisted of a 250 L fibreglass tank and constant aeration, but biofiltration in the recirculating system was provided with a 28 L (1 ft³) bubble‐washed bead filter. Water variables were measured either daily (dissolved oxygen, temperature, pH and salinity) or three times a week (total ammonia nitrogen, nitrate‐nitrogen, nitrite‐nitrogen and alkalinity). Shell length was measured every 2 weeks for 18 weeks. Only the alkalinity and pH were significantly different due to the addition of sodium bicarbonate to the recirculating system. Abalone growth rate was 26.1 ± 15.96 μm day^?1 in the recirculating systems and 22.21 ± 18.69 μm day^–1 in the flow‐through systems. The final survival was 78.74% in the recirculating systems and 71.82% in the flow‐through systems. Significant differences in the final size and survival of the abalones were found between systems (P<0.05). Therefore, recirculating aquaculture systems is a feasible alternative for juvenile red abalone culture.     

Optimal protein level in a semipurified diet for juvenile greenlip abalone Haliotis laevigata

To optimize dietary protein level in relation to growth, semipurified diets with an essential amino acid profile similar to that of the soft body profile were fed for 59 days to young greenlip abalone, Haliotis laevigata (initial shell length, 15–25 mm). Animals were housed in 10-L acrylic tanks, with flow-through seawater supplied at 1 L min^–1 (20 °C, salinity= 36 g L^–1). Protein level of feeds ranged from 122 g kg^–1 to 461 g kg^–1 crude protein (CP) on an 'as is' basis. Second-order polynomial regression analysis of specific growth rate indicated that maximal growth occurs at 270 g kg^–1 CP. The protein and energy components of the feed were estimated to have a digestibility of 71.7% and 55.6%, respectively.     

Histological evidence of accumulation of iron in postlarvae of red abalone,Haliotis rufescens

The effect of iron on abalone postlarvae (Haliotis rufescens) was investigated in a controlled‐culturing system. Three iron concentrations (0.15, 1.5 and 15 mg L^?1 of Fe) and a control (no iron added) were used to culture H. rufescens postlarvae while being fed the diatom Navicula inserta over 10 days. Results indicate that H. rufescens postlarvae accumulate iron granules in the stomach, digestive gland and mantle, but not in the gills or other tissues. The number and diameter of iron granules in tissues increased with increasing iron concentration in the culturing environment. The iron accumulation is assumed to have been acquired in the digestive system through the iron‐enriched diatom feed and in the mantle through subcutaneous iron transfer. The lack of iron granules in the gills suggests that iron is not absorbed through the respiratory system, as is the case for many filter feeding bivalves. Exposure to the highest iron concentration (15 mg L^?1) resulted in tissue abnormalities where granules accumulated, and may have significantly affected the health of H. rufescens postlarvae. These findings provide valuable information for the regulation of appropriate iron levels within aquaculture settings and highlights the importance of monitoring iron levels within abalone larval culturing environments.     

Growth performance of weaning red abalone (Haliotis rufescens) fed with Macrocystis pyrifera plantlets and Porphyra columbina compared with a formulated diet

A feeding experiment was carried out to evaluate two natural diets versus a formulated feed on the performance of weaning red abalone Haliotis rufescens . Four treatment diets were then investigated: a formulated diet; plantlets from culture Macrocystis pyrifera, Porphyra columbina from natural beds; and a mixed diet consisting of a blend of fresh P. columbina together with the formulated diet. This study was performed in a shallow aquaculture system with a horizontal water flow. After 90 days, significant differences were observed between treatments. The highest growth was obtained with Porphyra (3.3 mm month⁻¹), followed by the mixed diet (2.6 mm month⁻¹), then Macrocystis (2.1 mm month⁻¹) and lastly the formulated diet (1.4 mm month⁻¹). Moreover, after the diets were tested for stability and remnant nutrients after a 12-h water immersion, a positive trend appeared to be related to the protein/energy (P:E). It is concluded that fronds of P. columbina resulted in the best diet for weaning H. rufescens under horizontal water flow systems, even if apparently the water stability of the formulated diet had a negative impact on abalone performance, particularly due to a poor nutrient retention capacity. Therefore, formulated diets should be improved before being recommended for weaning red abalone.     

