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.     

Pharmacological and histological assessment of gut muscle movement in blacklip abalone, Haliotis rubra (Leach)

We examined the basic pharmacology of abalone gut tissues for adrenergic and peptide receptors using ligand binding and by determining the pharmacological effectors of abalone gut motility in a number of gut regions. Contractile responses could not be elicited, even though we could show the existence of α‐adrenergic and peptide receptors. Responses to any muscle contractile agents, including carbachol and potassium chloride, could not be found in blacklip abalone, Haliotis rubra (Leach). Histology confirmed the relative absence of muscular layers in all the tissues of the gut, indicating that previous literature reports of muscular involvement in contractile movement of gut contents did not appear to apply to this species.     

Food chain dynamics of abalone in a polyculture system

The food chain dynamics of three species of commercially important abalone — the red abalone (Haliotis rufescens), the green (H. fulgens) and the Japanese (H. discus) — were measured to determine their relative success in the multispecies aquaculture system at Woods Hole Oceanographic Institution. The abalone were fed sea lettuce (Ulva lactuca) used in the polyculture system to remove nutrients regenerated by the bivalves.This polyculture system was operated on a moderate-sized scale during the summer of 1973. Growth and feeding rates of the abalone were measured during a 112-day period and net growth efficiency was calculated (net production/food ingested). All three species grew during the experiment but H. rufescens had the lowest efficiency of 10.2%, and H. fulgens had the highest efficiency of 22.7%. These differences might be due to the temperature regime (16–23°C) during the experiment. H. rufescens is typically found in colder waters whereas H. fulgens is common in warmer Southern California waters. The results suggest the feasibility of the browsing role of abalone in polyculture systems.     

Terrestrial leaf meals or freshwater aquatic fern as potential feed ingredients for farmed abalone Haliotis asinina (Linnaeus 1758)

Three terrestrial leaf meals, Carica papaya, Leucaena leucocephala, Moringa oliefera and a freshwater aquatic fern, Azolla pinnata were evaluated as potential ingredients for farmed abalone diet. All diets were formulated to contain 27% crude protein, 13% of which was contributed by the various leaf meals. Fresh seaweed Gracilariopsis bailinae served as the control feed. Juvenile Haliotis asinina (mean body weight=13.4±1.6 g, mean shell length= 38.8±1.4 mm) were fed the diets at 2–3% of the body weight day^–1. Seaweed was given at 30% of body weight day^–1. After 120 days of feeding, abalone fed M. oliefera, A. pinnata‐based diets, and fresh G. bailinae had significantly higher (P<0.01) specific growth rates (SGR%) than abalone fed the L. leucocephala‐based diet. Abalone fed the M. oliefera‐based diet had a better growth rate in terms of shell length (P<0.05) compared with those fed the L. leucocephala‐based diet but not with those in other treatments. Furthermore, protein productive value (PPV) of H. asinina was significantly higher when fed the M. oliefera‐based diet compared with all other treatments (P<0.002). Survival was generally high (80–100%) with no significant differences among treatments. Abalone fed the M. oliefera‐based diet showed significantly higher carcass protein (70% dry weight) and lipid (5%) than the other treatments. Moringa oliefera leaf meal and freshwater aquatic fern (A. pinnata) are promising alternative feed ingredients for practical diet for farmed abalone as these are locally available year‐round in the Philippines.     

Growth performance and feed utilization of Haliotis tuberculata coccinea (Reeve) juveniles from the Azores (north-eastern Atlantic Ocean): An assay of three mono algae diets

We assessed the dietary value of available seaweeds (Ulva lactuca, Ulva intestinalis, and Sargassum vulgare) as food for juvenile abalone Haliotis tuberculata coccinea. During the 6 weeks that the experiment lasted, initial shell length and weight increased from 13.95 ± 0.32 mm to 15.30 ± 0.30 mm and from 0.39 ± 0.02 g to 0.47 ± 0.03 g. Ulva lactuca diet resulted in higher shell growth rate (58.47 ± 16.76 μm day^?1), SGR (1.15 ± 0.47%.day^?1), weight gain (65.5% ± 33.3), and PER (1.56 ± 0.31), possibly thanks to its higher crude protein content. Both green algae (U. lactuca, U. intestinalis) prove to support the growth of H. tuberculata coccinea. However, they might not be appropriate for an optimal intensive aquaculture production; further research should be conducted to find more efficient diets for juvenile abalone.     