Effects of temperature on growth of juvenile blackfoot abalone, Haliotis iris Gmelin

The effects of temperature on growth and survival of juvenile blackfoot abalone, Haliotis iris, were investigated. Animals of 10, 30 or 60 mm initial shell length were exposed to ambient (6–10°C), 14, 18, 22 and 26°C for 112 days in a flow‐through culture system. Maximum growth occurred at 22°C for the 10 and 30 mm size classes and at 18°C for the 60 mm size class. Regression analysis identified the optimal temperature for growth (T_optG) at around 21°C for the 10 and 30 mm size classes and at 17–18°C for the largest size class. In a second experiment, the critical thermal maximum of H. iris was determined as a measure of thermal tolerance. Abalone were subjected to increasing water temperatures at a rate of 2°C h^?1 until they detached from the substrate. Abalone of 10 mm displayed greater thermal tolerance than abalone of 30 and 60 mm in length. CT₅₀ temperatures were 28.8, 27.7 and 27.8°C, yielding deduced T_optG values of 19.7, 18.3 and 18.4°C for the 10, 30 and 60 mm size classes respectively. The size‐dependent nature of the relationship between growth and temperature could be capitalized upon in recirculating aquaculture systems.     

Evaluation of an in vitro digestibility technique for the prediction of protein digestibility in the South African abalone, Haliotis midae L.

A pH-stat multienzyme in vitro digestibility assay was investigated for its efficacy in predicting apparent protein digestibility coefficients in abalone diets. Linear regression analysis between in vitro digestibility estimates and in vivo digestibility coefficients obtained from abalone, revealed that the technique could be used to reliably predict apparent protein digestibility. Maximal predictability of the system was obtained when protein sources were analysed according to origin – animal or plant. The technique was used to assess the apparent protein digestibility of 34 potential protein sources for use in formulated feeds for Haliotis midae .     

Growth and feeding in juvenile triploid and diploid blacklip abalone, Haliotis rubra (Leach, 1814), at two temperatures

Growth and feeding of juvenile triploid and diploid blacklip abalone Haliotis rubra (Leach, 1814) were investigated at two temperatures of 17 and 21 °C over a 50‐day period. There were no differences in growth between triploid and diploid abalone as measured by shell length and body weight. Both triploid and diploid abalone increased in length but not in weight at 21 °C. Condition indices were similar for triploid abalone maintained at both temperatures; however, those for diploid abalone were significantly higher at 17 °C than at 21 °C. Food intake was significantly greater yet feed conversion efficiency was significantly lower in triploid than in diploid abalone. Both the feeding variables were independent of temperature. On average, diploid abalone were able to convert 1 g of dry food ingested to 0.58 g of body weight, but triploid abalone only 0.44 g.     

Growth and reproductive performance of triploid and diploid blacklip abalone, Haliotis rubra (Leach, 1814)

Growth and reproduction of triploid and diploid blacklip abalone Haliotis rubra (Leach, 1814) were compared in a 30-month study. Triploidy was induced by inhibition of the second polar body formation using 6-dimethylaminopurine (6-DMAP) or cytochalasin B (CB). There were no significant differences in growth and survivorship between triploid and diploid abalone. However, triploid abalone had a more elongated shell and greater foot muscles than diploid abalone. A slightly curvilinear growth in shell length was conformed to all treatments. While diploid abalone had reached sexual maturity and spawned during the study, gonadal development and gamete maturation were abnormal in triploids. Female triploids lacked an apparent gonad at the macroscopic level but microscopic examination revealed that they had a thin layer of oogonia development. In contrast, male triploids were able to form similar-sized gonads to diploids during most of the reproductive period, but with brown-yellow discolouration and stalled gametogenesis at spermatocyte formation. Sex ratio of triploid abalone did not deviate from 1:1. With the onset of sexual maturation, growth and gonadal maturation occurred concurrently in diploid abalone, and there was no indication that growth of (diploid) abalone was reduced.     