Thermal preference and tolerance of green abalone Haliotis fulgens (Philippi, 1845) and pink abalone Haliotis corrugata (Gray, 1828)

The thermoregulatory behaviour of green abalone Haliotis fulgens and pink abalone H. corrugata was investigated. Haliotis fulgens juveniles ranging in wet weight from 3.0 to 3.3 g and from 28.7 to 30.5 mm shell length and of H. corrugata 2.0 g and 25.7 mm in shell length were exposed to 19°C for 30 days in a flow‐through water system. Temperature preference was determined in a horizontal thermal gradient and was found to be 25.4°C for green abalone and 25.0°C for pink abalone. Displacement velocity was 4.3 cm h⁻¹ for H. fulgens and 12.8 cm h⁻¹ for H. corrugata. The optimum temperature for growth calculated for both abalone species was 24.6 and 24.5°C respectively. The critical thermal maxima (CTMax) of H. fulgens and H. corrugata were determined as a measure of thermal tolerance. Abalones were subjected to increasing water temperatures at a rate of 1°C on 30 min until they detached from the substrate. The CTMax at 50% were 33.6 and 32.0°C for green and pink abalone respectively. The results are discussed in relation to site selection and commercial rearing.     

The potential for utilizing fouling macroalgae as feed for abalone Haliotis discus hannai

In order to assess the potential of utilizing fouling macroalgae as feed for abalone Haliotis discus hannai, the species, biomass and total stock of fouling macroalgae attached on the aquaculture facility‐lines were investigated and assessed at Sungo Bay, China, in August 2007. The nutritional value of four fouling macroalgae (Codium fragile, Sargassum muticum, Ulva pertusa and Laminaria japonica) and one cultured macroalgae (Gracilaria lemaneiformis) as the diet of abalone was assessed by analysis of the chemical composition of the macroalgae and by measuring bioavailability and digestibility to abalone H. discus hannai under laboratory conditions in August 2007. The results showed that the biomass of fouling macroalgae at the two sites was 243.61 ± 26.23 and 1078.15 ± 50.28 g m^?1 respectively. The contents of protein of G. lemaneiformis and C. fragile were 18.30% and 17.60%, respectively, which were significantly higher than the others. The five macroalgae were accepted by the abalone, and growth energies (P) and growth efficiency (%) were positive. However, significant differences were found among the different macroalgae. The growth efficiencies of abalone show a negative relationship with the levels of dietary lipid and a positive relationship with the protein contents. These results suggest that fouling macroalgae have a great potential to be used as diets of abalone.     

Temperature and size range for the transport of juvenile donkey's ear abalone Haliotis asinina Linne

Live transport of hatchery‐produced juvenile donkey's ear abalone Haliotis asinina Linne was examined to evaluate the effect of transportation on the survival of juvenile abalone. Simulated transport experiments were conducted to determine the appropriate temperature using 5, 10 and 20 g L^?1 of ice to air volume for 8 h and the appropriate size using two size groups (Size A, 15–20 mm, 0.5–1.3 g, and Size B, 30–35 mm, 5.3–8.5 g) up to 24‐h out‐of‐water live transport. Survival was significantly higher (P<0.001) when 10 g L^?1 of ice was used to decrease the temperature to the range of 17–23 °C. At this temperature, both size groups subjected to simulated transport for 8 and 10 h had 100% survival after 48 h, while mortality occurred in abalones subjected to 16 and 24 h of simulated transport. The Size B abalone subjected to 24 h of transport had significantly higher survival (64.4 ± 2.9%) (P<0.001) than the Size A abalone (5.5 ± 1.6%) after 48 h. Live juvenile abalone were successfully transported to the field applying the protocols developed in the lab experiment. This study serves as a guide for handling and shipping live juvenile abalone.     

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.     

Production and sinking rates for bio‐deposits of abalone (Haliotis discus hannai Ino)

Particle size distribution, organic content and the sinking velocity of bio‐deposits of abalone Haliotis discus hannai Ino were evaluated. The abalone was divided into three size classes according to shell length: Group B: 68.8 ± 2.8 mm, group M: 56.1 ± 2.3 mm and group S: 42.4 ± 2.8 mm. The faecal pellets had an amorphous shape, with a cross‐sectional area ranging from 0.01 to 2.64 mm². The mean cross‐sectional area varied among the size classes: 0.59 (B), 0.21 (M) and 0.12 (S) mm². The bio‐deposit organic content was similar among the groups (B: 18.3 ± 1.1%; M: 19.9 ± 0.9%; S: 19.3 ± 1.0%). Sinking velocity ranged from 0.5 to 2.3 cm s^?1. Larger abalone tended to have larger and thus faster‐sinking faecal pellets. The median sinking velocity for groups B, M and S was 0.9, 0.5 and 0.7 cm s^?1 respectively. The mean daily production rates for groups S, M and B were 2.8 ± 1.2, 3.4 ± 1.3 and 4.3 ± 1.8 g ind.^?1 respectively. Bio‐deposit production rate estimated to be between 134.4 and 206.4 mg m^?2 day^?1 in the bay. Our results suggest that the pellet size was within the appropriate size range for filter‐feeding bivalves.     

Effect of Hybrid Abalone,Haliotis discus hannai × Haliotis discus discus,Cultivation on the Carbon Cycle: Carbon Source/Sink

Respiration, calcification, and bio‐deposition of hybrid abalone, Haliotis discus hannai × Haliotis discus discus, fed on different foodstuffs have been measured to evaluate the effect of hybrid abalone culture on carbon source/sink in coastal areas. Fed with Laminaria japonica, Undaria pinnatifida, Gracilaria lemaneiformis, U. pinnatifida, and Ulva pertusa, alternated mutually, the carbon bio‐deposition rate of hybrid abalone was 24.29 ± 6.39, 65.40 ± 10.55, 21.48 ± 5.99, and 29.28 ± 6.47 µg/g/h, respectively. Hybrid abalone fed on U. pinnatifida had a higher carbon bio‐deposition rate compared to that fed on other foodstuff (P < 5%). Rate of CO₂ released by respiration of hybrid abalone fed on the experimental foodstuff was 24.53 ± 8.57, 32.73 ± 7.99, 29.31 ± 6.39, and 33.67 ± 12.37 µg/g/h, respectively. Results indicated that calcification presented less relationship with body weight type of the foodstuff. The rate of CO₂ released by calcification into seawater and atmosphere was 2.77 ± 1.89 and 6.53 ± 3.36 µg/g/h, respectively. The total rate of CO₂ released because of bio‐deposition, respiration, and calcification processes was 16.19 ± 4.67 µg/g/h, while the total rate of carbon sequestered in shells and tissues was 8.94 ± 2.07 µg/g/h. The study revealed that hybrid abalone culture is a source of CO₂.     

Shell marking by artificial feeding of the tropical abalone Haliotis asinina Linne juveniles for sea ranching and stock enhancement

A method of marking abalone (Haliotis asinina Linne) for sea ranching and stock enhancement purposes was developed. Three‐month‐old abalone juveniles (11.8‐mm shell length, 0.28 g) were fed artificial diets for 1, 2, or 3 weeks. The width of the bluish‐green shell band produced by abalone juveniles was 1.7, 2.6, and 4.2 mm after 1, 2, or 3 weeks of feeding respectively. The growth and survival of juveniles fed artificial diets did not differ from that of juveniles fed the seaweed Gracilariopsis bailinae (control). Feeding the diet‐fed juveniles with the seaweed thereafter produced the natural brownish shell, thus forming a sandwiched bluish‐green band. An experimental release in outdoor tanks with natural growth of seaweeds and diatoms, and in a marine reserve showed that the shell band remained clear and distinct, indicating the usefulness of this shell marking method in sea ranching and stock enhancement of abalone.     

Hybridization between Haliotis rufescens and Haliotis discus hannai: evaluation of fertilization,larval development,growth and thermal tolerance

Two introduced abalone species are currently produced in Chile, red abalone Haliotis rufescens and Japanese abalone Haliotis discus hannai. However, red abalone accounts for 99% of total production, while the Japanese abalone has not adapted well to Chilean coastal waters. This study reports the hatching, growth and thermal tolerance performance in interspecific hybrids produced between red (R) and Japanese (J) abalone. Our results show that egg age and sperm concentration were critical factors to produce hybrids. The cross R♀ × J♂ showed a fertilization rate of 55.3 ± 3.5% using 20‐min‐old eggs and sperm concentrations of 14 × 10⁶ cells mL^?1, while the reciprocal cross (J♀ × R♂) was not successful. Further, larval development stages were similar in RR, JJ and RJ hybrid abalones. Among the experimental trials, settlement rate varied from 12.3% to 18.6% and final survival from 20.1% to 31.7%, being the RJ hybrid rates intermediate between parental species. The final shell lengths were similar between RR and RJ hybrids, but significantly higher in JJ abalones. In addition, thermal tolerance was ascertained due its pivotal role for the abalone physiology. Thus, RJ hybrids showed the highest HSP70 gene expression and offers new possibilities to expand Chilean abalone production in warm waters zones.     