Effects of oxygen supersaturation and temperature on juvenile greenlip, Haliotis laevigata Donovan, and blacklip, Haliotis rubra Leach, abalone

Growth and survival of juvenile greenlip (39.03 (SD 3.80) mm (n=524)) and blacklip (31.92 (SD 4.19) mm (n=531) abalone were investigated at high dissolved oxygen levels (95–120% saturation) between 17 and 19°C. Abalone were fed the same artificial diet and each species was contained in groups of approximately 30 individuals within triplicate tanks for each of six treatments and were exposed to flow through water adjusted to give experimental conditions for up to 75 days. Blacklip abalone held at 16.9°C and 97% oxygen saturation grew in shell length significantly faster than all other treatments of blacklip abalone held at 19°C, and significantly faster than blacklip abalone maintained at 111% oxygen saturation and 17.5°C. Both temperature and oxygen saturation significantly affected the survival of this species. Blacklip abalone held at 19°C had significantly lower survival for both 96% oxygen saturation and 120% oxygen saturation, compared with blacklip abalone maintained at either 110% oxygen saturation and 19°C, or for any 17°C treatment. No significant differences were noted for greenlip abalone within the range tested in terms of growth rate, food consumption rate or survival, indicating that greenlip abalone tolerated these conditions better than did blacklip abalone.     

Activity of digestive enzymes along the gut of juvenile red abalone, Haliotis rufescens, fed natural and balanced diets

Two carbohydrases (cellulase, lysozyme), three proteases (trypsin, aminopeptidase and non-specific protease), a non-specific lipase, and semiquantitative tests of 19 digestive enzymes were assayed in different gut sections of juvenile red abalone, Haliotis rufescens, in order to identify the regions where digestion takes place and investigate the extent to which diet composition can modify the digestive capacity of abalone. The abalone were fed either fresh kelp (K) or balanced diets containing 25 or 38% crude protein for 6 months. Enzyme assays were carried out on different sections of the abalone's gut at the end of this period. On a weight-specific basis, the digestive gland was the site containing most of the enzymes. On a protein-specific basis, two main digestion regions were identified: the digestive gland-stomach region that is characterized by high activities of cellulase and lysozyme, chymotrypsin and protease, and the mouth-intestine region with a typically high activity of lipase and amino peptidase. Significant dietary effects were observed on the activity of enzymes, especially in the digestive gland. Abalone fed with 25 and 38% crude protein diets exhibited higher cellulase (39.8 ± 4.6 and 14.2 ± 0.8 mU mg^− 1 protein, respectively) and lysozyme activities (88.0 ± 20.4 and 56.6 ± 15.7 U, respectively) than those fed with fresh kelp (5.5 ± 0.7 mU mg^− 1 protein and 17.1 ± 1.8 U). In contrast, higher protease activity was found in kelp-fed organisms (234.1 ± 20.4 μg product/mg protein) than those fed the 25 and 38% crude protein diets (109.5 ± 20.7 and 119.5 ± 20.5 μg product/mg protein, respectively). Semiquantitative API ZYM assays resulted in no clear food-specific effects on the activity of carbohydrases, proteases, ester hydrolases or phosphohydrolases, yet organ-specific differences were conspicuous in various cases, and generally agreed with quantitative results. It is suggested that the increased carbohydrase activity exhibited by organisms fed the balanced diets resulted from a combination of an increased number of resident bacteria in the abalone's gut and facilitated contact between dietary substrates and digestive cells. The present results indicate that H. rufescens can adjust their enzyme levels in order to maximize the acquisition of dietary protein and carbohydrates. This characteristic can be advantageously used to search for suitable diets in abalone aquaculture.     

光照强度对九孔鲍幼虫及幼鲍生长存活的影响

在自然光周期下，分别在高温长光照和低温短光照的两个节季节，采用控制光照强度的方法进行对九孔鲍幼虫及幼鲍50d左右培育的小型试验。结果表明，光照强度对附着基上摄食微藻的幼虫及幼鲍生长和存活等有重要的影响。对幼虫及幼鲍最适宜的光照强度，在高温长光照下为700－1200lx，在低温短光照下为1300－2000lx。选用适宜的光照强度在高温长光照和低温短光照下先后进行了3次经量育苗试验，获得了与小型试验相同的结果。     

Effects of alternate starvation and refeeding cycles on food consumption and compensatory growth of abalone, Haliotis asinina (Linnaeus)