Community and trophic structures of abalone <Emphasis Type="Italic">Haliotis diversicolor</Emphasis> habitat in Sagami Bay,Japan

Abstract:   The community and trophic structures of the subtidal habitat of the abalone Haliotis diversicolor were examined in Nagai, Sagami Bay, Japan. Conventional community indices showed no significant differences among three sampling stations. The overall continuum of stable isotope ratios was structured into three different trophic linkages: (i) brown algae-dependent benthic food chain; (ii) red algae-dependent benthic food chain; and (iii) planktonic food chain. Brown algae and red algae likely play different roles with respect to carbon sources in the habitat. Conventional fractionation values indicated that the abalone H. diversicolor (δ¹³C = −12.4 ± 1.0‰, δ¹⁵N = 9.3 ± 0.5‰) feeds on the lamina of Undaria pinnatifida during juvenile and adult stages (8.0–65.6 mm shell length). Stable isotope signatures suggested that the juveniles of other abalone species as well as some amphipods and a sea cucumber Holothuria decorata are competitors of H. diversicolor , whereas some Muricidae gastropods such as Thais bronni and Ergalatax contractus are predators. The isotopic differences among macroalgal species and the subsequent transfer to consumers indicate that stable isotopic analysis is an effective means of studying food webs in an open rocky shore community with little influence from external primary production such as terrestrial vegetation.     

Molecular identification of macroalgal fragments in gut contents of the sea urchin <Emphasis Type="Italic">Hemicentrotus pulcherrimus</Emphasis>

In order to improve the efficiency of stock enhancement programs for the sea urchin Hemicentrotus pulcherrimus, information on food algal species, which affect growth and gonad production greatly, is necessary. Since it is difficult to identify species from the macroalgal fragments within the gut contents of the sea urchin by microscopic observation, we tried to apply a DNA barcoding method for gut contents analysis. We used a partial rbcL gene sequence for taxonomic section and newly designed primer sets, respectively, for brown algae and for red algae. Direct sequencing of the PCR products was carried out. Species identification was based on the phylogenetic relationship. We could objectively identify four species and two taxonomic groups (genus or family) in brown algae, and two species and four taxonomic groups in red algae from the gut contents. Sargassum hemiphyllum was the most abundant brown alga in the gut contents but was not dominant in the study site. The result showed the importance of identification to the species level. In addition, red algal epiphytes were detected with brown algal fragments. The DNA barcoding method will enable the researchers to verify the important role of epiphytes as a potential food source.     

Digestive protease characterization, localization and adaptation in blacklip abalone (Haliotis rubra Leach)

Digestive protease in blacklip abalone (Haliotis rubra Leach) from whole gut extracts was found to have dual pH optima at pH 3 and 10 and an optimum temperature of 45 °C. Over the biologically relevant range of pH (5–8), protease activity dropped to a minimum at pH 5 (53% of the maximal rate at pH 3) rising gradually and continuously up to (and beyond) pH 8. Over the biologically relevant range of temperatures (9–24 °C), protease activity increased continuously with activity at 24 °C being 75% higher than activity at 9 °C. Protease digestion was relatively uniform in gut extracts from sections containing gut contents. Digestion thus appears to be significantly extracellular (in the lumen of the gut). Analysis of gut sections washed free of food matter suggests the anatomical origin of protease from activity of gut regions in the order: digestive gland » salivary gland ≈ stomach > crop > intestine > upper oesophagus. Whole gut protease levels did not alter in response to a high protein artificial diet.     

Impacts of the 2011 mega‐earthquake and tsunami on abalone Haliotis discus hannai and sea urchin Strongylocentrotus nudus populations at Oshika Peninsula,Miyagi, Japan

On 11 March 2011, a massive tsunami generated by a mega‐earthquake with a moment magnitude of 9.0 hit a wide area of Pacific coast of northeast Japan. We observed and analyzed the effects of the earthquake and tsunami event on populations of the abalone Haliotis discus hannai and sea urchin Strongylocentrotus nudus at Tomarihama in Miyagi, where we have carried out regular surveys since January 2008. Before the event, algal forests dominated by the brown macroalga Eisenia bicyclis had developed in the survey area shallower than 5 m in depth, where adult abalone >50 mm in shell length (SL) inhabited. Juvenile abalone <20 mm SL and juvenile and adult urchins inhabited the deeper area dominated by crustose coralline algae (CCA). After the event, although no apparent decrease was observed in the brown macroalgal population, the mean density of adult abalone >50 mm SL, mainly inhabiting the algal forests, was reduced by more than half. The impact of the tsunami was more profound in the CCA area than in the macroalgal forest. Juvenile abalone and urchins largely decreased to 14 and 5% of the densities just before the event, respectively. The distribution pattern of juvenile abalone and urchins could be a cause of the marked decrease, because most of these animals inhabited the CCA area where the disturbance by the massive water movement was not reduced by the effects of the macroalgal forest.     

Octopus as predators of abalone on a sea ranch

This study examined the occurrence of octopus across an abalone, Haliotis laevigata, Donovan, sea ranch in south‐western Australia, to understand how octopus may be impacting abalone production. Commercial divers removed 654 octopus and 17,666 empty abalone shells during regular, 2 to 4‐weekly surveys over 27 months. A negative binomial generalised linear model estimated a 78% increase in empty shells per artificial abalone habitat per day, when octopuses were present, after adjusting for location and season. Of the 408 shells examined for evidence of predation, 19% had a small, slightly ovoid hole consistent with those made by octopus. The mean (± 1 SE) length of shells with boreholes (70.3 ± 2.2 mm) was significantly longer than those without (59.8 ± 0.5 mm), and boreholes were concentrated over the adductor, respiratory organs and heart. This study provides important insights into the adaptable feeding regimes of octopus and their potential to impose strong top‐down controls on sea ranching operations.     

Trophic structure in natural habitats of the abalone Haliotis discus hannai with distinct algal vegetation of kelp and crustose coralline algae: implication of ontogenetic niche shifts

The abundance, species composition, and stable isotope ratios of benthic organisms were investigated to determine the trophic structures in abalone (Haliotis discus hannai) habitats, which are characterized by contrasting vegetation of crustose coralline algae (CCA) and kelp beds. A size–frequency analysis revealed that juvenile abalones with shell lengths (SLs) smaller than ~30 mm primarily inhabited CCA beds, whereas adults were abundant in kelp beds. Stable isotope analyses indicated that CCA beds were composed of a single food chain, whereas kelp beds supported multiple food chains. The abalone were divided into three size groups to estimate potential species interactions during their ontogeny. A small gastropod, Homalopoma sangarense, was the most abundant species, but is suspected to be less competitive with abalone, especially in CCA beds. An abundant starfish Asterina pectinifera appeared to function as a potential predator of juvenile abalones in both CCA and kelp beds. We concluded that CCA beds are essential for immediate post-settlement processes of abalones, whereas kelp beds are more important for providing refuge and food sources for adult abalones. The present study highlights that ontogenetic niche shifting can be a successful life-history strategy to sustain the abalone population in a subtidal rocky shore ecosystem.     

Evaluation of cryodamage in small abalone,Haliotis diversicolor,spermatozoa by using a flow cytometer

We used flow cytometry to determine the quality of small abalone, Haliotis diversicolor, sperm before freezing and after thawing. We investigated the effects of cryopreservation on the characteristics of small abalone semen and determined the motility and fertilization capacity of the pre-freezing and post-thawed sperm. The percentages of motility and fertility were 61 ± 2% and 67 ± 1%, respectively, for the post-thawed sperm and 90 ± 4% and 92 ± 0%, respectively, for the pre-freezing sperm. Sperm cells were stained with specific fluorescent dyes to measure plasma and acrosomal membrane integrity, mitochondrial status, oxidation level, DNA compaction, and viability through flow cytometry. The frozen–thawed sperm exhibited significantly higher mitopotential activity (p < 0.05, damaged mitochondria; 25.01 ± 1.18%) and oxidation value (p < 0.01, free radicals; 63.79 ± 3.93%) compared with the pre-freezing sperm. The oxidation level was the most sensitive signal of the cryopreservation-induced small abalone sperm damage. Flow cytometry is valuable for the objective, accurate, and rapid assessment of pre-freezing and post-thaw small abalone sperm quality.