The effects of alternate starvation and refeeding on food consumption and compensatory growth of hatchery‐bred abalone, Haliotis asinina (Linnaeus), were determined. Two groups of abalone juveniles (mean shell length = 29 mm, body weight = 5 g) were alternately starved and refed a macro‐alga, Gracilariopsis bailinae at equal duration (5/5 or 10/10) over 140 days. A control group (FR) was fed the seaweed ad libitum throughout a 200‐day experimental period. Starved and refed abalone showed slower growth rates (DGR, 63 and 70 mg/day in the 5/5 and 10/10 groups respectively), as a result of reduced food intake (DFI 15% and 16% day^?1 respectively), after repeated starvation and refeeding cycles. Percentage weight gains (5/5 = 196%, 10/10 = 177%) were significantly lower than that of the control (397%). When refed continuously over 60 days, the starved groups exhibited increased DFI and fed at the rate of 24% and 25% day^?1, which were not significantly different from that of the control at 26% day^?1. At the end of the experiment, no significant differences were observed among three treatments in terms of shell length (range: 46–48 mm), body weight (range 25–28 g), % weight gain (392–465%) and per cent survival (range 87–98%). The results indicated that H. asinina had a complete compensatory growth following a return to full rations after a series of intermittent starvation and refeeding cycles.     

Use of macroalgae supplemented with probiotics in the Haliotis rufescens (Swainson, 1822) culture in Northern Chile

In this study, we evaluate the use of macroalgae as vectors of probiotics bacteria into the digestive tract of abalone to improve their survival and growth. It is shown that when abalone Haliotis rufescens of different sizes were fed with a natural diet composed of fronds of the macroalga Macrocystis integrifolia supplemented with a mixture of Vibrio sp. C21‐UMA, Agarivorans albus F1‐UMA and Vibrio sp. F15‐UMA bacteria, there was a significant increase (P<0.05) in the average monthly growth rate and survival (%)in a period of 210 days, compared with the control without a probiotic supplement. The permanence of the probiotics in the digestive tract of the animals was monitored, and it was found that the number of culturable C21‐UMA and F1‐UMA bacteria decreased significantly (P<0.05) in recently weaned and adult abalone, and they almost disappeared completely on day 19 of the bioassay. However, the culturable Vibrio sp. F15‐UMA disappeared completely from the digestive tract on day 22 of the bioassay, and these were the bacteria that remained at the highest concentration compared with the other two bacterial strains over the experimental period. It is therefore shown that it is feasible to use a probiotic mixture to improve the profitability of the H. rufescens culture.     

Growth of the abalone Haliotis diversicolor (Reeve) fed with macroalgae in floating net cage and plastic tank

The abalone Haliotis diversicolor (Reeve) or ‘tokobushi’ is being cultured in Kagoshima, southern Japan for stock enhancement of fishing grounds. However, with decreasing catches from the wild some fishermen's group may eventually resort to rearing tokobushi until of marketable size. Growth experiments of tokobushi fed macroalgae in floating net cage and plastic tanks were conducted to evaluate the influence of culture system, macroalgal food, age and water temperature on their culture. In floating net cage, marketable sizes were obtained at the age of 457–822‐day old (15–27‐month old) with highest growth rate of 2.36% g day^?1 and 0.21 mm day^?1. One advantage of the plastic tank culture system was that the tokobushi shell remained clean throughout the culture period. Macroalgal food affected the growth rate; Sargassum fusiforme (Setchell)‐ and Ulva pertusa (Kjellman)‐fed tokobushi were better than Meristotheca papulosa (Montagne)‐fed. Further, 1‐year‐old tokobushi had better growth performance and condition index than the 2‐year‐old cohort, and growth rates were higher at 17°C than at 12°C. These findings should help tokobushi farmers design their mariculture scheme considering both economic and environmental aspects.     

Seasonal variations of chemical constituents in the muscle and viscera of small abalone fed different diets

SUMMARY: The seasonal changes in levels of chemical constituents in the muscle and viscera of small abalone Haliotis diversicolor fed gracilar and an artificial diet were investigated. Muscle yields were higher in winter and spring. In October specimens, total adenosine 5'-triphosphate (ATP)-related compounds (ARC), total free amino acids (FAA), and glycogen in both muscle and viscera decreased markedly. The artificial diet fed to small abalone had much higher glycogen in the muscle than those fed on gracilar, and showed a great seasonal change. Total amounts of ARC in the muscles were higher through March to July, while those in the viscera were maximal in January. Taurine, arginine, glycine, glutamic acid, and alanine were the major FAA in both tissues, accounting for 81–94% of the total FAA. Total amounts of FAA in the muscles were higher in the samples collected from winter and early spring than in other seasons. Glycine, glutamic acid, and adenosine monophosphate might be the most important taste components related to the palatability of small abalone. Their total amounts in the muscles of the two specimens were considerably high in December to March. This finding suggested that small abalone produced in winter and early spring might be more palatable.     

Changes of heritability and genetic correlations in production traits over time in red abalone (Haliotis rufescens) under culture

Red abalone Haliotis rufescens is one of the most valuable mollusks in the international market, but it has a low growth rate. A breeding program is being developed to increase its growth rate in Chile. We estimated the changes in direct heritability (h²), maternal/common environments heritability (m²) and genetic correlations (r_G) of growth traits (shell length and width, total mass, flesh mass and foot protein as an indicator of meat quality) measured during 2 years (every 4 months) from the juvenile stage (27 months) to the adult harvesting age (51 months), in 60 full‐sib red abalone families. Heritabilities for growth traits measured in juveniles and young adults (27–35 months of age), were low (0.07–0.17) and not significant. Initial low h² were associated with significant amounts of maternal/common environmental effects (m² = 0.4). In young adults and abalone near the harvest age (39–51 months of age) h² were much higher (0.32–0.75). These results emphasize the importance of multiple estimations of h² over time. Among meat quality traits, only the h² for the flesh mass for adults at harvesting age was significant (0.15). We observed strong positive r_G (>0.9) between shell sizes (easy to measure) and total and flesh masses (trait more related to market value than shell sizes but harder to measure) for adults at harvesting age. Thus, if the 5% largest 51 month old abalone were selected from the population as broodstock we expect a positively correlated response on flesh mass of 23.4%.     

Settling and raising postlarval abalone Haliotis diversicolor supertexta (Lischke) on microparticulate diets embedded in a layer of alginate

Two 21 day experiments were conducted to determine whether microparticulate diets could be used to substitute for diatoms for raising (Experiment 1) as well as settling and on‐growing (Experiment 2) Haliotis diversicolor supertexta postlarvae. The experiments were conducted in 10 L flow‐through tanks and each diet had three replicate tanks. In experiment 1, three experimental microparticulate diets supplied by a commercial feed company, labelled SF‐15, SF‐15 (T) and JFY‐13–1, were used, and in experiment 2, SF‐15 and SF‐15 (T). Artificial diets were compared to diatoms in terms of supporting growth and survival. The survival rate for experiment 1 was lowest for JFY‐13–1 (0%) and highest for diatoms (12.9±0.8%). The final size of postlarvae fed SF‐15 and SF‐15 (T) was significantly greater than that of postlarvae fed diatoms, 973±193 μm and 996±220 μm compared to 786±141 μm. In experiment 2, a total number of 501 larvae settled onto plates covered with SF‐15, 254 on SF‐15 (T) and 1085 on diatoms. The highest survival rate was 33.1±6.9% for SF‐15 and the lowest was 17.3±5.0% for diatoms. However, the final length of postlarvae fed diatoms was significantly greater than SF‐15 and SF‐15 (T); 1018±379 μm compared to 728±89 μm and 717±90 μm. The artificial micro diets used in this study were comparable to diatoms in terms of grow and survival of postlarval H. diversicolor supertexta.     

Shelter selection by juvenile Pacific abalone (Haliotis discus hannai Ino) as a function of food distribution and water flow velocity

Juvenile Pacific abalone (Haliotis discus hannai Ino) are currently reared in land‐based aquaculture systems until they reach the necessary size for seeding in the sea. One problem that this industry faces is that an uneven distribution of juveniles in tanks can lead to variations in the growth rate. Understanding the cues that affect the sheltering behaviours exhibited by juvenile Pacific abalone—namely, the food distribution and the water flow velocity—will help optimize abalone culture settings. In this study, a group of 1,000 juvenile Pacific abalone (distributed across three tanks) was visually observed and enumerated during six experiments that were conducted over a 5‐month study period. It was found that juvenile Pacific abalone preferred to shelter close to food sources when the food was unevenly distributed. When the food was evenly distributed, the juveniles tended to avoid areas of rapid water flow and distributed evenly across the sheltered areas receiving the equal water velocity. This distribution might be the confounding effects between water flow velocity and food stimulus. Based on these findings, it was recommend that the food and water velocity be evenly distributed in an abalone aquaculture system.